Identification of miRNAs And Their Target Genes In Genic Male Sterility Lines In Brassica Napus By Small RNA Sequencing

Identification of miRNAs And Their Target Genes In Genic Male Sterility Lines In Brassica Napus By Small RNA Sequencing | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Identification of miRNAs And Their Target Genes In Genic Male Sterility Lines In Brassica Napus By Small RNA Sequencing Jianxia Jiang, Pengfei Xu, Yajie Li, Yanli Li, Xirong Zhou, Meiyan Jiang, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-471166/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Background: Brassica napus is the third leading source of edible oil in the world. Genic male sterility (GMS) lines provide crucial material for harnessing heterosis for rapeseed. GMS lines have been used successfully for rapeseed hybrid production in China. MicroRNAs (miRNAs) play crucial regulatory roles in various plant growth, development, and stress response processes. However, reports on miRNAs that regulate the pollen development of GMS lines in B. napus are few. Results: In this study, 12 small RNA and transcriptome libraries were constructed and sequenced for the flower buds from the fertile and sterile lines of two recessive GMS (RGMS) lines, namely, “6251AB” and “6284AB”. At the same time, 12 small RNA and transcriptome libraries were also constructed and sequenced for the flower buds from the fertile and sterile lines of two dominant GMS (DGMS) lines, namely, “4001AB” and “4006AB”. Based on the results, 46 known miRNAs, 27 novel miRNAs on the other arm of known pre-miRNAs, and 44 new conserved miRNAs were identified. Thirty-five pairs of novel miRNA-3p/miRNA-5p were found. Among all the identified miRNAs, 21 differentially expressed miRNAs with over 1.5-fold change between flower buds of sterile and fertile lines were identified, including six differentially expressed miRNAs between “4001A” and “4001B”, two differentially expressed miRNAs between “4006A” and “4006B”, six differentially expressed miRNAs between “6251A” and “6251B”, and 14 differentially expressed miRNAs between “6284A” and “6284B”. The correlation analysis of small RNA and transcriptome sequencing was conducted. The analysis results indicated that 360 genes were predicted to be the candidate targets of 21 differentially expressed miRNAs. BnaA09g48720D , BnaA09g11120D , and BnaCnng51960D were demonstrated to be cleaved by bna-miR398a-3p, bna-miR158-3p and bna-miR159a, respectively. Among the differentially expressed miRNAs, miR159 was chosen to analyze its function in reproduction development and male sterility. Overexpression of bna-miR159 in Arabidopsis resulted in decreased seed setting rate, and shortened siliques, illustrating that miR159 may regulate the fertility and silique development in rapeseed. Conclusions: Our findings provide an overview of miRNAs that are potentially involved in GMS and pollen development. New information on miRNAs and their related target genes are provided to exploit the GMS mechanism and reveal the miRNA networks in B. napus. Plant Molecular Biology and Genetics Plant Physiology and Morphology Brassica napus miRNAs genic male sterility pollen development miR159 silique development Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Figure 8 Background Male sterility is largely due to the abnormal development of either the sporophytic or gametophytic anther tissues. According to its origin or genetic analysis, male sterility can be divided into cytoplasmic male sterility (CMS) and genic male sterility (GMS) [ 1 ]. CMS is caused by the interaction between the mitochondrial genome and nuclear genes [ 2 , 3 ]. GMS is derived from natural mutations in nuclear genes that control stamen development. GMS mutants always show advantages, such as complete and stable male sterility and no potential negative cytoplasmic effect, compared with most CMS mutants. GMS can be further divided into dominant GMS (DGMS) and recessive GMS (RGMS). In rapeseed, DGMS and RGMS are widely used for hybrid rapeseed production [ 4 ]. Multiple-allele DGMS was generally accepted and was proposed by Song et al. in 2005. This model presented multiple alleles in one locus inheritance to explain the fertility heredity of a newly reported DGMS line 609AB. In this model, Mf , Ms , and ms are three alleles at the same locus, with a relationship of Mf dominant over Ms and Ms over ms . The recessive allele is for normal fertility. The maintainers and restorers are easily screened. Thus, multiple-allele DGMS are widely used for hybrid rapeseed seed production through the construction of a three-line hybrid system [ 5 , 6 ]. Except for the advantages of GMS, the recessive GMS (RGMS) systems have another distinct advantage in that most inbred lines can restore their fertility, so that hybrids with strong heterosis are easily bred. Several RGMS lines have been successfully commercialized in China, including S45AB [ 7 ], 117AB [ 8 ], and 9012AB [ 9 ]. For S45AB and 117AB, approximately 50% of the fertile plants are required to be artificially removed before hybridizing the rest (50% sterile ones) with restorer lines in hybrid production, because no complete maintainer line is available [ 10 , 11 ]. However, a three-line hybrid production system was developed for 9012AB [ 12 ], and this system has been well documented [ 10 , 13 ]. The RGMS line 9012AB has been used successfully for rapeseed hybrid production in China. This male sterility was previously thought to be controlled by three independent genes ( BnMs3 , BnMs4 , and BnRf ). In 2012, Dong et al. demonstrated a major modification of the sterility inheritance model in 9012A. The modified inheritance model indicated that the male sterility was essentially controlled by two loci ( BnMs3 and BnRf ). The previously designated BnMs4 locus was just one allele of BnRf ; it was then designated as BnRf a , which was designated in addition to BnRf b (the allele from 9012A) and BnRf c (the allele from temporary maintainer). The dominance relationship of the three alleles is in the following order: BnRf a > BnRf b > BnRf c . The BnRf allele-specific molecular markers were identified; these markers would simplify the breeding process involving this RGMS line [ 14 ]. Oilseed rape ( Brassica napus , 2n = 38, AACC), which has low erucic acid and glucosinolate contents, is the third leading source of edible oil worldwide. In recent years, some conserved and novel miRNAs associated with silique length [ 15 ], thickness of pod canopy [ 16 ], cadmium stress [ 17 – 19 ], flower organ development [ 20 ], seed maturation [ 21 – 22 ], cold stress [ 23 ], seed development, and oil synthesis [ 24 ] have been widely identified in rapeseed. However, little information is available about the DGMS and RGMS occurrence at the post-transcriptional level in rapeseed. In this study, to systematically explore the roles of miRNAs and their targets involved in GMS occurrence during pollen development in rapeseed, 12 small RNA and transcriptome libraries were constructed and sequenced for the flower buds from the fertile and sterile lines of two RGMS lines (“6251AB” and “6284AB”). Meanwhile, 12 small RNA and transcriptome libraries were also constructed and sequenced for the flower buds from the fertile and sterile lines of two DGMS lines (“4001AB” and “4006AB”). The aims of this study were to identify known and potential novel miRNAs from the 24 libraries and to analyze the expression profiles of the miRNAs and their targets in relation to DGMS and RGMS during rapeseed microspore development. The results would provide a foundation for evaluating the important regulatory roles of miRNAs in pollen formation and GMS occurrence in rapeseed and other crops. Results Analysis of small RNA library data sets and the small RNA profile To identify miRNAs related to DGMS and RGMS during pollen development, the flower buds were collected from the sterile (6251A, 6284A) and fertile (6251B and 6284B) lines of the RGMS lines. Meanwhile, the flower buds were also respectively collected from the sterile (4001A, 4006A) and fertile (4001B and 4006B) lines of the DGMS lines. Three biological replicates were conducted for each of the eight kinds of samples. Thus, total of 24 sRNA libraries were constructed and deep-sequenced. The raw reads of the 24 sRNA libraries ranged from 20.58 to 42.68 million (Table 1 ). The raw reads of the 24 sRNA libraries were uploaded to SRA database of NCBI and 24 accession numbers were obtained, including SRX11350295, SRX11350296, SRX11350307, SRX11350312, SRX11350313, SRX11350315, and SRX11350316 ( https://dataview.ncbi.nlm.nih.gov/object/PRJNA743414?reviewer=t674c02cj415380e8oldre4s5a ). After removing the low-quality reads and contaminated adapter sequences, the clean reads of the 24 sRNA libraries ranged from 19.77 to 41.61 million. The mapped reads were further annotated against the Pfam database and subsequently divided into rRNAs, tRNAs, snRNAs, snoRNAs, ta-siRNA, and others. The endogenous sRNAs were identified as known and novel miRNAs. The average sRNA lengths of the three biological replicates for each sample were calculated, which showed the length distribution patterns of the sRNAs being similar to one another. In general, the majority of the small RNAs ranged from 21 nt to 24 nt in size. The 24 nt small RNAs were the most dominant, followed by 21 nt small RNAs (Figure. 1). Table 1 Overview of sRNA sequencing reads in Brassica napus Sample Raw reads N% > 10% Low quality 5 , adapter contamine 3 , adapter null or insert null With ployA/T/G/C Clean reads 4001B-1 24481718 41 16955 8590 761698 14436 23679998 4001B-2 27695084 70 36718 12896 698693 16785 26929922 4001B-3 20576690 38 16651 10078 768766 9577 19771580 4006B-1 28457208 131 44382 14481 721967 14232 27662015 4006B-2 27355185 45 7473 9466 895573 11463 26431165 4006B-3 35841966 57 25238 16152 1597289 18301 34184929 6251B-1 22307540 36 18341 18038 533864 14517 21722744 6251B-2 24409491 46 15603 27336 689062 7177 23670267 6251B-3 24485022 34 10518 24084 477366 16924 23956096 6284B-1 25552984 64 12764 21674 567970 12962 24937550 6284B-2 24763779 34 16161 23880 431298 15913 24276493 6284B-3 25378477 40 16388 25065 1546184 6044 23784756 4001A-1 21968633 56 16757 10355 723740 8680 21209045 4001A-2 26445702 23 5505 11592 1192819 7631 25228132 4001A-3 27397803 96 35289 12605 1483390 12457 25853966 4006A-1 23437578 357 10961 9403 612982 14712 22789163 4006A-2 42675389 50 10345 14879 1018244 21106 41610765 4006A-3 27452096 93 40026 12496 749916 14657 26634908 6251A-1 21484268 19 17038 20165 462483 17972 20966591 6251A-2 24127081 56 12533 20730 464608 24999 23604155 6251A-3 21608183 357 7763 13126 393936 10376 21182625 6284A-2 23525443 27 11236 20073 746946 12136 22735025 6284A-2 27605112 37 14315 23792 1211985 12779 26342204 6284A-3 29689151 56 30879 21998 753037 16533 28866648 Identification of known and novel miRNAs in B. napus To identify known miRNAs in B. napus , all mapped small RNA sequences were compared with the known mature bna-miRNA sequences deposited in the miRBase database 22.1. Forty-six small RNAs that have the same sequences with the known bna-miRNAs in miRBase were identified. The numbers of reads of the 46 known miRNAs in 24 libraries were listed in Additional Table S1. Among the 46 known miRNAs, bna-miR159a, bna-miR166a, and bna-miR167c showed very high expression levels. To predict novel miRNAs in B. napus , all the mapped small RNAs were blasted to the B. napus genome sequence in Brassica database and known plant miRNAs in miRBase. The small RNAs that exactly map to the genome sequence but not the known plant miRNAs were classified as candidate novel miRNAs. Five criteria described in the Materials and Methods were used to search for novel miRNAs. As a result, 35 pairs of novel miRNA-3p/miRNA-5p were identified. The mature sequences, reads numbers, positions in chromosomes, precursor sequences and minimum free energy are listed in Table 2 . The length distribution of the novel miRNAs was between 18 nt to 26 nt. The length of the novel miRNA precursors ranged from 51 nt to 300 nt with an average length of 154 nt. The minimum free energy ranged from − 240.26 to -9.9 kcal mol − 1 with an average of -70.14 kcal mol − 1 . The precursor sequences of the novel miRNA are shown in Additional Tables S2. Table 2 Novel miRNAs identified in the fertile and sterile lines of Brassica napus by high-throughput sequencing miR_ name Sequence Len Read pre-position pre-len MFE bna-novel_1-3p CUUCCUCCUAACACCAAUUGAUU 23 67 chrA09:27889716..27889818 102 -34.6 bna-novel_1-5p AUCAAUUGGUUUUAGGUUAAGAAGCC 26 124 bna-novel_2-3p UGGCAUUGGUAGUAAUGAGUGU 22 190 chrC04:42905032..42905108 76 -26.9 bna-novel_2-5p ACUCAUUACCAUCAGAGCCAC 21 7 bna-novel_3-3p UCAAUGUUGGCUCAAUUAUGU 21 120 chrC02:22515980..22516065 85 -29.5 bna-novel_3-5p UCAUUGAGUGCAGCGUUGAUGU 22 12 bna-novel_4-3p AUUAUCGACACUGAUCUCAUC 21 106 chrC08:2975781..2975915 134 -80.8 bna-novel_4-5p UAAGGUCACUGUGGUAAUCC 20 52 bna-novel_5-3p UCAAUGUUGGCUCAAUUAUG 20 49 chrC09:40959623..40959707 84 -31.6 bna-novel_5-5p UCAUUGAGUGCAGCGUUGAUGU 22 12 bna-novel_6-5p AAGGACUCUAAUCAGAAAUAUUGG 24 143 chrC06:35249139..35249190 51 -9.9 bna-novel_6-3p AAUGGUCUUAUCUGGAAUCCUUAA 24 11 bna-novel_7-5p UGCCUGGCUCCCUGUAUACCA 21 83 chrA08:8293061..8293144 83 -32.4 bna-novel_7-3p GUGUAUAGAGUAGUCAAGCAUG 22 2 bna-novel_8-5p AUCUCUAAUGUAUAACUCCAUUUU 24 24 chrA03:20065617..20065865 248 -95.7 bna-novel_8-3p AAUGGAGUAGAUAUGGAGAUGCCC 24 1 bna-novel_9-3p UUGGACUGAAGGGAACUCCCU 21 1527 chrA09:14645700..14645869 169 -64.4 bna-novel_9-5p AGAGUUUCCUUAAGUCCAUUC 21 17 bna-novel_10-5p UAAGAUCUUUGUACUUUCGGG 21 67 chrA10:15442817..15442916 99 -39.8 bna-novel_10-3p CGAAAGUACAAAGAUCUGAAA 21 3 bna-novel_11-5p AACAGUUGGAUUGGCUCUACGUGG 24 27 chrA09_random:3088720..3089020 300 -65.5 bna-novel_11-3p ACGAUGGAGGACAAAACUGAUGCA 24 2 bna-novel_12-5p UUUUCAGCAAUCUCUUUUCCAUU 23 44 chrA05_random:416209..416319 110 -29.5 bna-novel_12-3p AUGGGAAAGAUUGUUGAUCAGA 22 6 bna-novel_13-5p UAAAGUAGAGCUCGGUGACGG 21 1163 chrC03:20992442..20992727 285 -240.3 bna-novel_13-3p GUCACCGAGCUCUACUUUAUA 21 1058 bna-novel_14-5p UCGCUUCUGUUGAAUAAUUUUGAC 24 22 chrC04:45707769..45708016 247 -148 bna-novel_14-3p CAAAAUUAUUCAACAGAAGCGAAU 24 23 bna-novel_15-5p AUAUGAGGGUACAAUAGGAAG 21 137 chrAnn_random:33598332..33598540 208 -139.1 bna-novel_15-3p UAUUGUACCCUCAUAUAUAGC 21 89 bna-novel_16-3p CUAAGAGAUCUGUAAUAAACAUGC 24 30 chrC04:7124159..7124378 219 -117.5 bna-novel_16-5p AUGUUUAUUGUAGGUCUUUUAGGUU 25 7 bna-novel_17-5p ACGAACACUGAGUAAUAUCUG 21 15 chrC01_random:3928762..3929012 250 -164.8 bna-novel_17-3p GAUAUUACUCAGUGUUCGUUG 21 13 bna-novel_18-3p ACACUGCAGUGCACUGUACAUUGC 24 17 chrCnn_random:79954335..79954585 250 -113.8 bna-novel_18-5p GUUGUACAUUGUACACAGCGGUGUAC 26 3 bna-novel_19-5p UUGCAAACUGAAUUAUGAGUC 21 20 chrA09:30966144..30966235 91 -45.4 bna-novel_19-3p CUCAUAAUUCAGUUUGCAAUC 21 20 bna-novel_20-5p AAGAUACGGUCUCUUAACUUUUAG 24 259 chrC04_random:3818028..3818149 121 -67.8 bna-novel_20-3p GUUAAUAGACCGUAUCUUAUA 21 13 bna-novel_21-5p AACGAUCUUGUUUGGUUUUGAAGA 24 18 chrA05:21620032..21620188 156 -82.1 bna-novel_21-3p UUCAAAACCAUACAAGAUCGUUUU 24 24 bna-novel_22-3p GAUCAUGUUCGUAGUUUCACC 21 445 chrCnn_random:35712007..35712108 101 -47.3 bna-novel_22-5p UGAAGCUGCCAGCAUGAUCU 20 3 bna-novel_23-3p UUCUUGUGCGUUUAUAGGUAG 21 55 chrA06:23040126..23040236 110 -52.8 bna-novel_23-5p ACCUCUAAAACACACAAGAAGA 22 3 bna-novel_24-5p UGUUUCGCUGUUACUCAUGC 20 40 chrC02:8973302..8973545 243 -93.1 bna-novel_24-3p AUGAGUAACAGCGAAACAAA 20 26 bna-novel_25-3p AAACUGUGUGAACUCUCCAUGGAG 24 389 chrC02_random:2214651..2214881 230 -73.8 bna-novel_25-5p CCAUAAAAAGGGUUCACAAAGUAUUU 26 1 bna-novel_26-3p UUGAUACAUGUAGCUCUUUG 20 2089 chrA03:669098..669269 171 -83.8 bna-novel_26-5p AAGUGCUACCGGUAUCCACGUG 22 840 bna-novel_27-5p UUAAUCGUUUUGUGACUCUU 20 244 chrA07:19146929..19147018 89 -34.7 bna-novel_27-3p UAGUUACAAAACGAUUAGUGC 21 24 bna-novel_28-3p AUCAACGUUGGCUCAAUUAUG 21 453 chrA10_random:1861693..1861781 88 -33.2 bna-novel_28-5p UCAUUGAGUGCAGCGUUGAUGU 22 12 bna-novel_29-3p UCUUGUUACUGAGCUCGACG 20 308 chrA02:7209993..7210285 292 -99.9 bna-novel_29-5p UUCAGCUGGGUACGAGCCACC 21 710 bna-novel_30-5p AUCUGCAUCGAGUGAACUCUAUGG 24 426 chrCnn_random:65177977..65178226 249 -72.9 bna-novel_30-3p AUGGAAUUCACUGAUGCAGAUGCU 24 7 bna-novel_31-5p UUCUUGUGGUUGUAGAGUCUUG 22 367 chrA06:4069612..4069740 128 -56.1 bna-novel_31-3p AGACUCUACAACAUCAGAAAC 21 47 bna-novel_32-5p CGGAUUUUAGCUGCGUAGCUA 21 322 chrAnn_random:44030328..44030409 81 -42.5 bna-novel_32-3p GGCUACGCUGCUGAAUCCGC 20 2 bna-novel_33-3p UUGUAGAAUUUUGGGAAGGGC 21 289 chrC05_random:138762..138826 64 -32.6 bna-novel_33-5p CCUUCCCAAAAUUCUACAAUU 21 39 bna-novel_34-5p ACUUUGAAACUUUGAUCUAGA 21 5292 chrC06:5179422..5179524 102 -42.4 bna-novel_34-3p UAGAUCAAAGCUUUAAUGU 19 20 bna-novel_35-3p UUUUCGAUCUGUAAAUUU 18 4 chrA03:11978303..11978381 78 -30.5 bna-novel_35-5p CAUUUACAGAUCGAAGACAUU 21 3 miR_name, miRNA name; Len, length of mature miRNA; pre-position, the position of miRNA precursor sequences in chromosomes of Brassica napus , pre-len, length of miRNA precursor sequences; MFE, minimum folding free energy. Novel miRNA on the other arm of known pre-miRNA Through sRNA high-throughput sequencing, miRNA-3p and miRNA-5p were found to always be simultaneously present on the 5′ arm and 3′ arm of pre-miRNA secondary structures. To identify novel miRNAs on the other arm of known pre-miRNAs, all mapped small RNA sequences were compared to the known precursor sequences of bna-miRNAs in the miRBase database 22.1. Finally, 27 novel miRNAs on the other arm of known B. napus pre-miRNAs were identified. The miRNA sequences and the number of reads in 24 libraries are listed in Table 3 . Table 3 Identification of novel miRNAs on the other arm of known pre-miRNAs in Brassica napus bna-miRNA name mature miRNA sequence Length Read count bna-miR156f-3p GCUCACUGCUCUUUCUGUCAGA 22 749 bna-miR156e-3p UGCUCACCUCUCUUUCUGUCAGU 23 344 bna-miR160a-3p GCGUAUGAGGAGCCAUGCAUA 21 32 bna-miR160c-3p GCGUACAGAGUAGUCAAGCAUG 22 24 bna-miR160d-3p GCGUACAGAGUAGUCAAGCAUG 22 24 bna-miR161-3p GUCACUUUCAAUGCGUUGAUC 21 7 bna-miR164b/c/d-3p CACGUGUUCUACUACUCCAAC 21 21 bna-miR166d-5p GGACUGUUGUCUGGCUCGAGG 21 135 bna-miR166e-5p GGAAUGUUGUCUGGCACGAGG 21 10 bna-miR166f-5p GGAAUGUUGUCUGGAUCGAGG 21 202 bna-miR167a/b-3p GAUCAUGUUCGCAGUUUCACC 21 750 bna-miR167a/b-3p GAUCAUGUUCGCAGUUUCACC 21 750 bna-miR168a-3p CCCGCCUUGUAUCAAGUGAAU 21 104 bna-miR171a/b/c-5p AGAUAUUAGUGCGGUUCAAUC 21 51 bna-miR171d-5p AGAUAUUGGUGCGGUUCAAUC 21 12 bna-miR172a-5p GCAGCACCAUCAAGAUUCAC 20 48 bna-miR172b-5p GCAGCAUCAUUAAGAUUCACA 21 3 bna-miR172c-5p GCAGCAUCAUCAAGAUUCACA 21 9 bna-miR172d-5p GCAUCAUCAUCAAGAUUCAGA 21 218 bna-miR2111d-3p AUCCUCGGGAUACGGAUUACC 21 25 bna-miR390b-3p CGCUGUCCAUCCUGAGUUUCA 21 1109 bna-miR390c-3p CGCUAUCCAUCCUGAGUUCC 20 19 bna-miR395a/b/c-5p GUUCCUCUGAGCACUUCAUUG 21 61 bna-miR395d/f-5p GUUCCCUUUAACGCUUCAUUG 21 13 bna-miR399b-5p GGGCAAGAUCUCUAUUGGCAGG 22 12 bna-miR403-5p UGUUUUGUGCGUGAAUCUAAUU 22 287 bna-miR824-3p CCUUCUCAUCGAUGGUCUAGA 21 1640 Identification of new conserved miRNA families and new miRNA members To identify new conserved miRNAs in B. napus , all mapped small RNAs were mapped to known plant miRNAs in miRBase and B. napus genome sequences. If the small RNAs can match known plant miRNAs with no more than three mismatches and can exactly map to B. napus genome sequences, then these small RNAs were initially classified as candidate new conserved miRNAs. Five criteria described in the Materials and Methods were used to strictly screen the candidate conserved miRNAs. As a result, 44 miRNAs (22 pairs of miRNAs) belonging to 15 miRNA families were identified (Table 4 ). Among them, bna-miR159b was a new miRNA member of bna-miR159 family. The rest of the 36 miRNAs (14 pairs of miRNAs) have not been previously reported as bna-miRNAs in miRBase; they show high sequence similarity to some of the known plant miRNAs. The bna-miR158a.1 and bna-miR158a.2 were identified for bna-miR158a member. The two pairs of bna-miR158a shared the same mature sequences. Their precursor sequences were highly similar with each other, and these sequences were from different loci of the B. napus genome. These two pairs of miRNAs were called sub-members. This type of sub-member was also observed for bna-miR159b and bna-miR408a. Four sub-members (bna-miR159b.1, bna-miR159b.2, bna-miR159b.3, and bna-miR159b.4) were identified for bna-miR159b, and two sub-members (bna-miR408a.1 and bna-miR408a.2) were identified for bna-miR408a. This phenomenon suggests that some MIRNA genes might be produced through a replication event from one origin to another one, which results in more copies of the miRNA group. Two members were identified for bna-miR319 and bna-miR398 families. Except the above mentioned five miRNA families, the rest of 10 miRNA families had only one miRNA member (Table 4 ). Table 4 Identification of new conserved miRNA families in Brassica napus bna-miRNA Sequence Len Read pre-position bna-miR158a.1-5p CUUUGUCUAUCGUUUGGAAAAG 22 3884 chrA08:2748114..2748220 bna-miR158a.1-3p UUUCCAAAUGUAGACAAAGCA 21 32292 bna-miR158a.2-5p CUUUGUCUAUCGUUUGGAAAAG 22 3884 chrC08:3581242..3581348 bna-miR158a.2-3p UUUCCAAAUGUAGACAAAGCA 21 32292 bna-miR159b.1-5p AGCUGCUAAGCUAUGGAUCCC 21 258 chrA02:9865184..9865001 bna-miR159b.1-3p UUUGGAUUGAAGGGAGCUCUA 21 46073 bna-miR159b.2-5p AGCUGCUAAGCUAUGGAUCCC 21 258 chrA07_random:1944377..1944191 bna-miR159b.2-3p UUUGGAUUGAAGGGAGCUCUA 21 46073 bna-miR159b.3-5p AGCUGCUAAGCUAUGGAUCCC 21 258 chrC02:19215807..19215624 bna-miR159b.3-3p UUUGGAUUGAAGGGAGCUCUA 21 46073 bna-miR159b.4-5p AGCUGCUAAGCUAUGGAUCCC 21 258 chrC06:33954934..33954749 bna-miR159b.4-3p UUUGGAUUGAAGGGAGCUCUA 21 46073 bna-miR319a-5p AGAGCUUCCUUGAGUCCAUUC 21 27 chrC01:10651723.. 10651921 bna-miR319a-3p UUGGACUGAAGGGAGCUCCCU 21 4848 bna-miR319b-5p GGAGAUUCUUUCAGUCCAGUC 21 4 chrC04:46407584.. 46407846 bna-miR319b-3p UUGGACUGAAGGGAGCUCCUU 21 27901 bna-miR391-5p UUCGCAGGAGAGAUAGCGCCA 21 110 chrA10:10707678..10707812 bna-miR391-3p ACGGUAUCUCUCCUACGUAGC 21 237 bna-miR398a-5p GGGUCGACAUGAGAACACAUG 21 141 chrA03:2288822..2288945 bna-miR398a-3p UGUGUUCUCAGGUCACCCCUG 21 9870 bna-miR398b-5p GGAGUGUCAUGAGAACACGGA 21 25 chrC02:37793584..37793689 bna-miR398b-3p UGUGUUCUCAGGUCACCCCUU 21 145 bna-miR400-5p UAUGAGAGUAUUAUAAGUCAC 22 78 chrAnn_random:40582790..40582930 bna-miR400-3p GACUUAUAAUGAUCUCAUGAA 22 237 bna-miR408a.1-5p GGGAGCCAGGGAAGAGGCAGU 22 1232 chrA05:478954..479121 bna-miR408a.1-3p UGCUUGUUCCCUGUCUCUCUC 22 1002 bna-miR408a.2-5p GGGAGCCAGGGAAGAGGCAGU 22 1232 chrCnn_random:8448205..8448064 bna-miR408a.2-3p UGCUUGUUCCCUGUCUCUCUC 22 1002 bna-miR9554-5p GAAUGAUACUUGGAUAUAAUC 21 5 chrA06:19718101..19718250 bna-miR9554-3p UCAUAUCCAAGUAUCAUUCCU 21 81 bna-miR9558-5p AGAGAUGUCUGGCUUGCAACA 21 3 chrC03_random:1702602..1702746 bna-miR9558-3p UUGCAAGCCAGACAUUUCCUUU 22 8 bna-miR9559-5p UUUGGAUUUUGGUCAUUGUUG 21 5 chrAnn_random:36404086.. 36404194 bna-miR9559-3p ACAAUGAACGAAAUCCAAAUC 21 3 bna-miR9560a-5p ACAGGUGGUGGAACAAAUAUGAGU 25 30 chrA06:19552830..19552965 bna-miR9560a-3p UCAUAUUAGUUCUACCUCCUGCUG 25 2 bna-miR9562-5p ACUAUGCAAUUGUGAACAAAC 21 4 chrA02_random:1408210..1408358 bna-miR9562-3p UUAUUCACAACUGCAUAAUUC 21 3 bna-miR9563a-5p ACCCGUCUCUUAACUUUUAAC 22 15 chrAnn_random:9932700..9932850 bna-miR9563a-3p UAAAAGUUAAGAGACAAGUUA 22 17 bna-miR9568-5p UGCGGAUAUCUUAGGAUGAGGU 22 13 chrA03:13274664..13274813 bna-miR9568-3p UCAUCGUAAGAGAUCUGCAUU 21 2 bna-miR9569-5p UGAGUUAUCAUUGGUCUUGUG 21 1198 chrAnn_random:21855323..21855514 bna-miR9569-3p ACACAGGAACAAUACUAACUCAUU 24 3509 Len, length of mature miRNA; pre-position, the miRNA precursor sequences in chromosomes of Brassica napus Expression profiling of differentially expressed miRNAs in sterile and fertile lines The normalized expression levels of miRNAs were used for identifying differentially expressed miRNAs between the sterile line and the corresponding fertile line, such as “4001A” and “4001B”, “4006A” and “4006B”, “6251A” and “6251B”, and “6284A” and “6284B”. The known, identified conserved and novel miRNAs were followed to differential expression analysis criteria (qvalue 0.73). As a result, 6, 2, 6, and 14 differentially expressed miRNAs were obtained between the flower buds of “4001A” and “4001B”, “4006A” and “4006B”, “6251A” and “6251B”, and “6284A” and “6284B”, respectively (Fig. 2 , Tables 5 ). To further explore the miRNAs involved in the two DGMS lines, a Venn diagram analysis was conducted. The results indicated that two differentially expressed miRNAs were shared between the DGMS lines “4001AB” and “4006AB” (Fig. 3 A). In addition, four differentially expressed miRNAs were shared between the RGMS lines “6251AB” and “6284AB” (Fig. 3 B). The novel miRNA “bna-novel_34-5p” was the only miRNA that was simultaneously differentially expressed in the DGMS and RGMS lines. Table 5 Differentially expressed miRNAs in “4001A B”, “4006AB”, “6251AB” and “6284AB” libraries and their candidate targets by sRNA sequencing and transcriptome analysis in Brassica napus Sample miR _name A B log 2 (FoldChange) Candidate Targets 4001 bna-novel_4-3p 8.2 0.4 1.90 BnaC01g31810D,BnaA07g24350D,BnaC02g41520D,BnaA08g22150D,BnaA04g13020D,BnaA07g18960D,BnaA09g38810D,BnaC03g26470D,BnaC02g03200D,BnaA04g00990D,BnaA03g41600D,BnaA05g07140D,BnaA08g19190D bna-miR390a/b/c 26.8 11.1 1.32 BnaC03g02760D,BnaC01g03570D,BnaA05g28390D,BnaC03g53360D,BnaC01g22410D bna-novel_31-5p 10.7 55.2 -2.16 BnaC01g21190D,BnaA01g17940D,BnaC03g13660D,BnaA03g10950D,BnaA09g00170D,BnaA10g02710D,BnaC01g22250D,BnaA03g38770D,BnaA02g29970D,BnaA02g18470D bna-novel_34-5p 0.6 624.0 -8.16 BnaA09g14240D,BnaA05g30840D,BnaA01g24360D,BnaA07g34690D,BnaA03g14400D,BnaA05g09000D,BnaC01g09790D,BnaA08g26410D bna-novel_1-3p 0.1 8.1 -2.64 -- bna-miR158-3p 554.7 935.8 -0.74 BnaA05g08940D,BnaA04g29200D 4006 bna-novel_34-5p 0.2 125.1 -5.82 BnaA05g29360D,BnaA09g14240D,BnaA05g30840D,BnaA01g24360D,BnaA07g34690D,BnaC03g41430D,BnaA03g35600D bna-novel_31-5p 4.7 53.3 -2.81 BnaC01g21190D,BnaC03g13660D,BnaA09g19450D,BnaA05g09930D 6251 bna-novel_28-3p 5.2 38.3 -2.48 BnaA09g36810D,BnaA10g16750D bna-novel_73-5p 30.6 64.2 -1.19 BnaAnng19370D,BnaA06g16700D,BnaA05g05990D,BnaA10g17750D,BnaC03g50150D,BnaA10g19150D,BnaA03g58190D,BnaA05g28260D,BnaA07g15220D,BnaA01g18450D,BnaA09g50980D,BnaA10g21470D,BnaA09g13370D bna-miR408-5p 23.8 60.6 -1.15 BnaA02g05230D,BnaA10g18650D,BnaA08g27620D,BnaAnng21260D,BnaA10g02870D,BnaA08g00080D bna-novel_88-3p 214.1 511.8 -1.16 BnaA04g16310D,BnaA09g36970D,BnaA05g24870D,BnaC01g43200D bna-miR398a-3p 0 5.1 -2.10 BnaA05g02320D,BnaA06g14440D,BnaA08g08840D bna-novel_34-5p 23.2 57.5 -1.11 BnaA02g05230D,BnaA10g18650D,BnaC01g01790D,BnaA10g10960D,BnaAnng21260D,BnaA10g14890D,BnaA08g27620D,BnaA07g15260D,BnaAnng36200D,BnaA10g15730D,BnaA02g11840D,BnaA10g25360D,BnaA10g02870D,BnaA08g00080D,BnaA10g17820D,BnaA09g25870D 6284 bna-miR394a/b 195.2 87.5 1.04 BnaA05g16640D,BnaA01g24160D,BnaA05g11890D,BnaC02g17150D,BnaAnng09250D,BnaA08g10740D,BnaA07g36430D,BnaC02g29160D,BnaA06g08380D,BnaC01g07190D,BnaA05g00820D,BnaA05g13120D,BnaC01g39490D,BnaC01g16400D,BnaA02g34270D,BnaC02g43190D,BnaA01g33370D,BnaC01g39860D,BnaA08g27810D bna-novel_36-3p 1594.6 900.7 0.82 BnaA03g08820D,BnaA01g00910D,BnaC01g40540D,BnaA04g27250D,BnaA02g08190D,BnaC02g11600D,BnaA10g30150D,BnaA02g24120D,BnaC02g31870D,BnaA04g05090D,BnaA02g02340D bna-novel_67-3p 129.3 50.0 1.23 BnaA03g02150D,BnaC01g00150D,BnaA07g02160D,BnaC03g01120D,BnaA03g00810D,BnaC01g01370D,BnaA07g25630D,BnaA03g55940D,BnaA04g24770D,BnaC01g25070D,BnaA01g20170D,BnaA03g48520D,BnaA09g17190D,BnaA09g39530D,BnaAnng28310D,BnaA01g35930D,BnaAnng08400D,BnaC02g23160D,BnaC03g60340D,BnaA04g00950D,BnaA03g33460D,BnaA04g22260D,BnaA03g27270D,BnaA07g37210D,BnaA06g37910D,BnaC03g21760D,BnaA03g18230D,BnaA09g05960D,BnaA03g34840D,BnaA09g05960D bna-novel_33-3p 30.2 8.4 1.76 BnaA02g26940D,BnaA09g44810D,BnaA06g12040D,BnaA06g14250D,BnaA02g23840D,BnaA10g26020D,BnaA03g12030D,BnaA09g06400D,BnaC01g11370D,BnaA10g03030D,BnaA03g05700D,BnaA10g26690D,BnaA05g01020D,BnaA02g00710D bna-miR159a 2320.1 1295.7 0.82 BnaAnng27960D,BnaA04g18810D,BnaA03g15690D,BnaA07g18670D,BnaA07g12970D,BnaA03g22590D,BnaA07g25350D,BnaA06g20460D,BnaA01g18450D,BnaC01g19500D,BnaA01g16350D,BnaA09g27090D,BnaA06g13170D,BnaA09g42230D,BnaA09g30160D,BnaAnng05670D,BnaA06g18020D,BnaA08g20300D,BnaA02g05410D,BnaAnng21510D,BnaA09g08360D,BnaAnng14630D,BnaA09g02220D,BnaA04g25320D,BnaA09g10390D,BnaA02g30030D,BnaA09g44380D,BnaC01g19800D,BnaA01g35420D,BnaA08g27930D bna-novel_9-3p 114.4 68.3 0.75 BnaA03g33680D,BnaA08g01260D,BnaA01g05980D,BnaAnng27960D,BnaA03g22590D,BnaA07g12970D,BnaA04g09220D,BnaA07g12970D,BnaA02g05410D,BnaA09g55500D,BnaAnng13060D,BnaA01g23170D,BnaA09g13960D,BnaA09g47880D,BnaA04g01370D,BnaA08g17490D,BnaC02g08160D,BnaA02g33550D,BnaC02g42310D,BnaA10g14420D,BnaA05g16460D,BnaA03g42760D bna-miR398a-3p 436.2 972.6 -1.06 BnaA08g19040D,BnaC02g28060D,BnaC01g27860D,BnaA10g26450D,BnaC03g13330D,BnaA03g10640D,BnaC02g12070D,BnaC01g18280D,BnaA01g15390D,BnaC02g42090D,BnaAnng31090D,BnaC01g43200D,BnaA05g30940D,BnaA01g29240D,BnaC01g36670D bna-novel_3-3p 3.1 23.4 -2.43 BnaA05g24640D,BnaA09g38650D,BnaA06g39700D,BnaA09g39360D,BnaA09g54170D,BnaA06g10130D,BnaA10g14600D,BnaC01g22370D,BnaA10g24950D,BnaA10g23300D,BnaA04g14120D,BnaA09g41640D,BnaC01g09540D bna-novel_5-3p 1.5 13.5 -1.77 BnaA05g24640D,BnaA06g39700D,BnaC02g14840D,BnaA06g10130D,BnaA09g38650D,BnaC01g22370D,BnaA09g39360D,BnaA10g04390D,BnaA09g54170D,BnaA10g14600D,BnaA01g28170D,BnaA06g28940D,BnaA10g23460D,BnaA06g07690D,BnaA10g24950D,BnaA09g01710D,BnaA10g16750D,BnaA10g23300D,BnaC01g09540D bna-novel_73-5p 44.5 80.8 -0.84 BnaAnng19370D,BnaC02g44840D,BnaA06g16700D,BnaA05g05990D,BnaC03g50150D,BnaA10g19150D,BnaA05g06360D,BnaA03g26400D,BnaA09g49880D,BnaA10g04230D,BnaA03g28580D,BnaA06g06040D,BnaA03g29130D,BnaA07g15220D,BnaA10g27030D,BnaA04g10410D,BnaAnng07200D,BnaC01g14980D,BnaA04g10200D,BnaA08g03230D,BnaA10g23200D,BnaA10g21470D,BnaA03g07290D,BnaA07g21850D,BnaC02g16740D,BnaA10g09600D,BnaA03g21270D,BnaA08g29090D,BnaA09g13370D bna-novel_68-3p 0 7.1 -2.20 BnaA04g25570D,BnaA02g31820D,BnaC02g40450D,BnaAnng04800D,BnaA06g27210D,BnaA03g20670D,BnaC01g31630D,BnaA03g20660D,BnaA08g29130D,BnaA09g12560D,BnaA09g14580D,BnaA09g06270D,BnaA10g26450D,BnaA02g05300D,BnaA03g26340D,BnaA09g32850D bna-miR408-5p 41.3 94.4 -1.13 BnaA03g22490D,BnaA10g18290D,BnaA07g26450D,BnaA06g37530D,BnaC02g29240D,BnaA05g29880D,BnaA10g21900D,BnaC01g16210D,BnaA02g09770D,BnaA10g23140D,BnaA02g34310D,BnaC02g43230D,BnaA01g08900D,BnaA06g10230D,BnaA10g15210D,BnaA05g14430D,BnaC02g46010D,BnaA03g04360D,BnaA04g08040D,BnaAnng32010D,BnaA03g30060D bna-novel_1-3p 0.9 7.5 -1.72 BnaA02g30100D,BnaC02g38520D,BnaA10g21900D,BnaAnng29380D bna-novel_34-5p 1.0 456.9 -6.87 BnaA02g05300D,BnaA05g27100D,BnaA07g28780D,BnaA06g28070D,BnaA09g14240D,BnaA05g30840D,BnaA08g29690D,BnaC03g48340D,BnaA04g22150D,BnaC01g31340D,BnaA01g24360D,BnaA03g23950D,BnaC03g53270D,BnaA06g01440D,BnaA02g14960D,BnaA09g03120D,BnaA09g31030D,BnaC02g03920D,BnaA07g34690D,BnaA03g35600D,BnaA04g02140D,BnaC02g14950D,BnaA03g10460D,BnaAnng21410D A:fertile, B:sterile lines. qRT-PCR was conducted to verify the expression profiles of these differentially expressed miRNAs in deep sequencing. Five differentially expressed miRNAs were chosen for qRT-PCR analysis. The results were consistent with those of deep sequencing. In qRT-PCR, miR158 was up-regulated in “4001B” flower buds compared with that in “4001A”. Novel34 was greatly up-regulated in “4006B” flower buds (730-fold) compared with that in “4006A”. miR159 and miR827 were both up-regulated in “6284A” flower buds compared with that in “6284B”. miR398 was up-regulated in “6284B” flower buds compared with that in “6284A” (Fig. 4 ). Target prediction and identification of differentially expressed miRNAs in sterile and fertile lines To make clear the potential regulatory roles of differentially expressed miRNAs, a plant small RNA target analysis server psRNATarget ( http://plantgrn.noble.org/psRNATarget/ ) was performed to predict their target genes. A total of 707 transcripts were predicted to be targets of the 20 miRNAs (Additional Table S3). In addition, transcriptome sequencing was conducted using the same samples as sRNA sequencing (unpublished data). The differentially expressed and up-regulated mRNAs were predicted as the candidate targets for the differentially expressed and down-regulated miRNAs. At the same time, the differentially expressed and down-regulated mRNAs were predicted as the candidate targets for the differentially expressed and up-regulated miRNAs. As shown in Table 5 . Thirty eight candidate target genes were predicted for the six differentially expressed miRNAs between “4001A” and “4001B”. Eleven candidate genes were predicted for the two differentially expressed miRNAs between “4006A” and “4006B”. Forty-four candidate genes were predicted for the six differentially expressed miRNAs between “6251A” and “6251B”. Two hundred and sixty-seven candidate genes were predicted for the 14 differentially expressed miRNAs between “6284A” and “6284B”. To further demonstrate the potential target genes, 5’ modified RACE was performed using mixed samples from flower buds of the fertile lines (“6284B” and “4001B”). Three target genes were validated using 5’ modified RACE (Fig. 5 ). Bn.A09.CSD1 ( BnaA09g48720D ) was cleaved by bna-miR398a-3p. Bn.A09.PPR ( BnaA09g11120D ) was cleaved by bna-miR158-3p. Bn.Cnn.MYB ( BnaCnng51960D ) was cleaved by bna-miR159a. Overexpression of bna-miR159a affected seeds and siliques develpoment in Arabidopsis Among all the differentially expressed miRNAs in the two DGMS and RGMS lines, bna-miR159a had the highest expression level. Thus, bna-miR159a was chosen to analyze its function. Two vectors, namely, p35S::MIR159a-C6 and p35S::MIR159a-A7, which contained the precursor sequences of MIR159a located in C6 and A7 chromosomes of B. napus , were transformed into Arabidopsis , respectively. The corresponding Arabidopsis transgenic plants were named as MIR159OE-1 and MIR159OE-2. The expression levels of bna-miR159 and its target genes were analyzed in wild-type, MIR159OE-1, and MIR159OE-2. To reveal miR159 potential function in Arabidopsis , the expression level of miR159a was analyzed by qRT-PCR. In wild-type plants of Arabidopsis , the transcript of mature miR159a and its targets ( AtMYB33 and AtMYB65 ) were detected in root, stem, rosette leaf, stem leaf, flower, and silique through qRT-PCR. The expression level of mature miR159a was the highest in silique (5875-fold), relatively lower in stem, stem leaf, and flower compared with that in the root. The expression levels of AtMYB33 and AtMYB65 were very low and almost undetectable in silique, whereas they were relatively high in stem, stem leaf, and flower compared with that in the root (Fig. 6 ). In MIR159OE-1 and MIR159OE-2, the transcripts of mature miR159a and its targets were detected in stem leaf through qRT-PCR. The mature miR159a was overexpressed in MIR159OE-1 (4.04-fold) and MIR159OE-2 (13.6-fold) compared with that in WT. Meanwhile, the transcripts of AtMYB33 and AtMYB65 were suppressed in MIR159OE-1 and MIR159OE-2, especially in MIR159OE-2, compared with that in WT (Fig. 7 ). The morphological characters of MIR159OE-1, MIR159OE-2, and WT were observed along with their development processes, especially in the flowering and fruiting periods. No significant difference was observed between transgenic and WT plants in the vegetative growth phase. However, during the reproductive growth period, in the MIR159OE-1 and MIR159OE-2 transgenic plants, the seed setting rate decreased, and siliques became shorter compared with that in the WT (Fig. 8 ). The length of siliques from WT, MIR159OE-1, and MIR159OE-2 transgenic plants were measured. The results showed that the silique lengths of WT were approximately 13.4 mm, whereas the average silique lengths of MIR159OE-1 and MIR159OE-2 were 5.9 and 5.8 mm, respectively. The results indicated that the overexpression of MIR159 led to significantly shorter siliques and reduced seed setting rate (Fig. 8 E). Discussion MiRNAs, as the key post-transcriptional regulators, participate in various biological processes in plant. Recently, an increasing number of studies showed that plant miRNAs were also involved in pollen and fertility development. In 2014, Jiang et al. identified 54 new conserved miRNAs and 25 pairs of novel miRNA/miRNA* in a GMS system of B. campestris ssp. chinensis . Eighteen differentially expressed miRNAs with over two fold change between flower buds of male sterile and fertile lines were identified; they might be involved in the pollen development process [ 25 ]. In 2017, Ma et al. verified that the overexpression of MIR158 caused pollen abortion and reduced pollen vitality, which were caused by the degradation of pollen content from the binuclear microspore stage [ 26 ]. Dong et al. identified 85 known miRNAs and 120 novel miRNAs, which were expressed during rapeseed anther development in a novel recessive GMS system “CN12AB.” Moreover, 19 and 18 known miRNAs were found to be differentially expressed in 0.5–1.0 mm buds and in 2.5–3.0 mm buds between CN12A and CN12B, respectively. Among these, 14 miRNAs were up-regulated, and 23 miRNAs were down-regulated expressed in CN12A compared with those in CN12B [ 27 ]. In this study, to identify miRNAs and their targets involved in pollen development and GMS occurrence in rapeseed, 24 small RNA libraries and transcriptome libraries were constructed and sequenced for the flower buds from the fertile and sterile lines of two RGMS lines (“6251AB” and “6284AB”) and two DGMS lines (“4001AB” and “4006AB”). Based on the sequencing results, 21 differentially expressed miRNAs with over 1.5-fold change between flower buds of male sterile and fertile lines were identified, including six differentially expressed miRNAs between “4001A” and “4001B”, two differentially expressed miRNAs between “4006A” and “4006B”, six differentially expressed miRNAs between “6251A” and “6251B”, and 14 differentially expressed miRNAs between “6284A” and “6284B”. Among them, bna-novel_34-5p was common and differentially expressed between the fertile and sterile lines of “4001AB”, “4006AB”, “6251AB”, and “6284AB”. The results of previous studies and this study verified that miRNAs may play important regulatory roles in rapeseed pollen development and GMS occurrence. MiR159 is conserved in many plants and is involved in multiple growth and development processes of plants. Allen et al. obtained a mir159ab double mutant in Arabidopsis , which showed pleiotropic morphological defects, including altered growth habit, curled leaves, small siliques, and small seeds [ 28 ]. Millar and Gubler verified that miR159 regulated anther development by regulating the expression of its targets, such as MYB33 and MYB65 [ 29 ]. Overexpression of miR159 caused the down-regulation of MYB103 transcripts and earlier degeneration of the tapetum and aberrant pollen formation during anther development [ 30 ]. In radish, differential expression level of miR159 during anther development was observed among male sterile and maintainer lines. Increased levels of miR159 transcripts decreased the expression of MYB101 , thereby inhibiting tapetum development and exine formation [ 31 ]. Chen et al. identified 17 differentially expressed miRNAs between long and short siliques of rapeseed, including miR159. Correlation analysis of miR159 and its targets suggested that miR159 repressed cell proliferation to control silique length [ 15 ]. Hu et al. found that the overexpression of Bra- MIR159a caused pollen abortion and abnormal pollen germination [ 32 ]. In this study, the differentially expressed miR159 in “6284A” and “6284B” was chosen to analyze its function during anther development. The bna-miR159 was overexpressed in Arabidopsis and resulted in decreased seed setting rate, and shortened siliques, illustrating that miR159 may regulate the fertility and silique development of rapeseed. The results of previous reports and the results of the present study verified that miR159 and its target genes might be involved in the regulatory network of pollen development and male sterility and this module is conserved in plants. Conclusion A large number of miRNAs were identified during pollen development in the two DGMS and two RGMS lines by deep sequencing. These identified miRNAs included 27 novel miRNAs on the other arm of known pre-miRNAs, 44 new conserved miRNAs, and 35 pairs of novel miRNA-3p/miRNA-5p. Among all the identified miRNAs, 21 differentially expressed miRNAs with over 1.5-fold change between flower buds of male sterile and fertile lines were identified, including six differentially expressed miRNAs between “4001A” and “4001B”, two differentially expressed miRNAs between “4006A” and “4006B”, six differentially expressed miRNAs between “6251A” and “6251B”, and fourteen differentially expressed miRNAs between “6284A” and “6284B”. The qRT-PCR results of 5 differentially expressed miRNAs (miR158, novel34, miR159, miR827, and miR398) were consistent with deep sequencing results. The association analysis of small RNA and transcriptome sequencing was conducted, and the analysis results indicated that 360 genes were predicted to be the candidate targets of 21 differentially expressed miRNAs. The results of 5’ modified RACE verified that Bn.A09.CSD1 ( BnaA09g48720D ), Bn.A09.PPR ( BnaA09g11120D ), and Bn.Cnn.MYB ( BnaCnng51960D ) were cleaved by bna-miR398a-3p, bna-miR158-3p, and bna-miR159a. Additionally, overexpression of bna-miR159 in Arabidopsis resulted in decreased seed setting rate, and shortened siliques, illustrating that miR159 may regulate rapeseed fertility and silique development. All the results in our study would provide valuable clues for exploring miRNA-mediated regulatory networks in fertility development of GMS lines in B. napus . Methods Plant materials “6251AB” and “6284AB” are two recessive genic male sterile (RGMS) lines of B. napus . The “6251A” and “6284A” are male sterile lines and the “6251B” and “6284B” are the fertile lines. Their male sterility is controlled by two loci ( BnMs3 and BnRf ) [ 14 ]. The original source of the two RGMS lines was “9012A”, which was identified by Chen et al. [ 9 ] and Sun et al. [ 33 ]. “4001AB” and “4006AB” are two dominant genic male sterile (DGMS) lines of B. napus . The “4001A” and “4006A” are male sterile lines and the “4001B” and “4006B” are the fertile lines. Their male sterility is controlled by three alleles ( Mf , Ms , and ms ) at the same locus [ 5 ]. The original source of the two DGMS lines was “Yi3A”, which was identified by Li et al. [ 34 ]. The above RGMS lines and DGMS lines used in this study have been grown for several generations. The progenies of the two RGMS lines are both segregated into sterile and fertile types during reproduction at a ratio of 1:1. While the two DGMS lines are both segregated into sterile and fertile types at a ratio of 3:1. These plant materials were planted in the experimental farm of Zhuanghang comprehensive experimental station of Shanghai Academy of Agricultural Sciences. During flowering stage, mixed flower buds were respectively harvested from more than ten plants of the eight lines. The eight lines were “6251A”, “6251B”, “6284A”, “6284B”, “4001A”, “4001B”, “4006A”, and “4006B”. The eight kinds of samples were quickly frozen in liquid nitrogen and stored at -80°C. Three independent biological replicates were collected for each kind of sample. Small RNA library construction and sequencing Total RNA for each kind of sample was extracted in three biological replicates using Trizol reagent (Invitrogen, USA). RNA samples with an OD260/OD280 ratio of 2.0 and a total content of more than 2 µg were qualified for small RNA library construction. Then, 24 sequencing libraries (three biological replicates respectively for “6251A”, “6251B”, “6284A”, “6284B”, “4001A”, “4001B”, “4006A”, and “4006B”) were constructed using TruSeqSmall RNA Sample Preparation Kit (Illumina, USA) and then sequenced using Illumina Hiseq 2500/Miseq at Beijing Novogene Bioinformatics Technology Co. Ltd. Data analysis Clean reads were obtained by removing low-quality reads, N-containing fragments, and adapters. Then, the length of 18 to 30 nt clean reads were mapped to the B. napus genome sequence ( http://brassicadb.agridata.cn/brad/ ) using Bowtie2 with no mismatches allowed, more details were described in Niu et al [ 35 ]. Unmapped sequences were removed. The mapped small RNAs reads were aligned to known miRNAs in miRBase22.1. Modified software mirdeep2 [ 36 ] were used to predict the potential miRNAs and secondary structures. The ncRNAs includes rRNAs, tRNAs, snRNAs, snoRNAs, and small genome repeat sequences were removed. The rest of sRNA sequences were aligned to B. napus NAT-siRNAs in PlantNATsDB to remove NAT-siRNAs. Then miREvo [ 37 ] and mirdeep2 [ 36 ] were used to predict novel miRNAs in B. napus . To reveal the differentially expressed miRNAs, the miRNAs expression was analyzed using the DESeq2 [ 42 ]. The online software of Venny 2.1.0 was used ( https://bioinfogp.cnb.csic.es/tools/venny/index.html ) to draw Venn diagrams. Identification of conserved and novel miRNAs To identify conserved and novel miRNAs in B. napus , the candidate sRNAs were blasted against B. napus genome sequences. In addition, Mfold software was used to predict the secondary structures of pre-miRNAs with the flanking sequences of the small RNAs in the genome [ 43 ]. Five criteria must be met for identifying conserved and novel miRNAs [ 44 , 45 ]. The sequences and lengths, read counts, and positions in chromosome were further analyzed for conserved and novel miRNAs. qRT-PCR Total RNA was treated with DNase I (Takara, Japan) to remove residual genomic DNA. For the qRT-PCR analysis of AtMYB33 and MYB65 , AtACTIN2 gene was used as internal control. PrimeScript™ II reverse transcriptase (Takara) and oligo (dT) primers were used for first-strand cDNA synthesis. For the qRT-PCR analysis of mature miRNAs, U6 was used as internal control gene, and the first cDNA was synthetized using a miRNA First Strand cDNA Synthesis kit (Stem-loop Method) (Sangon Biotech). The qRT-PCR reactions were performed in a MyiQ2 qRT-PCR detection system (Bio-Rad, www.bio-rad.com/ ) using iQ SYBR Green supermix (Bio-Rad) [ 46 ]. Each experiment was conducted in three biological replicates, and the same sample was performed in three technical replicates. Relative expression levels of miRNAs and their target genes were quantified by using the 2 −ΔΔCt method [ 47 ]. All the primers used for qRT-PCR analysis are listed in Additional File 1: Table S4. 5’ modified RACE analysis A mixture of flower buds from the fertile lines (“6284B” and “4001B”) was used for total RNA isolation. The 5’ modified RACE was performed using a FirstChoice™ RLM-RACE Kit (Invitrogen, USA). Total RNA was directly ligated to the 5’RACE oligo. The first-strand cDNA synthesis and the two rounds of PCR reactions were conducted following the manufacturer’s instructions. The PCR products containing the target gene bands were ligated into pGEM-T Easy Vector (Promega, USA) for sequencing [ 25 ]. The primers are listed in Additional File 1: Table S4. Vector construction and plant transformation Two precursor sequences of bna-miR159a, which were located in A7 and C6 chromosomes of B. napus were designated as pre-miR159a-C6 and pre-miR159a-A7, respectively (Additional File 1: Table S5). The 411 nt and 470 nt genomic fragments containing pre-miR159a-C6 and pre-miR159a-A7 were amplified from B. napus using gene specific primers with endonuclease cleavage sites Sma I and Sal I. Then the fragments were cloned into pCAMBIA1301 binary vector with CaMV 35S promoter. The two vectors were designated as p35S::MIR159a-C6 and p35S::MIR159a-A7, which were introduced into Agrobacterium tumefaciens strains GV3101 and further transformed into Arabidopsis . The corresponding Arabidopsis transgenic plants were designated as MIR159OE-1 and MIR159OE-2. The transgenic and wild-type plants were cultivated in the greenhouse under the same environment. Their phenotypes were observed and recorded. The silique length data were collected from more than three plants for each transgenic line. Abbreviations GMS: Genic male sterility; RGMS: recessive genic male sterility; DGMS: dominant genic male sterility; CMS: cytoplasmic male sterility Declarations Ethics approval and consent to participate Not applicable Consent for publication Not applicable. Availability of Data and Materials The raw reads of the 24 sRNA libraries were uploaded to SRA database of NCBI and 24 accession numbers were obtained, including SRX11350295, SRX11350296, SRX11350307, SRX11350312, SRX11350313, SRX11350315, and SRX11350316 (https://dataview.ncbi.nlm.nih.gov/object/PRJNA743414?reviewer=t674c02cj415380e8oldre4s5a). The raw transcriptome data will also be further utilized to excavate differentially expressed mRNAs in the fertile and sterile lines of RGMS and DGMS line. Competing interests The authors declare that they have no competing interests. Funding This work was supported by grants from the Natural Science Foundation of China (Grant No. 31901502), and Shanghai Agriculture Applied Technology Development Program, China (Grant No.Z20160105). The former fund supported the design of the study and collection, including deep sequencing and data analysis. The second fund supported the writing and publishing of this manuscript. Authors' contributions J-XJ and L-YY conceived the project and research plan. J-XJ finished the manuscript. J-XJ, P-FX and Y-JL analyzed the sequencing data, performed the experiments and drafted the manuscript. X-RZ, J-YZ, and J-FZ planted and observed the DGMS plants. Y-LL, M-YJ, and W-RW planted and observed the RGMS plants. All authors have read and approved the final manuscript. Acknowledgements Not applicable Authors' information 1 Crop Breeding and Cultivation Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China. 2 National Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China. 3 School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China. References Kaul MLH. Male sterility in higher plants. Springer, Berlin Heidelberg, 1988 Hanson MR, Bentolila S. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-471166","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":38451121,"identity":"c11a5d1a-4fb5-4b56-8c6b-2cdf27c37bcd","order_by":0,"name":"Jianxia Jiang","email":"","orcid":"","institution":"Shanghai Academy of Agricultural Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jianxia","middleName":"","lastName":"Jiang","suffix":""},{"id":38451122,"identity":"5a7ba07a-8981-4b1b-aa97-5bf7585a7a92","order_by":1,"name":"Pengfei Xu","email":"","orcid":"","institution":"Chinese Academy of Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Pengfei","middleName":"","lastName":"Xu","suffix":""},{"id":38451123,"identity":"9d34a89a-825a-4afa-bb6d-1c7c837d6683","order_by":2,"name":"Yajie Li","email":"","orcid":"","institution":"Shanghai Jiao Tong University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yajie","middleName":"","lastName":"Li","suffix":""},{"id":38451124,"identity":"e3ea413c-1dba-4704-9c16-034912437726","order_by":3,"name":"Yanli Li","email":"","orcid":"","institution":"Shanghai Academy of Agricultural Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yanli","middleName":"","lastName":"Li","suffix":""},{"id":38451125,"identity":"515e0644-c582-4e35-bb3c-a1ff49e60628","order_by":4,"name":"Xirong Zhou","email":"","orcid":"","institution":"Shanghai Academy of Agricultural Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xirong","middleName":"","lastName":"Zhou","suffix":""},{"id":38451126,"identity":"351b0b7e-3501-44d2-aac0-ba29b8eab212","order_by":5,"name":"Meiyan Jiang","email":"","orcid":"","institution":"Shanghai Academy of Agricultural Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Meiyan","middleName":"","lastName":"Jiang","suffix":""},{"id":38451127,"identity":"1b4c1f00-4eb7-4eae-b024-83538bf6120a","order_by":6,"name":"Junying Zhang","email":"","orcid":"","institution":"Shanghai Academy of Agricultural Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Junying","middleName":"","lastName":"Zhang","suffix":""},{"id":38451128,"identity":"d8ba4d50-7cf1-4c69-a35c-4e9209d2cd6a","order_by":7,"name":"Jifeng Zhu","email":"","orcid":"","institution":"Shanghai Academy of Agricultural Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jifeng","middleName":"","lastName":"Zhu","suffix":""},{"id":38451129,"identity":"fd44f3a7-3902-4600-a334-ce049985c261","order_by":8,"name":"Weirong Wang","email":"","orcid":"","institution":"Shanghai Academy of Agricultural Sciences","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Weirong","middleName":"","lastName":"Wang","suffix":""},{"id":38451130,"identity":"39b1d359-fd07-43da-b3ee-7b0ea418eb7e","order_by":9,"name":"Liyong Yang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA+UlEQVRIiWNgGAWjYLCCBwYgkvkAhCdBjJYEsBa2BFK0gEkeA+K0GBw/e/hFQsFhOXP+NZ8//KipY5Cf3cD4uQCfljN5aRYJBoeNLWe83SbZc+wwg8GdA8zSM/BpOZBjZgDUkrjhxtltDDxsBxgMJBLYmHnwaTn/BqblzOOPf/4BHTaDkJYbOcYPwFrO9zBI87YxMzDcIKBF8sYbM2Agpxsb3GAzk5btO8xjcCOxWRqfFr7zOcYfPvyxljM4f/jxxzff6uTkZyQf/IxPi8IBBjZgRDQDoyMBLABUzNiARwMDg3wDA/MHBoY6Bgb+A3gVjoJRMApGwQgGAEdnUjf1Mh68AAAAAElFTkSuQmCC","orcid":"","institution":"Shanghai Academy of Agricultural Sciences","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Liyong","middleName":"","lastName":"Yang","suffix":""}],"badges":[],"createdAt":"2021-04-28 13:14:15","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-471166/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-471166/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":11392742,"identity":"cef86036-76bf-4f45-b9af-73232d1d7653","added_by":"auto","created_at":"2021-07-12 20:12:21","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":546341,"visible":true,"origin":"","legend":"Length distribution of sRNAs in flower buds of A line and B line libraries of DGMS and RGMS in Brassica napus. (A) Length distribution of sRNAs in 4001A and 4001B. (B) Length distribution of sRNAs in 4006A and 4006B. (C) Length distribution of sRNAs in 6251A and 6251B. (D) Length distribution of sRNAs in 6284A and 6284B.","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/88d1a3a711c545787e032c51.png"},{"id":11392746,"identity":"0493bc93-cf25-4aa7-9b9a-a01824ea6455","added_by":"auto","created_at":"2021-07-12 20:12:22","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":1615897,"visible":true,"origin":"","legend":"Relative expression analysis of miRNAs in the flower buds of the A line and B line of RGMS and DGMS in Brassica napus by high-throughput sequencing. Six miRNAs (bule columns) showed significant expression differences between ‘4001A’ and ‘4001B’ flower buds. Two miRNAs (orange columns) showed significant expression differences between ‘4006A’ and ‘4006B’ flower buds. Six miRNAs (red columns) showed significant expression differences between ‘6251A’ and ‘6251B’ flower buds. Fourteen miRNAs (gray columns) showed significant expression differences between ‘6284A’ and ‘6284B’ flower buds. The Y-axis represents the differentially expressed miRNAs. The X-axis represents the fold change of relative abundance between the flower buds of A line and B line. Left indicates the miRNAs enriched in B line flower buds. Right indicates the miRNAs enriched in A line flower buds.","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/8b1cf51dedf3fa60ebd0fb91.png"},{"id":11392749,"identity":"ca9b9733-7db1-4e58-9a32-c7e76d9aca4a","added_by":"auto","created_at":"2021-07-12 20:12:22","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":348853,"visible":true,"origin":"","legend":"Venn diagram showing overlaps of significantly differential expressed miRNAs between different genic male sterility (GMS) lines of Brassica napus. (A) Two differentially expressed miRNAs were shared between 4001AB and 4006AB lines. (B) Four differentially expressed miRNAs were shared between 6251AB and 6284AB lines.","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/929241c10cd81187d21428eb.png"},{"id":11392763,"identity":"da20aca2-57bf-4571-abc1-f94b58db5647","added_by":"auto","created_at":"2021-07-12 20:15:21","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1000785,"visible":true,"origin":"","legend":"The qRT-PCR analysis of differentially expressed miRNAs between the flower buds of A lines and B lines.","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/f86a1259ca6b806d0f502b42.png"},{"id":11392747,"identity":"29da976b-2806-46c7-a236-6340df82cd63","added_by":"auto","created_at":"2021-07-12 20:12:22","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":424842,"visible":true,"origin":"","legend":"Identification of miRNA target genes by 5’ modified RACE.","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/1e5e094cb2be2a8e2f73a3b9.png"},{"id":11392745,"identity":"402dd3ec-95ca-422a-989f-b3a6ced482b5","added_by":"auto","created_at":"2021-07-12 20:12:22","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":1153844,"visible":true,"origin":"","legend":"The relative expression levels of miR159 and its targets in wild type plants of Arabidopsis.","description":"","filename":"Figure6.png","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/7884254b95574edb898d4b74.png"},{"id":11392748,"identity":"dcfd0efa-7cab-456c-af59-d3fdb6f0cf38","added_by":"auto","created_at":"2021-07-12 20:12:22","extension":"png","order_by":7,"title":"Figure 7","display":"","copyAsset":false,"role":"figure","size":1197711,"visible":true,"origin":"","legend":"The relative expression levels of miR159 and its targets in miR159-OE transgenic lines of Arabidopsis.","description":"","filename":"Figure7.png","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/eacd37523b321b5485fd4a84.png"},{"id":11392750,"identity":"885cf762-a14f-4720-abd3-3fd00ea64667","added_by":"auto","created_at":"2021-07-12 20:12:22","extension":"png","order_by":8,"title":"Figure 8","display":"","copyAsset":false,"role":"figure","size":7668804,"visible":true,"origin":"","legend":"The phenotypes of miR159-OE transgenic lines in Arabidopsis.","description":"","filename":"Figure8.png","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/ff282f6ede18c969c2a39540.png"},{"id":15673944,"identity":"41a87f7d-73e3-4953-b2db-0336d36b03c8","added_by":"auto","created_at":"2021-11-18 14:20:15","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3332574,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/a7b59869-9bf6-4fbb-9e98-c199dfa37797.pdf"},{"id":11392743,"identity":"aa45b916-34c2-406e-a7e5-20c47b8cd618","added_by":"auto","created_at":"2021-07-12 20:12:21","extension":"docx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":37315,"visible":true,"origin":"","legend":"","description":"","filename":"Additionalfile1.docx","url":"https://assets-eu.researchsquare.com/files/rs-471166/v1/39991ec150ee2adeabbe3889.docx"}],"financialInterests":"No competing interests reported.","formattedTitle":"\u003cp\u003eIdentification of miRNAs And Their Target Genes In Genic Male Sterility Lines In \u003cem\u003eBrassica Napus\u003c/em\u003e By Small RNA Sequencing\u003c/p\u003e","fulltext":[{"header":"Background","content":"\u003cp\u003eMale sterility is largely due to the abnormal development of either the sporophytic or gametophytic anther tissues. According to its origin or genetic analysis, male sterility can be divided into cytoplasmic male sterility (CMS) and genic male sterility (GMS) [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. CMS is caused by the interaction between the mitochondrial genome and nuclear genes [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. GMS is derived from natural mutations in nuclear genes that control stamen development. GMS mutants always show advantages, such as complete and stable male sterility and no potential negative cytoplasmic effect, compared with most CMS mutants. GMS can be further divided into dominant GMS (DGMS) and recessive GMS (RGMS). In rapeseed, DGMS and RGMS are widely used for hybrid rapeseed production [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. Multiple-allele DGMS was generally accepted and was proposed by Song et al. in 2005. This model presented multiple alleles in one locus inheritance to explain the fertility heredity of a newly reported DGMS line 609AB. In this model, \u003cem\u003eMf\u003c/em\u003e, \u003cem\u003eMs\u003c/em\u003e, and \u003cem\u003ems\u003c/em\u003e are three alleles at the same locus, with a relationship of \u003cem\u003eMf\u003c/em\u003e dominant over \u003cem\u003eMs\u003c/em\u003e and \u003cem\u003eMs\u003c/em\u003e over \u003cem\u003ems\u003c/em\u003e. The recessive allele is for normal fertility. The maintainers and restorers are easily screened. Thus, multiple-allele DGMS are widely used for hybrid rapeseed seed production through the construction of a three-line hybrid system [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eExcept for the advantages of GMS, the recessive GMS (RGMS) systems have another distinct advantage in that most inbred lines can restore their fertility, so that hybrids with strong heterosis are easily bred. Several RGMS lines have been successfully commercialized in China, including S45AB [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e], 117AB [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], and 9012AB [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. For S45AB and 117AB, approximately 50% of the fertile plants are required to be artificially removed before hybridizing the rest (50% sterile ones) with restorer lines in hybrid production, because no complete maintainer line is available [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. However, a three-line hybrid production system was developed for 9012AB [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], and this system has been well documented [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. The RGMS line 9012AB has been used successfully for rapeseed hybrid production in China. This male sterility was previously thought to be controlled by three independent genes (\u003cem\u003eBnMs3\u003c/em\u003e, \u003cem\u003eBnMs4\u003c/em\u003e, and \u003cem\u003eBnRf\u003c/em\u003e). In 2012, Dong et al. demonstrated a major modification of the sterility inheritance model in 9012A. The modified inheritance model indicated that the male sterility was essentially controlled by two loci (\u003cem\u003eBnMs3\u003c/em\u003e and \u003cem\u003eBnRf\u003c/em\u003e). The previously designated \u003cem\u003eBnMs4\u003c/em\u003e locus was just one allele of \u003cem\u003eBnRf\u003c/em\u003e; it was then designated as \u003cem\u003eBnRf\u003c/em\u003e\u003csup\u003e\u003cem\u003ea\u003c/em\u003e\u003c/sup\u003e, which was designated in addition to \u003cem\u003eBnRf\u003c/em\u003e\u003csup\u003e\u003cem\u003eb\u003c/em\u003e\u003c/sup\u003e (the allele from 9012A) and \u003cem\u003eBnRf\u003c/em\u003e\u003csup\u003e\u003cem\u003ec\u003c/em\u003e\u003c/sup\u003e (the allele from temporary maintainer). The dominance relationship of the three alleles is in the following order: \u003cem\u003eBnRf\u003c/em\u003e\u003csup\u003e\u003cem\u003ea\u003c/em\u003e\u003c/sup\u003e\u0026gt;\u003cem\u003eBnRf\u003c/em\u003e\u003csup\u003e\u003cem\u003eb\u003c/em\u003e\u003c/sup\u003e\u0026gt;\u003cem\u003eBnRf\u003c/em\u003e\u003csup\u003e\u003cem\u003ec\u003c/em\u003e\u003c/sup\u003e. The \u003cem\u003eBnRf\u003c/em\u003e allele-specific molecular markers were identified; these markers would simplify the breeding process involving this RGMS line [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eOilseed rape (\u003cem\u003eBrassica napus\u003c/em\u003e, 2n\u0026thinsp;=\u0026thinsp;38, AACC), which has low erucic acid and glucosinolate contents, is the third leading source of edible oil worldwide. In recent years, some conserved and novel miRNAs associated with silique length [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e], thickness of pod canopy [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], cadmium stress [\u003cspan additionalcitationids=\"CR18\" citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e], flower organ development [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e], seed maturation [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e], cold stress [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e], seed development, and oil synthesis [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] have been widely identified in rapeseed. However, little information is available about the DGMS and RGMS occurrence at the post-transcriptional level in rapeseed.\u003c/p\u003e \u003cp\u003eIn this study, to systematically explore the roles of miRNAs and their targets involved in GMS occurrence during pollen development in rapeseed, 12 small RNA and transcriptome libraries were constructed and sequenced for the flower buds from the fertile and sterile lines of two RGMS lines (\u0026ldquo;6251AB\u0026rdquo; and \u0026ldquo;6284AB\u0026rdquo;). Meanwhile, 12 small RNA and transcriptome libraries were also constructed and sequenced for the flower buds from the fertile and sterile lines of two DGMS lines (\u0026ldquo;4001AB\u0026rdquo; and \u0026ldquo;4006AB\u0026rdquo;). The aims of this study were to identify known and potential novel miRNAs from the 24 libraries and to analyze the expression profiles of the miRNAs and their targets in relation to DGMS and RGMS during rapeseed microspore development. The results would provide a foundation for evaluating the important regulatory roles of miRNAs in pollen formation and GMS occurrence in rapeseed and other crops.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv class=\"Section2\" id=\"Sec3\"\u003e\n \u003cp\u003e\u003cstrong\u003eAnalysis of small RNA library data sets and the small RNA profile\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eTo identify miRNAs related to DGMS and RGMS during pollen development, the flower buds were collected from the sterile (6251A, 6284A) and fertile (6251B and 6284B) lines of the RGMS lines. Meanwhile, the flower buds were also respectively collected from the sterile (4001A, 4006A) and fertile (4001B and 4006B) lines of the DGMS lines. Three biological replicates were conducted for each of the eight kinds of samples. Thus, total of 24 sRNA libraries were constructed and deep-sequenced. The raw reads of the 24 sRNA libraries ranged from 20.58 to 42.68 million (Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e). The raw reads of the 24 sRNA libraries were uploaded to SRA database of NCBI and 24 accession numbers were obtained, including SRX11350295, SRX11350296, SRX11350307, SRX11350312, SRX11350313, SRX11350315, and SRX11350316 (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://dataview.ncbi.nlm.nih.gov/object/PRJNA743414?reviewer=t674c02cj415380e8oldre4s5a\u003c/span\u003e\u003c/span\u003e). After removing the low-quality reads and contaminated adapter sequences, the clean reads of the 24 sRNA libraries ranged from 19.77 to 41.61 million. The mapped reads were further annotated against the Pfam database and subsequently divided into rRNAs, tRNAs, snRNAs, snoRNAs, ta-siRNA, and others. The endogenous sRNAs were identified as known and novel miRNAs. The average sRNA lengths of the three biological replicates for each sample were calculated, which showed the length distribution patterns of the sRNAs being similar to one another. In general, the majority of the small RNAs ranged from 21 nt to 24 nt in size. The 24 nt small RNAs were the most dominant, followed by 21 nt small RNAs (Figure. 1).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 1 Overview of sRNA sequencing reads in \u003cem\u003eBrassica napus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003ctable align=\"left\" border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003eSample\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003eRaw reads\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003eN% \u0026gt; 10%\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003eLow quality\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e5\u003csup\u003e,\u0026nbsp;\u003c/sup\u003eadapter contamine\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e3\u003csup\u003e,\u003c/sup\u003e adapter null or insert null\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003eWith ployA/T/G/C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003eClean reads\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e4001B-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e24481718\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e41\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e16955\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e8590\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e761698\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e14436\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e23679998\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e4001B-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e27695084\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e70\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e36718\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e12896\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e698693\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e16785\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e26929922\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e4001B-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e20576690\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e16651\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e10078\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e768766\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e9577\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e19771580\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e4006B-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e28457208\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e131\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e44382\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e14481\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e721967\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e14232\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e27662015\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e4006B-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e27355185\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e7473\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e9466\u003c/p\u003e\n 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width=\"13.68421052631579%\"\u003e\n \u003cp\u003e1597289\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e18301\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e34184929\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e6251B-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e22307540\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e18341\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e18038\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n 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width=\"9.473684210526315%\"\u003e\n \u003cp\u003e16924\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e23956096\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e6284B-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e25552984\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e12764\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e21674\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e567970\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n 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width=\"13.68421052631579%\"\u003e\n \u003cp\u003e23784756\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e4001A-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e21968633\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e16757\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e10355\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e723740\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e8680\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n 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width=\"12.631578947368421%\"\u003e\n \u003cp\u003e4006A-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e42675389\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e50\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e10345\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e14879\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e1018244\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e21106\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e41610765\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e4006A-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e27452096\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e93\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e40026\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e12496\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e749916\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e14657\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e26634908\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e6251A-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e21484268\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e17038\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e20165\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e462483\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e17972\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e20966591\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e6251A-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e24127081\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e12533\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e20730\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e464608\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e24999\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e23604155\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e6251A-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e21608183\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e357\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e7763\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e13126\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e393936\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e10376\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e21182625\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e6284A-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e23525443\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e11236\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e20073\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e746946\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e12136\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e22735025\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e6284A-2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e27605112\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e14315\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e23792\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e1211985\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e12779\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e26342204\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"12.631578947368421%\"\u003e\n \u003cp\u003e6284A-3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.736842105263158%\"\u003e\n \u003cp\u003e29689151\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.421052631578947%\"\u003e\n \u003cp\u003e56\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.578947368421053%\"\u003e\n \u003cp\u003e30879\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.789473684210526%\"\u003e\n \u003cp\u003e21998\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e753037\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.473684210526315%\"\u003e\n \u003cp\u003e16533\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.68421052631579%\"\u003e\n \u003cp\u003e28866648\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003cstrong\u003eIdentification of known and novel miRNAs in\u003c/strong\u003e \u003cspan class=\"BoldItalic\" name=\"Emphasis\" type=\"BoldItalic\"\u003e\u003cstrong\u003e\u003cem\u003eB. napus\u003c/em\u003e\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp\u003eTo identify known miRNAs in \u003cem\u003eB. napus\u003c/em\u003e, all mapped small RNA sequences were compared with the known mature bna-miRNA sequences deposited in the miRBase database 22.1. Forty-six small RNAs that have the same sequences with the known bna-miRNAs in miRBase were identified. The numbers of reads of the 46 known miRNAs in 24 libraries were listed in Additional Table S1. Among the 46 known miRNAs, bna-miR159a, bna-miR166a, and bna-miR167c showed very high expression levels.\u003c/p\u003e\n \u003cp\u003eTo predict novel miRNAs in \u003cem\u003eB. napus\u003c/em\u003e, all the mapped small RNAs were blasted to the \u003cem\u003eB. napus\u003c/em\u003e genome sequence in \u003cem\u003eBrassica\u003c/em\u003e database and known plant miRNAs in miRBase. The small RNAs that exactly map to the genome sequence but not the known plant miRNAs were classified as candidate novel miRNAs. Five criteria described in the Materials and Methods were used to search for novel miRNAs. As a result, 35 pairs of novel miRNA-3p/miRNA-5p were identified. The mature sequences, reads numbers, positions in chromosomes, precursor sequences and minimum free energy are listed in Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e. The length distribution of the novel miRNAs was between 18 nt to 26 nt. The length of the novel miRNA precursors ranged from 51 nt to 300 nt with an average length of 154 nt. The minimum free energy ranged from \u0026minus;\u0026thinsp;240.26 to -9.9 kcal mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e with an average of -70.14 kcal mol\u003csup\u003e\u0026minus;\u0026thinsp;1\u003c/sup\u003e. The precursor sequences of the novel miRNA are shown in Additional Tables S2.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 2 Novel miRNAs identified in the fertile and sterile lines of \u003cem\u003eBrassica napus\u003c/em\u003e by high-throughput sequencing\u003c/strong\u003e\u003c/p\u003e\n \u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003emiR_ name\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eSequence\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003eLen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003eRead\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003epre-position\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003epre-len\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003eMFE\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_1-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eCUUCCUCCUAACACCAAUUGAUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA09:27889716..27889818\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e102\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-34.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_1-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUCAAUUGGUUUUAGGUUAAGAAGCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e124\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_2-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUGGCAUUGGUAGUAAUGAGUGU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e190\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC04:42905032..42905108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-26.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_2-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eACUCAUUACCAUCAGAGCCAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_3-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUCAAUGUUGGCUCAAUUAUGU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e120\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC02:22515980..22516065\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e85\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-29.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_3-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUCAUUGAGUGCAGCGUUGAUGU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_4-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUUAUCGACACUGAUCUCAUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e106\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC08:2975781..2975915\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e134\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-80.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_4-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUAAGGUCACUGUGGUAAUCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_5-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUCAAUGUUGGCUCAAUUAUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e49\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC09:40959623..40959707\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-31.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_5-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUCAUUGAGUGCAGCGUUGAUGU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_6-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAAGGACUCUAAUCAGAAAUAUUGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e143\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC06:35249139..35249190\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e51\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-9.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_6-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAAUGGUCUUAUCUGGAAUCCUUAA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_7-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUGCCUGGCUCCCUGUAUACCA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA08:8293061..8293144\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e83\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-32.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_7-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eGUGUAUAGAGUAGUCAAGCAUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_8-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUCUCUAAUGUAUAACUCCAUUUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA03:20065617..20065865\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e248\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-95.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_8-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAAUGGAGUAGAUAUGGAGAUGCCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_9-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUGGACUGAAGGGAACUCCCU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e1527\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA09:14645700..14645869\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e169\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-64.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_9-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAGAGUUUCCUUAAGUCCAUUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_10-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUAAGAUCUUUGUACUUUCGGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e67\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA10:15442817..15442916\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e99\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-39.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_10-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eCGAAAGUACAAAGAUCUGAAA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_11-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAACAGUUGGAUUGGCUCUACGUGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA09_random:3088720..3089020\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e300\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-65.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_11-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eACGAUGGAGGACAAAACUGAUGCA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_12-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUUUCAGCAAUCUCUUUUCCAUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA05_random:416209..416319\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e110\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-29.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_12-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUGGGAAAGAUUGUUGAUCAGA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_13-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUAAAGUAGAGCUCGGUGACGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e1163\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC03:20992442..20992727\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e285\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-240.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_13-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eGUCACCGAGCUCUACUUUAUA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e1058\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_14-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUCGCUUCUGUUGAAUAAUUUUGAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC04:45707769..45708016\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e247\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-148\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_14-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eCAAAAUUAUUCAACAGAAGCGAAU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_15-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUAUGAGGGUACAAUAGGAAG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e137\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrAnn_random:33598332..33598540\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e208\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-139.1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_15-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUAUUGUACCCUCAUAUAUAGC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_16-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eCUAAGAGAUCUGUAAUAAACAUGC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC04:7124159..7124378\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e219\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-117.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_16-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUGUUUAUUGUAGGUCUUUUAGGUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_17-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eACGAACACUGAGUAAUAUCUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC01_random:3928762..3929012\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e250\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-164.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_17-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eGAUAUUACUCAGUGUUCGUUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_18-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eACACUGCAGUGCACUGUACAUUGC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrCnn_random:79954335..79954585\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e250\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-113.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_18-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eGUUGUACAUUGUACACAGCGGUGUAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_19-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUGCAAACUGAAUUAUGAGUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA09:30966144..30966235\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-45.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_19-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eCUCAUAAUUCAGUUUGCAAUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_20-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAAGAUACGGUCUCUUAACUUUUAG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e259\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC04_random:3818028..3818149\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e121\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-67.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_20-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eGUUAAUAGACCGUAUCUUAUA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_21-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAACGAUCUUGUUUGGUUUUGAAGA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA05:21620032..21620188\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e156\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-82.1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_21-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUCAAAACCAUACAAGAUCGUUUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_22-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eGAUCAUGUUCGUAGUUUCACC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e445\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrCnn_random:35712007..35712108\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e101\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-47.3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_22-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUGAAGCUGCCAGCAUGAUCU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_23-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUCUUGUGCGUUUAUAGGUAG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e55\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA06:23040126..23040236\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e110\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-52.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_23-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eACCUCUAAAACACACAAGAAGA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_24-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUGUUUCGCUGUUACUCAUGC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e40\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC02:8973302..8973545\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e243\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-93.1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_24-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUGAGUAACAGCGAAACAAA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_25-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAAACUGUGUGAACUCUCCAUGGAG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e389\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC02_random:2214651..2214881\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e230\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-73.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_25-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eCCAUAAAAAGGGUUCACAAAGUAUUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_26-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUGAUACAUGUAGCUCUUUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e2089\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA03:669098..669269\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e171\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-83.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_26-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAAGUGCUACCGGUAUCCACGUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e840\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_27-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUAAUCGUUUUGUGACUCUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e244\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA07:19146929..19147018\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e89\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-34.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_27-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUAGUUACAAAACGAUUAGUGC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_28-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUCAACGUUGGCUCAAUUAUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e453\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA10_random:1861693..1861781\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e88\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-33.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_28-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUCAUUGAGUGCAGCGUUGAUGU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_29-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUCUUGUUACUGAGCUCGACG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e308\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA02:7209993..7210285\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e292\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-99.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_29-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUCAGCUGGGUACGAGCCACC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e710\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_30-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUCUGCAUCGAGUGAACUCUAUGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e426\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrCnn_random:65177977..65178226\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e249\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-72.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_30-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAUGGAAUUCACUGAUGCAGAUGCU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_31-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUCUUGUGGUUGUAGAGUCUUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e367\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA06:4069612..4069740\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e128\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-56.1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_31-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eAGACUCUACAACAUCAGAAAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e47\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_32-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eCGGAUUUUAGCUGCGUAGCUA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e322\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrAnn_random:44030328..44030409\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-42.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_32-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eGGCUACGCUGCUGAAUCCGC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_33-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUGUAGAAUUUUGGGAAGGGC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e289\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC05_random:138762..138826\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-32.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_33-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eCCUUCCCAAAAUUCUACAAUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_34-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eACUUUGAAACUUUGAUCUAGA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e5292\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrC06:5179422..5179524\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e102\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-42.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_34-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUAGAUCAAAGCUUUAAUGU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_35-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eUUUUCGAUCUGUAAAUUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"8.333333333333334%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"18.75%\"\u003e\n \u003cp\u003echrA03:11978303..11978381\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"9.375%\"\u003e\n \u003cp\u003e78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"10.416666666666666%\"\u003e\n \u003cp\u003e-30.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"22.916666666666668%\"\u003e\n \u003cp\u003ebna-novel_35-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"23.958333333333332%\"\u003e\n \u003cp\u003eCAUUUACAGAUCGAAGACAUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"5.208333333333333%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"28.125%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"7.291666666666667%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"6.25%\"\u003e\n \u003cp\u003e \u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003emiR_name, miRNA name; Len, length of mature miRNA; pre-position, the position of miRNA precursor sequences in chromosomes of \u003cem\u003eBrassica napus\u003c/em\u003e, pre-len, length of miRNA precursor sequences; MFE, minimum folding free energy.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eNovel miRNA on the other arm of known pre-miRNA\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThrough sRNA high-throughput sequencing, miRNA-3p and miRNA-5p were found to always be simultaneously present on the 5\u0026prime; arm and 3\u0026prime; arm of pre-miRNA secondary structures. To identify novel miRNAs on the other arm of known pre-miRNAs, all mapped small RNA sequences were compared to the known precursor sequences of bna-miRNAs in the miRBase database 22.1. Finally, 27 novel miRNAs on the other arm of known \u003cem\u003eB. napus\u003c/em\u003e pre-miRNAs were identified. The miRNA sequences and the number of reads in 24 libraries are listed in Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 3 Identification of novel miRNAs on the other arm of known pre-miRNAs in \u003cem\u003eBrassica napus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"97%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003e\u003cstrong\u003ebna-miRNA name\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003e\u003cstrong\u003emature miRNA sequence\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e\u003cstrong\u003eLength\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e\u003cstrong\u003eRead count\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR156f-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGCUCACUGCUCUUUCUGUCAGA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e749\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR156e-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eUGCUCACCUCUCUUUCUGUCAGU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e344\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR160a-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGCGUAUGAGGAGCCAUGCAUA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e32\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR160c-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGCGUACAGAGUAGUCAAGCAUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR160d-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGCGUACAGAGUAGUCAAGCAUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR161-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGUCACUUUCAAUGCGUUGAUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR164b/c/d-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eCACGUGUUCUACUACUCCAAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR166d-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGGACUGUUGUCUGGCUCGAGG\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e135\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR166e-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGGAAUGUUGUCUGGCACGAGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR166f-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGGAAUGUUGUCUGGAUCGAGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e202\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR167a/b-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGAUCAUGUUCGCAGUUUCACC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e750\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR167a/b-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGAUCAUGUUCGCAGUUUCACC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e750\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR168a-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eCCCGCCUUGUAUCAAGUGAAU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e104\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR171a/b/c-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eAGAUAUUAGUGCGGUUCAAUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e51\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR171d-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eAGAUAUUGGUGCGGUUCAAUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR172a-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGCAGCACCAUCAAGAUUCAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e48\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR172b-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGCAGCAUCAUUAAGAUUCACA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR172c-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGCAGCAUCAUCAAGAUUCACA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR172d-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGCAUCAUCAUCAAGAUUCAGA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e218\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR2111d-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eAUCCUCGGGAUACGGAUUACC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR390b-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eCGCUGUCCAUCCUGAGUUUCA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e1109\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR390c-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eCGCUAUCCAUCCUGAGUUCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR395a/b/c-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGUUCCUCUGAGCACUUCAUUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e61\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR395d/f-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGUUCCCUUUAACGCUUCAUUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR399b-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eGGGCAAGAUCUCUAUUGGCAGG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR403-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eUGUUUUGUGCGUGAAUCUAAUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e287\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"26.804123711340207%\"\u003e\n \u003cp\u003ebna-miR824-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"50.51546391752577%\"\u003e\n \u003cp\u003eCCUUCUCAUCGAUGGUCUAGA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.34020618556701%\"\u003e\n \u003cp\u003e1640\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003e\u003cstrong\u003eIdentification of new conserved miRNA families and new miRNA members\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eTo identify new conserved miRNAs in \u003cem\u003eB. napus\u003c/em\u003e, all mapped small RNAs were mapped to known plant miRNAs in miRBase and \u003cem\u003eB. napus\u003c/em\u003e genome sequences. If the small RNAs can match known plant miRNAs with no more than three mismatches and can exactly map to \u003cem\u003eB. napus\u003c/em\u003e genome sequences, then these small RNAs were initially classified as candidate new conserved miRNAs. Five criteria described in the Materials and Methods were used to strictly screen the candidate conserved miRNAs. As a result, 44 miRNAs (22 pairs of miRNAs) belonging to 15 miRNA families were identified (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e). Among them, bna-miR159b was a new miRNA member of bna-miR159 family. The rest of the 36 miRNAs (14 pairs of miRNAs) have not been previously reported as bna-miRNAs in miRBase; they show high sequence similarity to some of the known plant miRNAs. The bna-miR158a.1 and bna-miR158a.2 were identified for bna-miR158a member. The two pairs of bna-miR158a shared the same mature sequences. Their precursor sequences were highly similar with each other, and these sequences were from different loci of the \u003cem\u003eB. napus\u003c/em\u003e genome. These two pairs of miRNAs were called sub-members. This type of sub-member was also observed for bna-miR159b and bna-miR408a. Four sub-members (bna-miR159b.1, bna-miR159b.2, bna-miR159b.3, and bna-miR159b.4) were identified for bna-miR159b, and two sub-members (bna-miR408a.1 and bna-miR408a.2) were identified for bna-miR408a. This phenomenon suggests that some \u003cem\u003eMIRNA\u003c/em\u003e genes might be produced through a replication event from one origin to another one, which results in more copies of the miRNA group. Two members were identified for bna-miR319 and bna-miR398 families. Except the above mentioned five miRNA families, the rest of 10 miRNA families had only one miRNA member (Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u0026nbsp;\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 4 Identification of new conserved miRNA families in \u003cem\u003eBrassica napus\u003c/em\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"100%\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003e\u003cstrong\u003ebna-miRNA\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSequence\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e\u003cstrong\u003eLen\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e\u003cstrong\u003eRead\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003e\u003cstrong\u003epre-position\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR158a.1-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eCUUUGUCUAUCGUUUGGAAAAG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e3884\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA08:2748114..2748220\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR158a.1-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUUCCAAAUGUAGACAAAGCA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e32292\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR158a.2-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eCUUUGUCUAUCGUUUGGAAAAG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e3884\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrC08:3581242..3581348\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR158a.2-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUUCCAAAUGUAGACAAAGCA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e32292\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR159b.1-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eAGCUGCUAAGCUAUGGAUCCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e258\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA02:9865184..9865001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR159b.1-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUUGGAUUGAAGGGAGCUCUA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e46073\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR159b.2-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eAGCUGCUAAGCUAUGGAUCCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e258\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA07_random:1944377..1944191\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR159b.2-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUUGGAUUGAAGGGAGCUCUA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e46073\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR159b.3-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eAGCUGCUAAGCUAUGGAUCCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e258\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrC02:19215807..19215624\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR159b.3-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUUGGAUUGAAGGGAGCUCUA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e46073\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR159b.4-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eAGCUGCUAAGCUAUGGAUCCC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e258\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrC06:33954934..33954749\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR159b.4-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUUGGAUUGAAGGGAGCUCUA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e46073\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR319a-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eAGAGCUUCCUUGAGUCCAUUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrC01:10651723.. 10651921\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR319a-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUGGACUGAAGGGAGCUCCCU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e4848\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR319b-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eGGAGAUUCUUUCAGUCCAGUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrC04:46407584.. 46407846\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR319b-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUGGACUGAAGGGAGCUCCUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e27901\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR391-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUCGCAGGAGAGAUAGCGCCA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e110\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA10:10707678..10707812\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR391-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eACGGUAUCUCUCCUACGUAGC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e237\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR398a-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eGGGUCGACAUGAGAACACAUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e141\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA03:2288822..2288945\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR398a-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUGUGUUCUCAGGUCACCCCUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e9870\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR398b-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eGGAGUGUCAUGAGAACACGGA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrC02:37793584..37793689\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR398b-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUGUGUUCUCAGGUCACCCCUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e145\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003e\u0026nbsp;bna-miR400-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUAUGAGAGUAUUAUAAGUCAC\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e78\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrAnn_random:40582790..40582930\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003e\u0026nbsp;bna-miR400-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eGACUUAUAAUGAUCUCAUGAA\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e237\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003e\u0026nbsp;bna-miR408a.1-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eGGGAGCCAGGGAAGAGGCAGU\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e1232\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA05:478954..479121\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003e\u0026nbsp;bna-miR408a.1-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUGCUUGUUCCCUGUCUCUCUC\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e1002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003e\u0026nbsp;bna-miR408a.2-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eGGGAGCCAGGGAAGAGGCAGU\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e1232\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrCnn_random:8448205..8448064\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003e\u0026nbsp;bna-miR408a.2-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUGCUUGUUCCCUGUCUCUCUC\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e1002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9554-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eGAAUGAUACUUGGAUAUAAUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA06:19718101..19718250\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9554-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUCAUAUCCAAGUAUCAUUCCU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9558-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eAGAGAUGUCUGGCUUGCAACA\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrC03_random:1702602..1702746\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9558-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUGCAAGCCAGACAUUUCCUUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9559-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUUGGAUUUUGGUCAUUGUUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrAnn_random:36404086.. 36404194\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9559-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eACAAUGAACGAAAUCCAAAUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9560a-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eACAGGUGGUGGAACAAAUAUGAGU\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA06:19552830..19552965\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9560a-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUCAUAUUAGUUCUACCUCCUGCUG\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9562-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eACUAUGCAAUUGUGAACAAAC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA02_random:1408210..1408358\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9562-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUUAUUCACAACUGCAUAAUUC\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9563a-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eACCCGUCUCUUAACUUUUAAC\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrAnn_random:9932700..9932850\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9563a-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUAAAAGUUAAGAGACAAGUUA\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9568-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUGCGGAUAUCUUAGGAUGAGGU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrA03:13274664..13274813\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9568-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUCAUCGUAAGAGAUCUGCAUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9569-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eUGAGUUAUCAUUGGUCUUGUG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e1198\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\n \u003cp\u003echrAnn_random:21855323..21855514\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"24.742268041237114%\"\u003e\n \u003cp\u003ebna-miR9569-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"31.95876288659794%\"\u003e\n \u003cp\u003eACACAGGAACAAUACUAACUCAUU\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"8.24742268041237%\"\u003e\n \u003cp\u003e24\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.278350515463918%\"\u003e\n \u003cp\u003e3509\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"25.77319587628866%\"\u003e\u003cbr\u003e\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003eLen, length of mature miRNA;\u0026nbsp;pre-position, the miRNA precursor sequences in chromosomes of \u003cem\u003eBrassica napus\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eExpression profiling of differentially expressed miRNAs in sterile and fertile lines\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eThe normalized expression levels of miRNAs were used for identifying differentially expressed miRNAs between the sterile line and the corresponding fertile line, such as \u0026ldquo;4001A\u0026rdquo; and \u0026ldquo;4001B\u0026rdquo;, \u0026ldquo;4006A\u0026rdquo; and \u0026ldquo;4006B\u0026rdquo;, \u0026ldquo;6251A\u0026rdquo; and \u0026ldquo;6251B\u0026rdquo;, and \u0026ldquo;6284A\u0026rdquo; and \u0026ldquo;6284B\u0026rdquo;. The known, identified conserved and novel miRNAs were followed to differential expression analysis criteria (qvalue\u0026thinsp;\u0026lt;\u0026thinsp;0.01 and |log2 (fold change)| \u0026gt; 0.73). As a result, 6, 2, 6, and 14 differentially expressed miRNAs were obtained between the flower buds of \u0026ldquo;4001A\u0026rdquo; and \u0026ldquo;4001B\u0026rdquo;, \u0026ldquo;4006A\u0026rdquo; and \u0026ldquo;4006B\u0026rdquo;, \u0026ldquo;6251A\u0026rdquo; and \u0026ldquo;6251B\u0026rdquo;, and \u0026ldquo;6284A\u0026rdquo; and \u0026ldquo;6284B\u0026rdquo;, respectively (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, Tables \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e). To further explore the miRNAs involved in the two DGMS lines, a Venn diagram analysis was conducted. The results indicated that two differentially expressed miRNAs were shared between the DGMS lines \u0026ldquo;4001AB\u0026rdquo; and \u0026ldquo;4006AB\u0026rdquo; (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eA). In addition, four differentially expressed miRNAs were shared between the RGMS lines \u0026ldquo;6251AB\u0026rdquo; and \u0026ldquo;6284AB\u0026rdquo; (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eB). The novel miRNA \u0026ldquo;bna-novel_34-5p\u0026rdquo; was the only miRNA that was simultaneously differentially expressed in the DGMS and RGMS lines.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTable 5 Differentially expressed miRNAs in \u0026ldquo;4001A\u003c/strong\u003e\u003cstrong\u003eB\u0026rdquo;, \u0026ldquo;4006AB\u0026rdquo;, \u0026ldquo;6251AB\u0026rdquo; and \u0026ldquo;6284AB\u0026rdquo; libraries and their candidate targets by sRNA sequencing and transcriptome analysis in \u003cem\u003eBrassica napus\u003c/em\u003e\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"2.6975863700899194%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSample\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"7.288215806909607%\"\u003e\n \u003cp\u003e\u003cstrong\u003emiR\u003c/strong\u003e\u003cstrong\u003e_name\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e\u003cstrong\u003eA\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e\u003cstrong\u003eB\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.685281590156176%\"\u003e\n \u003cp\u003e\u003cstrong\u003elog\u003csub\u003e2\u003c/sub\u003e\u003csup\u003e(FoldChange)\u003c/sup\u003e\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"80.88026502602935%\"\u003e\n \u003cp\u003e\u003cstrong\u003eCandidate\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eTargets\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"5\" valign=\"top\" width=\"2.6975863700899194%\"\u003e\n \u003cp\u003e4001\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"7.288215806909607%\"\u003e\n \u003cp\u003ebna-novel_4-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e8.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.685281590156176%\"\u003e\n \u003cp\u003e1.90\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"80.88026502602935%\"\u003e\n \u003cp\u003eBnaC01g31810D,BnaA07g24350D,BnaC02g41520D,BnaA08g22150D,BnaA04g13020D,BnaA07g18960D,BnaA09g38810D,BnaC03g26470D,BnaC02g03200D,BnaA04g00990D,BnaA03g41600D,BnaA05g07140D,BnaA08g19190D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"7.490272373540856%\"\u003e\n \u003cp\u003ebna-miR390a/b/c\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2859922178988326%\"\u003e\n \u003cp\u003e26.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2859922178988326%\"\u003e\n \u003cp\u003e11.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.815175097276264%\"\u003e\n \u003cp\u003e1.32\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"83.12256809338521%\"\u003e\n \u003cp\u003eBnaC03g02760D,BnaC01g03570D,BnaA05g28390D,BnaC03g53360D,BnaC01g22410D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"7.490272373540856%\"\u003e\n \u003cp\u003ebna-novel_31-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2859922178988326%\"\u003e\n \u003cp\u003e10.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2859922178988326%\"\u003e\n \u003cp\u003e55.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.815175097276264%\"\u003e\n \u003cp\u003e-2.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"83.12256809338521%\"\u003e\n \u003cp\u003eBnaC01g21190D,BnaA01g17940D,BnaC03g13660D,BnaA03g10950D,BnaA09g00170D,BnaA10g02710D,BnaC01g22250D,BnaA03g38770D,BnaA02g29970D,BnaA02g18470D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"7.490272373540856%\"\u003e\n \u003cp\u003ebna-novel_34-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2859922178988326%\"\u003e\n \u003cp\u003e0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2859922178988326%\"\u003e\n \u003cp\u003e624.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.815175097276264%\"\u003e\n \u003cp\u003e-8.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"83.12256809338521%\"\u003e\n \u003cp\u003eBnaA09g14240D,BnaA05g30840D,BnaA01g24360D,BnaA07g34690D,BnaA03g14400D,BnaA05g09000D,BnaC01g09790D,BnaA08g26410D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"7.490272373540856%\"\u003e\n \u003cp\u003ebna-novel_1-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2859922178988326%\"\u003e\n \u003cp\u003e0.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2859922178988326%\"\u003e\n \u003cp\u003e8.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.815175097276264%\"\u003e\n \u003cp\u003e-2.64\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"83.12256809338521%\"\u003e\n \u003cp\u003e--\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"2.6975863700899194%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"7.288215806909607%\"\u003e\n \u003cp\u003ebna-miR158-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e554.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e935.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.685281590156176%\"\u003e\n \u003cp\u003e-0.74\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"80.88026502602935%\"\u003e\n \u003cp\u003eBnaA05g08940D,BnaA04g29200D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"2.6975863700899194%\"\u003e\n \u003cp\u003e4006\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"7.288215806909607%\"\u003e\n \u003cp\u003ebna-novel_34-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e125.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.685281590156176%\"\u003e\n \u003cp\u003e-5.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"80.88026502602935%\"\u003e\n \u003cp\u003eBnaA05g29360D,BnaA09g14240D,BnaA05g30840D,BnaA01g24360D,BnaA07g34690D,BnaC03g41430D,BnaA03g35600D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"2.6975863700899194%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"6\" valign=\"top\" width=\"7.288215806909607%\"\u003e\n \u003cp\u003ebna-novel_31-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e4.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.2243256034074776%\"\u003e\n \u003cp\u003e53.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.685281590156176%\"\u003e\n \u003cp\u003e-2.81\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"80.88026502602935%\"\u003e\n \u003cp\u003eBnaC01g21190D,BnaC03g13660D,BnaA09g19450D,BnaA05g09930D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.308712121212121%\"\u003e\n \u003cp\u003e6251\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"4.59280303030303%\"\u003e\n \u003cp\u003ebna-novel_28-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"2.9829545454545454%\"\u003e\n \u003cp\u003e5.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.2670454545454546%\"\u003e\n \u003cp\u003e38.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.119318181818182%\"\u003e\n \u003cp\u003e-2.48\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"80.72916666666667%\"\u003e\n \u003cp\u003eBnaA09g36810D,BnaA10g16750D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.308712121212121%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"4.59280303030303%\"\u003e\n \u003cp\u003ebna-novel_73-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"2.9829545454545454%\"\u003e\n \u003cp\u003e30.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.2670454545454546%\"\u003e\n \u003cp\u003e64.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.119318181818182%\"\u003e\n \u003cp\u003e-1.19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"80.72916666666667%\"\u003e\n \u003cp\u003eBnaAnng19370D,BnaA06g16700D,BnaA05g05990D,BnaA10g17750D,BnaC03g50150D,BnaA10g19150D,BnaA03g58190D,BnaA05g28260D,BnaA07g15220D,BnaA01g18450D,BnaA09g50980D,BnaA10g21470D,BnaA09g13370D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.308712121212121%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"4.59280303030303%\"\u003e\n \u003cp\u003ebna-miR408-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"2.9829545454545454%\"\u003e\n \u003cp\u003e23.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.2670454545454546%\"\u003e\n \u003cp\u003e60.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.119318181818182%\"\u003e\n \u003cp\u003e-1.15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"80.72916666666667%\"\u003e\n \u003cp\u003eBnaA02g05230D,BnaA10g18650D,BnaA08g27620D,BnaAnng21260D,BnaA10g02870D,BnaA08g00080D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.308712121212121%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"4.59280303030303%\"\u003e\n \u003cp\u003ebna-novel_88-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"2.9829545454545454%\"\u003e\n \u003cp\u003e214.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.2670454545454546%\"\u003e\n \u003cp\u003e511.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.119318181818182%\"\u003e\n \u003cp\u003e-1.16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"80.72916666666667%\"\u003e\n \u003cp\u003eBnaA04g16310D,BnaA09g36970D,BnaA05g24870D,BnaC01g43200D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.308712121212121%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"4.59280303030303%\"\u003e\n \u003cp\u003ebna-miR398a-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"2.9829545454545454%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.2670454545454546%\"\u003e\n \u003cp\u003e5.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.119318181818182%\"\u003e\n \u003cp\u003e-2.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"80.72916666666667%\"\u003e\n \u003cp\u003eBnaA05g02320D,BnaA06g14440D,BnaA08g08840D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.308712121212121%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"4.59280303030303%\"\u003e\n \u003cp\u003ebna-novel_34-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"2.9829545454545454%\"\u003e\n \u003cp\u003e23.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.2670454545454546%\"\u003e\n \u003cp\u003e57.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"4.119318181818182%\"\u003e\n \u003cp\u003e-1.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"80.72916666666667%\"\u003e\n \u003cp\u003eBnaA02g05230D,BnaA10g18650D,BnaC01g01790D,BnaA10g10960D,BnaAnng21260D,BnaA10g14890D,BnaA08g27620D,BnaA07g15260D,BnaAnng36200D,BnaA10g15730D,BnaA02g11840D,BnaA10g25360D,BnaA10g02870D,BnaA08g00080D,BnaA10g17820D,BnaA09g25870D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e6284\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-miR394a/b\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e195.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e87.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e1.04\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA05g16640D,BnaA01g24160D,BnaA05g11890D,BnaC02g17150D,BnaAnng09250D,BnaA08g10740D,BnaA07g36430D,BnaC02g29160D,BnaA06g08380D,BnaC01g07190D,BnaA05g00820D,BnaA05g13120D,BnaC01g39490D,BnaC01g16400D,BnaA02g34270D,BnaC02g43190D,BnaA01g33370D,BnaC01g39860D,BnaA08g27810D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_36-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e1594.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e900.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e0.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA03g08820D,BnaA01g00910D,BnaC01g40540D,BnaA04g27250D,BnaA02g08190D,BnaC02g11600D,BnaA10g30150D,BnaA02g24120D,BnaC02g31870D,BnaA04g05090D,BnaA02g02340D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_67-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e129.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e50.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e1.23\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA03g02150D,BnaC01g00150D,BnaA07g02160D,BnaC03g01120D,BnaA03g00810D,BnaC01g01370D,BnaA07g25630D,BnaA03g55940D,BnaA04g24770D,BnaC01g25070D,BnaA01g20170D,BnaA03g48520D,BnaA09g17190D,BnaA09g39530D,BnaAnng28310D,BnaA01g35930D,BnaAnng08400D,BnaC02g23160D,BnaC03g60340D,BnaA04g00950D,BnaA03g33460D,BnaA04g22260D,BnaA03g27270D,BnaA07g37210D,BnaA06g37910D,BnaC03g21760D,BnaA03g18230D,BnaA09g05960D,BnaA03g34840D,BnaA09g05960D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_33-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e30.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e8.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e1.76\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA02g26940D,BnaA09g44810D,BnaA06g12040D,BnaA06g14250D,BnaA02g23840D,BnaA10g26020D,BnaA03g12030D,BnaA09g06400D,BnaC01g11370D,BnaA10g03030D,BnaA03g05700D,BnaA10g26690D,BnaA05g01020D,BnaA02g00710D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-miR159a\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e2320.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e1295.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e0.82\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaAnng27960D,BnaA04g18810D,BnaA03g15690D,BnaA07g18670D,BnaA07g12970D,BnaA03g22590D,BnaA07g25350D,BnaA06g20460D,BnaA01g18450D,BnaC01g19500D,BnaA01g16350D,BnaA09g27090D,BnaA06g13170D,BnaA09g42230D,BnaA09g30160D,BnaAnng05670D,BnaA06g18020D,BnaA08g20300D,BnaA02g05410D,BnaAnng21510D,BnaA09g08360D,BnaAnng14630D,BnaA09g02220D,BnaA04g25320D,BnaA09g10390D,BnaA02g30030D,BnaA09g44380D,BnaC01g19800D,BnaA01g35420D,BnaA08g27930D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_9-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e114.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e68.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e0.75\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA03g33680D,BnaA08g01260D,BnaA01g05980D,BnaAnng27960D,BnaA03g22590D,BnaA07g12970D,BnaA04g09220D,BnaA07g12970D,BnaA02g05410D,BnaA09g55500D,BnaAnng13060D,BnaA01g23170D,BnaA09g13960D,BnaA09g47880D,BnaA04g01370D,BnaA08g17490D,BnaC02g08160D,BnaA02g33550D,BnaC02g42310D,BnaA10g14420D,BnaA05g16460D,BnaA03g42760D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-miR398a-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e436.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e972.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e-1.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA08g19040D,BnaC02g28060D,BnaC01g27860D,BnaA10g26450D,BnaC03g13330D,BnaA03g10640D,BnaC02g12070D,BnaC01g18280D,BnaA01g15390D,BnaC02g42090D,BnaAnng31090D,BnaC01g43200D,BnaA05g30940D,BnaA01g29240D,BnaC01g36670D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_3-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e3.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e23.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e-2.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA05g24640D,BnaA09g38650D,BnaA06g39700D,BnaA09g39360D,BnaA09g54170D,BnaA06g10130D,BnaA10g14600D,BnaC01g22370D,BnaA10g24950D,BnaA10g23300D,BnaA04g14120D,BnaA09g41640D,BnaC01g09540D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_5-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e1.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e13.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e-1.77\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA05g24640D,BnaA06g39700D,BnaC02g14840D,BnaA06g10130D,BnaA09g38650D,BnaC01g22370D,BnaA09g39360D,BnaA10g04390D,BnaA09g54170D,BnaA10g14600D,BnaA01g28170D,BnaA06g28940D,BnaA10g23460D,BnaA06g07690D,BnaA10g24950D,BnaA09g01710D,BnaA10g16750D,BnaA10g23300D,BnaC01g09540D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_73-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e44.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e80.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e-0.84\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaAnng19370D,BnaC02g44840D,BnaA06g16700D,BnaA05g05990D,BnaC03g50150D,BnaA10g19150D,BnaA05g06360D,BnaA03g26400D,BnaA09g49880D,BnaA10g04230D,BnaA03g28580D,BnaA06g06040D,BnaA03g29130D,BnaA07g15220D,BnaA10g27030D,BnaA04g10410D,BnaAnng07200D,BnaC01g14980D,BnaA04g10200D,BnaA08g03230D,BnaA10g23200D,BnaA10g21470D,BnaA03g07290D,BnaA07g21850D,BnaC02g16740D,BnaA10g09600D,BnaA03g21270D,BnaA08g29090D,BnaA09g13370D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_68-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e7.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e-2.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA04g25570D,BnaA02g31820D,BnaC02g40450D,BnaAnng04800D,BnaA06g27210D,BnaA03g20670D,BnaC01g31630D,BnaA03g20660D,BnaA08g29130D,BnaA09g12560D,BnaA09g14580D,BnaA09g06270D,BnaA10g26450D,BnaA02g05300D,BnaA03g26340D,BnaA09g32850D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003e\u0026nbsp;bna-miR408-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e41.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e94.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e-1.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA03g22490D,BnaA10g18290D,BnaA07g26450D,BnaA06g37530D,BnaC02g29240D,BnaA05g29880D,BnaA10g21900D,BnaC01g16210D,BnaA02g09770D,BnaA10g23140D,BnaA02g34310D,BnaC02g43230D,BnaA01g08900D,BnaA06g10230D,BnaA10g15210D,BnaA05g14430D,BnaC02g46010D,BnaA03g04360D,BnaA04g08040D,BnaAnng32010D,BnaA03g30060D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_1-3p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e7.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e-1.72\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA02g30100D,BnaC02g38520D,BnaA10g21900D,BnaAnng29380D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.074739829706717%\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"3.642384105960265%\"\u003e\n \u003cp\u003ebna-novel_34-5p\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"2.554399243140965%\"\u003e\n \u003cp\u003e1.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"4\" valign=\"top\" width=\"3.3112582781456954%\"\u003e\n \u003cp\u003e456.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"3.1693472090823085%\"\u003e\n \u003cp\u003e-6.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"3\" valign=\"top\" width=\"84.24787133396404%\"\u003e\n \u003cp\u003eBnaA02g05300D,BnaA05g27100D,BnaA07g28780D,BnaA06g28070D,BnaA09g14240D,BnaA05g30840D,BnaA08g29690D,BnaC03g48340D,BnaA04g22150D,BnaC01g31340D,BnaA01g24360D,BnaA03g23950D,BnaC03g53270D,BnaA06g01440D,BnaA02g14960D,BnaA09g03120D,BnaA09g31030D,BnaC02g03920D,BnaA07g34690D,BnaA03g35600D,BnaA04g02140D,BnaC02g14950D,BnaA03g10460D,BnaAnng21410D\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n \u003cp\u003eA:fertile, B:sterile lines.\u0026nbsp;\u003c/p\u003e\n \u003cp\u003eqRT-PCR was conducted to verify the expression profiles of these differentially expressed miRNAs in deep sequencing. Five differentially expressed miRNAs were chosen for qRT-PCR analysis. The results were consistent with those of deep sequencing. In qRT-PCR, miR158 was up-regulated in \u0026ldquo;4001B\u0026rdquo; flower buds compared with that in \u0026ldquo;4001A\u0026rdquo;. Novel34 was greatly up-regulated in \u0026ldquo;4006B\u0026rdquo; flower buds (730-fold) compared with that in \u0026ldquo;4006A\u0026rdquo;. miR159 and miR827 were both up-regulated in \u0026ldquo;6284A\u0026rdquo; flower buds compared with that in \u0026ldquo;6284B\u0026rdquo;. miR398 was up-regulated in \u0026ldquo;6284B\u0026rdquo; flower buds compared with that in \u0026ldquo;6284A\u0026rdquo; (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eTarget prediction and identification of differentially expressed miRNAs in sterile and fertile lines\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eTo make clear the potential regulatory roles of differentially expressed miRNAs, a plant small RNA target analysis server psRNATarget (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://plantgrn.noble.org/psRNATarget/\u003c/span\u003e\u003c/span\u003e) was performed to predict their target genes. A total of 707 transcripts were predicted to be targets of the 20 miRNAs (Additional Table S3). In addition, transcriptome sequencing was conducted using the same samples as sRNA sequencing (unpublished data). The differentially expressed and up-regulated mRNAs were predicted as the candidate targets for the differentially expressed and down-regulated miRNAs. At the same time, the differentially expressed and down-regulated mRNAs were predicted as the candidate targets for the differentially expressed and up-regulated miRNAs. As shown in Table \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e. Thirty eight candidate target genes were predicted for the six differentially expressed miRNAs between \u0026ldquo;4001A\u0026rdquo; and \u0026ldquo;4001B\u0026rdquo;. Eleven candidate genes were predicted for the two differentially expressed miRNAs between \u0026ldquo;4006A\u0026rdquo; and \u0026ldquo;4006B\u0026rdquo;. Forty-four candidate genes were predicted for the six differentially expressed miRNAs between \u0026ldquo;6251A\u0026rdquo; and \u0026ldquo;6251B\u0026rdquo;. Two hundred and sixty-seven candidate genes were predicted for the 14 differentially expressed miRNAs between \u0026ldquo;6284A\u0026rdquo; and \u0026ldquo;6284B\u0026rdquo;.\u003c/p\u003e\n \u003cp\u003eTo further demonstrate the potential target genes, 5\u0026rsquo; modified RACE was performed using mixed samples from flower buds of the fertile lines (\u0026ldquo;6284B\u0026rdquo; and \u0026ldquo;4001B\u0026rdquo;). Three target genes were validated using 5\u0026rsquo; modified RACE (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e). \u003cem\u003eBn.A09.CSD1\u003c/em\u003e (\u003cem\u003eBnaA09g48720D\u003c/em\u003e) was cleaved by bna-miR398a-3p. \u003cem\u003eBn.A09.PPR\u003c/em\u003e (\u003cem\u003eBnaA09g11120D\u003c/em\u003e) was cleaved by bna-miR158-3p. \u003cem\u003eBn.Cnn.MYB\u003c/em\u003e (\u003cem\u003eBnaCnng51960D\u003c/em\u003e) was cleaved by bna-miR159a.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eOverexpression of bna-miR159a affected seeds and siliques develpoment in\u003c/strong\u003e \u003cspan class=\"BoldItalic\" name=\"Emphasis\" type=\"BoldItalic\"\u003e\u003cstrong\u003eArabidopsis\u003c/strong\u003e\u003c/span\u003e\u003c/p\u003e\n \u003cp\u003eAmong all the differentially expressed miRNAs in the two DGMS and RGMS lines, bna-miR159a had the highest expression level. Thus, bna-miR159a was chosen to analyze its function. Two vectors, namely, p35S::MIR159a-C6 and p35S::MIR159a-A7, which contained the precursor sequences of \u003cem\u003eMIR159a\u003c/em\u003e located in C6 and A7 chromosomes of \u003cem\u003eB. napus\u003c/em\u003e, were transformed into \u003cem\u003eArabidopsis\u003c/em\u003e, respectively. The corresponding \u003cem\u003eArabidopsis\u003c/em\u003e transgenic plants were named as MIR159OE-1 and MIR159OE-2. The expression levels of bna-miR159 and its target genes were analyzed in wild-type, MIR159OE-1, and MIR159OE-2.\u003c/p\u003e\n \u003cp\u003eTo reveal miR159 potential function in \u003cem\u003eArabidopsis\u003c/em\u003e, the expression level of miR159a was analyzed by qRT-PCR. In wild-type plants of \u003cem\u003eArabidopsis\u003c/em\u003e, the transcript of mature miR159a and its targets (\u003cem\u003eAtMYB33\u003c/em\u003e and \u003cem\u003eAtMYB65\u003c/em\u003e) were detected in root, stem, rosette leaf, stem leaf, flower, and silique through qRT-PCR. The expression level of mature miR159a was the highest in silique (5875-fold), relatively lower in stem, stem leaf, and flower compared with that in the root. The expression levels of \u003cem\u003eAtMYB33\u003c/em\u003e and \u003cem\u003eAtMYB65\u003c/em\u003e were very low and almost undetectable in silique, whereas they were relatively high in stem, stem leaf, and flower compared with that in the root (Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e). In MIR159OE-1 and MIR159OE-2, the transcripts of mature miR159a and its targets were detected in stem leaf through qRT-PCR. The mature miR159a was overexpressed in MIR159OE-1 (4.04-fold) and MIR159OE-2 (13.6-fold) compared with that in WT. Meanwhile, the transcripts of \u003cem\u003eAtMYB33\u003c/em\u003e and \u003cem\u003eAtMYB65\u003c/em\u003e were suppressed in MIR159OE-1 and MIR159OE-2, especially in MIR159OE-2, compared with that in WT (Fig. \u003cspan class=\"InternalRef\"\u003e7\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eThe morphological characters of MIR159OE-1, MIR159OE-2, and WT were observed along with their development processes, especially in the flowering and fruiting periods. No significant difference was observed between transgenic and WT plants in the vegetative growth phase. However, during the reproductive growth period, in the MIR159OE-1 and MIR159OE-2 transgenic plants, the seed setting rate decreased, and siliques became shorter compared with that in the WT (Fig. \u003cspan class=\"InternalRef\"\u003e8\u003c/span\u003e). The length of siliques from WT, MIR159OE-1, and MIR159OE-2 transgenic plants were measured. The results showed that the silique lengths of WT were approximately 13.4 mm, whereas the average silique lengths of MIR159OE-1 and MIR159OE-2 were 5.9 and 5.8 mm, respectively. The results indicated that the overexpression of \u003cem\u003eMIR159\u003c/em\u003e led to significantly shorter siliques and reduced seed setting rate (Fig. \u003cspan class=\"InternalRef\"\u003e8\u003c/span\u003eE).\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eMiRNAs, as the key post-transcriptional regulators, participate in various biological processes in plant. Recently, an increasing number of studies showed that plant miRNAs were also involved in pollen and fertility development. In 2014, Jiang et al. identified 54 new conserved miRNAs and 25 pairs of novel miRNA/miRNA* in a GMS system of \u003cem\u003eB. campestris\u003c/em\u003e ssp. \u003cem\u003echinensis\u003c/em\u003e. Eighteen differentially expressed miRNAs with over two fold change between flower buds of male sterile and fertile lines were identified; they might be involved in the pollen development process [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. In 2017, Ma et al. verified that the overexpression of \u003cem\u003eMIR158\u003c/em\u003e caused pollen abortion and reduced pollen vitality, which were caused by the degradation of pollen content from the binuclear microspore stage [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. Dong et al. identified 85 known miRNAs and 120 novel miRNAs, which were expressed during rapeseed anther development in a novel recessive GMS system \u0026ldquo;CN12AB.\u0026rdquo; Moreover, 19 and 18 known miRNAs were found to be differentially expressed in 0.5\u0026ndash;1.0 mm buds and in 2.5\u0026ndash;3.0 mm buds between CN12A and CN12B, respectively. Among these, 14 miRNAs were up-regulated, and 23 miRNAs were down-regulated expressed in CN12A compared with those in CN12B [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]. In this study, to identify miRNAs and their targets involved in pollen development and GMS occurrence in rapeseed, 24 small RNA libraries and transcriptome libraries were constructed and sequenced for the flower buds from the fertile and sterile lines of two RGMS lines (\u0026ldquo;6251AB\u0026rdquo; and \u0026ldquo;6284AB\u0026rdquo;) and two DGMS lines (\u0026ldquo;4001AB\u0026rdquo; and \u0026ldquo;4006AB\u0026rdquo;). Based on the sequencing results, 21 differentially expressed miRNAs with over 1.5-fold change between flower buds of male sterile and fertile lines were identified, including six differentially expressed miRNAs between \u0026ldquo;4001A\u0026rdquo; and \u0026ldquo;4001B\u0026rdquo;, two differentially expressed miRNAs between \u0026ldquo;4006A\u0026rdquo; and \u0026ldquo;4006B\u0026rdquo;, six differentially expressed miRNAs between \u0026ldquo;6251A\u0026rdquo; and \u0026ldquo;6251B\u0026rdquo;, and 14 differentially expressed miRNAs between \u0026ldquo;6284A\u0026rdquo; and \u0026ldquo;6284B\u0026rdquo;. Among them, bna-novel_34-5p was common and differentially expressed between the fertile and sterile lines of \u0026ldquo;4001AB\u0026rdquo;, \u0026ldquo;4006AB\u0026rdquo;, \u0026ldquo;6251AB\u0026rdquo;, and \u0026ldquo;6284AB\u0026rdquo;. The results of previous studies and this study verified that miRNAs may play important regulatory roles in rapeseed pollen development and GMS occurrence.\u003c/p\u003e \u003cp\u003eMiR159 is conserved in many plants and is involved in multiple growth and development processes of plants. Allen et al. obtained a \u003cem\u003emir159ab\u003c/em\u003e double mutant in \u003cem\u003eArabidopsis\u003c/em\u003e, which showed pleiotropic morphological defects, including altered growth habit, curled leaves, small siliques, and small seeds [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Millar and Gubler verified that miR159 regulated anther development by regulating the expression of its targets, such as \u003cem\u003eMYB33\u003c/em\u003e and \u003cem\u003eMYB65\u003c/em\u003e [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]. Overexpression of miR159 caused the down-regulation of \u003cem\u003eMYB103\u003c/em\u003e transcripts and earlier degeneration of the tapetum and aberrant pollen formation during anther development [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. In radish, differential expression level of miR159 during anther development was observed among male sterile and maintainer lines. Increased levels of miR159 transcripts decreased the expression of \u003cem\u003eMYB101\u003c/em\u003e, thereby inhibiting tapetum development and exine formation [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. Chen et al. identified 17 differentially expressed miRNAs between long and short siliques of rapeseed, including miR159. Correlation analysis of miR159 and its targets suggested that miR159 repressed cell proliferation to control silique length [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]. Hu et al. found that the overexpression of Bra-\u003cem\u003eMIR159a\u003c/em\u003e caused pollen abortion and abnormal pollen germination [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. In this study, the differentially expressed miR159 in \u0026ldquo;6284A\u0026rdquo; and \u0026ldquo;6284B\u0026rdquo; was chosen to analyze its function during anther development. The bna-miR159 was overexpressed in \u003cem\u003eArabidopsis\u003c/em\u003e and resulted in decreased seed setting rate, and shortened siliques, illustrating that miR159 may regulate the fertility and silique development of rapeseed. The results of previous reports and the results of the present study verified that miR159 and its target genes might be involved in the regulatory network of pollen development and male sterility and this module is conserved in plants.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eA large number of miRNAs were identified during pollen development in the two DGMS and two RGMS lines by deep sequencing. These identified miRNAs included 27 novel miRNAs on the other arm of known pre-miRNAs, 44 new conserved miRNAs, and 35 pairs of novel miRNA-3p/miRNA-5p. Among all the identified miRNAs, 21 differentially expressed miRNAs with over 1.5-fold change between flower buds of male sterile and fertile lines were identified, including six differentially expressed miRNAs between \u0026ldquo;4001A\u0026rdquo; and \u0026ldquo;4001B\u0026rdquo;, two differentially expressed miRNAs between \u0026ldquo;4006A\u0026rdquo; and \u0026ldquo;4006B\u0026rdquo;, six differentially expressed miRNAs between \u0026ldquo;6251A\u0026rdquo; and \u0026ldquo;6251B\u0026rdquo;, and fourteen differentially expressed miRNAs between \u0026ldquo;6284A\u0026rdquo; and \u0026ldquo;6284B\u0026rdquo;. The qRT-PCR results of 5 differentially expressed miRNAs (miR158, novel34, miR159, miR827, and miR398) were consistent with deep sequencing results. The association analysis of small RNA and transcriptome sequencing was conducted, and the analysis results indicated that 360 genes were predicted to be the candidate targets of 21 differentially expressed miRNAs. The results of 5\u0026rsquo; modified RACE verified that \u003cem\u003eBn.A09.CSD1\u003c/em\u003e (\u003cem\u003eBnaA09g48720D\u003c/em\u003e), \u003cem\u003eBn.A09.PPR\u003c/em\u003e (\u003cem\u003eBnaA09g11120D\u003c/em\u003e), and \u003cem\u003eBn.Cnn.MYB\u003c/em\u003e (\u003cem\u003eBnaCnng51960D\u003c/em\u003e) were cleaved by bna-miR398a-3p, bna-miR158-3p, and bna-miR159a. Additionally, overexpression of bna-miR159 in \u003cem\u003eArabidopsis\u003c/em\u003e resulted in decreased seed setting rate, and shortened siliques, illustrating that miR159 may regulate rapeseed fertility and silique development. All the results in our study would provide valuable clues for exploring miRNA-mediated regulatory networks in fertility development of GMS lines in \u003cem\u003eB. napus\u003c/em\u003e.\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv class=\"Section2\" id=\"Sec8\"\u003e\n \u003cp\u003e\u003cstrong\u003ePlant materials\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003e\u0026ldquo;6251AB\u0026rdquo; and \u0026ldquo;6284AB\u0026rdquo; are two recessive genic male sterile (RGMS) lines of \u003cem\u003eB. napus\u003c/em\u003e. The \u0026ldquo;6251A\u0026rdquo; and \u0026ldquo;6284A\u0026rdquo; are male sterile lines and the \u0026ldquo;6251B\u0026rdquo; and \u0026ldquo;6284B\u0026rdquo; are the fertile lines. Their male sterility is controlled by two loci (\u003cem\u003eBnMs3\u003c/em\u003e and \u003cem\u003eBnRf\u003c/em\u003e) [\u003cspan class=\"CitationRef\"\u003e14\u003c/span\u003e]. The original source of the two RGMS lines was \u0026ldquo;9012A\u0026rdquo;, which was identified by Chen et al. [\u003cspan class=\"CitationRef\"\u003e9\u003c/span\u003e] and Sun et al. [\u003cspan class=\"CitationRef\"\u003e33\u003c/span\u003e]. \u0026ldquo;4001AB\u0026rdquo; and \u0026ldquo;4006AB\u0026rdquo; are two dominant genic male sterile (DGMS) lines of \u003cem\u003eB. napus\u003c/em\u003e. The \u0026ldquo;4001A\u0026rdquo; and \u0026ldquo;4006A\u0026rdquo; are male sterile lines and the \u0026ldquo;4001B\u0026rdquo; and \u0026ldquo;4006B\u0026rdquo; are the fertile lines. Their male sterility is controlled by three alleles (\u003cem\u003eMf\u003c/em\u003e, \u003cem\u003eMs\u003c/em\u003e, and \u003cem\u003ems\u003c/em\u003e) at the same locus [\u003cspan class=\"CitationRef\"\u003e5\u003c/span\u003e]. The original source of the two DGMS lines was \u0026ldquo;Yi3A\u0026rdquo;, which was identified by Li et al. [\u003cspan class=\"CitationRef\"\u003e34\u003c/span\u003e]. The above RGMS lines and DGMS lines used in this study have been grown for several generations. The progenies of the two RGMS lines are both segregated into sterile and fertile types during reproduction at a ratio of 1:1. While the two DGMS lines are both segregated into sterile and fertile types at a ratio of 3:1. These plant materials were planted in the experimental farm of Zhuanghang comprehensive experimental station of Shanghai Academy of Agricultural Sciences. During flowering stage, mixed flower buds were respectively harvested from more than ten plants of the eight lines. The eight lines were \u0026ldquo;6251A\u0026rdquo;, \u0026ldquo;6251B\u0026rdquo;, \u0026ldquo;6284A\u0026rdquo;, \u0026ldquo;6284B\u0026rdquo;, \u0026ldquo;4001A\u0026rdquo;, \u0026ldquo;4001B\u0026rdquo;, \u0026ldquo;4006A\u0026rdquo;, and \u0026ldquo;4006B\u0026rdquo;. The eight kinds of samples were quickly frozen in liquid nitrogen and stored at -80\u0026deg;C. Three independent biological replicates were collected for each kind of sample.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eSmall RNA library construction and sequencing\u003c/strong\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003eTotal RNA for each kind of sample was extracted in three biological replicates using Trizol reagent (Invitrogen, USA). RNA samples with an OD260/OD280 ratio of 2.0 and a total content of more than 2 \u0026micro;g were qualified for small RNA library construction. Then, 24 sequencing libraries (three biological replicates respectively for \u0026ldquo;6251A\u0026rdquo;, \u0026ldquo;6251B\u0026rdquo;, \u0026ldquo;6284A\u0026rdquo;, \u0026ldquo;6284B\u0026rdquo;, \u0026ldquo;4001A\u0026rdquo;, \u0026ldquo;4001B\u0026rdquo;, \u0026ldquo;4006A\u0026rdquo;, and \u0026ldquo;4006B\u0026rdquo;) were constructed using TruSeqSmall RNA Sample Preparation Kit (Illumina, USA) and then sequenced using Illumina Hiseq 2500/Miseq at Beijing Novogene Bioinformatics Technology Co. Ltd.\u003c/p\u003e\n\u003cdiv class=\"Section2\" id=\"Sec10\"\u003e\n \u003cp\u003e\u003cstrong\u003eData analysis\u003c/strong\u003e\u003c/p\u003e\n \u003cp\u003eClean reads were obtained by removing low-quality reads, N-containing fragments, and adapters. Then, the length of 18 to 30 nt clean reads were mapped to the \u003cem\u003eB. napus\u003c/em\u003e genome sequence (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://brassicadb.agridata.cn/brad/\u003c/span\u003e\u003c/span\u003e) using Bowtie2 with no mismatches allowed, more details were described in Niu et al [\u003cspan class=\"CitationRef\"\u003e35\u003c/span\u003e]. Unmapped sequences were removed.\u003c/p\u003e\n \u003cp\u003eThe mapped small RNAs reads were aligned to known miRNAs in miRBase22.1. Modified software mirdeep2 [\u003cspan class=\"CitationRef\"\u003e36\u003c/span\u003e] were used to predict the potential miRNAs and secondary structures. The ncRNAs includes rRNAs, tRNAs, snRNAs, snoRNAs, and small genome repeat sequences were removed. The rest of sRNA sequences were aligned to \u003cem\u003eB. napus\u003c/em\u003e NAT-siRNAs in PlantNATsDB to remove NAT-siRNAs. Then miREvo [\u003cspan class=\"CitationRef\"\u003e37\u003c/span\u003e] and mirdeep2 [\u003cspan class=\"CitationRef\"\u003e36\u003c/span\u003e] were used to predict novel miRNAs in \u003cem\u003eB. napus\u003c/em\u003e. To reveal the differentially expressed miRNAs, the miRNAs expression was analyzed using the DESeq2 [\u003cspan class=\"CitationRef\"\u003e42\u003c/span\u003e]. The online software of Venny 2.1.0 was used (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://bioinfogp.cnb.csic.es/tools/venny/index.html\u003c/span\u003e\u003c/span\u003e) to draw Venn diagrams.\u003c/p\u003e\n \u003cp\u003e\u003cstrong\u003eIdentification of conserved and novel miRNAs\u003c/strong\u003e\u003c/p\u003e\n\u003c/div\u003e\n\u003cp\u003eTo identify conserved and novel miRNAs in \u003cem\u003eB. napus\u003c/em\u003e, the candidate sRNAs were blasted against \u003cem\u003eB. napus\u003c/em\u003e genome sequences. In addition, Mfold software was used to predict the secondary structures of pre-miRNAs with the flanking sequences of the small RNAs in the genome [\u003cspan class=\"CitationRef\"\u003e43\u003c/span\u003e]. Five criteria must be met for identifying conserved and novel miRNAs [\u003cspan class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan class=\"CitationRef\"\u003e45\u003c/span\u003e]. The sequences and lengths, read counts, and positions in chromosome were further analyzed for conserved and novel miRNAs.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eqRT-PCR\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTotal RNA was treated with DNase I (Takara, Japan) to remove residual genomic DNA. For the qRT-PCR analysis of \u003cem\u003eAtMYB33\u003c/em\u003e and \u003cem\u003eMYB65\u003c/em\u003e, \u003cem\u003eAtACTIN2\u003c/em\u003e gene was used as internal control. PrimeScript\u0026trade; II reverse transcriptase (Takara) and oligo (dT) primers were used for first-strand cDNA synthesis. For the qRT-PCR analysis of mature miRNAs, U6 was used as internal control gene, and the first cDNA was synthetized using a miRNA First Strand cDNA Synthesis kit (Stem-loop Method) (Sangon Biotech). The qRT-PCR reactions were performed in a MyiQ2 qRT-PCR detection system (Bio-Rad, \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e\u003ca href=\"https://dataview.ncbi.nlm.nih.gov/object/PRJNA743414?reviewer=t674c02cj415380e8oldre4s5a\" target=\"_blank\"\u003ewww.bio-rad.com/\u003c/a\u003e\u003c/span\u003e\u003c/span\u003e) using iQ SYBR Green supermix (Bio-Rad) [\u003cspan class=\"CitationRef\"\u003e46\u003c/span\u003e]. Each experiment was conducted in three biological replicates, and the same sample was performed in three technical replicates. Relative expression levels of miRNAs and their target genes were quantified by using the 2\u003csup\u003e\u0026minus;\u0026Delta;\u0026Delta;Ct\u003c/sup\u003e method [\u003cspan class=\"CitationRef\"\u003e47\u003c/span\u003e]. All the primers used for qRT-PCR analysis are listed in Additional File 1: Table S4.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e5\u0026rsquo; modified RACE analysis\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eA mixture of flower buds from the fertile lines (\u0026ldquo;6284B\u0026rdquo; and \u0026ldquo;4001B\u0026rdquo;) was used for total RNA isolation. The 5\u0026rsquo; modified RACE was performed using a FirstChoice\u0026trade; RLM-RACE Kit (Invitrogen, USA). Total RNA was directly ligated to the 5\u0026rsquo;RACE oligo. The first-strand cDNA synthesis and the two rounds of PCR reactions were conducted following the manufacturer\u0026rsquo;s instructions. The PCR products containing the target gene bands were ligated into pGEM-T Easy Vector (Promega, USA) for sequencing [\u003cspan class=\"CitationRef\"\u003e25\u003c/span\u003e]. The primers are listed in Additional File 1: Table S4.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eVector construction and plant transformation\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTwo precursor sequences of bna-miR159a, which were located in A7 and C6 chromosomes of \u003cem\u003eB. napus\u003c/em\u003e were designated as pre-miR159a-C6 and pre-miR159a-A7, respectively (Additional File 1: Table S5). The 411 nt and 470 nt genomic fragments containing pre-miR159a-C6 and pre-miR159a-A7 were amplified from \u003cem\u003eB. napus\u003c/em\u003e using gene specific primers with endonuclease cleavage sites \u003cem\u003eSma\u003c/em\u003e I and \u003cem\u003eSal\u003c/em\u003e I. Then the fragments were cloned into pCAMBIA1301 binary vector with CaMV 35S promoter. The two vectors were designated as p35S::MIR159a-C6 and p35S::MIR159a-A7, which were introduced into \u003cem\u003eAgrobacterium tumefaciens\u003c/em\u003e strains GV3101 and further transformed into \u003cem\u003eArabidopsis\u003c/em\u003e. The corresponding \u003cem\u003eArabidopsis\u003c/em\u003e transgenic plants were designated as MIR159OE-1 and MIR159OE-2. The transgenic and wild-type plants were cultivated in the greenhouse under the same environment. Their phenotypes were observed and recorded. The silique length data were collected from more than three plants for each transgenic line.\u003c/p\u003e"},{"header":"Abbreviations","content":"\u003cp\u003eGMS: Genic male sterility; RGMS: recessive genic male sterility; DGMS: dominant genic male sterility; CMS: cytoplasmic male sterility\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/p\u003e\u003cp\u003e\u003c/strong\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of Data and Materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe raw reads of the 24 sRNA libraries were uploaded to SRA database of NCBI and 24 accession numbers were obtained, including SRX11350295, SRX11350296, SRX11350307, SRX11350312, SRX11350313, SRX11350315, and SRX11350316 (https://dataview.ncbi.nlm.nih.gov/object/PRJNA743414?reviewer=t674c02cj415380e8oldre4s5a).\u0026nbsp;The raw transcriptome data will also be further utilized to excavate differentially expressed mRNAs in the fertile and sterile lines of RGMS and DGMS line.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\u003cp\u003eThe authors declare that they have no competing interests.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by grants from the Natural Science Foundation of China (Grant No. 31901502), and Shanghai Agriculture Applied Technology Development Program, China (Grant No.Z20160105). The former fund supported the design of the study and collection, including deep sequencing and data analysis. The second fund supported the writing and publishing of this manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eJ-XJ and L-YY conceived the project and research plan. J-XJ finished the manuscript. J-XJ, P-FX and Y-JL analyzed the sequencing data, performed the experiments and drafted the manuscript. X-RZ, J-YZ, and J-FZ planted and observed the DGMS plants. Y-LL, M-YJ, and W-RW planted and observed the RGMS plants. All authors have read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; information\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003csup\u003e1\u003c/sup\u003eCrop Breeding and Cultivation Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201403, China. \u003csup\u003e2\u003c/sup\u003eNational Key Laboratory of Plant Molecular Genetics, Shanghai Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China. \u003csup\u003e3\u003c/sup\u003eSchool of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, 200240, China.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eKaul MLH. Male sterility in higher plants. 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BMC Plant Biology. 2018; 18:127\u0026ndash;35.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLivak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2\u003csup\u003e\u0026ndash;∆∆CT\u003c/sup\u003e method. Methods. 2001; 25(4): 402\u0026ndash;8.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"bmc-plant-biology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pbio","sideBox":"Learn more about [BMC Plant Biology](http://bmcplantbiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/pbio/default.aspx","title":"BMC Plant Biology","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"Brassica napus, miRNAs, genic male sterility, pollen development, miR159, silique development","lastPublishedDoi":"10.21203/rs.3.rs-471166/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-471166/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e \u003cem\u003eBrassica napus\u003c/em\u003e is the third leading source of edible oil in the world. Genic male sterility (GMS) lines provide crucial material for harnessing heterosis for rapeseed. GMS lines have been used successfully for rapeseed hybrid production in China. MicroRNAs (miRNAs) play crucial regulatory roles in various plant growth, development, and stress response processes. However, reports on miRNAs that regulate the pollen development of GMS lines in \u003cem\u003eB. napus \u003c/em\u003eare few.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eResults: \u003c/strong\u003eIn this study, 12 small RNA and transcriptome libraries were constructed and sequenced for the flower buds from the fertile and sterile lines of two recessive GMS (RGMS) lines, namely, “6251AB” and “6284AB”. At the same time, 12 small RNA and transcriptome libraries were also constructed and sequenced for the flower buds from the fertile and sterile lines of two dominant GMS (DGMS) lines, namely, “4001AB” and “4006AB”. Based on the results, 46 known miRNAs, 27 novel miRNAs on the other arm of known pre-miRNAs, and 44 new conserved miRNAs were identified. Thirty-five pairs of novel miRNA-3p/miRNA-5p were found. Among all the identified miRNAs, 21 differentially expressed miRNAs with over 1.5-fold change between flower buds of sterile and fertile lines were identified, including six differentially expressed miRNAs between “4001A” and “4001B”, two differentially expressed miRNAs between “4006A” and “4006B”, six differentially expressed miRNAs between “6251A” and “6251B”, and 14 differentially expressed miRNAs between “6284A” and “6284B”. The correlation analysis of small RNA and transcriptome sequencing was conducted. The analysis results indicated that 360 genes were predicted to be the candidate targets of 21 differentially expressed miRNAs. \u003cem\u003eBnaA09g48720D\u003c/em\u003e, \u003cem\u003eBnaA09g11120D\u003c/em\u003e, and \u003cem\u003eBnaCnng51960D\u003c/em\u003e were demonstrated to be cleaved by bna-miR398a-3p, bna-miR158-3p and bna-miR159a, respectively. Among the differentially expressed miRNAs, miR159 was chosen to analyze its function in reproduction development and male sterility. Overexpression of bna-miR159 in \u003cem\u003eArabidopsis\u003c/em\u003e resulted in decreased seed setting rate, and shortened siliques, illustrating that miR159 may regulate the fertility and silique development in rapeseed.\u003c/p\u003e\u003cp\u003e\u003cstrong\u003eConclusions: \u003c/strong\u003eOur findings provide an overview of miRNAs that are potentially involved in GMS and pollen development. New information on miRNAs and their related target genes are provided to exploit the GMS mechanism and reveal the miRNA networks in \u003cem\u003eB. napus.\u003c/em\u003e\u003c/p\u003e","manuscriptTitle":"Identification of miRNAs And Their Target Genes In Genic Male Sterility Lines In Brassica Napus By Small RNA Sequencing","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-07-12 20:12:19","doi":"10.21203/rs.3.rs-471166/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2021-08-17T05:40:54+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2021-08-04T19:48:56+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"b0791fd3-9e7a-4d7c-8683-e872c7dbf9d7","date":"2021-07-22T10:01:22+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"4f211a06-442e-4f12-b668-19e343893660","date":"2021-07-20T15:35:22+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2021-07-13T10:09:59+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2021-07-12T06:43:58+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2021-07-10T03:45:46+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2021-07-10T03:44:03+00:00","index":"","fulltext":""},{"type":"submitted","content":"BMC Plant Biology","date":"2021-04-28T13:07:29+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"bmc-plant-biology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"pbio","sideBox":"Learn more about [BMC Plant Biology](http://bmcplantbiol.biomedcentral.com/)","snPcode":"","submissionUrl":"https://www.editorialmanager.com/pbio/default.aspx","title":"BMC Plant Biology","twitterHandle":"BMC_series","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"em","reportingPortfolio":"BMC Series","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"5dd8315c-36b2-49fc-aa9a-1a936650ba6f","owner":[],"postedDate":"July 12th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"under-review","subjectAreas":[{"id":5638969,"name":"Plant Molecular Biology and Genetics"},{"id":5638970,"name":"Plant Physiology and Morphology"}],"tags":[],"updatedAt":"2021-10-30T04:14:02+00:00","versionOfRecord":[],"versionCreatedAt":"2021-07-12 20:12:19","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-471166","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-471166","identity":"rs-471166","version":["v1"]},"buildId":"rHA-KDH7Qsr4HCuvH75dn","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}