Morphological And Molecular Identification of Double Flowered Stock (Matthiola Incana R. Br) Cultivars With High Fertility

Morphological And Molecular Identification of Double Flowered Stock (Matthiola Incana R. Br) Cultivars With High Fertility | 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 Morphological And Molecular Identification of Double Flowered Stock ( Matthiola Incana R. Br) Cultivars With High Fertility Un-Hyang Ho, Jong-Hyang Ri, Chol-Jun Ri This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-578473/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Garden stock ( Matthiola incana R. Br.) is a commercially important horticultural crop owing to its ornamental effect. There are different stock cultivars varied in color and shape, especially flowered phenotype is an essential index evaluating its commercial value, because double flowered cultivars have more brilliant flowers compared to single flowered one. The present work aimed: (1) to make superior cultivars with different colors, high fertility, being capable of early selecting only double flowered seedlings by leaf color and to investigate morphological characteristics and (2) to select RAPD and ISSR primers for the cultivar certification and identification to culture and produce good commercial stock cultivars. Here we obtained new double flowered stock cultivars with different colors including pink, pale pink and white, through outcrossing between “white” cultivar (high fertile but unable to select double flower phenotype) and “pink” cultivar (vice. versa). Among newly obtained stock cultivars, single and double flower seedlings are distinguishable from each other by leaf color, having about 70% of fertility. Moreover RAPD and ISSR markers selected in this study can be applied to identify different stock cultivars in seed production, culture and to establish cultivar certification system. Agronomy Molecular Genetics Matthiola incana RAPD ISSR cultivar identification fertility Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Garden stock ( Matthiola incana R. Br.) is a commercially important horticultural crop having different types of flowers varied in color and flowered phenotype, belonging to the family Brassicaceae (Tatsuzawa et al. 2012 ). Due to high ornamental value, many studies associated in culture and breeding of the garden stock have focused on improvement of morphological and physiological traits such as flower color, double flower phenotype, inflorescence length, and fertility (Sima 2019a, b; Daozong et al. 2018). There are single and double flowered M. incana cultivars, and latter is more widely used for floriculture because it has more beautiful flowers. Seed production is only possible in single flowered individuals. It is impossible in double flowered plants due to degenerative pistils and stamens. Therefore many researchers and breeders have aimed to produce of more double flowered plants than single flowered ones in M. incana and to develop markers to select them. To date, several morphological traits such as cotyledon shape, serrate leaf, seed color, and leaf color have been used in early selection of double flowered stock individuals, because these traits were linked to double flowers (Saunders 1911 ; 1915 ; 1921 ; 1928 ; Ecker et al. 1993 ). Takashi and Kanae (2018) identified the gene related to double flowers and developed a molecular marker to select double flowered plants efficiently, based on the study of double flower-linked molecular markers in Japanese gentian (Tasaki et al. 2017 ). In general, the double flower trait is inherited by a single locus, s and it is linked to leaf color. In some stock cultivars, leaf color is a simple morphological marker, that is, green color indicates single flowered individual while pale green color represents double flowered individual (Ecker et al. 1993 ). And also the fertility of plant is an important trait in cultivation and breeding of stock as well as other plants. Consequently, it is necessary to make a good stock cultivar with high fertility, different colors, and being capable of early discrimination of double flowered phenotype, in culture and production for commercial use. Random amplified polymorphic DNA technique has been widely applied to evaluate genetic diversity on intraspecific level such as Ocimum ( Lamiaceae ) (Tanmay et al. 2016) and Swertia ( Gentianaceae ) (Prabhjot et al. 2019) and interspecific level including Citrullus colocynthis (Kumar et al. 2017), Lactuca sativa L. (Shubhangi et al. 2018), Nilgirianthus ciliates , Artemisia herba -alba (Khaled et al. 2019). Moreover, RAPD method has also been used to assess genetic diversity on the level of variety or cultivar in following plants, for example, common beans ( Phaseolus vulgaris L.) (Maciel et al. 2001 ), ginger (Siddharth et al. 2007), apricot ( Prunus armeniaca L.) (Chroboková et al. 2011 ), cowpea (Masvodza et al. 2014 ), Prosopis cineraria (L.) Druce (Palaiyur et al. 2016 ), flax ( Linumu sitatissimum L.) (Rozhmina et al. 2016 ), and Penthorum chinense Pursh (Zhiqiang et al. 2017). And also ISSR method has been widely used in genetic diversity analysis because of its high reproducibility than RAPD method (Domenyuk et al. 2002 ; Galvan et al. 2003 ; Dogan et al. 2007 ). However, in stock ( M. incana ), there are few reports on the study using molecular markers (Amaal 2009; Bekir et al. 2016 ; Takashi and Kanae 2018). The present work aimed to: (1) make superior cultivars with different colors, high fertility, being capable of early selecting only double flowered seedlings by leaf color and examine morphological characteristics and (2) select RAPD and ISSR primers for cultivar certification and identification to culture and maintain good commercial stock cultivars. The white cultivar is highly fertile, but it is not possible to distinguish between single flowered individuals and double flowered ones. Otherwise, in case of the pink cultivar, although it shows very low fertility, it is possible to select double flowered seedlings according to leaf color; single flower phenotype with green color and double flower phenotype with pale green color. Here we evaluated genetic diversity among new double flowered stock cultivars obtained in cross “white” cultivar × “pink” cultivar by using morphological traits including leaf color, leaf shape, flower color and double flower phenotype, and RAPD and ISSR markers. In this study we made some good stock cultivars with different colors including pink, pale pink and white (high fertile and capable of distinguishing between single and double flowered seedlings) through outcrossing between white cultivar (high fertile but impossible to select double flower phenotype) and pink cultivar (vis. versa). In newly obtained stock cultivars, it is possible to distinguish between single and double flower seedlings by leaf color, having about 70 ~ 75% of fertility, thereby they are efficient good cultivars than both parent cultivars in seed production and culture. Additionally, our cultivars were classified into two main groups, including the pink and the white cultivars respectively, the genetic distances among them varied from 0.03 to 0.24, with similarity to the previous research results in apricot and cowpea (Chroboková et al. 2011 ; Masvodza et al. 2014 ). In the future, by using our selected RAPD and ISSR primers, the present method might be contributed to establish the cultivar certification, identification, and conservation system in culture and management of double flowered stock cultivars with high fertility and different colors. Materials And Methods Plant material and cross Matthiola incana R. Br. cultivar “white” (DPR Korea) as a maternal parent and “pink” (DPR Korea) as a paternal parent were grown in a greenhouse at Pyongyang floriculture institute. In the cultivar “white”, plant height is high with no hairy leaves, white flowers, and high ratio of seed production (approximately 95%) otherwise, in the cultivar “pink”, plant height is lower than “white” with hairy leaves, pink flowers, and very low ratio of seed production (about 2.2%), besides with different leaf color between single and double flowered individuals. We fertilized a “white” plant’s ovules using pollen produced by “pink” plant. We obtained F 1 offspring by growing 320 seeds resulted from a hybridized plant, and mixed the seeds by self-fertilizing F 1 heterozygous plants. In the F 2 progeny produced from 10 plants of them, the selection was performed according to floral and leaf’s traits among 1757 plants. Subsequently, we conducted a family selection in the F 3 generation and selection of fixed line in the F 4 generation, finally obtained 4 pink, 2 pale pink and 2 white stock cultivars fixed in targeting traits in the F 5 generation. Morphological analysis Observations were made on each plant for phenotypic traits such as leaf color, leaf shape and flower color. The following plant measurements were recorded: plant height, stem length, number of leaves per plant, leaf length and width, flower diameter, inflorescence length; numbers of flowers in inflorescence and pod (silique) length. The data obtained from the research study was statistically analyzed by using ANOVA in Statistica ver. 6.0 and means were compared using Tukey’ s tests at the 5% level of significance. All means are presented with standard error. Molecular analysis Genomic DNA isolation Fresh leaf samples were collected from individuals of all cultivars according to flower type (single or double) and leaf shape (hairy and no hairy). Genomic DNA were isolated from 500 mg of fresh leaf material using CTAB method (Cetyl Trimethyl Ammonium Bromide) developed by Doyle and Doyle ( 1990 ). Qualitative and Quantitative estimation of DNA DNA quality was assessed by using Nanodrop Spectrophotometer (Thermoscientific Nanodrop 1000, USA) at the absorbance ratio of 260 and 280 nm providing a value of 1.8 which determines pure DNA preparation. Quality of DNA fragment was electrophoretically analyzed through 0.8% agarose gel using 1X TAE buffer at 50 V for 45 mins. RAPD PCR amplification RAPD PCR amplification was performed to amplify randomly unknown target sequences by using arbitary random primers according to the protocol described by Williams et al. ( 1990 ). PCR was carried out in a 25µl reaction volume containing 2.5µl of 10X Taq buffer, 2.5µl of dNTPs, 2.5µL of MgCl 2, 1µl of primer, 0.1µL 0.1U/µL Taq polymerase, 1µl of temple DNA (100 ng/ µl) and 15.4 µl of ddH 2 O for each sample in a Mastercycler (nexus gradient). We initially tested 25 primers from RAPD primer set (Opéron, sets D, H, I, N and P) on two individuals for each of the eight cultivars. We selected 10 primers (OPD3, OPH18, OPI14, OPN4, OPP19, OPH8, OPH19, OPP9, OPP17 and OPP20) with both reproducible and polymorphic variation with well defined and darkly staining bands. Mastercycler (nexus gradient) programmed for an initial denaturation step of 94°C for 5 min, followed by 40 cycles of 30 s at 94°C, 60 s at 37°C and 90 s at 72°C, a final extension step of 72°C for 2 min. Amplification products were separated on 1.5% agarose gels in 1×TAE buffer at 80 V for 1h 30 mins. Gels were stained with ethidium bromide and photographed under UV light by using Geldoc-It™ (USA). ISSR PCR amplification ISSR PCR amplification was performed by using 7 primers with high GC content according to the protocol described by Bekir et al. ( 2016 ). Data analysis Only clear and polymorphic DNA bands were used for data analysis. The bands were scored as present (1) or absent (0) and a binary data matrix was constructed. DNA fragments of identical size amplified with the same primer were considered to be the same DNA marker. To examine the genetic relationship among populations, a dendrogram was constructed using the unweighted paired group method of cluster analysis using arithmetic averages (UPGMA) and principal coordinate analysis (PCA) was performed with NTSYSpc version 2.20 (Rohlf 2008 ). Based on RAPD and ISSR data, the POPGENE ver. 1.32 (Yeh et al. 2000 ) was used to estimate values meaning genetic diversity: number of polymorphic band, observed number of alleles, effective number of alleles (Kimura and Crow 1964 ), Nei’s (1973) gene diversity (h), Shannon’s information index (I) (Lewontin 1972 ) and genetic distances among populations (Nei 1978 ). A dendrogram of the populations was constructed which has been based on the unweighed pair group method (UPGMA) analysis of Nei’s (1978) genetic distances. And also, G ST representing population differentiation and gene flow (Nm) among populations were estimated. Using the Arlequin ver. 3.5.2.2 (Excoffier and Lischer 2010 ), an analysis of molecular variance (AMOVA) was performed to evaluate the distribution of genetic variation within and among stock cultivars, and the Fixation index (Fst) was estimated too. Results Morpological diversity of new double flowered stock cultivars We compared and analysed the morphological traits in new double flowered stock cultivars obtained from outcrossing including 4 pink, 2 pale pink and 2 white cultivars (Fig. 1 ). Table 1 Characteristics of morphological traits in stock cultivas No cultivar percent of double flowered plants /% Flower color double flower phenotype Population ID Flower diameter /cm inflorescence length/cm number of flowers in inflorescence Leaf color Leaf shape Plant height /cm number of leaves per plant Leaf length /cm Leaf width /cm Pod length/ cm percent of fertile plants/% 1 《pink》 (♂) 54.3 ± 5.8 pink double 1 5.0 ± 0.5 20.5 ± 2.8 16.5 ± 2.4 pale green no hairy 50.8 ± 2.4 17.5 ± 2.4 12.8 ± 1.4 3.6 ± 0.3 2.8 ± 0.3 2.12 ± 0.8 single 2 3.6 ± 0.3 15.3 ± 2.5 15.3 ± 1.8 green 46.1 ± 1.8 18.5 ± 2.1 12.1 ± 1.8 2.5 ± 0.7 2 《white》 53.4 ± 5.6 white double 3 5.2 ± 0.7 27.5 ± 1.9 21.0 ± 1.5 pale green no hairy 80.8 ± 3.4 27.7 ± 0.9 15.4 ± 2.2 2.8 ± 0.2 2.7 ± 0.2 2.12 ± 0.7 single 4 4.0 ± 1.1 17.6 ± 2.2 12.4 ± 0.9 green 79.8 ± 5.9 28.9 ± 1.6 12.1 ± 2.3 2.4 ± 0.6 3 《white》 (♀) 47.4 ± 4.4 white double 4.7 ± 0.7 19.8 ± 2.6 18.3 ± 1.1 green hairy 53.8 ± 6.2 26.2 ± 2.3 12.4 ± 1.9 2.7 ± 0.9 7.6 ± 0.6 99.7 ± 5.6 single 5 3.6 ± 0.4 18.3 ± 3.3 17.3 ± 1.3 green 50.7 ± 3.8 28.8 ± 2.1 11.4 ± 2.4 2.3 ± 0.7 4 《pink 1》 49.3 ± 2.8 Pink double 6 4.4 ± 0.7 28.8 ± 1.7 22.8 ± 2.4 pale green no hairy 80.8 ± 7.6 27.7 ± 2.7 15.4 ± 1.9 2.8 ± 0.5 5.4 ± 0.4 75.5 ± 7.2 single 7 3.6 ± 0.2 18.9 ± 2.3 16.5 ± 1.7 green 79.8 ± 4.5 28.9 ± 1.1 12.1 ± 2.4 2.4 ± 0.4 5 《pink 2》 52.8 ± 3.6 Pink double 8 4.7 ± 0.8 27.9 ± 3.4 22.4 ± 2.7 pale green hairy 80.8 ± 6.3 27.7 ± 2.5 15.4 ± 2.3 2.8 ± 0.3 5.3 ± 0.9 72.3 ± 6.7 single 9 4.0 ± 0.4 17.7 ± 2.9 16.2 ± 1.9 green 79.8 ± 4.2 28.9 ± 1.8 12.1 ± 1.2 2.4 ± 0.5 6 《pink 3》 53.2 ± 3.8 Pink double 10 5.2 ± 0.9 28.2 ± 2.4 21.0 ± 2.6 pale green no hairy 80.8 ± 5.3 27.7 ± 3.2 15.4 ± 2.4 2.8 ± 0.4 5.5 ± 0.6 73.5 ± 8.5 single 11 4.0 ± 0.3 18.5 ± 3.2 15.4 ± 1.7 green 79.8 ± 2.9 28.9 ± 2.9 12.1 ± 2.9 2.4 ± 0.4 7 《pink 4》 46.4 ± 2.7 Pink double 12 5.2 ± 0.5 27.3 ± 2.6 22.0 ± 2.6 pale green hairy 80.8 ± 3.4 27.7 ± 2.2 15.4 ± 1.8 2.8 ± 0.8 5.4 ± 0.7 75.1 ± 9.4 single 13 4.0 ± 0.2 17.8 ± 2.1 14.8 ± 1.7 green 79.8 ± 2.8 28.9 ± 1.9 12.1 ± 1.3 2.4 ± 0.5 8 《pale pink 1》 51.7 ± 4.2 Pink double 14 5.0 ± 0.4 20.9 ± 3.8 18.3 ± 2.4 pale green no hairy 45.4 ± 3.6 26.8 ± 2.0 12.2 ± 1.1 3.0 ± 0.3 5.4 ± 0.3 73.5 ± 8.2 single 15 3.6 ± 0.7 17.8 ± 2.7 17.3 ± 1.7 green 47.8 ± 3.8 24.8 ± 2.3 11.2 ± 1.4 2.6 ± 0.6 9 《pale pink 2》 49.4 ± 2.8 Pink double 16 4.4 ± 0.6 19.9 ± 1.2 18.3 ± 2.1 pale green hairy 44.4 ± 4.2 15.5 ± 2.8 12.6 ± 1.9 3.4 ± 0.5 5.3 ± 0.7 70.5 ± 9.2 single 17 3.6 ± 0.8 15.9 ± 2.4 17.3 ± 1.6 green 43.8 ± 3.9 16.5 ± 2.6 12.4 ± 1.6 3.2 ± 0.2 10 《white 1》 48.4 ± 4.6 White double 18 4.7 ± 0.3 28.5 ± 2.1 20.8 ± 1.4 pale green no hairy 80.8 ± 2.8 27.7 ± 3.1 15.4 ± 2.8 2.8 ± 0.7 5.6 ± 0.9 75.5 ± 7.2 single 19 4.0 ± 0.4 18.1 ± 1.7 14.8 ± 2.3 green 79.8 ± 4.2 28.9 ± 1.9 12.1 ± 0.4 2.4 ± 0.4 11 《white 2》 53.2 ± 4.1 White double 20 5.2 ± 0.5 27.9 ± 2.9 20.5 ± 2.0 pale green no hairy 80.8 ± 3.9 27.7 ± 2.3 15.4 ± 1.7 2.8 ± 0.6 5.5 ± 0.5 71.9 ± 6.9 single 21 4.0 ± 0.9 18.0 ± 1.9 14.4 ± 1.6 green 79.8 ± 2.7 28.9 ± 3.1 12.1 ± 1.6 2.4 ± 0.9 As shown in Table 1 , in newly obtained 8 stock cultivars, the ratio of double flowered plants was about 50%, and some traits such as flower diameter, leaf length and leaf width, showed no significant differences (P > 0.05) in all 11 cultivars including both parent. Conversely, in other traits including inflorescence length, number of flowers per inflorescence, plant height, number of leaves per plant, “white” cultivar was markedly bigger than “pink” cultivar (P 0.05), “white”, whereas pale pink cultivars were not significantly different from paternal parent, “pink” (P > 0.05). Besides, in pod length and the percent of fertile plants, new eight cultivars had medium values between maternal and paternal parent cultivars, not showing a significant difference (P > 0.05) according to cultivar. Molecular identification of new stock cultivars using RAPD and ISSR markers Genetic relationship among twenty one stock individuals In the present study the genetic relationship among and within cultivars of M. incana were analysed by ten informative RAPD and ISSR primers (Table 2 ). Table 2 Primers used for RAPD and ISSR analysis, total number of scored fragments and their size range Primer Sequence (5’→ 3’) Fragment size range (bp) Total number of bands OPD3 5’ GTCGCCGTCA 3’ 100–4000 10 OPH18 5’ GAATCGGCCA 3’ 300–2000 6 OPI14 5’ TGACGGCGGT 3’ 100–2500 11 OPN4 5’ GACCGACCCA 3’ 100–3000 12 OPP19 5’ GGGAAGGCA 3’ 200–2000 5 ISSRM1 (AGC) 6 -G 200–3000 10 ISSRM2 (ACC) 6 -G 150–2000 10 ISSRM3 (AGC) 6 -C 100–4000 12 ISSRM5 (GA) 9 -C 200–3000 11 ISSRF3 (AG) 8 -CG 150–2000 10 Based on above obtained RAPD and ISSR profiles (Fig. 3), genetic relationship among 21 stock samples was estimated using NTSYSpc2.11. We obtained data matrix based on Nei ( 1973 )’s genetic distance and constructed a dendrogram using UPGMA method. As shown in Fig. 4 , at genetic distance of 0.15, dendrogram grouped all individuals in two clusters, and principal coordinate analysis result (Fig. 5 ) was agreed with cluster analysis (Fig. 4 ). First cluster, Ⅰ contained No. 1, 9, 10, 12, 6, 14, 16 individuals, being pink and single flowered. Second cluster, Ⅱ contained No. 2, 13, 8, 11, 7, 15, 17 individuals, being pink and double flowered. Third cluster, Ⅲ contained No. 3, 4, 18, 20 individuals, being white and single flowered. Final fourth cluster, Ⅳ contained No. 5, 21, 19 individuals, being white and double flowered. This suggests that our selected RAPD and ISSR primers might reflect not only flower color but also double flower phenotype. Genetic relationship among eleven stock cultivars Genetic relationship among eleven stock cultivars distinguished by flower color and leaf shape was evaluated using PopGene.version 1.31. Table 3 Nei’s (1978) genetic distance among eleven studied stock populations. The abbreviated population names are given according to Table 1 . pop ID 1 2 3 4 5 6 7 8 9 10 11 1 **** 0.8741 0.8612 0.9480 0.9694 0.9604 0.9636 0.9440 0.9393 0.8437 0.8638 2 0.1346 **** 0.9439 0.8031 0.8296 0.8219 0.8282 0.8124 0.8152 0.8826 0.8918 3 0.1494 0.0577 **** 0.8280 0.8150 0.8169 0.8132 0.7925 0.8066 0.9424 0.9514 4 0.0534 0.2193 0.1888 **** 0.9153 0.9163 0.9144 0.8845 0.8964 0.8055 0.8501 5 0.0311 0.1868 0.2015 0.0885 **** 0.9549 0.9597 0.9209 0.9393 0.7928 0.8044 6 0.0404 0.1961 0.2022 0.0874 0.0461 **** 0.9507 0.9124 0.9169 0.7855 0.7969 7 0.0371 0.1885 0.2068 0.0895 0.0412 0.0505 **** 0.9104 0.9048 0.7947 0.8160 8 0.0576 0.2077 0.2326 0.1227 0.0824 0.0917 0.0939 **** 0.9681 0.8112 0.8092 9 0.0626 0.2044 0.2149 0.1094 0.0626 0.0867 0.1000 0.0324 **** 0.8074 0.8155 10 0.1699 0.1249 0.0594 0.2163 0.2322 0.2415 0.2298 0.2092 0.2140 **** 0.9684 11 0.1464 0.1146 0.0498 0.1624 0.2177 0.2270 0.2033 0.2118 0.2040 0.0321 **** Nei’s genetic identity (above diagonal) and genetic distance (below diagonal). As shown in Fig. 6 , eleven stock cultivars were classified into two main groups; pink and white cultiivar group. The genetic distance values for all 11 cultivars ranged from 0.03 to 0.25 showing the lowest genetic distance (0.02415) between “pink 3” and “white 1”, while the highest genetic distance (0.0321) between “white 1” and “white 2” (Table 3 ). In the group of white cultivars, new cultivar “white 2” was genetically closest to control cultivar “white” (genetic distance- 0.0498), and new cultivar “white 1” was the most distant to maternal cultivar “white” (genetic distance- 0.1062). Otherwise, in the group of pink cultivars, new cultivar “pink 2” was the genetically closest to paternal cultivar “pink” (genetic distance- 0.0534), and new cultivar “pink 1” was the most distant to new cultivar “pale pink 1” (genetic distance- 0.0965). Among 11 newly obtained stock cultivars, differentiation index (G ST) and gene flow index (Nm) were 0.639(0.1309-1.000) and 0.2817(0.0430–3.3195) respectively. The values of genetic diversity indices for studied stock cultivars are given in Table 4 . The highest n a was observed for “pale pink 2” (1.26) and the least for “pink 3” (1.07), showing the lowest Shannon’s index (0.04) and Nei’s gene diversity (0.03) as well. Nei’s gene diversity were estimated to be 0.03 and 0.11 and the Shannon indices of the M. incana cultivars ranged from 0.04 to 0.16. Table 4 Genetic diversity indices for eleven stock cultivars. N PB - number of polymorphic bands, P PB -percentage of polymorphic bands, n a – observed number of alleles, n e –effective number of alleles, h - Nei’s (1987) gene diversity, I - Shannon’s information index Cultivar N PB P PB n a n e h I 《pink》(paternal) 5 10.87 1.11 ± 0.31 1.08 ± 0.22 0.05 0.07 《white》 0 0 1 1 0 0 《white》(maternal) 8 17.39 1.17 ± 0.38 1.23 ± 0.27 0.07 0.11 《pink 1》 8 17.39 1.17 ± 0.38 1.23 ± 0.27 0.07 0.11 《pink 2》 5 10.87 1.11 ± 0.31 1.08 ± 0.22 0.05 0.07 《pink 3》 3 6.52 1.07 ± 0.25 1.05 ± 0.18 0.03 0.04 《pink 4》 7 15.22 1.15 ± 0.36 1.11 ± 0.26 0.06 0.09 《pale pink 1》 8 17.39 1.17 ± 0.38 1.23 ± 0.27 0.07 0.11 《pale pink 2》 12 26.09 1.26 ± 0.44 1.18 ± 0.31 0.11 0.16 《white 1》 11 23.91 1.24 ± 0.43 1.17 ± 0.31 0.10 0.14 《white 2》 7 15.22 1.15 ± 0.36 1.11 ± 0.26 0.06 0.09 Table 5 Analysis of molecular variance (AMOVA) based on RAPD profiless in eleven stock cultivars Source of variation degrees of freedom Sum of squares Variance components Percentage of variation (%) P Among cultivars 10 283.562 2.584 67.27 < 0.0001 Within cultivars 20 105.600 1.257 32.73 < 0.0001 Total 30 389.162 3.841 Genetic variation among cultivars was more than that within cultivars, as indicated by the sum of squares values calculated by AMOVA using Arlequin ver. 3.5.2.2 (Table 5 ). Of the total genetic diversity, 67.27 % was attributable to differences among culivars and 32.73% was to differences within culivars, showing a significant varietal differentiation in newly obtained stock cultivars. The fixation index (Fst) was 0.673. Discussion Because double flowered cultivars have superior ornamental value to single flowered ones, stock breeders have attempted to make double flowered cultivars with different colors. Additionally, in breeding of M. incana , the studies also have been reported to identify the morphological and molecular markers which could be used to discriminate the double flowered individuals. Ecker et al. ( 1993 ) founded that the c allele for sinuate leaf shape might be recessive to the C allele for normal entire leaf and it was tightly linked to the s recessive allele for double flowering, suggesting its role as a double flowering marker in the cultivation and the breeding of M. incana. Takashi and Kanae (2018) revealed that co-dominant marker, MiAG could be used for early discrimination of double flowered individuals among seedlings not showing phenotypic differences. He examined that the single flowered individuals of cultivars being distinguished double flowered ones according to cotyledon shape (‘Kiss me white’, ‘Iron white’, ‘Quartet cherry’), serrated leaf (‘Pygmy white’), seed color (‘White wonder no.2′) were heterozygotes MiAG/miag1 , whereas the individuals correlated with leaf color (in ‘Aida’ and ‘Opera’) were heterozygotes MiAG/miag2. In this study we made the garden stock varieties with different colors (pink, pale pink and white) and high fertility which were capable of discriminating double flowered individuals in cross “white” cultivar (high fertile but impossible in selection of double flowered plants) × “pink” cultivar (very low fertile but possible in selection of double flowered plants). Considering with Takashi and Kanae et al. (2018)’s result, our newly raised stock varieties are likely MiAG/miag2 heterozygotes being capable of selecting double flowered ones among the seedlings according to leaf color (Fig. 2 ), and harboring high fertility, therefore they might be superior to both parents cultivars in the cultivation and seed production for commercial use. To date, random amplified polymorphic DNA (RAPD) method have been widely applied to assess the intra- and inter-specific genetic diversity in many plants. Tanmay et al. (2016) described the diversity of nine genotypes in the genus Ocimum from India based on RAPD markers combining with morphological and chemical markers. Prabhjot et al. (2019) assessed the molecular diversity among 48 accessions of five Swertia species collected from Western Himalayas, India using 18 RAPD markers. RAPD primer based genetic diversity within the species were reported earlier in several plant species such as Citrullus colocynthis (Kumar et al. 2017), lettuce ( Lactuca sativa L.) (Shubhangi et al. 2018), Nilgirianthus ciliates (Ramakrishnan et al. 2019) and Artemisia herba -alba (Khaled et al. 2019). Besides, RAPD technique has also been used to examine the genetic diversity on the level of variety or cultivar (Maciel et al. 2001 ; Siddharth et al. 2007; Chroboková et al. 2011 ; Masvodza et al. 2014 ; Palaiyur et al. 2016 ; Rozhmina et al. 2016 ; Zhiqiang et al. 2017). Palaiyur et al. ( 2016 ) showed that the similarity coefficients ranged from 0.57 to 0.78 by using nine of 80 RAPD primers and they could be applied to identify different varieties in Prosopis cineraria (L.) Druce. Rozhmina et al. ( 2016 ) revealed genetic similarity between cultivars of fiber flax from Russia and other the European countries using RAPD markers. On the other hand, ISSR method is more reproducible than RAPD method, so it is more widely used in the analysis of genetic diversity in many plants such as maize (Domenyuk et al. 2002 ), common bean (Galvan et al. 2003 ) and Jurinea (Dogan et al. 2007 ). And also, recent researchers (Ramakrishnan et al. 2019; Joyashree et al. 2019; Zhiqiang et al. 2017; Kumar et al. 2017) reported that RAPD and ISSR markers could be applied together to reveal genetic relationship between germplasm lines and to establish the cultivar identification system using cluster analysis and PCA. However, there are few reports in stock plant ( Matthiola incana ) to analyze genetic relationship using molecular markers. Amaal (2009) performed molecular systematic analysis for twenty two taxa belonging to Brassicaceae including Matthiola incana and Matthiola longipetala subsp. livida using RAPD markers and suggested its role for varietal identification and assessment of genetic relationships even between closely related species. And Bekir et al. ( 2016 ) studied phylogenetic relationship of Turkish species of Matthiola based on only ISSR markers. In contrast to previous study, the present method might be applied to identify new double flowered stock plant varieties obtained through outcrossing, by using RAPD and ISSR markers together. By combining new selected RAPD and ISSR primers, our analysis method grouped stock plant varieties into two main clusters, one including pink varieties and the other including white ones. Besides single and double flowered individuals were classified in each cluster, and RAPD based genetic distances varied from 0.03 to 0.24 among these varieties which is similar to Chroboková et al. ( 2011 ) (0.02 ~ 0.45), Masvodza et al. ( 2014 ) (0.1 ~ 0.22)’ s research result and has somewhat smaller scale than Palaiyur et al. ( 2016 ) (0.22 ~ 0.43), Maciel et al. ( 2001 ) (0.2 ~ 0.75), and Siddharth et al. (2007) (0.34 ~ 0.74)’ report. Conclusion We bred new double flowered stock cultivars with different colors and high fertility in cross white cultivar that was high fertile but impossible to select double flower phenotype × pink cultivar (vice. versa). RAPD and ISSR markers selected in this study can be used to identify different stock cultivars according to flower color and flowered phenotype, therefore in addition to morphological traits, they will be applied to cultivar certification, identification, and establishing conservation system in their culture and management. Abbreviations RAPD (Random Amplified Polymorphic DNA), ISSR (Inter Simple Sequence Repeat) Declarations Conflict of interest The authors declare that there are no conflicts of interest. Acknowledgements We thank all horticulturists of Pyongyang floriculture institute who helped culturation during stock breeding. References Amaal Hasan Mohamed (2009) Molecular Systematic of Some Brassicaceae Taxa in Egypt Based on Electrophoretic Isoenzymes Pattern and RAPD Markers. Aust J Basic & Appl Sci 3(3): 1499-1511 Bekir DOĞAN, Mustafa ÇELİK, Murat ÜNAL, Abdurrahman SEFALI, Esra MARTİN, Ayla KAYA (2016) Study of phylogenetic relationship of Turkish species of Matthiola (Brassicaceae) based on ISSR amplification. Turk J Bot 40: 130-136 Chroboková E, Raddová J, Vachůn M, Krška B, Pidra M (2011) An analysis of apricot cultivars by random amplified polymorphic DNA and microsatellite primers. Hort Sci (Prague) 38(4): 125–133 Daozong Chen, Yi Liu, Qi Pan, Fengfeng Li, Qinghua Zhang, Xianhong Ge, Zaiyun Li (2018) De novo transcriptome assembly, gene expressions and metabolites for flower color variation of two garden species in Brassicaceae. Sci Hortic 240: 592 – 602 Dogan B, Duran A, Hakki E E (2007) Phylogenetic analysis of Jurinea ( Asteraceae ) species from Turkey based on ISSR amplifiation. Ann Bot Fenn 44: 353–358 Domenyuk V P, Verbitskaya T G, Belousov A A, Sivolap Y M (2002) Marker analysis of quantitative traits in maize by ISSR-PCR. Russian J Genet 38: 1161–1168 Doyle J J, Doyle J L (1990) Isolation of plant DNA from fresh tissue. Focus 12: 13-15 Ecker R, Barzilay A, Osherenko E (1993) Linkage relationships of genes for leaf morphology and double flowering in Matthiola incana . Euphytica 74: 133 –136 Excoffier L, Lischer H E L (2010) Arlequin suite ver 3.5: a new series of programs to perform population genetics analyses under Linux and Windows. Mol Ecol Resour 10: 564–567 Galvan M Z, Bornet B, Balatti P A, Branchard M (2003) Inter simple sequence repeat (ISSR) markers as a tool for the assessment of both genetic diversity and gene pool origin in common bean ( Phaseolus vulgaris L.). Euphytica 132: 297–301 Joyashree Baruah, Sudin Kumar Pandey, Twahira Begum, Neelav Sarma, Manabi Paw, Mohan Lal (2019) Molecualr diversity assessed amongst high dry rhizome recovery Ginger germplasm ( Zinger officinale Roscoe) from NE-India using RAPD and ISSR markers, Industrial Crops & Products 129: 463-471 Khaled Elmeer, Abdelrezak Elkhgkheg (2019) Genetic Diversity of Artemisia herba-alba in Libyan Green Mountain. Iranian Journal of Science and Technology, Transactions A: Science 43(4): 1507–1512 Kimura M, Crow J F (1964) The number of alleles that can be maintained in a finite population. Genetics 49: 725-738 Kumar Sambhav Verma, Shamshad ul Haq, Sumita Kachhwaha, Kothari S L (2017) RAPD and ISSR marker assessment of genetic diversity in Citrullus colocynthis (L.) Schrad: a unique source of germplasm highly adapted to drought and high-temperature stress. 3 Biotech 7: 288 Lewontin R C (1972) The apportionment of human diversity. Evol Biol 6: 381–398 Maciel F L, Gerald L T S, Echeverrigaray S (2001) Random amplified polymorphic DNA (RAPD) markers variability among cultivars and landraces of common beans ( Phaseolus vulgaris L.) of south-Brazil. Euphytica 120: 257–263 Masvodza D R, Mazhude N, Mugabe J, Musango R (2014) Preliminary genetic diversity Analysis of introduced and local Zimbabwe cowpea landraces. African Journal of Agricultural Research 9(49): 3571-3580 Nei M (1973) Analysis of gene diversity in subdivided populations. P Natl Acad Sci USA 70: 3321-3323 Nei M (1978) Estimation of average heterozygosity and genetic distance from a small number of individuals. Genetics 89: 583-590 Palaiyur N, Sivalingam Dilip K, Samadia Dhurendra Singh, Sarita Chauhan (2016) Molecular markers to distinguish ‘Thar Shoba’, a variety of khejri [Prosopis cineraria (L.) Druce], from trees in natural populations. J Hortic Sci Biotech 91(4): 353-361 Prabhjot Kaur, Devendra Kumar Pandey, Gupta R C, Abhijit Dey (2019) Assessment of genetic diversity among different population of five Swertia species by using molecular and phytochemical markers. Ind Crop Prod 138: 111569 Ramakrishnan Rameshkumar, Subramani Pandian, Periyasamy Rathinapriya, Chinnar Tamil Selvi, Lakkakula Satish, Shanmugaraj Gowrishankar, David W.M. Leung, Manikandan Ramesh (2019) Genetic diversity and phylogenetic relationship of Nilgirianthus ciliatus populations using ISSR and RAPD markers: Implications for conservation of an endemic and vulnerable medicinal plant. Biocatalysis and Agricultural Biotechnology 18: 101072 Rohlf F J (2008) NTSYSpc: Numerical taxonomy system, ver. 2.20. (Exerter Publishing Ltd., Setauket, New York). Rozhmina T A, Fu Y-B, Diederichsen A, Richards K W, Pavelek M, Miroslava Vrbova Ms (2016) Research of Genetic Polymorphism Species Linumu sitatissimum L. on a Basis a RAPD-Method. J Nat Fibers 15(2): 155-161 Saunders E S (1911) Further experiments on the inheritance of doubleness and other characters in stock. J Genet 1: 303–376 Saunders E S (1915) A suggested explanation of the abnormally high records of doubles quoted by growers of stocks ( Matthiola ). J Genet 5: 137–143 Saunders E S (1921) Note on the evolution of the double stock ( Matthiola incana ). J Genet 11: 69–74. Saunders E S (1928) Further studies on inheritance in Matthiola incana . II. Plastid colour and doubing. J Genet 20: 53–77. Shubhangi Sharma, Pankaj Kumar, Geetika Gambhir, Ramesh Kumar, Srivastava D K (2018) Assessment of genetic diversity in lettuce ( Lactuca sativa L.) germplasm using RAPD markers. 3 Biotech 8: 9 Siddharth Kumar Palai, Gyana Ranjan Rout (2007) Identification and genetic variation among eight varieties of ginger by using random amplified polymorphic DNA markers. Plant Biotechnol 24: 417–420 Sima Jafari a , Seyyed Ebrahim Hashemi Garmdareh, Behzad Azadegan (2019) Effects of drought stress on morphological, physiological, and biochemical characteristics of stock plant ( Matthiola incana L.). Sci Hortic 253: 128-133 Sima Jafari b , Seyyed Ebrahim Hashemi Garmdareh (2019) Effects of salinity on morpho-physiological, and biochemical characteristics of stock plant ( Matthiola incana L.). Sci Hortic 257: 108731 Takashi Nakatsukaa, Kanae Koishi (2018) Molecular characterization of a double-flower mutation in Matthiola incana . Plant Sci 268: 39–46 Tanmay Chowdhury, Amitava Mandal, Subhas Chandra Roy, Dilip De Sarker (2016) Diversity of the genus Ocimum ( Lamiaceae ) through morpho-molecular (RAPD) and chemical (GC–MS) analysis. Journal of Genetic Engineering and Biotechnology 15(1): 275–286. Tasaki K, Higuchi A, Fujita K, Watanabe A, Sasaki N, Fujiwara K, Abe H, Naito Z, Takahashi R, Hikage T, Nishihara M (2017) Development of molecular markers for breeding of double flowers in Japanese gentian. Mol Breed 37. Tatsuzawa F, Saito N, Toki K, Shinoda K, Honda T (2012) Flower colors and their anthocyanins in Matthiola incana cultivars ( Brassicaceae ). J Jpn Soc Hortic Sci 81: 91–100 Williams J G K, Kubelik A R, Livak K J, Rafalski J A, Tingey S V (1990) DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res 18: 653l-6535 Yeh F C, Boyle T, Yang R-C, Ye Z M J X (2000) POPGENE, version 1.32. Department of Renewable Resources, University of Alberta, Edmonton, AB, Canada. Zhiqiang Mei, Xianqin Zhang, Md Asaduzzaman Khan, Saber Imani, Xiaoyan Liu, Hui Zou, Chunli Wei, Junjiang Fu (2017) Genetic analysis of Penthorum chinense Pursh by improved RAPD and ISSR in China. Electron J Biotechn 30: 6–11 Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. 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-578473","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":33536244,"identity":"293a1230-3092-4e29-84a3-a8ff0fe7081b","order_by":0,"name":"Un-Hyang Ho","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABAElEQVRIiWNgGAWjYDACCQY2BoYKDEGGBAJazpCshbGNFC0GtxvYHnycdy+xX7r92oOfOXXy5gzMB2/zMKTl4dRy5wC74cxtxYkz55wpN+zddthwZwNbsjUPQ04xTi03EtikebclJG64kZMmwbvtAOOGAzxm0jwMFYkNeLXMgWiR/Lutzn7DAf5vRGhpAGlJPwa0jjkRaAsbUEsOTi2SNxLbJGccSzCeOSOHTVp22+HkDYfZjC3nGKTh9AvfjeRjEh9qEmT7JdKfSb7dVme74XjzwxtvKpJxhhgDAyPYAY4NDDwGEAFmsIPxxSUE2DMwsD9AESGoZRSMglEwCkYMAAAin1kcbZeJ2wAAAABJRU5ErkJggg==","orcid":"","institution":"Kim Il Sung University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Un-Hyang","middleName":"","lastName":"Ho","suffix":""},{"id":33536245,"identity":"6ccc7be9-6412-4a13-8e6c-4bdffb8772c1","order_by":1,"name":"Jong-Hyang Ri","email":"","orcid":"","institution":"Kim Il Sung University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Jong-Hyang","middleName":"","lastName":"Ri","suffix":""},{"id":33536246,"identity":"f7dde404-cc69-43a1-9584-51b2398c8e23","order_by":2,"name":"Chol-Jun Ri","email":"","orcid":"","institution":"Pyongyang Floriculture Institute","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Chol-Jun","middleName":"","lastName":"Ri","suffix":""}],"badges":[],"createdAt":"2021-05-31 19:20:08","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-578473/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-578473/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":10506454,"identity":"3d344d3b-84bb-45b4-8529-7ac61c1cd567","added_by":"auto","created_at":"2021-06-17 20:05:48","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2663461,"visible":true,"origin":"","legend":"Phenotypes of newly obtained stock plants and both parent plants\n“-s” means single flowered phenotype and “-d” means double flowered phenotype.\n","description":"","filename":"fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-578473/v1/e81fd27dd6341b3d2196408d.png"},{"id":10506588,"identity":"72ab04ca-683a-408f-ad5e-5fd6472d7ffe","added_by":"auto","created_at":"2021-06-17 20:08:48","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":867097,"visible":true,"origin":"","legend":"Leaf color in young seedlings in newly obtained stock plants (green: double flowered seeding, pale green: single flowered seeding)","description":"","filename":"fig2.png","url":"https://assets-eu.researchsquare.com/files/rs-578473/v1/4829b37d5e5bc3ac3697171e.png"},{"id":10506652,"identity":"2111a365-d362-4740-afe8-2d7c3d782445","added_by":"auto","created_at":"2021-06-17 20:11:48","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":361333,"visible":true,"origin":"","legend":"RAPD and ISSR agarose gel electrophoresis profiles of eleven stock cultivars using OPD3(left) and ISSRM3(right) primers.\nLines marked by numbers (1, 2, 3…, 21) represent the individuals that belong to cultivars listed in table 1. M –1 kb ladder\n","description":"","filename":"fig3.png","url":"https://assets-eu.researchsquare.com/files/rs-578473/v1/5942e6f21d990e2e136edc08.png"},{"id":10506449,"identity":"90f71384-7a3a-43a8-8dfb-a05643cb13d0","added_by":"auto","created_at":"2021-06-17 20:05:48","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":29630,"visible":true,"origin":"","legend":"UPGMA dendrogram illustrating the genetic relationships between M. incana genotypes based on Nei's genetic distance. Individual Id is as population Id in table 1. 1,2 are male parents, 5 is the female parent.","description":"","filename":"fig4.png","url":"https://assets-eu.researchsquare.com/files/rs-578473/v1/366aa026481c0fd89b90ed44.png"},{"id":10506651,"identity":"50de2920-589b-4bf6-a783-2043bfb00692","added_by":"auto","created_at":"2021-06-17 20:11:48","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":85851,"visible":true,"origin":"","legend":"2-D(left) and 3-D(right) PCA plot based on RAPD and ISSR data in M. incana genotypes.","description":"","filename":"fig5.png","url":"https://assets-eu.researchsquare.com/files/rs-578473/v1/19a8154a722cb95202e22bf7.png"},{"id":10506586,"identity":"cfe98efa-5ad8-403f-abaa-605b938367c3","added_by":"auto","created_at":"2021-06-17 20:08:48","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":42791,"visible":true,"origin":"","legend":"UPGMA dendrogram illustrating the genetic relationships between eleven cultivars of M. incana.","description":"","filename":"fig6.png","url":"https://assets-eu.researchsquare.com/files/rs-578473/v1/0a47835b68ebf6f392e5713f.png"},{"id":15673118,"identity":"c8c3b6f9-e6d4-42c9-aebb-0a7d551ba8f8","added_by":"auto","created_at":"2021-11-18 14:16:22","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3069773,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-578473/v1/e736dad8-fa90-4e80-b730-2a295326f157.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eMorphological And Molecular Identification of Double Flowered Stock (\u003cem\u003eMatthiola Incana\u003c/em\u003e R. Br) Cultivars With High Fertility\u003c/p\u003e","fulltext":[{"header":"Introduction","content":" \u003cp\u003eGarden stock (\u003cem\u003eMatthiola incana\u003c/em\u003e R. Br.) is a commercially important horticultural crop having different types of flowers varied in color and flowered phenotype, belonging to the family \u003cem\u003eBrassicaceae\u003c/em\u003e (Tatsuzawa et al. \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2012\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eDue to high ornamental value, many studies associated in culture and breeding of the garden stock have focused on improvement of morphological and physiological traits such as flower color, double flower phenotype, inflorescence length, and fertility (Sima 2019a, b; Daozong et al. 2018).\u003c/p\u003e \u003cp\u003eThere are single and double flowered \u003cem\u003eM. incana\u003c/em\u003e cultivars, and latter is more widely used for floriculture because it has more beautiful flowers. Seed production is only possible in single flowered individuals. It is impossible in double flowered plants due to degenerative pistils and stamens.\u003c/p\u003e \u003cp\u003eTherefore many researchers and breeders have aimed to produce of more double flowered plants than single flowered ones in \u003cem\u003eM. incana\u003c/em\u003e and to develop markers to select them.\u003c/p\u003e \u003cp\u003eTo date, several morphological traits such as cotyledon shape, serrate leaf, seed color, and leaf color have been used in early selection of double flowered stock individuals, because these traits were linked to double flowers (Saunders \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e1911\u003c/span\u003e; \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e1915\u003c/span\u003e; \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1921\u003c/span\u003e; \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e1928\u003c/span\u003e; Ecker et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1993\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eTakashi and Kanae (2018) identified the gene related to double flowers and developed a molecular marker to select double flowered plants efficiently, based on the study of double flower-linked molecular markers in Japanese gentian (Tasaki et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn general, the double flower trait is inherited by a single locus, \u003cem\u003es\u003c/em\u003e and it is linked to leaf color. In some stock cultivars, leaf color is a simple morphological marker, that is, green color indicates single flowered individual while pale green color represents double flowered individual (Ecker et al. \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1993\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAnd also the fertility of plant is an important trait in cultivation and breeding of stock as well as other plants. Consequently, it is necessary to make a good stock cultivar with high fertility, different colors, and being capable of early discrimination of double flowered phenotype, in culture and production for commercial use.\u003c/p\u003e \u003cp\u003eRandom amplified polymorphic DNA technique has been widely applied to evaluate genetic diversity on intraspecific level such as Ocimum (\u003cem\u003eLamiaceae\u003c/em\u003e) (Tanmay et al. 2016) and Swertia (\u003cem\u003eGentianaceae\u003c/em\u003e) (Prabhjot et al. 2019) and interspecific level including \u003cem\u003eCitrullus colocynthis\u003c/em\u003e (Kumar et al. 2017), \u003cem\u003eLactuca sativa\u003c/em\u003e L. (Shubhangi et al. 2018), \u003cem\u003eNilgirianthus ciliates\u003c/em\u003e, \u003cem\u003eArtemisia herba\u003c/em\u003e-alba (Khaled et al. 2019).\u003c/p\u003e \u003cp\u003eMoreover, RAPD method has also been used to assess genetic diversity on the level of variety or cultivar in following plants, for example, common beans (\u003cem\u003ePhaseolus vulgaris\u003c/em\u003e L.) (Maciel et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2001\u003c/span\u003e), ginger (Siddharth et al. 2007), apricot (\u003cem\u003ePrunus armeniaca\u003c/em\u003e L.) (Chrobokov\u0026aacute; et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2011\u003c/span\u003e), cowpea (Masvodza et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2014\u003c/span\u003e), \u003cem\u003eProsopis cineraria\u003c/em\u003e (L.) Druce (Palaiyur et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), flax (\u003cem\u003eLinumu sitatissimum\u003c/em\u003e L.) (Rozhmina et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e), and \u003cem\u003ePenthorum chinense\u003c/em\u003e Pursh (Zhiqiang et al. 2017).\u003c/p\u003e \u003cp\u003eAnd also ISSR method has been widely used in genetic diversity analysis because of its high reproducibility than RAPD method (Domenyuk et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Galvan et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Dogan et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eHowever, in stock (\u003cem\u003eM. incana\u003c/em\u003e), there are few reports on the study using molecular markers (Amaal 2009; Bekir et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Takashi and Kanae 2018).\u003c/p\u003e \u003cp\u003eThe present work aimed to: (1) make superior cultivars with different colors, high fertility, being capable of early selecting only double flowered seedlings by leaf color and examine morphological characteristics and (2) select RAPD and ISSR primers for cultivar certification and identification to culture and maintain good commercial stock cultivars.\u003c/p\u003e \u003cp\u003eThe white cultivar is highly fertile, but it is not possible to distinguish between single flowered individuals and double flowered ones. Otherwise, in case of the pink cultivar, although it shows very low fertility, it is possible to select double flowered seedlings according to leaf color; single flower phenotype with green color and double flower phenotype with pale green color.\u003c/p\u003e \u003cp\u003eHere we evaluated genetic diversity among new double flowered stock cultivars obtained in cross \u0026ldquo;white\u0026rdquo; cultivar \u0026times; \u0026ldquo;pink\u0026rdquo; cultivar by using morphological traits including leaf color, leaf shape, flower color and double flower phenotype, and RAPD and ISSR markers.\u003c/p\u003e \u003cp\u003eIn this study we made some good stock cultivars with different colors including pink, pale pink and white (high fertile and capable of distinguishing between single and double flowered seedlings) through outcrossing between white cultivar (high fertile but impossible to select double flower phenotype) and pink cultivar (vis. versa). In newly obtained stock cultivars, it is possible to distinguish between single and double flower seedlings by leaf color, having about 70\u0026thinsp;~\u0026thinsp;75% of fertility, thereby they are efficient good cultivars than both parent cultivars in seed production and culture.\u003c/p\u003e \u003cp\u003eAdditionally, our cultivars were classified into two main groups, including the pink and the white cultivars respectively, the genetic distances among them varied from 0.03 to 0.24, with similarity to the previous research results in apricot and cowpea (Chrobokov\u0026aacute; et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Masvodza et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn the future, by using our selected RAPD and ISSR primers, the present method might be contributed to establish the cultivar certification, identification, and conservation system in culture and management of double flowered stock cultivars with high fertility and different colors.\u003c/p\u003e "},{"header":"Materials And Methods","content":"\u003cdiv class=\"Section2\" id=\"Sec3\"\u003e\n \u003cp\u003e\u003cem\u003ePlant material and cross\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003e\u003cem\u003eMatthiola incana\u003c/em\u003e R. Br. cultivar \u0026ldquo;white\u0026rdquo; (DPR Korea) as a maternal parent and \u0026ldquo;pink\u0026rdquo; (DPR Korea) as a paternal parent were grown in a greenhouse at Pyongyang floriculture institute. In the cultivar \u0026ldquo;white\u0026rdquo;, plant height is high with no hairy leaves, white flowers, and high ratio of seed production (approximately 95%) otherwise, in the cultivar \u0026ldquo;pink\u0026rdquo;, plant height is lower than \u0026ldquo;white\u0026rdquo; with hairy leaves, pink flowers, and very low ratio of seed production (about 2.2%), besides with different leaf color between single and double flowered individuals.\u003c/p\u003e\n \u003cp\u003eWe fertilized a \u0026ldquo;white\u0026rdquo; plant\u0026rsquo;s ovules using pollen produced by \u0026ldquo;pink\u0026rdquo; plant.\u003c/p\u003e\n \u003cp\u003eWe obtained F\u003csub\u003e1\u003c/sub\u003e offspring by growing 320 seeds resulted from a hybridized plant, and mixed the seeds by self-fertilizing F\u003csub\u003e1\u003c/sub\u003e heterozygous plants. In the F\u003csub\u003e2\u003c/sub\u003e progeny produced from 10 plants of them, the selection was performed according to floral and leaf\u0026rsquo;s traits among 1757 plants. Subsequently, we conducted a family selection in the F\u003csub\u003e3\u003c/sub\u003e generation and selection of fixed line in the F\u003csub\u003e4\u003c/sub\u003e generation, finally obtained 4 pink, 2 pale pink and 2 white stock cultivars fixed in targeting traits in the F\u003csub\u003e5\u003c/sub\u003e generation.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec4\"\u003e\n \u003cp\u003e\u003cem\u003eMorphological analysis\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eObservations were made on each plant for phenotypic traits such as leaf color, leaf shape and flower color.\u003c/p\u003e\n \u003cp\u003eThe following plant measurements were recorded: plant height, stem length, number of leaves per plant, leaf length and width, flower diameter, inflorescence length; numbers of flowers in inflorescence and pod (silique) length.\u003c/p\u003e\n \u003cp\u003eThe data obtained from the research study was statistically analyzed by using ANOVA in Statistica ver. 6.0 and means were compared using Tukey\u0026rsquo; s tests at the 5% level of significance. All means are presented with standard error.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec5\"\u003e\n \u003cp\u003e\u003cem\u003eMolecular analysis\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eGenomic DNA isolation\u003c/p\u003e\n \u003cp\u003eFresh leaf samples were collected from individuals of all cultivars according to flower type (single or double) and leaf shape (hairy and no hairy). Genomic DNA were isolated from 500 mg of fresh leaf material using CTAB method (Cetyl Trimethyl Ammonium Bromide) developed by Doyle and Doyle (\u003cspan class=\"CitationRef\"\u003e1990\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eQualitative and Quantitative estimation of DNA\u003c/p\u003e\n \u003cp\u003eDNA quality was assessed by using Nanodrop Spectrophotometer (Thermoscientific Nanodrop 1000, USA) at the absorbance ratio of 260 and 280 nm providing a value of 1.8 which determines pure DNA preparation. Quality of DNA fragment was electrophoretically analyzed through 0.8% agarose gel using 1X TAE buffer at 50 V for 45 mins.\u003c/p\u003e\n \u003cp\u003eRAPD PCR amplification\u003c/p\u003e\n \u003cp\u003eRAPD PCR amplification was performed to amplify randomly unknown target sequences by using arbitary random primers according to the protocol described by Williams et al. (\u003cspan class=\"CitationRef\"\u003e1990\u003c/span\u003e). PCR was carried out in a 25\u0026micro;l reaction volume containing 2.5\u0026micro;l of 10X Taq buffer, 2.5\u0026micro;l of dNTPs, 2.5\u0026micro;L of MgCl\u003csub\u003e2,\u003c/sub\u003e 1\u0026micro;l of primer, 0.1\u0026micro;L 0.1U/\u0026micro;L \u003cem\u003eTaq\u003c/em\u003e polymerase, 1\u0026micro;l of temple DNA (100 ng/ \u0026micro;l) and 15.4 \u0026micro;l of ddH\u003csub\u003e2\u003c/sub\u003eO for each sample in a Mastercycler (nexus gradient).\u003c/p\u003e\n \u003cp\u003eWe initially tested 25 primers from RAPD primer set (Op\u0026eacute;ron, sets D, H, I, N and P) on two individuals for each of the eight cultivars. We selected 10 primers (OPD3, OPH18, OPI14, OPN4, OPP19, OPH8, OPH19, OPP9, OPP17 and OPP20) with both reproducible and polymorphic variation with well defined and darkly staining bands.\u003c/p\u003e\n \u003cp\u003eMastercycler (nexus gradient) programmed for an initial denaturation step of 94\u0026deg;C for 5 min, followed by 40 cycles of 30 s at 94\u0026deg;C, 60 s at 37\u0026deg;C and 90 s at 72\u0026deg;C, a final extension step of 72\u0026deg;C for 2 min.\u003c/p\u003e\n \u003cp\u003eAmplification products were separated on 1.5% agarose gels in 1\u0026times;TAE buffer at 80 V for 1h 30 mins.\u003c/p\u003e\n \u003cp\u003eGels were stained with ethidium bromide and photographed under UV light by using Geldoc-It\u0026trade; (USA).\u003c/p\u003e\n \u003cp\u003eISSR PCR amplification\u003c/p\u003e\n \u003cp\u003eISSR PCR amplification was performed by using 7 primers with high GC content according to the protocol described by Bekir et al. (\u003cspan class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec6\"\u003e\n \u003cp\u003e\u003cem\u003eData analysis\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eOnly clear and polymorphic DNA bands were used for data analysis. The bands were scored as present (1) or absent (0) and a binary data matrix was constructed. DNA fragments of identical size amplified with the same primer were considered to be the same DNA marker.\u003c/p\u003e\n \u003cp\u003eTo examine the genetic relationship among populations, a dendrogram was constructed using the unweighted paired group method of cluster analysis using arithmetic averages (UPGMA) and principal coordinate analysis (PCA) was performed with NTSYSpc version 2.20 (Rohlf \u003cspan class=\"CitationRef\"\u003e2008\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eBased on RAPD and ISSR data, the POPGENE ver. 1.32 (Yeh et al. \u003cspan class=\"CitationRef\"\u003e2000\u003c/span\u003e) was used to estimate values meaning genetic diversity: number of polymorphic band, observed number of alleles, effective number of alleles (Kimura and Crow \u003cspan class=\"CitationRef\"\u003e1964\u003c/span\u003e), Nei\u0026rsquo;s (1973) gene diversity (h), Shannon\u0026rsquo;s information index (I) (Lewontin \u003cspan class=\"CitationRef\"\u003e1972\u003c/span\u003e) and genetic distances among populations (Nei \u003cspan class=\"CitationRef\"\u003e1978\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eA dendrogram of the populations was constructed which has been based on the unweighed pair group method (UPGMA) analysis of Nei\u0026rsquo;s (1978) genetic distances. And also, G\u003csub\u003eST\u003c/sub\u003e representing population differentiation and gene flow (Nm) among populations were estimated.\u003c/p\u003e\n \u003cp\u003eUsing the Arlequin ver. 3.5.2.2 (Excoffier and Lischer \u003cspan class=\"CitationRef\"\u003e2010\u003c/span\u003e), an analysis of molecular variance (AMOVA) was performed to evaluate the distribution of genetic variation within and among stock cultivars, and the Fixation index (Fst) was estimated too.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Results","content":"\u003cdiv class=\"Section2\" id=\"Sec8\"\u003e\n \u003cp\u003e\u003cem\u003eMorpological diversity of new double flowered stock cultivars\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eWe compared and analysed the morphological traits in new double flowered stock cultivars obtained from outcrossing including 4 pink, 2 pale pink and 2 white cultivars (Fig. \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\n \u003cdiv class=\"gridtable\"\u003e\u003ctable border=\"1\" id=\"Tab1\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eCharacteristics of morphological traits in stock cultivas\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ecultivar\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003epercent of double flowered plants /%\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eFlower color\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003edouble flower phenotype\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePopulation ID\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eFlower diameter\u003c/p\u003e\n \u003cp\u003e/cm\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003einflorescence length/cm\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003enumber of flowers in inflorescence\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLeaf color\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLeaf shape\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePlant height\u003c/p\u003e\n \u003cp\u003e/cm\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003enumber of leaves per plant\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLeaf length\u003c/p\u003e\n \u003cp\u003e/cm\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eLeaf width\u003c/p\u003e\n \u003cp\u003e/cm\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePod length/ cm\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003epercent of fertile plants/%\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《pink》\u003c/p\u003e\n \u003cp\u003e(♂)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e54.3\u0026thinsp;\u0026plusmn;\u0026thinsp;5.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003epink\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eno hairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e50.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e46.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《white》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e53.4\u0026thinsp;\u0026plusmn;\u0026thinsp;5.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ewhite\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.5\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e21.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eno hairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e80.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e2.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.6\u0026thinsp;\u0026plusmn;\u0026thinsp;2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e79.8\u0026thinsp;\u0026plusmn;\u0026thinsp;5.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《white》\u003c/p\u003e\n \u003cp\u003e(♀)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e47.4\u0026thinsp;\u0026plusmn;\u0026thinsp;4.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ewhite\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e19.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ehairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e53.8\u0026thinsp;\u0026plusmn;\u0026thinsp;6.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e26.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e7.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e99.7\u0026thinsp;\u0026plusmn;\u0026thinsp;5.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.3\u0026thinsp;\u0026plusmn;\u0026thinsp;3.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e50.7\u0026thinsp;\u0026plusmn;\u0026thinsp;3.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《pink 1》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e49.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ePink\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e22.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eno hairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e80.8\u0026thinsp;\u0026plusmn;\u0026thinsp;7.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e5.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e75.5\u0026thinsp;\u0026plusmn;\u0026thinsp;7.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.5\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e79.8\u0026thinsp;\u0026plusmn;\u0026thinsp;4.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《pink 2》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e52.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ePink\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.9\u0026thinsp;\u0026plusmn;\u0026thinsp;3.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e22.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ehairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e80.8\u0026thinsp;\u0026plusmn;\u0026thinsp;6.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e5.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e72.3\u0026thinsp;\u0026plusmn;\u0026thinsp;6.7\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e79.8\u0026thinsp;\u0026plusmn;\u0026thinsp;4.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《pink 3》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e53.2\u0026thinsp;\u0026plusmn;\u0026thinsp;3.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ePink\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.2\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e21.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eno hairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e80.8\u0026thinsp;\u0026plusmn;\u0026thinsp;5.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.7\u0026thinsp;\u0026plusmn;\u0026thinsp;3.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e5.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e73.5\u0026thinsp;\u0026plusmn;\u0026thinsp;8.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.5\u0026thinsp;\u0026plusmn;\u0026thinsp;3.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e79.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《pink 4》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e46.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ePink\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e22.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ehairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e80.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e5.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e75.1\u0026thinsp;\u0026plusmn;\u0026thinsp;9.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e79.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《pale pink 1》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e51.7\u0026thinsp;\u0026plusmn;\u0026thinsp;4.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ePink\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20.9\u0026thinsp;\u0026plusmn;\u0026thinsp;3.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eno hairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e45.4\u0026thinsp;\u0026plusmn;\u0026thinsp;3.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e26.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e5.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e73.5\u0026thinsp;\u0026plusmn;\u0026thinsp;8.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e47.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e24.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11.2\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《pale pink 2》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e49.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ePink\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e19.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.3\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003ehairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e44.4\u0026thinsp;\u0026plusmn;\u0026thinsp;4.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.6\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e5.3\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e70.5\u0026thinsp;\u0026plusmn;\u0026thinsp;9.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e17.3\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e43.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e16.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《white 1》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e48.4\u0026thinsp;\u0026plusmn;\u0026thinsp;4.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eWhite\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.7\u0026thinsp;\u0026plusmn;\u0026thinsp;0.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20.8\u0026thinsp;\u0026plusmn;\u0026thinsp;1.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eno hairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e80.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.7\u0026thinsp;\u0026plusmn;\u0026thinsp;3.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e5.6\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e75.5\u0026thinsp;\u0026plusmn;\u0026thinsp;7.2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e79.8\u0026thinsp;\u0026plusmn;\u0026thinsp;4.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.9\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003e《white 2》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e53.2\u0026thinsp;\u0026plusmn;\u0026thinsp;4.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eWhite\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003edouble\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5.2\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.9\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003epale green\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" rowspan=\"2\"\u003e\n \u003cp\u003eno hairy\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e80.8\u0026thinsp;\u0026plusmn;\u0026thinsp;3.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e27.7\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e15.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.8\u0026thinsp;\u0026plusmn;\u0026thinsp;0.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e5.5\u0026thinsp;\u0026plusmn;\u0026thinsp;0.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\" rowspan=\"2\"\u003e\n \u003cp\u003e71.9\u0026thinsp;\u0026plusmn;\u0026thinsp;6.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003esingle\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e4.0\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e18.0\u0026thinsp;\u0026plusmn;\u0026thinsp;1.9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e14.4\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003egreen\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e79.8\u0026thinsp;\u0026plusmn;\u0026thinsp;2.7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e28.9\u0026thinsp;\u0026plusmn;\u0026thinsp;3.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.4\u0026thinsp;\u0026plusmn;\u0026thinsp;0.9\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cp\u003eAs shown in Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e, in newly obtained 8 stock cultivars, the ratio of double flowered plants was about 50%, and some traits such as flower diameter, leaf length and leaf width, showed no significant differences (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) in all 11 cultivars including both parent. Conversely, in other traits including inflorescence length, number of flowers per inflorescence, plant height, number of leaves per plant, \u0026ldquo;white\u0026rdquo; cultivar was markedly bigger than \u0026ldquo;pink\u0026rdquo; cultivar (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05), newly obtained pink and white cultivars were not significantly different from maternal parent (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05), \u0026ldquo;white\u0026rdquo;, whereas pale pink cultivars were not significantly different from paternal parent, \u0026ldquo;pink\u0026rdquo; (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Besides, in pod length and the percent of fertile plants, new eight cultivars had medium values between maternal and paternal parent cultivars, not showing a significant difference (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05) according to cultivar.\u003c/p\u003e\n\u003c/div\u003e\n\u003cdiv class=\"Section2\" id=\"Sec9\"\u003e\n \u003cp\u003e\u003cem\u003eMolecular identification of new stock cultivars using RAPD and ISSR markers\u003c/em\u003e\u003c/p\u003e\n \u003cp\u003eGenetic relationship among twenty one stock individuals\u003c/p\u003e\n \u003cp\u003eIn the present study the genetic relationship among and within cultivars of \u003cem\u003eM. incana\u003c/em\u003e were analysed by ten informative RAPD and ISSR primers (Table \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). \u0026nbsp;\u003c/p\u003e\n \u003ctable border=\"1\" id=\"Tab2\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003ePrimers used for RAPD and ISSR analysis, total number of scored fragments and their size range\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePrimer\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSequence\u003c/p\u003e\n \u003cp\u003e(5\u0026rsquo;\u0026rarr; 3\u0026rsquo;)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eFragment size\u003c/p\u003e\n \u003cp\u003erange (bp)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eTotal number\u003c/p\u003e\n \u003cp\u003eof bands\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOPD3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u0026rsquo; GTCGCCGTCA 3\u0026rsquo;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u0026ndash;4000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOPH18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u0026rsquo; GAATCGGCCA 3\u0026rsquo;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e300\u0026ndash;2000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOPI14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u0026rsquo; TGACGGCGGT 3\u0026rsquo;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u0026ndash;2500\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOPN4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u0026rsquo; GACCGACCCA 3\u0026rsquo;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u0026ndash;3000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eOPP19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u0026rsquo; GGGAAGGCA 3\u0026rsquo;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e200\u0026ndash;2000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eISSRM1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e(AGC)\u003csup\u003e6\u003c/sup\u003e-G\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e200\u0026ndash;3000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eISSRM2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e(ACC)\u003csup\u003e6\u003c/sup\u003e-G\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e150\u0026ndash;2000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eISSRM3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e(AGC)\u003csup\u003e6\u003c/sup\u003e-C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e100\u0026ndash;4000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eISSRM5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e(GA)\u003csup\u003e9\u003c/sup\u003e-C\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e200\u0026ndash;3000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eISSRF3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e(AG)\u003csup\u003e8\u003c/sup\u003e-CG\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e150\u0026ndash;2000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003eBased on above obtained RAPD and ISSR profiles (Fig. 3), genetic relationship among 21 stock samples was estimated using NTSYSpc2.11.\u003c/p\u003e\n \u003cp\u003eWe obtained data matrix based on Nei (\u003cspan class=\"CitationRef\"\u003e1973\u003c/span\u003e)\u0026rsquo;s genetic distance and constructed a dendrogram using UPGMA method.\u003c/p\u003e\n \u003cp\u003eAs shown in Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e, at genetic distance of 0.15, dendrogram grouped all individuals in two clusters, and principal coordinate analysis result (Fig. \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e) was agreed with cluster analysis (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eFirst cluster, Ⅰ contained No. 1, 9, 10, 12, 6, 14, 16 individuals, being pink and single flowered. Second cluster, Ⅱ contained No. 2, 13, 8, 11, 7, 15, 17 individuals, being pink and double flowered. Third cluster, Ⅲ contained No. 3, 4, 18, 20 individuals, being white and single flowered. Final fourth cluster, Ⅳ contained No. 5, 21, 19 individuals, being white and double flowered. This suggests that our selected RAPD and ISSR primers might reflect not only flower color but also double flower phenotype.\u003c/p\u003e\n \u003cp\u003eGenetic relationship among eleven stock cultivars\u003c/p\u003e\n \u003cp\u003eGenetic relationship among eleven stock cultivars distinguished by flower color and leaf shape was evaluated using PopGene.version 1.31.\u0026nbsp;\u003c/p\u003e\n \u003ctable border=\"1\" id=\"Tab3\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eNei\u0026rsquo;s (1978) genetic distance among eleven studied stock populations. The abbreviated population names are given according to Table \u003cspan class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003epop ID\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8741\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8612\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9480\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9694\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9604\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9636\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9440\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9393\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8437\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8638\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1346\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9439\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8031\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8296\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8219\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8282\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8124\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8152\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8826\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8918\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1494\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0577\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8280\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8150\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8169\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8132\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7925\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8066\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9424\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9514\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0534\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2193\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1888\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9153\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9163\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9144\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8845\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8964\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8055\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8501\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0311\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1868\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2015\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0885\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9549\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9597\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9209\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9393\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7928\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8044\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0404\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1961\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0874\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0461\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9507\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9124\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9169\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7855\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7969\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0371\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1885\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2068\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0895\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0412\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0505\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9104\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9048\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.7947\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8160\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0576\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2077\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2326\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1227\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0824\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0917\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0939\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9681\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8112\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8092\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0626\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2044\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2149\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1094\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0626\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0867\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0324\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8074\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.8155\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1699\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1249\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0594\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2163\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2322\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2415\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2298\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2092\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2140\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e****\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.9684\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1464\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1146\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0498\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.1624\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2177\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2270\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2033\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2118\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.2040\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.0321\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\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\u003eNei\u0026rsquo;s genetic identity (above diagonal) and genetic distance (below diagonal).\u003c/p\u003e\n \u003cp\u003eAs shown in Fig. \u003cspan class=\"InternalRef\"\u003e6\u003c/span\u003e, eleven stock cultivars were classified into two main groups; pink and white cultiivar group.\u003c/p\u003e\n \u003cp\u003eThe genetic distance values for all 11 cultivars ranged from 0.03 to 0.25 showing the lowest genetic distance (0.02415) between \u0026ldquo;pink 3\u0026rdquo; and \u0026ldquo;white 1\u0026rdquo;, while the highest genetic distance (0.0321) between \u0026ldquo;white 1\u0026rdquo; and \u0026ldquo;white 2\u0026rdquo; (Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e\n \u003cp\u003eIn the group of white cultivars, new cultivar \u0026ldquo;white 2\u0026rdquo; was genetically closest to control cultivar \u0026ldquo;white\u0026rdquo; (genetic distance- 0.0498), and new cultivar \u0026ldquo;white 1\u0026rdquo; was the most distant to maternal cultivar \u0026ldquo;white\u0026rdquo; (genetic distance- 0.1062). Otherwise, in the group of pink cultivars, new cultivar \u0026ldquo;pink 2\u0026rdquo; was the genetically closest to paternal cultivar \u0026ldquo;pink\u0026rdquo; (genetic distance- 0.0534), and new cultivar \u0026ldquo;pink 1\u0026rdquo; was the most distant to new cultivar \u0026ldquo;pale pink 1\u0026rdquo; (genetic distance- 0.0965).\u003c/p\u003e\n \u003cp\u003eAmong 11 newly obtained stock cultivars, differentiation index (G\u003csub\u003eST)\u003c/sub\u003e and gene flow index (Nm) were 0.639(0.1309-1.000) and 0.2817(0.0430\u0026ndash;3.3195) respectively.\u003c/p\u003e\n \u003cp\u003eThe values of genetic diversity indices for studied stock cultivars are given in Table \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003e. The highest n\u003csub\u003ea\u003c/sub\u003e was observed for \u0026ldquo;pale pink 2\u0026rdquo; (1.26) and the least for \u0026ldquo;pink 3\u0026rdquo; (1.07), showing the lowest Shannon\u0026rsquo;s index (0.04) and Nei\u0026rsquo;s gene diversity (0.03) as well.\u003c/p\u003e\n \u003cp\u003eNei\u0026rsquo;s gene diversity were estimated to be 0.03 and 0.11 and the Shannon indices of the \u003cem\u003eM. incana\u003c/em\u003e cultivars ranged from 0.04 to 0.16.\u003c/p\u003e\n \u003ctable border=\"1\" id=\"Tab4\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eGenetic diversity indices for eleven stock cultivars. N\u003csub\u003ePB\u003c/sub\u003e- number of polymorphic bands, P\u003csub\u003ePB\u003c/sub\u003e-percentage of polymorphic bands, n\u003csub\u003ea\u003c/sub\u003e \u0026ndash; observed number of alleles, n\u003csub\u003ee\u003c/sub\u003e \u0026ndash;effective number of alleles, h - Nei\u0026rsquo;s (1987) gene diversity, I - Shannon\u0026rsquo;s information index\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eCultivar\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eN\u003csub\u003ePB\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP\u003csub\u003ePB\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en\u003csub\u003ea\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003en\u003csub\u003ee\u003c/sub\u003e\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eh\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eI\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《pink》(paternal)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《white》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《white》(maternal)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《pink 1》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《pink 2》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e10.87\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《pink 3》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e6.52\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.07\u0026thinsp;\u0026plusmn;\u0026thinsp;0.25\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.05\u0026thinsp;\u0026plusmn;\u0026thinsp;0.18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.03\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.04\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《pink 4》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《pale pink 1》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e17.39\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.38\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.23\u0026thinsp;\u0026plusmn;\u0026thinsp;0.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.07\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《pale pink 2》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e26.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.44\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.18\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.16\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《white 1》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e23.91\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.24\u0026thinsp;\u0026plusmn;\u0026thinsp;0.43\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.17\u0026thinsp;\u0026plusmn;\u0026thinsp;0.31\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.14\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e《white 2》\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e7\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.15\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.11\u0026thinsp;\u0026plusmn;\u0026thinsp;0.26\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.06\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.09\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003ctable border=\"1\" id=\"Tab5\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eAnalysis of molecular variance (AMOVA) based on RAPD profiless in eleven stock cultivars\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSource of variation\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003edegrees of freedom\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSum of squares\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eVariance components\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ePercentage of variation (%)\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eP\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAmong cultivars\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e283.562\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e2.584\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e67.27\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eWithin cultivars\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e105.600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e1.257\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e32.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e\u0026lt;\u0026thinsp;0.0001\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eTotal\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e30\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e389.162\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"char\"\u003e\n \u003cp\u003e3.841\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003cp\u003eGenetic variation among cultivars was more than that within cultivars, as indicated by the sum of squares values calculated by AMOVA using Arlequin ver. 3.5.2.2 (Table \u003cspan class=\"InternalRef\"\u003e5\u003c/span\u003e). Of the total genetic diversity, 67.27 % was attributable to differences among culivars and 32.73% was to differences within culivars, showing a significant varietal differentiation in newly obtained stock cultivars. The fixation index (Fst) was 0.673.\u003c/p\u003e\n\u003c/div\u003e"},{"header":"Discussion","content":" \u003cp\u003eBecause double flowered cultivars have superior ornamental value to single flowered ones, stock breeders have attempted to make double flowered cultivars with different colors.\u003c/p\u003e \u003cp\u003eAdditionally, in breeding of \u003cem\u003eM. incana\u003c/em\u003e, the studies also have been reported to identify the morphological and molecular markers which could be used to discriminate the double flowered individuals.\u003c/p\u003e \u003cp\u003eEcker et al. (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1993\u003c/span\u003e) founded that the \u003cem\u003ec\u003c/em\u003e allele for sinuate leaf shape might be recessive to the \u003cem\u003eC\u003c/em\u003e allele for normal entire leaf and it was tightly linked to the \u003cem\u003es\u003c/em\u003e recessive allele for double flowering, suggesting its role as a double flowering marker in the cultivation and the breeding of \u003cem\u003eM. incana.\u003c/em\u003e\u003c/p\u003e \u003cp\u003eTakashi and Kanae (2018) revealed that co-dominant marker, \u003cem\u003eMiAG\u003c/em\u003e could be used for early discrimination of double flowered individuals among seedlings not showing phenotypic differences. He examined that the single flowered individuals of cultivars being distinguished double flowered ones according to cotyledon shape (\u0026lsquo;Kiss me white\u0026rsquo;, \u0026lsquo;Iron white\u0026rsquo;, \u0026lsquo;Quartet cherry\u0026rsquo;), serrated leaf (\u0026lsquo;Pygmy white\u0026rsquo;), seed color (\u0026lsquo;White wonder no.2\u0026prime;) were heterozygotes \u003cem\u003eMiAG/miag1\u003c/em\u003e, whereas the individuals correlated with leaf color (in \u0026lsquo;Aida\u0026rsquo; and \u0026lsquo;Opera\u0026rsquo;) were heterozygotes \u003cem\u003eMiAG/miag2.\u003c/em\u003e\u003c/p\u003e \u003cp\u003eIn this study we made the garden stock varieties with different colors (pink, pale pink and white) and high fertility which were capable of discriminating double flowered individuals in cross \u0026ldquo;white\u0026rdquo; cultivar (high fertile but impossible in selection of double flowered plants) \u0026times; \u0026ldquo;pink\u0026rdquo; cultivar (very low fertile but possible in selection of double flowered plants). Considering with Takashi and Kanae et al. (2018)\u0026rsquo;s result, our newly raised stock varieties are likely \u003cem\u003eMiAG/miag2\u003c/em\u003e heterozygotes being capable of selecting double flowered ones among the seedlings according to leaf color (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), and harboring high fertility, therefore they might be superior to both parents cultivars in the cultivation and seed production for commercial use.\u003c/p\u003e \u003cp\u003eTo date, random amplified polymorphic DNA (RAPD) method have been widely applied to assess the intra- and inter-specific genetic diversity in many plants.\u003c/p\u003e \u003cp\u003eTanmay et al. (2016) described the diversity of nine genotypes in the genus \u003cem\u003eOcimum\u003c/em\u003e from India based on RAPD markers combining with morphological and chemical markers.\u003c/p\u003e \u003cp\u003ePrabhjot et al. (2019) assessed the molecular diversity among 48 accessions of five \u003cem\u003eSwertia\u003c/em\u003e species collected from Western Himalayas, India using 18 RAPD markers.\u003c/p\u003e \u003cp\u003eRAPD primer based genetic diversity within the species were reported earlier in several plant species such as \u003cem\u003eCitrullus colocynthis\u003c/em\u003e (Kumar et al. 2017), lettuce (\u003cem\u003eLactuca sativa\u003c/em\u003e L.) (Shubhangi et al. 2018), \u003cem\u003eNilgirianthus ciliates\u003c/em\u003e (Ramakrishnan et al. 2019) and \u003cem\u003eArtemisia herba\u003c/em\u003e-alba (Khaled et al. 2019).\u003c/p\u003e \u003cp\u003eBesides, RAPD technique has also been used to examine the genetic diversity on the level of variety or cultivar (Maciel et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Siddharth et al. 2007; Chrobokov\u0026aacute; et al. \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Masvodza et al. \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Palaiyur et al. \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Rozhmina et al. \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Zhiqiang et al. 2017).\u003c/p\u003e \u003cp\u003ePalaiyur et al. (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) showed that the similarity coefficients ranged from 0.57 to 0.78 by using nine of 80 RAPD primers and they could be applied to identify different varieties in \u003cem\u003eProsopis cineraria\u003c/em\u003e (L.) Druce.\u003c/p\u003e \u003cp\u003eRozhmina et al. (\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) revealed genetic similarity between cultivars of fiber flax from Russia and other the European countries using RAPD markers. On the other hand, ISSR method is more reproducible than RAPD method, so it is more widely used in the analysis of genetic diversity in many plants such as maize (Domenyuk et al. \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2002\u003c/span\u003e), common bean (Galvan et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2003\u003c/span\u003e) and \u003cem\u003eJurinea\u003c/em\u003e (Dogan et al. \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAnd also, recent researchers (Ramakrishnan et al. 2019; Joyashree et al. 2019; Zhiqiang et al. 2017; Kumar et al. 2017) reported that RAPD and ISSR markers could be applied together to reveal genetic relationship between germplasm lines and to establish the cultivar identification system using cluster analysis and PCA.\u003c/p\u003e \u003cp\u003eHowever, there are few reports in stock plant (\u003cem\u003eMatthiola incana\u003c/em\u003e) to analyze genetic relationship using molecular markers.\u003c/p\u003e \u003cp\u003eAmaal (2009) performed molecular systematic analysis for twenty two taxa belonging to \u003cem\u003eBrassicaceae\u003c/em\u003e including \u003cem\u003eMatthiola incana\u003c/em\u003e and \u003cem\u003eMatthiola longipetala\u003c/em\u003e subsp. \u003cem\u003elivida\u003c/em\u003e using RAPD markers and suggested its role for varietal identification and assessment of genetic relationships even between closely related species.\u003c/p\u003e \u003cp\u003eAnd Bekir et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) studied phylogenetic relationship of Turkish species of \u003cem\u003eMatthiola\u003c/em\u003e based on only ISSR markers.\u003c/p\u003e \u003cp\u003eIn contrast to previous study, the present method might be applied to identify new double flowered stock plant varieties obtained through outcrossing, by using RAPD and ISSR markers together.\u003c/p\u003e \u003cp\u003eBy combining new selected RAPD and ISSR primers, our analysis method grouped stock plant varieties into two main clusters, one including pink varieties and the other including white ones. Besides single and double flowered individuals were classified in each cluster, and RAPD based genetic distances varied from 0.03 to 0.24 among these varieties which is similar to Chrobokov\u0026aacute; et al. (\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) (0.02\u0026thinsp;~\u0026thinsp;0.45), Masvodza et al. (\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2014\u003c/span\u003e) (0.1\u0026thinsp;~\u0026thinsp;0.22)\u0026rsquo; s research result and has somewhat smaller scale than Palaiyur et al. (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) (0.22\u0026thinsp;~\u0026thinsp;0.43), Maciel et al. (\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2001\u003c/span\u003e) (0.2\u0026thinsp;~\u0026thinsp;0.75), and Siddharth et al. (2007) (0.34\u0026thinsp;~\u0026thinsp;0.74)\u0026rsquo; report.\u003c/p\u003e "},{"header":"Conclusion","content":" \u003cp\u003eWe bred new double flowered stock cultivars with different colors and high fertility in cross white cultivar that was high fertile but impossible to select double flower phenotype \u0026times; pink cultivar (vice. versa). RAPD and ISSR markers selected in this study can be used to identify different stock cultivars according to flower color and flowered phenotype, therefore in addition to morphological traits, they will be applied to cultivar certification, identification, and establishing conservation system in their culture and management.\u003c/p\u003e "},{"header":"Abbreviations","content":"\u003cp\u003eRAPD (Random Amplified Polymorphic DNA), ISSR (Inter Simple Sequence Repeat)\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eConflict of interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that there are no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank all horticulturists of Pyongyang floriculture institute who helped culturation during stock breeding.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eAmaal Hasan Mohamed (2009) Molecular Systematic of Some \u003cem\u003eBrassicaceae\u003c/em\u003e Taxa in Egypt Based on Electrophoretic Isoenzymes Pattern and RAPD Markers. 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J Genet 1: 303\u0026ndash;376\u003c/li\u003e\n \u003cli\u003eSaunders E S (1915) A suggested explanation of the abnormally high records of doubles quoted by growers of stocks (\u003cem\u003eMatthiola\u003c/em\u003e). J Genet 5: 137\u0026ndash;143\u003c/li\u003e\n \u003cli\u003eSaunders E S (1921) Note on the evolution of the double stock (\u003cem\u003eMatthiola incana\u003c/em\u003e). J Genet 11: 69\u0026ndash;74.\u003c/li\u003e\n \u003cli\u003eSaunders E S (1928) Further studies on inheritance in \u003cem\u003eMatthiola incana\u003c/em\u003e. II. Plastid colour and doubing. 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Department of Renewable Resources, University of Alberta, Edmonton, AB, Canada.\u003c/li\u003e\n \u003cli\u003eZhiqiang\u0026nbsp;Mei,\u0026nbsp;Xianqin\u0026nbsp;Zhang,\u0026nbsp;Md\u0026nbsp;Asaduzzaman\u0026nbsp;Khan,\u0026nbsp;Saber\u0026nbsp;Imani,\u0026nbsp;Xiaoyan\u0026nbsp;Liu,\u0026nbsp;Hui\u0026nbsp;Zou,\u0026nbsp;Chunli\u0026nbsp;Wei,\u0026nbsp;Junjiang\u0026nbsp;Fu\u0026nbsp;(2017)\u0026nbsp;Genetic analysis of \u003cem\u003ePenthorum chinense\u003c/em\u003e Pursh by improved RAPD and ISSR in China. Electron J Biotechn 30: 6\u0026ndash;11\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Matthiola incana, RAPD, ISSR, cultivar, identification, fertility","lastPublishedDoi":"10.21203/rs.3.rs-578473/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-578473/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eGarden stock (\u003cem\u003eMatthiola incana\u003c/em\u003e R. Br.) is a commercially important horticultural crop owing to its ornamental effect. There are different stock cultivars varied in color and shape, especially flowered phenotype is an essential index evaluating its commercial value, because double flowered cultivars have more brilliant flowers compared to single flowered one. The present work aimed: (1) to make superior cultivars with different colors, high fertility, being capable of early selecting only double flowered seedlings by leaf color and to investigate morphological characteristics and (2) to select RAPD and ISSR primers for the cultivar certification and identification to culture and produce good commercial stock cultivars. Here we obtained new double flowered stock cultivars with different colors including pink, pale pink and white, through outcrossing between “white” cultivar (high fertile but unable to select double flower phenotype) and “pink” cultivar (vice. versa). Among newly obtained stock cultivars, single and double flower seedlings are distinguishable from each other by leaf color, having about 70% of fertility. Moreover RAPD and ISSR markers selected in this study can be applied to identify different stock cultivars in seed production, culture and to establish cultivar certification system.\u003c/p\u003e","manuscriptTitle":"Morphological And Molecular Identification of Double Flowered Stock (Matthiola Incana R. Br) Cultivars With High Fertility","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2021-06-17 20:05:46","doi":"10.21203/rs.3.rs-578473/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"9134b407-a968-48f2-9d3b-98e698a77e5c","owner":[],"postedDate":"June 17th, 2021","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":5097811,"name":"Agronomy"},{"id":5097812,"name":"Molecular Genetics"}],"tags":[],"updatedAt":"2021-09-17T11:45:45+00:00","versionOfRecord":[],"versionCreatedAt":"2021-06-17 20:05:46","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-578473","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-578473","identity":"rs-578473","version":["v1"]},"buildId":"WrCJVZZCHTDjtuVLN7oU0","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}