FveTRM5 plays a critical role in regulating fruit shape in woodland strawberry

FveTRM5 plays a critical role in regulating fruit shape in woodland strawberry | bioRxiv /* */ /* */ <!-- <!-- /*! * yepnope1.5.4 * (c) WTFPL, GPLv2 */ (function(a,b,c){function d(a){return"[object Function]"==o.call(a)}function e(a){return"string"==typeof a}function f(){}function g(a){return!a||"loaded"==a||"complete"==a||"uninitialized"==a}function h(){var a=p.shift();q=1,a?a.t?m(function(){("c"==a.t?B.injectCss:B.injectJs)(a.s,0,a.a,a.x,a.e,1)},0):(a(),h()):q=0}function i(a,c,d,e,f,i,j){function k(b){if(!o&&g(l.readyState)&&(u.r=o=1,!q&&h(),l.onload=l.onreadystatechange=null,b)){"img"!=a&&m(function(){t.removeChild(l)},50);for(var d in y[c])y[c].hasOwnProperty(d)&&y[c][d].onload()}}var j=j||B.errorTimeout,l=b.createElement(a),o=0,r=0,u={t:d,s:c,e:f,a:i,x:j};1===y[c]&&(r=1,y[c]=[]),"object"==a?l.data=c:(l.src=c,l.type=a),l.width=l.height="0",l.onerror=l.onload=l.onreadystatechange=function(){k.call(this,r)},p.splice(e,0,u),"img"!=a&&(r||2===y[c]?(t.insertBefore(l,s?null:n),m(k,j)):y[c].push(l))}function j(a,b,c,d,f){return q=0,b=b||"j",e(a)?i("c"==b?v:u,a,b,this.i++,c,d,f):(p.splice(this.i++,0,a),1==p.length&&h()),this}function k(){var a=B;return a.loader={load:j,i:0},a}var l=b.documentElement,m=a.setTimeout,n=b.getElementsByTagName("script")[0],o={}.toString,p=[],q=0,r="MozAppearance"in l.style,s=r&&!!b.createRange().compareNode,t=s?l:n.parentNode,l=a.opera&&"[object Opera]"==o.call(a.opera),l=!!b.attachEvent&&!l,u=r?"object":l?"script":"img",v=l?"script":u,w=Array.isArray||function(a){return"[object Array]"==o.call(a)},x=[],y={},z={timeout:function(a,b){return b.length&&(a.timeout=b[0]),a}},A,B;B=function(a){function b(a){var a=a.split("!"),b=x.length,c=a.pop(),d=a.length,c={url:c,origUrl:c,prefixes:a},e,f,g;for(f=0;f<d;f++)g=a[f].split("="),(e=z[g.shift()])&&(c=e(c,g));for(f=0;f<b;f++)c=x[f](c);return c}function g(a,e,f,g,h){var i=b(a),j=i.autoCallback;i.url.split(".").pop().split("?").shift(),i.bypass||(e&&(e=d(e)?e:e[a]||e[g]||e[a.split("/").pop().split("?")[0]]),i.instead?i.instead(a,e,f,g,h):(y[i.url]?i.noexec=!0:y[i.url]=1,f.load(i.url,i.forceCSS||!i.forceJS&&"css"==i.url.split(".").pop().split("?").shift()?"c":c,i.noexec,i.attrs,i.timeout),(d(e)||d(j))&&f.load(function(){k(),e&&e(i.origUrl,h,g),j&&j(i.origUrl,h,g),y[i.url]=2})))}function h(a,b){function c(a,c){if(a){if(e(a))c||(j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}),g(a,j,b,0,h);else if(Object(a)===a)for(n in m=function(){var b=0,c;for(c in a)a.hasOwnProperty(c)&&b++;return b}(),a)a.hasOwnProperty(n)&&(!c&&!--m&&(d(j)?j=function(){var a=[].slice.call(arguments);k.apply(this,a),l()}:j[n]=function(a){return function(){var b=[].slice.call(arguments);a&&a.apply(this,b),l()}}(k[n])),g(a[n],j,b,n,h))}else!c&&l()}var h=!!a.test,i=a.load||a.both,j=a.callback||f,k=j,l=a.complete||f,m,n;c(h?a.yep:a.nope,!!i),i&&c(i)}var i,j,l=this.yepnope.loader;if(e(a))g(a,0,l,0);else if(w(a))for(i=0;i (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];var j=d.createElement(s);var dl=l!='dataLayer'?'&l='+l:'';j.src='//www.googletagmanager.com/gtm.js?id='+i+dl;j.type='text/javascript';j.async=true;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-M677548'); Skip to main content Home About Submit ALERTS / RSS Search for this keyword Advanced Search New Results FveTRM5 plays a critical role in regulating fruit shape in woodland strawberry Zhenzhen Zheng , Liyang Wang , Qi Gao , Shaoqiang Hu , View ORCID Profile Chunying Kang doi: https://doi.org/10.1101/2025.03.10.642363 Zhenzhen Zheng 1 National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, Huazhong Agricultural University , Wuhan, 430070, China 2 Hubei Hongshan Laboratory , Wuhan, 430070, China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Liyang Wang 1 National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, Huazhong Agricultural University , Wuhan, 430070, China 2 Hubei Hongshan Laboratory , Wuhan, 430070, China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Qi Gao 1 National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, Huazhong Agricultural University , Wuhan, 430070, China 2 Hubei Hongshan Laboratory , Wuhan, 430070, China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Shaoqiang Hu 1 National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, Huazhong Agricultural University , Wuhan, 430070, China 2 Hubei Hongshan Laboratory , Wuhan, 430070, China Find this author on Google Scholar Find this author on PubMed Search for this author on this site Chunying Kang 1 National Key Laboratory for Germplasm Innovation & Utilization of Horticultural Crops, Huazhong Agricultural University , Wuhan, 430070, China 2 Hubei Hongshan Laboratory , Wuhan, 430070, China Find this author on Google Scholar Find this author on PubMed Search for this author on this site ORCID record for Chunying Kang For correspondence: ckang{at}mail.hzau.edu.cn Abstract Full Text Info/History Metrics Supplementary material Preview PDF Abstract Cultivated strawberry is a globally important fruit crop with high economic value. Fruit shape contributes to fruit quality and diversity and is a target for breeding, but very few regulatory genes have been reported in strawberry. Here, we identified an ethyl methanesulfonate (EMS) round fruit ( rf ) mutant that produces round or flat fruits in woodland strawberry. The primary candidate point mutation is located in the second exon of FvH4_2g22810, causing a premature stop codon at residue 266. This gene encodes a protein with a high similarity to TON1 RECRUITING MOTIF 5 (TRM5) and has therefore been named FveTRM5 . Transformation of FveTRM5 pro: FveTRM5 into rf could rescue the round fruit phenotype, suggesting that FveTRM5 is responsible for rf . Overexpression of FveTRM5 produced elongated organs in both Arabidopsis and woodland strawberry, suggesting a conserved role in different species. FveTRM5 is ubiquitously expressed with higher levels in developing organs. Observation of cell shape showed that FveTRM5 promotes cell elongation and inhibits cell division in the medial-lateral direction in the receptacle. The FveTRM5 protein localized to microtubules. In conclusion, our results suggest that FveTRM5 plays an essential role in regulating strawberry fruit shape by influencing cell elongation and cell division, providing an excellent target gene for breeding new fruit shape cultivars. 1. Introduction Fruit shape is an important external quality trait that strongly influences consumer preference. The organization of floral organs and which part can develop into fruit flesh has evolved differently among species [ 1 , 2 ]. The diversity of fruit shapes arises from the meristem activity and the growth patterns along the adaxial-abaxial, proximal-distal, and mediolateral axes during ovary and fruit development [ 1 , 3 , 4 ]. Understanding the regulation of fruit shape is of great importance for the genetic improvement of fruit crops to create new cultivars. Three gene families are known to play key roles in the organ shape control in different plant species, including the TONNEAU1 Recruiting Motif proteins (TRMs), the Ovate Family Proteins (OFPs), and the IQ67 domain-containing proteins (IQDs) [ 5 - 7 ]. Mutation or overexpression of TRMs can cause altered shapes of various organs, including seeds/grains/fruits in Arabidopsis [ 8 , 9 ], rice [ 10 , 11 ], wheat [ 12 ], maize [ 13 ], tomato and cucumber [ 14 , 15 ]. In the OFP family, OVATE and SlOFP20 in tomato are negative regulators of fruit elongation [ 14 , 16 ], and PpOFP1 overexpression due to genomic inversion results in flat fruits in peach [ 17 - 19 ]. The IQD protein SUN1, a calmodulin-binding protein, promotes fruit elongation in tomato [ 20 ]. A number of other OFPs or IQDs have also been reported to influence organ shape [ 21 ]. Fruit shape is closely related to cell division and cell expansion in different directions. The TRM proteins affect both cell division and cell expansion [ 9 , 10 , 12 , 14 , 15 ]. A common mechanism underlying this biological process is the modulation of the microtubular cytoskeleton. TRMs can recruit TON1 and phosphatase 2A to the microtubule arrays and the preprophase bands to control the direction of cell division [ 8 , 22 - 25 ]. TRMs can also interact with OFPs to antagonistically modulate microtubule arrangement [ 6 , 14 ]. Tomato SlTRM5 localizes to the microtubules, whereas SlOFP20 localizes to the cytoplasm and nucleus; when SlTRM5 and SlOFP20 were transiently expressed together in Nicotiana benthamiana leaves, SlOFP20 was largely translocated to the microtubules [ 14 ]. SUN1/SlIQD12 also localizes to microtubules to alter their structure, leading to changes in cell division patterns [ 26 ]. In this process, IQDs may interact with calmodulin (CaM) to sense and respond to the calcium signals [ 27 ]. The TRM-OFP module may coordinate with IQDs on the microtubular cytoskeleton to regulate organ shape [ 5 , 6 ]. Cultivated strawberry ( Fragaria × ananassa , octoploid) is a globally important fruit crop with high economic value. Fruit shape contributes to fruit quality and diversity and is therefore a selected trait in strawberry breeding. Unlike most fruit crops, strawberry flesh develops from the enlarged tip of the stem below the floral organs, known as the receptacle [ 2 ]. Both wild and cultivated strawberries have different fruit shapes, ranging from flat to long conical [ 28 , 29 ]. Fruit shape in strawberries responds to environmental stimuli and endogenous factors such as auxin and gibberellin (GA) levels [ 30 , 31 ]. Some quantitative trait loci (QTLs) responsible for natural shape variation in strawberry have been identified [ 32 , 33 ], but few regulatory genes were reported. Woodland strawberry ( F. vesca ) is a wild diploid species that typically produces small conical fruits. In this study, we isolated the fruit shape control gene, FveTRM5 , from the ethyl methanesulfonate (EMS) mutant round fruit ( rf ) in woodland strawberry. Genetic complementation and overexpression assays validated the important functions of FveTRM5 in promoting organ elongation. We showed that FveTRM5 regulates fruit shape by influencing both cell elongation and cell division. Our findings provide an excellent target gene for breeding new fruit shape cultivars and exploring fruit shape regulatory mechanisms in strawberry. 2. Materials and methods 2.1 Plant materials and growth conditions Three F. vesca accessions, Yellow Wonder (white-fruited), Ruegen (red-fruited) and Hawaii 4 (H4, white-fruited), were used as the WT in this study. All plants were grown in a growth room under a light intensity of 100 μmol m −2 s −1 with a photoperiod of 16 h light and 8 h dark at 22 °C. For EMS mutagenesis, 0.4% EMS (Sigma-Aldrich; cat no. M0880) was used for seed treatment for 8 h at room temperature with gentle shaking. Mutants were screened in the M 2 generation. 2.2 Phenotypic analysis Flowers or fruits from three to five plants of each genotype were analyzed. The maximum length and width of each tissue were measured using Image J software, and the shape index was calculated as the ratio of length to width. Three fruits at 0 DAP of Ruegen, H4, rf and FveTRM5 -OE were used for paraffin sectioning. The samples were quickly placed in a mixed fixative of formalin-acetic acid-ethanol (FAA) (70 % ethanol, 5 % formaldehyde and 5 % acetic acid). Paraffin embedding and sectioning were performed as previously described [ 34 ]. The section thickness was 10 μm for each fruit. For cell shape analysis, 20 cells in the pith with clear shapes were selected from one section of each fruit, resulting in 60 cells for each genotype. For cell number analysis, the cells in the widest part of the receptacle were counted over the entire receptacle from two sections of each fruit, resulting in 6 samples for each genotype. 2.3 Gene isolation of the rf mutant The rf mutant was crossed with wild-type Ruegen to generate an F 2 population. Equal amounts of young leaves from 11 F 2 mutant and 20 F 2 wild type plants were pooled. Genomic DNA was extracted using a CTAB method. Genome sequencing was performed on the Illumina HiSeq X Ten platform (Novogene, Beijing) and analyzed as previously described [ 35 ]. DNA-seq reads from the two pools were aligned to the woodland strawberry reference genome ver. 4 using BWA-MEM with default parameters [ 36 , 37 ]. The SAM file was further transformed into a sorted bam file and duplicate reads were removed using SAMtools [ 38 ]. The SNPs in F. vesca were called using GATK HaplotypeCaller [ 39 ] with ‘--min-base-quality-score 20; --minimum-mapping-quality 20’ and then filtered with the following parameters: QD 60.0 || MQ < 40.0 || MQRankSum < -12.5 || ReadPosRankSum < -8.0 || -cluster 2 -window 20. Only G-A or C-T sites were retained for further analysis. The ΔSNP index was first calculated by subtracting the SNP frequency between two pools, and then a sliding window analysis with a 300-kb window and 100-kb step was performed to calculate the average ΔSNP index. SNPs located in the CDS region were extracted using BEDTools [ 40 ]. The candidate mutation was examined in individual F 2 mutants by PCR amplification and Sanger sequencing. 2.4 Phylogenetic analysis The FveTRM proteins in woodland strawberry were identified from the PLAZA website ( https://bioinformatics.psb.ugent.be/plaza/versions/plaza_v5_dicots/ ) grouped together with AtTRMs. Protein sequences were obtained from TAIR for Arabidopsis ( Arabidopsis.org ), Sol Genomics Network for tomato ( solgenomics.net ), and Cucurbit Genomics Data for cucumber (cucurbit genomics.org ). An unrooted phylogenetic tree was constructed using MEGA7 with the neighbor-joining statistical method and bootstrap analysis (1000 replicates). 2.5 Plasmid construction Genomic DNA or total RNA was extracted from fruits or young leaves of Ruegen and used for gene amplification. For overexpression in woodland strawberry, the full-length coding sequence of FveTRM5 was cloned into pENTR1A and inserted into the binary vector pK7WG2D. For overexpression in Arabidopsis, the full-length coding sequence of FveTRM5 was inserted into pRI101 at the Sal I and BamH I site and fused with GFP using the ClonExpress II One Step Cloning Kit (Vazyme, C112-01). For FveTRM5pro : FveTRM5 , 304 bp upstream of the translation start site, 3,219 bp of the gene body (exons and introns), and 999 bp downstream of the stop codon of FveTRM5 were inserted into the binary vector pCAMBIA1300 at the Sal I and Bam HI sites. For subcellular localization, the coding sequence of FveTRM5 was cloned into the binary vector pH7LIC5.0 fused to N-terminal GFP at the Sal I and Stu I sites using the ClonExpress II One Step Cloning Kit (Vazyme, C112-01). Primers are listed in Table S1. 2.6 Stable transformation in woodland strawberry Transformation in woodland strawberry was performed as previously described [ 41 ]. The overexpression and complementation constructs were transformed separately into woodland strawberry variety H4. During transformation, positive overexpression transgenic calli and regenerated plants were selected using 10 mg L −1 kanamycin and GFP fluorescence as examined under a fluorescence dissecting microscope (Microshot Technology Ltd, Guangzhou, China, MZX81). Positive transgenic calli of FveTRM5 pro: FveTRM5 were selected on the medium containing 4 mg L −1 hygromycin. 2.7 Stable transformation in Arabidopsis Arabidopsis Col-0 was transformed with Agrobacterium tumefaciens GV3101 using the floral-dip method. The transgenic lines were selected on half strength MS (M5524, Sigma) with 100 mg L -1 kanamycin in the T 1 generation. 2.8 Subcellular localization analysis Nicotiana benthamiana leaves were infiltrated on the abaxial side with the A. tumefaciens strain GV3101 cells containing the construct and the silencing suppressor p19 in a 1:1 ratio. The 35S : RFP-AtTUA5 construct was co-infiltrated as a microtubule marker [ 42 ]. Agrobacterium cells were harvested and resuspended in the buffer (10 mM MgCl 2 , 10 mM MES, pH 5.6 and 150 mM acetosyringone at pH 5.6) and adjusted to an OD 600 of 0.6. The solutions were incubated for 2 hours at room temperature without shaking before infiltration. Fluorescence signals were acquired four days after infiltration using a Leica TCS SP8 inverted microscope. Excitation wavelengths were 488 nm for GFP and 552 nm for RFP, and emission was detected at 505-550 nm for GFP and 590-640 nm for RFP. 2.9 Quantitative RT-PCR Total RNA was extracted by using a HiPure Plant RNA Mini Kit (Magen, Guangzhou, China; cat no. R4151) and reverse transcribed to cDNA using HiScript III RT SuperMix for qPCR (+gDNA/wispr, Vazyme, Najing, China; cat no. R323). qPCR was performed using a Quant Studio 7 Flex system (Applied Biosystems, Waltham, MA, USA). The expression level of each gene was calculated using the 2 −ΔΔCT method. FvH4_1g05910 was used as an internal control. Three biological replicates were used for each sample, and three technical replicates were analyzed for each biological replicate. Primers are listed in Table S1. 2.10 RNA in situ hybridization Shoot tips and flower buds at different developmental stages of wild-type H4 were collected and fixed in the 50% FAA fixative solution at 4 °C. A 200 bp fragment of the FveTRM5 gene (215 to 414 bp in the coding sequence) was inserted into the pGEM-T vector at the Nco I and Sal I sites. The DIG RNA Labeling Kit (SP6/T7; Roche, Cat#11175025910) was used to synthesize DIG-labeled RNA probes by in vitro transcription. The sense probe was synthesized by T7 polymerase, and the antisense probe was synthesized by SP6 polymerase. The hybridization signals were detected using the DIG Nucleic Acid Detection Kit (Roche, Cat#11175041910). Slides were developed for 18 h in a dark moist container and stored in 1× TE after the reaction was stopped. Images were captured using a Leica microscope (DM6B) with a 10× or 20× optical adapter. Primers are listed in Table S1. 2.11 Statistical analyses Statistical analyses were performed using the GraphPad Prism 8 software. Pairwise comparisons were made using Student’s t -test (ns, not significant; *, P < 0.05; **, P < 0.01). 3. Results 3.1 The rf mutant produces round or flat fruits in woodland strawberry To identify genes regulating strawberry fruit shape, we found a round fruit ( rf ) mutant from the ethyl methanesulfonate (EMS)-mutagenized population of the woodland strawberry variety Yellow Wonder (YW, white-fruited). The rf mutant with red fruits was generated after crossing with the red-fruited variety Ruegen (Fig. S1A) [ 43 ]. The wild-type fruits were conical, whereas the rf fruits were nearly round or flat ( Fig. 1A ). Fruit measurements showed a significant reduction in fruit length and a significant increase in fruit width, resulting in a significantly reduced fruit shape index (length/width) in rf compared to wild type, but no difference in fruit weight ( Fig. 1B ). Other rf organs, such as petals, sepals, receptacles, stamens, and leaves, were also shorter or rounder than the wild type ( Fig. 1C, D ; Fig. S1B, C). Further observation showed that the rf receptacle was always shorter than the wild type from anthesis to fruit ripening ( Fig. 1E ). These results indicate that the RF gene plays an important role in regulating the shape of both fruit and other organs in woodland strawberry. Download figure Open in new tab Figure 1. Fruit morphologies in round fruit mutants in in woodland strawberry. (A) Mature fruits of wild-type Ruegen and the rf mutant. Scale bars: 1 cm. (B) Fruit width, length, shape index and weight of wild-type Ruegen and rf. n > 20. (C) Images showing the floral tissues of wild-type Ruegen and the rf mutant. Scale bars: 2 mm for stamens and 1 cm for others. (D) Petal shape index of wild-type Ruegen and the rf mutant. n > 20. (E) Fruits of wild-type Ruegen and rf at different developmental stages from anthesis to ripening. DAP, day after pollination. Scale bars: 1 cm. For statistical analysis, data are the mean ±SD; ** , P < 0.01; ns, not significant, Student’s t -test. 3.2 FveTRM5 is the causal gene of the rf mutant To identify the causal gene, the rf mutant was crossed with the wild-type Ruegen, resulting in F 1 progeny that all exhibited the wild-type phenotype. These F 1 plants were then self-pollinated to produce a segregating F 2 population. In this F 2 population, 81 plants produced conical fruits, and 27 plants produced round fruits, suggesting a monogenic recessive mutation responsible for rf . Bulked genome resequencing and data analysis identified a highly linked peak on chromosome 2 ( Fig. 2A ). The following criteria were used to filter the SNPs: (1) G-to-A or C-to-T transition; (2) with an index of 100% in the mutant library, < 50% in the WT library; (3) located in the coding sequence and causing nonsynonymous mutations in the protein [ 35 ]. After filtering, seven candidates were identified in the region from 16.48 to 20.03 Mb (Table S2). One of the candidates is the C-to-T mutation in the second exon of FvH4_2g22810 (annotation ver. 4), which causes a premature stop codon at amino acid residue 266 ( Fig. 2B ). This was further confirmed to be homozygous in 52 F 2 mutants by Sanger sequencing, in which the mutation in FvH4_2g22810 co-segregated with the round fruit phenotype. The protein sequence of FvH4_2g22810 shared a high similarity with Arabidopsis TON1 RECRUITING MOTIF 5 (AtTRM5), tomato SlTRM5, and cucumber CsTRM5 ( Fig. 2C ) [ 8 , 14 , 15 ]. Therefore, this gene was named FveTRM5 . We found a total of 18 TRM family members in the woodland strawberry genome with distinct expression patterns (Fig. S2A, B). The AtTRM1-5 subclade contains three woodland strawberry genes, including FveTRM1 (FvH4_6g46710), FveTRM4 (FvH4_5g13690) and FveTRM5 ( Fig. 2C ). When examined by RT-qPCR, FveTRM5 was significantly downregulated in the rf fruits at 7 DAP (days after pollination) compared to wild type ( Fig. 2D ), suggesting nonsense-mediated RNA decay. Download figure Open in new tab Figure 2. Isolation and characterization of FveTRM5 in woodland strawberry. (A) Diagram showing the SNPs associated with rf along the chromosomes. The X axis represents seven chromosomes in woodland strawberry. The Y axis indicates the differences in allele frequencies between the wild type and rf mutant pools. The arrow indicates the linked peak on chromosome 2. (B) Diagram showing the candidate causal mutation in FvH4_2g22810 for rf . The red letters indicate the candidate point mutation. (C) Phylogenetic tree of the TRM proteins in woodland strawberry (Fve), Arabidopsis (At), tomato (Sl), and cucumber (Cs) in the AtTRM1-5 subclade using full-length sequences. Bootstrap values at the nodes are percentages of 1,000 replicates. The branch length indicates the number of substitutions per site. (D) Relative expression level of FveTRM5 in wild-type Ruegen and rf fruits (7 DAP) examined by RT-qPCR. n = 3. (E) Images showing the mature fruits of wild-type H4, rf , and two complementation lines ( Comp -L1 and Comp -L2) in woodland strawberry. Scale bars: 1 cm. (F) Relative expression level of FveTRM5 in wild-type H4, rf, Comp -L1 and Comp -L2 leaves examined by RT-qPCR. n = 3. (G) Fruit shape index of H4, rf, Comp -L1 and Comp -L2. n > 10. For statistical analysis, data are the mean ±SD; * , P < 0.05; ** , P < 0.01; Student’s t -test. To genetically validate the gene cloning result, the complementation construct FveTRM5pro : FveTRM5 was generated and stably transformed into the wild-type Hawaii 4 (H4) for higher transformation efficiency. We obtained a total of 28 transgenic lines with either wild-type or elongated fruits in the T 0 generation. We selected 2 lines (L1 and L2) with elongated fruits to cross with the rf mutant. In the F 2 generation, the transgenic plants with the homozygous rf mutation were identified and named Comp -L1 and Comp -L2 ( Fig. 2E ). RT-qPCR showed that the expression level of FveTRM5 in Comp -L1 and Comp -L2 was 28 to 30 times higher than in the wild type ( Fig. 2F ). The fruit shape index of Comp -L1 and Comp -L2 was significantly higher than that of rf and wild type ( Fig. 2G ), indicating that FveTRM5 could rescue the fruit shape defect in rf . Taken together, these results indicate that FveTRM5 is the causal gene of the rf mutant. 3.3 Overexpression of FveTRM5 results in elongated organs in Arabidopsis and woodland strawberry To determine the function of FveTRM5, FveTRM 5 driven by the 35S promoter was first transformed into WT Arabidopsis. Twenty-five independent FveTRM5 -OE lines were obtained in the T 1 generation with much narrower and longer leaves (Fig. S3A). FveTRM5 expression was confirmed in two transgenic Arabidopsis lines (Fig. S3B). Compared to the wild type, overexpression of FveTRM5 resulted in sufficiently elongated organs, including petals, stamens, gynoecia, siliques, and seeds (Fig. S3C, D). These results suggest that FveTRM5 could promote organ elongation in alternative species. To further clarify the function of FveTRM5 in strawberry, the FveTRM5 -overexpression construct was also stably transformed into the wild-type woodland strawberry strain H4. We obtained 10 independent FveTRM5 overexpressing lines. Two lines (L2 and L3) were carefully characterized at T 0 . Both lines produced much thinner mature fruits and floral organs, such as petals and receptacles ( Fig. 3A ). RT-qPCR confirmed that FveTRM5 was expressed at 177-fold and 100-fold higher levels in these two lines than in the wild type ( Fig. 3B ). Fruit shape measurements showed that fruit length was significantly greater and fruit width was significantly smaller in FveTRM5 -OE than in the wild type, resulting in a higher fruit shape index ( Fig. 3C ). It should be noted that the fruit weight also decreased significantly in FveTRM5 -OE, mainly due to the extreme reduction in fruit width. The change in fruit shape in FveTRM5 -OE started at anthesis and remained the same until ripening ( Fig. 3D ). FveTRM5 -OE also showed an increase in the petal shape index (length/width) ( Fig. 3E ). In addition, FveTRM5 overexpression resulted in narrower leaves (Fig. S4A-C). Taken together, FveTRM5 is an important regulator of organ shape in woodland strawberry and may exert similar phenotypes in other plants. Download figure Open in new tab Figure 3. Phenotypic characterization of the FveTRM5-OE transgenic lines in woodland strawberry. (A) Images showing the fruits and floral organs of two FveTRM5 -OE transgenic lines (L2 and L3) in woodland strawberry. Scale bars: 1 cm. (B) Relative expression levels of FveTRM5 in the leaves of wild-type H4 and FveTRM5 -OE (L2 and L3) examined by RT-qPCR. n = 3. (C) Fruit length, width, shape index and weight of H4 and FveTRM5 -OE (L2 and L3). n > 15. (D) Fruits of wild type H4 and two independent FveTRM5 -OE lines (L2 and L3) at different developmental stages from anthesis to ripening. DAP, days after pollination. Scale bars: 1 cm. (E) Petal shape index of H4 and FveTRM5 -OE (L2 and L3). n > 20. For statistical analysis, data are the mean ±SD; * , P < 0.05; ** , P < 0.01, Student’s t -test. 3.4 FveTRM5 is highly expressed in developing organs Based on the woodland strawberry transcriptome database [ 44 ], FveTRM5 was widely expressed in flower and fruit tissues, such as flower meristems (FM), carpels, embryos, and receptacles (cortex and pith), as well as shoot apical meristems (SAM), leaves and roots ( Fig. 4A ). When examined by RT-qPCR, FveTRM5 expression was indeed higher in leaves, flower buds and young fruits, moderate in roots, but strongly decreased in mature fruits at 30 DAP ( Fig. 4B ). We further examined the expression pattern of FveTRM5 in shoot tips and floral buds by RNA in situ hybridization. Flower stages were designated as previously described [ 45 ]. Strong signals of FveTRM5 were detected in the SAM, leaf primordia, FM, receptacle meristem and the four whorls of floral organs at flower stages 1-13, no signal was detected by hybridization with the FveTRM5 sense probe ( Fig. 4C ). Download figure Open in new tab Figure 4. Expression pattern of FveTRM5 in woodland strawberry. (A) Expression pattern of FveTRM5 in woodland strawberry as indicated by transcripts per million (TPM) values obtained from the transcriptome database (Li et al., 2019). (B) Relative expression levels of FveTRM5 in the tissues of wild-type Ruegen examined by RT-qPCR. n = 3. DAP, days after pollination. (C) Expression pattern of FveTRM5 in longitudinal sections of wild-type shoot tips and flower buds at different stages by RNA in situ hybridization. SAM, shoot apical meristem; L, leaf; FM, floral meristem; Se, sepal; St, stamen; Pe, petal; Ca, carpel; An, anther. Scale bars: 100 μm. 3.5 FveTRM5 regulates cell expansion and cell division in strawberry fruit As the change in fruit shape remains the same throughout development, the cell morphology in the receptacles of rf, FveTRM5 -OE and the wild types (Ruegen and H4) at anthesis was examined by paraffin sectioning. Initial observations showed that the receptacle cells appeared shorter and smaller in rf and longer in FveTRM5 -OE ( Fig. 5A ). Cell measurements showed that rf receptacle cells were significantly shorter than wild type, whereas FveTRM5 -OE receptacle cells were significantly longer than wild type ( Fig. 5B ), suggesting that FveTRM5 positively regulates cell elongation in the longitudinal direction. In contrast, cell width was significantly decreased in rf and the two FveTRM5 -OE transgenic lines, resulting in the significant change of cell shape index. In addition, there were more cell layers in rf and fewer cell layers in FveTRM5 -OE in the transverse direction, suggesting that FveTRM5 could inhibit cell division in the medial-lateral direction of the receptacle ( Fig. 5B ). Download figure Open in new tab Figure 5. Changes in cell morphology in different materials and subcellular localization of FveTRM5. (A) Images showing cell shapes in the receptacle of wild-type Ruegen and H4, rf , and FveTRM5-OE (L2 and L3) at anthesis. The blue arrow indicates the longitudinal direction of the fruit. Scale bars: 100 μm. (B) Cell shape and number of cell layers in the transverse direction at the widest part of wild-type Ruegen and H4, rf , and FveTRM5-OE (L2 and L3) receptacles. For statistical analysis, data are the mean ±SD; ** , P < 0.01; student’s t -test. n = 60 for cell shapes and 6 for cell layers. (C) Subcellular localization of GFP-FveTRM5 in Nicotiana benthamiana leaves. Green indicates GFP fluorescence, and red indicates RFP fluorescence of the microtubule marker RFP-AtTUB6. Scale bars: 10 μm. Previous studies have shown that some TRM proteins co-localize with microtubules, a major component of the cytoskeleton [ 14 ]. To test this possibility, GFP-FveTRM5 and the microtubule marker RFP-AtTUB6 were simultaneously infiltrated into the Nicotiana benthamiana leaves for transient expression. The RFP-AtTUB6 signals were shown as lines, indicating the localization of microtubules ( Fig. 5C ). GFP-FveTRM5 signals colocalized with RFP-AtTUB6 on microtubules ( Fig. 5C ). This localization is similar to that of SlTRM5, which has been implicated in fruit shape regulation [ 14 , 46 ]. Taken together, these results indicate that FveTRM5 regulates strawberry fruit shape by controlling both cell expansion and cell division. 4. Discussion Cultivated strawberries produce fruits with a wide variety of shapes, but those with round fruits are relatively rare. In this study, we identified an EMS mutant called round fruit ( rf ) in woodland strawberry. Gene cloning and genetic analysis revealed that FveTRM5 is the causal gene for this mutant. Overexpression of FveTRM5 resulted in elongated organs in both woodland strawberry and Arabidopsis, suggesting a conserved role in different species. Furthermore, FveTRM5 can inhibit transverse cell division and promote longitudinal cell elongation in the fruit receptacle. This work provides compelling evidence for the role of FveTRM5 in fruit shape control and makes it a strong candidate for fruit shape manipulation in strawberry and other fruit crops. The TRM proteins in the AtTRM1-5 subclade promote elongation of fruits as well as other organs in a wide range of plant species [ 9 - 11 , 14 , 15 ]. There are three members in this subclade in the woodland strawberry genome ( Fig. 2C ). In this study, we have demonstrated the important functions of FveTRM5 in fruit shape regulation. It remains to be tested whether FveTRM1 and FveTRM4 have similar and redundant functions. This possibility is supported by the tomato mutants of SlTRM3/4 and SlTRM5 , which show additive effects in shortening the ovate Slofp20 fruits [ 46 ]. In addition, mutations in SlTRM19 and SlTRM17/20a from another subclade could enhance fruit elongation in the ovate Slofp20 double mutant, suggesting that the TRM proteins from different subclades may also have redundant or opposing functions. This finding extends the range of potential TRMs involved in the fruit shape control. The TRM proteins promote organ elongation by increasing cell division in the longitudinal direction and decreasing cell division in the transverse direction [ 9 , 14 , 15 ]. Our results showed that FveTRM5 not only inhibited cell division in the transverse direction, but also decreased cell width and increased cell length, a result of anisotropic cell expansion ( Fig. 5 ). Although TRM proteins can be localized on microtubules or in the cytosol [ 8 ], SlTRM5 is particularly localized on microtubules [ 14 , 46 ]. FveTRM5 also localizes to microtubules, similar to SlTRM5. The TRM proteins often interact with OFPs to regulate organ shape [ 6 , 14 ]. Thus, protein interaction assays may help to find the FveOFPs involved in the fruit shape regulation. Fruit shape is finely regulated by plant hormones, such as auxin and gibberellic acid (GA), as well as other stimuli. In tomato, whole-plant application of exogenous auxin before anthesis results in elongated ovaries and fruits, with an increased number of cells along the longitudinal axis and enlarged cells in most tissues of the ovary [ 47 ]. GA 3 treatment in tomato results in elongated fruits, whereas application of the GA inhibitor paclobutrazol results in more flattened fruits [ 48 , 49 ]. In woodland strawberry, auxin application results in rounder fruits, whereas GA treatment leads to thinner fruits [ 30 ]. The hormone pathways are known to cross-talk with the OFP and IQD family genes [ 6 , 47 , 50 , 51 ], which may indirectly interfere with TRM functions. In addition to hormones, mutation of the red photoreceptor FvePhyB results in fruits with higher shape index [ 52 ]. Some upstream regulators of the TRM genes have been reported. For example, the bHLH transcription factors Leaf-related Protein 1 (LP1) and LP2 can directly regulate the expression levels of AtTRM1 and AtTRM2 to induce longitudinal cell elongation in Arabidopsis [ 53 ]. OsSPL16 can directly bind to the GW7 (ortholog of AtTRM1 ) promoter to inhibit its expression in rice [ 10 ]. Whether and how FveTRM5 is regulated by hormones and light signaling remains to be investigated. In conclusion, we have reported a key player in the regulation of strawberry fruit shape, FveTRM5, which influences cell division and cell elongation in fruit tissues. Based on the genetic studies, we believe that it is possible to knock out FveTRM5 to produce round fruits. However, FveTRM5 cannot be overexpressed to high levels as this will reduce the biomass. These findings can provide a theoretical basis for breeding new fruit shape cultivars in strawberry and enrich our knowledge of fruit shape control in fruit crops. Data Availability All relevant data and vectors that support the findings of this study are available from the corresponding author, upon request. Author Contributions CK conceived and designed the experiments; ZZ and LW performed the experiments; QG created the F 2 population of rf ; SH analyzed the sequencing data; ZZ and CK wrote the manuscript. All the authors have read and approved the manuscript. Funding This work was supported by the National Natural Science Foundation of China (32172539). Declaration of Competing Interest Patent entitled “The RF gene regulating strawberry fruit shape and its application (ZL202210171639.0)” has been authorized in China. CK and LW are listed as inventors. Supplementary data Figure S1 . Leaf phenotypic characterization of the rf mutant in woodland strawberry. Figure S2 . Phylogenetic and expression pattern analysis of the FveTRM genes. Figure S3 . Phenotypic characterization of the FveTRM5 -OE transgenic lines in Arabidopsis . Figure S4 . Leaf phenotypic characterization of the FveTRM5 -OE transgenic lines in woodland strawberry. Table S1 . Summary of primers used in this study. Table S2 . The list of candidate SNPs in the EMS mutant rf . Acknowledgments The authors thank Dr. Pengwei Wang (Huazhong Agricultural University) for providing the microtubule marker used for subcellular localization. References [1]. ↵ Y. Dong , L. Østergaard , Fruit development and diversification , Current Biology , 29 ( 2019 ) R781 – R785 . OpenUrl CrossRef PubMed [2]. ↵ Z. Liu , H. Ma , S. Jung , D. Main , L. Guo , Developmental Mechanisms of Fleshy Fruit Diversity in Rosaceae , Annual Review of Plant Biology , 71 ( 2020 ) 547 – 573 . OpenUrl CrossRef PubMed [3]. ↵ Y. Pan , Y. Wang , C. McGregor , S. Liu , F. Luan , M. Gao , Y. Weng , Genetic architecture of fruit size and shape variation in cucurbits: a comparative perspective , Theoretical and Applied Genetics , 133 ( 2020 ) 1 – 21 . OpenUrl CrossRef PubMed [4]. ↵ J.-P. Mauxion , C. Chevalier , N. Gonzalez , Complex cellular and molecular events determining fruit size , Trends in Plant Science , 26 ( 2021 ) 1023 – 1038 . OpenUrl CrossRef PubMed [5]. ↵ M.D. Lazzaro , S. Wu , A. Snouffer , Y. Wang , E. van der Knaap , Plant Organ Shapes Are Regulated by Protein Interactions and Associations With Microtubules , Frontiers in Plant Science , 9 ( 2018 ). [6]. ↵ A. Snouffer , C. Kraus , E. van der Knaap , The shape of things to come: ovate family proteins regulate plant organ shape , Current Opinion in Plant Biology , 53 ( 2020 ) 98 – 105 . OpenUrl CrossRef PubMed [7]. ↵ Q. Li , S. Luo , L. Zhang , Q. Feng , L. Song , M. Sapkota , S. Xuan , Y. Wang , J. Zhao , E. van der Knaap , X. Chen , S. Shen , Molecular and genetic regulations of fleshy fruit shape and lessons from Arabidopsis and rice , Horticulture Research , 10 ( 2023 ). [8]. ↵ S. Drevensek , M. Goussot , Y. Duroc , A. Christodoulidou , S. Steyaert , E. Schaefer , E. Duvernois , O. Grandjean , M. Vantard , D. Bouchez , M. Pastuglia , The Arabidopsis TRM1–TON1 Interaction Reveals a Recruitment Network Common to Plant Cortical Microtubule Arrays and Eukaryotic Centrosomes The Plant Cell , 24 ( 2012 ) 178 – 191 . OpenUrl [9]. ↵ Y.K. Lee , G.-T. Kim , I.-J. Kim , J. Park , S.-S. Kwak , G. Choi , W.-I. Chung , LONGIFOLIA1 and LONGIFOLIA2, two homologous genes,regulate longitudinal cell elongation in Arabidopsis , Development , 133 ( 2006 ) 4305 – 4314 . OpenUrl Abstract / FREE Full Text [10]. ↵ S. Wang , S. Li , Q. Liu , K. Wu , J. Zhang , S. Wang , Y. Wang , X. Chen , Y. Zhang , C. Gao , F. Wang , H. Huang , X. Fu , The OsSPL16-GW7 regulatory module determines grain shape and simultaneously improves rice yield and grain quality , Nature Genetics , 47 ( 2015 ) 949 – 954 . OpenUrl CrossRef PubMed [11]. ↵ Y. Wang , G. Xiong , J. Hu , L. Jiang , H. Yu , J. Xu , Y. Fang , L. Zeng , E. Xu , J. Xu , W. Ye , X. Meng , R. Liu , H. Chen , Y. Jing , Y. Wang , X. Zhu , J. Li , Q. Qian , Copy number variation at the GL7 locus contributes to grain size diversity in rice , Nature Genetics , 47 ( 2015 ) 944 – 948 . OpenUrl CrossRef PubMed [12]. ↵ W. Wang , Q. Pan , B. Tian , F. He , Y. Chen , G. Bai , A. Akhunova , H.N. Trick , E. Akhunov , Gene editing of the wheat homologs of TONNEAU1-recruiting motif encoding gene affects grain shape and weight in wheat , The Plant Journal , 100 ( 2019 ) 251 – 264 . OpenUrl CrossRef PubMed [13]. ↵ Q. Wang , J. Fan , J. Cong , M. Chen , J. Qiu , J. Liu , X. Zhao , R. Huang , H. Liu , X. Huang , Natural variation of ZmLNG1 alters organ shapes in maize , New Phytologist , 237 ( 2023 ) 471 – 482 . OpenUrl CrossRef PubMed [14]. ↵ S. Wu , B. Zhang , N. Keyhaninejad , G.R. Rodríguez , H.J. Kim , M. Chakrabarti , E. Illa-Berenguer , N.K. Taitano , M.J. Gonzalo , A. Díaz , Y. Pan , C.P. Leisner , D. Halterman , C.R. Buell , Y. Weng , S.H. Jansky , H. van Eck , J. Willemsen , A.J. Monforte , T. Meulia , E. van der Knaap , A common genetic mechanism underlies morphological diversity in fruits and other plant organs , Nature Communications , 9 ( 2018 ) 4734 . OpenUrl CrossRef PubMed [15]. ↵ Y. Xie , X. Liu , C. Sun , X. Song , X. Li , H. Cui , J. Guo , L. Liu , A. Ying , Z. Zhang , X. Zhu , L. Yan , X. Zhang , CsTRM5 regulates fruit shape via mediating cell division direction and cell expansion in cucumber , Horticulture Research , 10 ( 2023 ). [16]. ↵ J. Liu , J. Van Eck , B. Cong , S.D. Tanksley , A new class of regulatory genes underlying the cause of pear-shaped tomato fruit , Proceedings of the National Academy of Sciences of the United States of America , 99 ( 2002 ) 13302 – 13306 . OpenUrl Abstract / FREE Full Text [17]. ↵ H. Zhou , R. Ma , L. Gao , J. Zhang , A. Zhang , X. Zhang , F. Ren , W. Zhang , L. Liao , Q. Yang , S. Xu , C. Otieno Ogutu , J. Zhao , M. Yu , Q. Jiang , S.S. Korban , Y. Han , A 1.7-Mb chromosomal inversion downstream of a PpOFP1 gene is responsible for flat fruit shape in peach , Plant Biotechnology Journal , 19 ( 2021 ) 192 – 205 . OpenUrl CrossRef PubMed [18]. J. Guan , Y. Xu , Y. Yu , J. Fu , F. Ren , J. Guo , J. Zhao , Q. Jiang , J. Wei , H. Xie , Genome structure variation analyses of peach reveal population dynamics and a 1.67 Mb causal inversion for fruit shape , Genome Biology , 22 ( 2021 ) 13 . OpenUrl CrossRef PubMed [19]. ↵ J. Guo , K. Cao , C. Deng , Y. Li , G. Zhu , W. Fang , C. Chen , X. Wang , J. Wu , L. Guan , S. Wu , W. Guo , J.-L. Yao , Z. Fei , L. Wang , An integrated peach genome structural variation map uncovers genes associated with fruit traits , Genome Biology , 21 ( 2020 ) 258 . OpenUrl CrossRef PubMed [20]. ↵ H. Xiao , N. Jiang , E. Schaffner , E.J. Stockinger , E. van der Knaap , A Retrotransposon-Mediated Gene Duplication Underlies Morphological Variation of Tomato Fruit , Science , 319 ( 2008 ) 1527 – 1530 . OpenUrl Abstract / FREE Full Text [21]. ↵ I.L. Goldman , Y. Wang , A.V. Alfaro , S. Brainard , M.W. Oravec , C.E. McGregor , E. van der Knaap , Form and contour: breeding and genetics of organ shape from wild relatives to modern vegetable crops , Frontiers in Plant Science , 14 ( 2023 ). [22]. ↵ J. Azimzadeh , P. Nacry , A. Christodoulidou , S.p. Drevensek , C. Camilleri , N. Amiour , F.o. Parcy , M. Pastuglia , D. Bouchez , Arabidopsis TONNEAU1 Proteins Are Essential for Preprophase Band Formation and Interact with Centrin , The Plant Cell , 20 ( 2008 ) 2146 – 2159 . OpenUrl Abstract / FREE Full Text [23]. L. Spinner , A. Gadeyne , K. Belcram , M. Goussot , M. Moison , Y. Duroc , D. Eeckhout , N. De Winne , E. Schaefer , E. Van De Slijke , G. Persiau , E. Witters , K. Gevaert , G. De Jaeger , D. Bouchez , D. Van Damme , M. Pastuglia , A protein phosphatase 2A complex spatially controls plant cell division , Nature Communications , 4 ( 2013 ) 1863 . OpenUrl CrossRef PubMed [24]. C.G. Rasmussen , M. Bellinger , An overview of plant division-plane orientation , New Phytologist , 219 ( 2018 ) 505 – 512 . OpenUrl CrossRef PubMed [25]. ↵ E. Schaefer , K. Belcram , M. Uyttewaal , Y. Duroc , M. Goussot , D. Legland , E. Laruelle , M.-L. de Tauzia-Moreau , M. Pastuglia , D. Bouchez , The preprophase band of microtubules controls the robustness of division orientation in plants , Science , 356 ( 2017 ) 186 – 189 . OpenUrl Abstract / FREE Full Text [26]. ↵ S. Wu , H. Xiao , A. Cabrera , T. Meulia , E. van der Knaap , SUN Regulates Vegetative and Reproductive Organ Shape by Changing Cell Division Patterns Plant Physiology , 157 ( 2011 ) 1175 – 1186 . OpenUrl PubMed [27]. ↵ K. Bürstenbinder , B. Möller , R. Plötner , G. Stamm , G. Hause , D. Mitra , S. Abel , The IQD Family of Calmodulin-Binding Proteins Links Calcium Signaling to Microtubules, Membrane Subdomains, and the Nucleus , Plant Physiology , 173 ( 2017 ) 1692 – 1708 . OpenUrl Abstract / FREE Full Text [28]. ↵ M. Urrutia , V. Meco , J.L. Rambla , C. Martín-Pizarro , J. Pillet , J. Andrés , J.F. Sánchez-Sevilla , A. Granell , T. Hytönen , D. Posé , Diversity of the volatilome and the fruit size and shape in European woodland strawberry (Fragaria vesca) , The Plant Journal , 116 ( 2023 ) 1201 – 1217 . OpenUrl CrossRef PubMed [29]. ↵ S. Nagamatsu , M. Tsubone , T. Wada , K. Oku , M. Mori , C. Hirata , A. Hayashi , T. Tanabata , S. Isobe , K. Takata , K. Shimomura , Strawberry fruit shape: quantification by image analysis and QTL detection by genome-wide association analysis , Breeding science , 71 ( 2021 ) 167 – 175 . OpenUrl CrossRef PubMed [30]. ↵ X. Liao , M. Li , B. Liu , M. Yan , X. Yu , H. Zi , R. Liu , C. Yamamuro , Interlinked regulatory loops of ABA catabolism and biosynthesis coordinate fruit growth and ripening in woodland strawberry , Proceedings of the National Academy of Sciences , 115 ( 2018 ) E11542 – E11550 . OpenUrl Abstract / FREE Full Text [31]. ↵ S.-m. Wang , W.-j. Li , Y.-x. Liu , H. Li , Y. Ma , Z.-h. Zhang , Comparative transcriptome analysis of shortened fruit mutant in woodland strawberry (Fragaria vesca) using RNA-Seq , Journal of Integrative Agriculture , 16 ( 2017 ) 828 – 844 . OpenUrl CrossRef [32]. ↵ H.M. Cockerton , A. Karlström , A.W. Johnson , B. Li , E. Stavridou , K.J. Hopso n, A.B. Whitehouse , R.J. Harrison , Genomic Informed Breeding Strategies for Strawberry Yield and Fruit Quality Traits , Frontiers in Plant Science , 12 ( 2021 ). [33]. ↵ P. Rey-Serra , M. Mnejja , A. Monfort , Shape, firmness and fruit quality QTLs shared in two non-related strawberry populations , Plant Science , 311 ( 2021 ) 111010 . OpenUrl CrossRef PubMed [34]. ↵ Y. Li , P. Zhang , G. Wang , W. Zhao , Z. Bao , F. Ma , FvUVI4 inhibits cell division and cell expansion to modulate fruit development in Fragaria vesca , Plant Physiology and Biochemistry , 213 ( 2024 ) 108804 . OpenUrl CrossRef PubMed [35]. ↵ M. Pi , R. Zhong , S. Hu , Z. Cai , M. Plunkert , W. Zhang , Z. Liu , C. Kang , A GT-1 and PKc domain-containing transcription regulator SIMPLE LEAF1 controls compound leaf development in woodland strawberry , New Phytologist , 237 ( 2023 ) 1391 – 1404 . OpenUrl CrossRef PubMed [36]. ↵ H. Li , R. Durbin , Fast and accurate short read alignment with Burrows–Wheeler transform , Bioinformatics , 25 ( 2009 ) 1754 – 1760 . OpenUrl CrossRef PubMed Web of Science [37]. ↵ P.P. Edger , R. VanBuren , M. Colle , T.J. Poorten , C.M. Wai , C.E. Niederhuth , E.I. Alger , S. Ou , C.B. Acharya , J. Wang , P. Callow , M.R. McKain , J. Shi , C. Collier , Z. Xiong , J.P. Mower , J.P. Slovin , T. Hytönen , N. Jiang , K.L. Childs , S.J. Knapp , Single-molecule sequencing and optical mapping yields an improved genome of woodland strawberry (Fragaria vesca) with chromosome-scale contiguity , GigaScience , 7 ( 2018 ). [38]. ↵ H. Li , B. Handsaker , A. Wysoker , T. Fennell , J. Ruan , N. Homer , G. Marth , G. Abecasis , R. Durbin , G.P.D.P. Subgroup , The Sequence Alignment/Map format and SAMtools , Bioinformatics , 25 ( 2009 ) 2078 – 2079 . OpenUrl CrossRef PubMed Web of Science [39]. ↵ A. McKenna , M. Hanna , E. Banks , A. Sivachenko , K. Cibulskis , A. Kernytsky , K. Garimella , D. Altshuler , S. Gabriel , M. Daly , M.A. DePristo , The Genome Analysis Toolkit: A MapReduce framework for analyzing next-generation DNA sequencing data , Genome Research , 20 ( 2010 ) 1297 – 1303 . OpenUrl Abstract / FREE Full Text [40]. ↵ A.R. Quinlan , I.M. Hall , BEDTools: a flexible suite of utilities for comparing genomic features , Bioinformatics , 26 ( 2010 ) 841 – 842 . OpenUrl CrossRef PubMed Web of Science [41]. ↵ J. Feng , C. Dai , H. Luo , Y. Han , Z. Liu , C. Kang , Reporter gene expression reveals precise auxin synthesis sites during fruit and root development in wild strawberry , Journal of Experimental Botany , 70 ( 2019 ) 563 – 574 . OpenUrl CrossRef PubMed [42]. ↵ Z. Bao , Y. Guo , Y. Deng , J. Zang , J. Zhang , Y. Deng , B. Ouyang , X. Qu , K. Bürstenbinder , P. Wang , Microtubule-associated protein SlMAP70 interacts with IQ67-domain protein SlIQD21a to regulate fruit shape in tomato , The Plant Cell , 35 ( 2023 ) 4266 – 4283 . OpenUrl CrossRef PubMed [43]. ↵ C. Hawkins , J. Caruana , E. Schiksnis , Z. Liu , Genome-scale DNA variant analysis and functional validation of a SNP underlying yellow fruit color in wild strawberry , Scientific Reports , 6 ( 2016 ) 29017 . OpenUrl CrossRef PubMed [44]. ↵ Y. Li , M. Pi , Q. Gao , Z. Liu , C. Kang , Updated annotation of the wild strawberry Fragaria vesca V4 genome , Horticulture Research , 6 ( 2019 ). [45]. ↵ C.A. Hollender , A.C. Geretz , J.P. Slovin , Z. Liu , Flower and early fruit development in a diploid strawberry , Fragaria vesca, Planta , 235 ( 2012 ) 1123 – 1139 . OpenUrl PubMed [46]. ↵ B. Zhang , Q. Li , N. Keyhaninejad , N. Taitano , M. Sapkota , A. Snouffer , E. van der Knaap , A combinatorial TRM-OFP module bilaterally fine-tunes tomato fruit shape , New Phytologist , 238 ( 2023 ) 2393 – 2409 . OpenUrl CrossRef PubMed [47]. ↵ Y. Wang , J.P. Clevenger , E. Illa-Berenguer , T. Meulia , E. van der Knaap , L. Sun , A Comparison of sun, ovate, fs8.1 and Auxin Application on Tomato Fruit Shape and Gene Expression , Plant and Cell Physiology , 60 ( 2019 ) 1067 – 1081 . OpenUrl CrossRef PubMed [48]. ↵ P. Zhao , F. Wang , Y. Deng , F. Zhong , P. Tian , D. Lin , J. Deng , Y. Zhang , T. Huang , Sly-miR159 regulates fruit morphology by modulating GA biosynthesis in tomato , Plant Biotechnology Journal , 20 ( 2022 ) 833 – 845 . OpenUrl CrossRef PubMed [49]. ↵ S. Chen , X.-J. Wang , G.-F. Tan , W.-Q. Zhou , G.-L. Wang , Gibberellin and the plant growth retardant Paclobutrazol altered fruit shape and ripening in tomato , Protoplasma , 257 ( 2020 ) 853 – 861 . OpenUrl CrossRef PubMed [50]. ↵ A. Carvalho , M.H. Vicente , L.F. Ferigolo , E.M. Silva , M. Levy , L.E.P. Peres , R. Sablowski , C. Schommer , N. Ori , F.T.S. Nogueira , miR319-targeted TCP4/LANCEOLATE directly regulates OVATE and auxin responses to modulate tomato gynoecium patterning and fruit morphology , bioRxiv , ( 2022 ) 2022.2003.2015.484181. [51]. ↵ J.R. Wendrich , B.-J. Yang , P. Mijnhout , H.-W. Xue , B.D. Rybel , D. Weijers , IQD proteins integrate auxin and calcium signaling to regulate microtubule dynamics during Arabidopsis development , bioRxiv , ( 2018 ) 275560 . [52]. ↵ Q. Gao , S. Hu , X. Wang , F. Han , H. Luo , Z. Liu , C. Kang , The red/far-red light photoreceptor FvePhyB regulates tissue elongation and anthocyanin accumulation in woodland strawberry , Horticulture Research , 10 ( 2023 ). [53]. ↵ R. Lu , J. Zhang , Y.-W. Wu , Y. Wang , J. Zhang , Y. Zheng , Y. Li , X.-B. Li , bHLH transcription factors LP1 and LP2 regulate longitudinal cell elongation , Plant Physiology , 187 ( 2021 ) 2577 – 2591 . OpenUrl CrossRef PubMed View the discussion thread. Back to top Previous Next Posted March 13, 2025. Download PDF Supplementary Material Email Thank you for your interest in spreading the word about bioRxiv. NOTE: Your email address is requested solely to identify you as the sender of this article. Your Email * Your Name * Send To * Enter multiple addresses on separate lines or separate them with commas. You are going to email the following FveTRM5 plays a critical role in regulating fruit shape in woodland strawberry Message Subject (Your Name) has forwarded a page to you from bioRxiv Message Body (Your Name) thought you would like to see this page from the bioRxiv website. Your Personal Message CAPTCHA This question is for testing whether or not you are a human visitor and to prevent automated spam submissions. Share FveTRM5 plays a critical role in regulating fruit shape in woodland strawberry Zhenzhen Zheng , Liyang Wang , Qi Gao , Shaoqiang Hu , Chunying Kang bioRxiv 2025.03.10.642363; doi: https://doi.org/10.1101/2025.03.10.642363 Share This Article: Copy Citation Tools FveTRM5 plays a critical role in regulating fruit shape in woodland strawberry Zhenzhen Zheng , Liyang Wang , Qi Gao , Shaoqiang Hu , Chunying Kang bioRxiv 2025.03.10.642363; doi: https://doi.org/10.1101/2025.03.10.642363 Citation Manager Formats BibTeX Bookends EasyBib EndNote (tagged) EndNote 8 (xml) Medlars Mendeley Papers RefWorks Tagged Ref Manager RIS Zotero Tweet Widget Facebook Like Google Plus One Subject Area Plant Biology Subject Areas All Articles Animal Behavior and Cognition (7635) Biochemistry (17691) Bioengineering (13892) Bioinformatics (41937) Biophysics (21452) Cancer Biology (18588) Cell Biology (25504) Clinical Trials (138) Developmental Biology (13378) Ecology (19899) Epidemiology (2067) Evolutionary Biology (24320) Genetics (15609) Genomics (22506) Immunology (17736) Microbiology (40394) Molecular Biology (17181) Neuroscience (88605) Paleontology (666) Pathology (2832) Pharmacology and Toxicology (4824) Physiology (7641) Plant Biology (15156) Scientific Communication and Education (2045) Synthetic Biology (4294) Systems Biology (9825) Zoology (2271)