Genetic mapping reveals a candidate gene CmoFL1 controlling fruit length in pumpkin

Yimei Zhou, Meng Zhao, Qinghui Shen, Mengyi Zhang, Chenhao Wang, Yutong Zhang, Qinrong Yang, Yongming Bo, Zhongyuan Hu, Jinghua Yang, Mingfang Zhang, Xiaolong Lyu
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Abstract

Fruit length (FL) is an important economical trait that affects fruit yield and appearance. Pumpkin (Cucurbita moschata Duch) contains a wealth genetic variation in fruit length. However, the natural variation underlying differences in pumpkin fruit length remains unclear. In this study, we constructed a F2 segregate population using KG1 producing long fruit and MBF producing short fruit as parents to identify the candidate gene for fruit length. By bulked segregant analysis (BSA-seq) and Kompetitive Allele-Specific PCR (KASP) approach of fine mapping, we obtained a 50.77 kb candidate region on chromosome 14 associated with the fruit length. Then, based on sequence variation, gene expression and promoter activity analyses, we identified a candidate gene (CmoFL1) encoding E3 ubiquitin ligase in this region may account for the variation of fruit length. One SNP variation in promoter of CmoFL1 changed the GT1CONSENSUS, and DUAL-LUC assay revealed that this variation significantly affected the promoter activity of CmoFL1. RNA-seq analysis indicated that CmoFL1 might associated with the cell division process and negatively regulate fruit length. Collectively, our work identifies an important allelic affecting fruit length, and provides a target gene manipulating fruit length in future pumpkin breeding.
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基因图谱揭示了控制南瓜果实长度的候选基因 CmoFL1
果实长度(FL)是影响果实产量和外观的重要经济性状。南瓜(Cucurbita moschata Duch)果实长度的遗传变异非常丰富。然而,南瓜果实长度差异背后的自然变异仍不清楚。在本研究中,我们以结长果实的 KG1 和结短果实的 MBF 为亲本,构建了一个 F2 分离群体,以确定果实长度的候选基因。通过批量分离分析(BSA-seq)和竞争性等位基因特异性 PCR(KASP)精细作图方法,我们在 14 号染色体上获得了一个与果实长度相关的 50.77 kb 候选区域。然后,基于序列变异、基因表达和启动子活性分析,我们确定了该区域编码 E3 泛素连接酶的候选基因(CmoFL1)可能是果实长度变异的原因。CmoFL1启动子中的一个SNP变异改变了GT1CONSENSUS,DUAL-LUC检测发现该变异显著影响了CmoFL1的启动子活性。RNA-seq分析表明,CmoFL1可能与细胞分裂过程有关,并负向调控果实长度。总之,我们的研究发现了一个影响果实长度的重要等位基因,并为未来南瓜育种提供了一个操纵果实长度的目标基因。
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