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通过跨群体关联作图和基因组变异筛选鉴定控制芝麻叶片卷曲和蒴果不开裂的 SiCL1 基因。

Identification of a SiCL1 gene controlling leaf curling and capsule indehiscence in sesame via cross-population association mapping and genomic variants screening.

机构信息

Henan Sesame Research Center, Henan Academy of Agricultural Sciences, Zhengzhou, 450002, Henan, China.

出版信息

BMC Plant Biol. 2018 Nov 22;18(1):296. doi: 10.1186/s12870-018-1503-2.

Abstract

BACKGROUND

Leaf shape can affect plantlet development and seed yield in sesame. The morphological, histological and genetic analyses of a sesame mutant cl1 (cl) with curly leaf and indehiscent capsule traits were performed in this study. In order to clone the cl1 gene for breeding selection, genome re-sequencing of the 130 individuals of cl1 × USA (0)-26 F population and a bulked segregation analysis (BSA) pool was carried out. The genome re-sequencing data of the 822 germplasm with normal leaf shape were applied.

RESULTS

For cl1 mutant, the adaxial/abaxial character of the parenchyma cells in the leaf blades is reduced. Results proved that the leaf curling trait is controlled by a recessive gene (Sicl1). Cross- population association of the F population of cl1 × USA (0)-26 indicated that the target cl locus was located on the interval C29 between C29_6522236 and C29_6918901 of SiChr. 1. Further regional genome variants screening determined the 6 candidate variants using genomic variants data of 822 natural germplasm and a BSA pool data. Of which, 5 markers C29_6717525, C29_6721553, C29_6721558, C29_6721563, and C29_6721565 existed in the same gene (C29.460). With the aid of the validation in the test F population of cl1 × Yuzhi 11 and natural germplasm, the integrated marker SiCLInDel1 (C29: 6721553-6721572) was determined as the target marker, and C29.460 was the target gene SiCL1 in sesame. SiCL1 is a KAN1 homolog with the full length of 6835 bp. In cl1, the 20 nucleic acids (CAGGTAGCTATGTATATGCA) of SiCLInDel1 marker were mutagenized into 6 nucleic acids (TCTTTG). The deletion led to a frameshift mutation and resulted in the earlier translation termination of the CL gene. The Sicl1 allele was shortened to 1829 bp. SiCL1 gene was expressed mainly in the tissues of stem, leaf, bud, capsule and seed.

CONCLUSIONS

SiCL1 encodes a transcription repressor KAN1 protein and controls leaf curling and capsule indehiscence in sesame. The findings provided an example of high-efficient gene cloning in sesame. The SiCL1 gene and the cl1 mutant supply the opportunity to explore the development regulation of leaf and capsule, and would improve the new variety breeding with high harvest mechanization adaption in sesame.

摘要

背景

叶片形状会影响芝麻的植株发育和种子产量。本研究对具有卷曲叶和不开裂蒴果性状的芝麻突变体 cl1(cl)进行了形态学、组织学和遗传学分析。为了克隆 cl1 基因进行选育,对 cl1×USA(0)-26 F 群体的 130 个个体和一个分离群体分析(BSA)池进行了基因组重测序。应用了 822 份正常叶形种质资源的基因组重测序数据。

结果

对于 cl1 突变体,叶片中叶肉细胞的背腹特征减少。结果表明,卷曲叶性状受隐性基因(Sicl1)控制。cl1×USA(0)-26 的 F 群体的跨群体关联表明,目标 cl 基因座位于 SiChr.1 的 C29 区间内,位于 C29_6522236 和 C29_6918901 之间。进一步的区域基因组变异筛选使用 822 份自然种质资源的基因组变异数据和 BSA 池数据确定了 6 个候选变异。其中,5 个标记 C29_6717525、C29_6721553、C29_6721558、C29_6721563 和 C29_6721565 存在于同一基因(C29.460)中。借助 cl1×Yuzhi 11 和自然种质资源的测试 F 群体的验证,确定了整合标记 SiCLInDel1(C29:6721553-6721572)为目标标记,C29.460 为芝麻中的目标基因 SiCL1。SiCL1 是一个全长为 6835 bp 的 KAN1 同源物。在 cl1 中,SiCLInDel1 标记的 20 个核苷酸(CAGGTAGCTATGTATATGCA)突变为 6 个核苷酸(TCTTTG)。缺失导致移码突变,导致 CL 基因的翻译提前终止。Sicl1 等位基因缩短至 1829 bp。SiCL1 基因主要在茎、叶、芽、蒴果和种子组织中表达。

结论

SiCL1 编码一个转录抑制子 KAN1 蛋白,控制芝麻的卷曲叶和蒴果不开裂。该研究为芝麻中高效基因克隆提供了一个范例。SiCL1 基因和 cl1 突变体为探索叶片和蒴果的发育调控提供了机会,并将提高芝麻高收获机械化适应性的新品种选育。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7a0d/6251216/0b9f7f517b97/12870_2018_1503_Fig1_HTML.jpg

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