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棉花学报 ›› 2021, Vol. 33 ›› Issue (5): 412-421.doi: 10.11963/cs20210013

• 研究简报 • 上一篇    下一篇

陆地棉L-D1等位基因特异性分子标记的开发及应用

姜辉1(),郑锦秀2,王永翠1,张超1,王秀丽1,陈莹1,高明伟1,王家宝1,柴启超1,赵军胜1,*()   

  1. 1.山东省农业科学院经济作物研究所,济南 250100
    2.汶上县第一中学,山东 济宁 272500
  • 收稿日期:2021-02-06 出版日期:2021-09-15 发布日期:2022-01-27
  • 通讯作者: 赵军胜 E-mail:jianghui-agro@163.com;zhaojunshengsd@163.com
  • 作者简介:姜辉(1984―),男,博士, jianghui-agro@163.com
  • 基金资助:
    山东省农业科学院青年基金(2016YQN09);山东省良种工程(2020LZGC002);国家现代农业产业技术体系(CARS-15-05)

Development and application of specific molecular markers for L-D1 alleles in upland cotton

Jiang Hui1(),Zheng Jinxiu2,Wang Yongcui1,Zhang Chao1,Wang Xiuli1,Chen Ying1,Gao Mingwei1,Wang Jiabao1,Chai Qichao1,Zhao Junsheng1,*()   

  1. 1. Institute of Industrial Crops, Shandong Academy of Agricultural Sciences, Jinan 250100, China
    2. Wenshang County No. 1 Middle School, Jining, Shandong 272500, China
  • Received:2021-02-06 Online:2021-09-15 Published:2022-01-27
  • Contact: Zhao Junsheng E-mail:jianghui-agro@163.com;zhaojunshengsd@163.com

摘要:

【目的】L-D1基因调控陆地棉叶形。本研究设计特异分子标记精准鉴定L-D1等位基因,为其在陆地棉冠层结构改良中的应用提供支撑。【方法】利用具有鲁棉研28号遗传背景的L-D1等位基因近等基因系,分析不同等位基因组合对叶形的影响。根据4个等位基因的启动子和编码区的多态性设计特异分子标记,并对不同叶形中的等位基因进行检测。【结果】L-D1基因从3叶期开始调控叶片叶裂的形成,4~8叶期叶裂不断加深,从9叶期开始基本稳定;L-D1等位基因不同组合可形成相似叶形,仅从形态上难以准确辨别。克隆获得L-D1位点4个等位基因起始密码子前约4 kb的启动子片段,发现24个SNPs(Single nucleotide polymorphism,单核苷酸多态性)、1个133 bp及1个14 bp的插入缺失。根据等位基因启动子及编码区的SNP和缺失插入开发了1个Indel分子标记InDel_8和2个衍生酶切扩增多态性序列标记dCAPS_192、dCAPS_519,分别为l2L2oL2s的特异分子标记。【结论】根据陆地棉中控制叶形的L-D1等位基因间的差异开发了3个特异性分子标记,可用来鉴定不同的L-D1等位基因。

关键词: 冠层改良; 叶形; L-D1位点; 特异分子标记; 陆地棉

Abstract:

[Objective] L-D1 regulates leaf shape in upland cotton. This study develops specific molecular markers for L-D1 alleles and provides a basis for the application of L-D1 alleles in improving canopy structure. [Method] Morphological traits were investigated to analyze the effect of different combinations of L-D1 alleles with the genetic background of Lumianyan 28 (LMY28) on leaf shape. Specific molecular markers based on polymorphism of alleles' promoters and CDS (Coding sequence) were used to check the combinations of L-D1 alleles in different materials. [Result] L-D1 alleles regulated the formation of leaf lobes from the 3-leaf stage. The lobes deepened from the 4-leaf stage to 8-leaf stage and stabilized from the 9-leaf stage. It was difficult to distinguish the combination of L-D1 alleles from similar leaf shapes. A total of 24 SNPs (Single nucleotide polymorphism) and a 14 bp and a 133 bp insertion-deletion(InDel) were identified in the promoter of 4 alleles. Based on the SNPs and InDels in the promoter and coding sequences, three specific molecular markers, InDel_8, dCAPS_192, and dCAPS_519, were developed for l2, L2o, and L2s, respectively. [Conclusion] Specific molecular markers were developed and could be used to identify L-D1 alleles, which controlled leaf shape of cotton.

Key words: canopy improvement; leaf shape; L-D1 locus; specific molecular marker; upland cotton