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大豆FUL基因家族进化规律分析及基因编辑靶点鉴定
黎永力, 杜浩, 李泰, 甘卓然, 侯智红, 董利东, 刘宝辉, 程群
0
(广州大学生命科学学院)
摘要:
栽培大豆起源于我国黄淮海地区,我国具有悠久的大豆种植历史和丰富的大豆遗传资源。然而,近年来我国大豆进口依赖度极高,严重威胁到我国的粮食安全,因此培育高产优质的大豆品种至关重要。株高,籽粒大小等是影响大豆产量的重要农艺性状,因此,挖掘调控这些农艺性状的基因,及解析其分子机制具有重要意义。FRUITFULL(FUL)基因属于MADS-box类转录因子,在植物的开花,生长发育,果实成熟等方面起着重要的作用。本研究通过生物信息学分析发现,在大豆中含有6个FUL基因,它们均具有一个保守的MADS-box和一个较为保守的K-box结构域,并主要在豆荚中表达,说明该基因家族可能能够调控大豆种子相关性状。其中,只有GmFUL3b基因在叶片中高度表达,说明该基因在进化过程中功能可能产生了分化。此外,还发现6个FUL之间的进化规律不同。为进一步确定其生物学功能,利用CRISPR/Cas9技术,分别构建6个FUL基因的敲除突变体载体,转化大豆毛根进行靶点验证,并成功鉴定出其中5个基因的有效靶点,为进一步获得稳定的大豆突变体材料及解析GmFULs家族基因的功能提供了重要的理论基础。
关键词:  大豆  FUL基因  生物信息学分析  CRISPR/Cas9技术
DOI:10.13430/j.cnki.jpgr.20201230001
投稿时间:2020-12-30修订日期:2021-02-18
基金项目:国家自然科学基金项目(31901568,32001508)
Molecular Evolution of FUL Family Genes and Identification of Gene Editing Targets in Soybean
LI Yong-li, DU Hao, LI Tai, GAN Zhuo-ran, HOU Zhi-hong, DONG Li-dong, LIU Bao-hui, CHENG Qun
(School of Life Sciences, Guangzhou University)
Abstract:
Cultivated soybean originated from temperate regions of China, and has a long history of planting in the country, with rich genetic resources. In recent years, however, the high degree of dependance on imported soyabean is a serious potential threat to the food security in China. Therefore, it is crucial to breed high-yield and high-quality soybean varieties, for which the exploration of genes that regulate major yield-correlated agronomic traits such as plant height and bean size, and the analysis of their molecular mechanisms are highly significant. FRITFULL (FUL) genes belong to the MADS box transcription factor, which play important roles in flowering, growth, development, and fruit ripening of plants. By bioinformatics analysis we found six FUL genes in soybean, all of which had a conservative MADS-box and a relatively conservative K-Box domain, and were mainly expressed in the pod, indicating that the gene family might be able to regulate seed-related traits of soybean. Among them, only GmFUL3b gene was highly expressed in the leaves, indicating that its function might have been differentiated in the process of evolution. In addition, we found that the six FUL genes followed different evolutionary rules. In order to further study their biological functions, knockout mutant vectors of the six FUL genes were constructed by CRISPR/Cas9 technology. Positive clones were transformed into soybean hairy roots for test target verification, and five of them were successfully identified as effective targets. It provided an important theoretical basis for obtaining stable soybean mutant materials and analyzing the function of GmFUL family genes.
Key words:  soybean  FUL genes  bioinformatics analysis  CRISPR/Cas9 technology

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