CRISPR/Cas9编辑芥菜型多心皮油菜BjROP10基因的研究  

Preliminary study of gene editing of BjROP10 by CRISPR/Cas9 in Brassica juncea L.

作　　者：徐珂 李爽 赵歌 孙振宇 李海源 李珂欣 王效华 徐平 沈金雄 XU Ke;LI Shuang;ZHAO Ge;SUN Zhen-yu;LI Hai-yuan;LI Ke-xin;WANG Xiao-hua;XU Ping;SHEN Jin-xiong(College of Agriculture and Forestry Sciences,Linyi University,Linyi 276000,China;National Key Laboratory of Crop Genetic Improvement,National Engineering Research Center of Rapeseed,Huazhong Agricultural University,Wuhan 430070,China)

机构地区：[1]临沂大学农林科学学院,山东临沂276000 [2]华中农业大学作物遗传改良国家重点实验室/国家油菜工程技术研究中心,湖北武汉430070

出　　处：《中国油料作物学报》2022年第4期770-779,共10页Chinese Journal of Oil Crop Sciences

基　　金：国家自然科学基金青年项目(32001575);国家自然科学基金面上项目(31571698);山东省自然科学基金项目(ZR2019PC055);大学生创新创业训练计划项目(s202110452093)。

摘　　要：油菜多心皮性状可显著增加油菜的每角果粒数,在油菜高产育种中有潜在的应用价值。前期研究发现,芥菜型多心皮油菜J163-4可形成稳定的四心皮性状,且小G蛋白基因BjROP10可能参与调控其多心皮的形成,但其分子机制尚不明确。本研究利用CRISPR/Cas9技术,构建了BjROP10的双靶点载体,并遗传转化芥菜型多心皮油菜J163-4。在T_(0)代,通过筛选鉴定获得了20株阳性苗,其中13株心皮数目发生改变。通过测序分析了7株心皮数发生变化的转基因阳性植株的BjROP10序列突变情况,结果显示其目标基因序列发生变异,导致无法编码形成正常的BjROP10蛋白。本研究对进一步解析BjROP10基因响应CLV信号调控油菜心皮数发育的功能提供了材料基础,也为研究油菜多心皮性状发育的分子机制奠定理论基础。The multi-carpel trait of rapeseed can significantly increase the number of seeds per silique of rapeseed,which has a potential application value for high-yield breeding.Previous studies have found that the multilocualr Brassica juncea,J163-4,can form a stable four-carpel trait,and the small G protein gene BjROP10 migh tbe involved in regulating the formation of the multiloculus,but the molecular mechanism is still unclear.In this study,using CRISPR/Cas9 technology,a dual-target vector of BjROP10 was constructed and genetically transformed into J163-4.In the T_(0) generation,20 transgenetic positive seedlings were obtained,13 of which showed changes of the number of carpels.The mutation sites of the target gene in 7 transgenic positive plants which showed the carpel number variation were analyzed by sequencing technology.The gene sequences of BjROP10 in the 7 transgenic positive plants were mutated,resulting in the inability of encoding a normal BjROP10 protein.This study provides a material basis for further analysis of the function of BjROP10 gene in regulating the development of carpel number of rapeseed in response to CLV signals,and provides experimental basis and experimental materials for studying the molecular mechanism of the development of multilocualr trait in rapeseed too.

关 键 词：芥菜型油菜 BjROP10 CRISPR/Cas9 多心皮 

分 类 号：S565.4[农业科学—作物学]