利用基因编辑技术研究BRANCHED1参与油菜分枝过程的调控  

Regulation of Shoot Branching by BRANCHED1 in Brassica napus Based on Gene Editing Technology

作　　者：Olalekan Amoo 胡利民 翟云孤 范楚川[1,2] 周永明 Olalekan Amoo;HU Li-min;ZHAI Yun-gu;FAN Chu-chuan;ZHOU Yong-ming(National Kay Laboratory of Crop Genetic Improvement,Huazhong Agricultural University,Wuhan 430070;Hubei Hongshan Laboratory,Wuhan 430070)

机构地区：[1]华中农业大学作物遗传改良国家重点实验室,武汉430070 [2]湖北洪山实验室,武汉430070

出　　处：《生物技术通报》2022年第4期97-105,共9页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31671279,31371240)。

摘　　要：植物的株型决定于其分枝方式,而改良分枝方式是提高作物产量的有效方式之一。目前关于油菜分枝调控的分子机理研究较少,缺乏用于株型改良的种质资源。本研究以油菜BRANCHED1(BRC1)为目的基因,利用CRISPR/Cas9基因编辑技术靶向突变油菜BRC1基因,通过农杆菌介导的遗传转化获得再生苗,PCR技术筛选出转基因阳性单株,再利用Hi-TOM方法进行靶点突变基因型的测序分析,证实基因编辑产生的定点突变可稳定遗传。最后对获得的突变体进行表型观察发现,BRC1的BnaC01g34090D和BnaA01g26700D的双拷贝纯合突变体表现出明显的多分枝表型,研究结果表明BnaBRC1参与油菜中的分枝和株型调控,为后续研究油菜的株型调控分子机理提供了重要研究材料。Plant architecture depends on shoot branching patterns,thus the improvement of shoot branching pattern is an efficient way of increasing crop yield.However,the molecular mechanism of shoot branching in rapeseed(Brassica napus L.)is not properly understood yet.Also,genetic resources for the improvement of plant architecture in agricultural crops are lacking.In the present study,we conducted the successful application of CRISPR/Cas9 genome editing technology for the targeted mutation of BRANCHED1(BRC1)in rapeseed.The transformed seedlings were acquired using the Agrobacterium-mediated transformation and the transgenic positive plants were selected using PCR.Hi-TOM sequencing analysis was performed to detect the sequences site-targeted mutated genes.The results demonstrated that the induced mutations were heritably transmitted to successive generations.Phenotypic observation of the obtained mutants showed that the double-copy homologous mutants from the BnaC01g34090 and BnaA01g26700D of BRC1 gene presented significant much shoot branching.Collectively,our results revealed that BnaBRC1 is involved in the shoot branching and in the regulation of plant architecture,thus providing the critical study materials to understant the mechanisms involved in the regulation of plant architecture in B.napus.

关 键 词：CRISPR/Cas9 BRC1 分枝 株型 油菜 

分 类 号：S565.4[农业科学—作物学] Q943.2[生物学—植物学]