摘要
通过CRISPR/Cas9技术敲除SRK基因或者SP11基因可以直接将甘蓝类蔬菜自交不亲和材料改造成自交亲和材料。本研究中根据全基因组重测序获得青花菜(Brassica oleracea var.italic) BoSP11基因序列,构建双靶位点的CRISPR/Cas9基因编辑载体,利用农杆菌介导的遗传转化法对BoSP11进行敲除。结果显示,获得的29株转基因植株中有23株发生基因突变,编辑效率为79.3%。TA克隆测序结果表明有4株为纯合突变,19株为杂合突变。统计纯合突变T0代自交结籽情况发现花期自交亲和指数高达6.82,为目前所报道的能获得的亲和性最高指数。T1代植株在隔离大棚中通过蜜蜂授粉平均单株采收种子24.8 g,完全实现不需要人工干预完成亲本材料的自交繁育。
By using CRISPR/Cas9 technology to knock out SRK gene or SP11 gene,the self-incompatible Brassica oleracea vegetables can be directly transformed into self-compatible materials.In this study,a double target CRISPR/Cas9 gene editing vector was constructed based on the sequence of broccoli(Brassica oleracea var.italic)BoSP11 gene obtained by whole genome sequencing,and then used Agrobacterium mediated genetic transformation to knock out the BoSP11 gene.The results showed that 23 of the 29 transgenic plants had gene mutation,and the editing efficiency was 79.3%.The results of TA clone sequencing showed that there were four homozygous mutations and 19 heterozygous mutations.The statistics of the seed setting of T 0 generation of homozygous mutant plants showed that the self-compatibility index at flowering stage was as high as 6.82,which was the highest compatibility index increase reported so far.The T 1 generation plants were pollinated by bees in the isolated greenhouses,and the average seed yield per plant was 24.8 g,which completely conducted the self-pollination of parent materials for seed production without any manual involvement.
作者
马存发
武婷
赵辉
张天培
赵立坚
余永辉
肖建成
李军
巫水钦
MA Cunfa;WU Ting;ZHAO Hui;ZHANG Tianpei;ZHAO Lijian;YU Yonghui;XIAO Jiancheng;LI Jun;WU Shuiqin(Zhejiang MITSUO Seed Co.,Ltd,Jiaxing,Zhejiang 314023,China;School of Modern Agriculture,Jiaxing Vocational&Technical College,Jiaxing,Zhejiang 314036,China)
出处
《园艺学报》
CAS
CSCD
北大核心
2024年第3期509-519,共11页
Acta Horticulturae Sinica
基金
农业农村部国家西兰花良种重大科研联合攻关项目(2018-2023)
浙江省基础公益研究计划项目(LGN21C150004)
浙江省蔬菜新品种选育重大科技专项(2021C02065-4-1)。
