利用CRISPR/Cas9技术创制镉低积累香型水稻  

Using CRISPR/Cas9 Technology to Create Low Cadmium Accumulation and Aromatic Rice

作　　者：胡黎明 彭彦[2] 郑文杰 周凯 毛慧 唐丽 毛毕刚[1,2] HU Li-ming;PENG Yan;ZHENG Wen-jie;ZHOU Kai;MAO Hui;TANG Li;MAO Bi-gang(Longping Branch of Graduate School,Hunan University,Changsha,Hunan 410125,China;Hunan Hybrid Rice Research Center/State Key Laboratory of Hybrid Rice,Changsha,Hunan 410125,China)

机构地区：[1]湖南大学研究生院隆平分院,湖南长沙410125 [2]湖南杂交水稻研究中心/杂交水稻国家重点实验室,湖南长沙410125

出　　处：《杂交水稻》2021年第6期77-83,共7页Hybrid Rice

基　　金：国家水稻现代农业产业技术体系(CARS-01-13);湖南省科技重大专项(2018NK1020-2);湖南省自然科学基金(2018JJ3377);长沙市科技计划项目(kq2004100)。

摘　　要：尝试利用CRISPR/Cas9基因编辑技术对镉吸收转运基因OsNramp5和香味基因OsBadh2同时进行编辑,培育出后代性状稳定的优质低镉水稻品种。选用华南地区推广面积较大的常规稻华航31为受体材料,通过CRISPR/Cas9技术创制OsNramp5和OsBadh2双基因敲除植株,在镉污染土壤中种植并测定野生型和双基因敲除植株糙米的镉、锰、铁元素含量以及香味物质2-乙酰-1-吡咯啉(2-AP)的含量,并考察野生型与双基因敲除植株的农艺性状。成功获得了5种类型的华航31双基因敲除植株。与野生型华航31相比,大部分双基因敲除植株的镉和锰含量显著下降,铁含量有一定程度的升高;所有双基因敲除植株中的香味物质2-AP含量均显著提高;大部分双基因敲除植株的株高、每穗实粒数显著下降;全部双基因敲除植株的单株有效穗数、每穗总粒数、结实率均无显著变化。通过CRISPR/Cas9技术可快速创制具有镉低积累和香味的水稻种质,有利于加快优质、安全水稻品种的育种进程。It is tried to use the CRISPR/Cas9 technology to edit the cadmium absorption and transport gene OsNramp5 and the fragrance gene OsBadh2 at the same time to develop genetically stable rice varieties with the characteristics of low cadmium and aroma.In this study,using a widely-used conventional rice variety Huahang 31 in south China as the receptor,a series of double gene knockout plants of OsNramp5 and OsBadh2 were created through the CRISPR/Cas9 technology.The double gene knockout lines were planted in the cadmium-polluted field,and the contents of cadmium,manganese,iron and 2-acetyl-1-pyrroline(2-AP)in brown rice were detected and the agronomic traits were surveyed in the wild type and double gene knockout plants of OsNramp5 and OsBadh2.Five types of double gene knockout plants of OsNramp5 and OsBadh2 were successfully obtained.Compared with the wild type Huahang 31,most of double gene knockout plants had a significant decrease in the contents of cadmium and manganese but a significant increase in the content of iron;all double gene knockout plants had a significant increase in the content of 2-AP;most of double gene knockout plants significantly decreased in plant height and filled grains per panicle;and all double gene knockout plants had not a significant change in effective panicles per plant,total spikelets per panicle and seed-setting rate.The CRISPR/Cas9 technology can be used to create new rice germplasms with the characteristics of low cadmium and aroma,in favor of speeding up the breeding process of safe and high quality rice varieties.

关 键 词：水稻 镉 香味 基因编辑 CRISPR/Cas9技术 育种 

分 类 号：S511.01[农业科学—作物学] Q75[生物学—分子生物学]