利用CRISPR/Cas9对大豆种子贮藏蛋白基因进行诱变。
Mutagenesis of seed storage protein genes in Soybean using CRISPR/Cas9.
作者信息
Li Chenlong, Nguyen Vi, Liu Jun, Fu Wenqun, Chen Chen, Yu Kangfu, Cui Yuhai
机构信息
London Research and Development Center, Agriculture and Agri-Food Canada, London, ON, Canada.
Department of Biology, Western University, London, ON, Canada.
出版信息
BMC Res Notes. 2019 Mar 27;12(1):176. doi: 10.1186/s13104-019-4207-2.
OBJECTIVE
Soybean seeds are an important source of vegetable proteins for both food and industry worldwide. Conglycinins (7S) and glycinins (11S), which are two major families of storage proteins encoded by a small family of genes, account for about 70% of total soy seed protein. Mutant alleles of these genes are often necessary in certain breeding programs, as the relative abundance of these protein subunits affect amino acid composition and soy food properties. In this study, we set out to test the efficiency of the CRISPR/Cas9 system in editing soybean storage protein genes using Agrobacterium rhizogenes-mediated hairy root transformation system.
RESULTS
We designed and tested sgRNAs to target nine different major storage protein genes and detected DNA mutations in three storage protein genes in soybean hairy roots, at a ratio ranging from 3.8 to 43.7%. Our work provides a useful resource for future soybean breeders to engineer/develop varieties with mutations in seed storage proteins.
目的
大豆种子是全球食品和工业领域重要的植物蛋白来源。伴大豆球蛋白（7S）和大豆球蛋白（11S）是由一小家族基因编码的两个主要贮藏蛋白家族，约占大豆种子总蛋白的70%。在某些育种计划中，这些基因的突变等位基因通常是必需的，因为这些蛋白质亚基的相对丰度会影响氨基酸组成和大豆食品特性。在本研究中，我们着手利用发根农杆菌介导的毛状根转化系统测试CRISPR/Cas9系统编辑大豆贮藏蛋白基因的效率。
结果
我们设计并测试了靶向九个不同主要贮藏蛋白基因的sgRNA，并在大豆毛状根中的三个贮藏蛋白基因中检测到DNA突变，比例在3.8%至43.7%之间。我们的工作为未来大豆育种者培育/开发种子贮藏蛋白发生突变的品种提供了有用的资源。
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