近期，东北农业大学刘珊珊/张淑珍团队合作在Journal of AGRICULTURAL and FOOD CHEMISTRY杂志在线发表题为‘Generation of New β‑Conglycinin-Deficient Soybean Lines by Editing the lincRNA lincCG1 Using the CRISPR/Cas9 System’的封面论文。
该研究利用双靶点CRISPR/Case9载体系统，定向编辑‘大豆致敏蛋白相关’基因间长链非编码RNA（long intergenic non-coding RNA，lincRNAs），lincCG1。 双靶点敲除lincCG1基因导致大片段缺失，产生lincCG1基因功能丧失型（LincCG1-type）突变体，lincCG1功能丧失可导致大豆7S（β–conglycinin） α′-, α-, 与β-三个主要致敏亚基同时缺失，LincCG1-type突变体主要农艺及品质性状分析结果表明：T4代Cas9-free、LincCG1-type突变品系（CF-LincCG1）的产量性状相对于野生型对照品种‘东农50’无明显差异，但在含硫氨基酸、游离精氨酸及种子蛋白总量上显著高于‘东农50’。
This study provided an excellent new allele (LincCG1) and an excellent new line (CF-LincCG1) for the improved breeding of soybean protein components. It opens up a new chapter in the functional research of soybean long-chain non-coding RNA, and has important reference value for further in-depth study of the function and mechanism of soybean non-coding RNA.
7S (β–conglycinin) and 11S (glycinin) globulins are the main components of soybean seed storage protein, accounting for about 70% of the total soybean seed storage protein. The current study shows that the nutritional and processing quality of 7S globulin is inferior to that of 11S globulin, and soybean allergenic protein mainly exists in 7S globulin, and its three main subunits a¢-(76kD), a- (72kD) and b- (52-54kD) are soybean allergens. 7S globulin subunit-deficient soybean varieties can effectively improve the nutritional quality and processing characteristics of soybean protein while reducing or removing allergenic proteins. However, since 7S globulin is encoded by a polygenic family, the 'a¢-,-and b-three subunits are deleted at the same time' mutant germplasm is rarely reported.
Long non-coding RNAs (linRNAs) are a group of newly discovered RNA molecules in recent years, which are generally larger than 200 nucleotides in length and lack coding potential. They can regulate the expression of genes at the transcriptional and post-transcriptional levels in four ways: signaling molecules, capture molecules, guide molecules, and scaffold molecules. According to the location of lncRNAs in the genome, lncRNAs can be subdivided into lncRNAs in the intergenic region (lincRNAs), lncRNAs in introns (intron lncRNAs), and antisense lncRNAs (antisense lncRNAs). Compared with animal lncRNAs, there are relatively few studies on lncRNAs in plants, and the current results show that plant lncRNAs play an important regulatory role in growth and development and stress response, and the functions of most plant lncRNAs have not yet been resolved, and the research prospects are broad.
In this study, lincCG1, a functional gene of 'intergenic long non-coding RNA', was identified and cloned in soybean for the first time, and the two-way verification of lincCG1 function from gene to phenotype was completed, proving that: (1) lincCG1 is a lincRNA with important regulatory function in affecting the composition of protein subunits in soybean seed storage; (2) dual-target CRISPR/Cas9 technology can realize the loss of lincCG1 gene function, so as to achieve efficient and efficient Precise regulation of soy protein fractions. The above results prove that the design of a dual-target CRISPR/Case9 vector system for soybean lncRNAs gene is an efficient and feasible method to overcome the shortcomings of low expression and low conservation of linRNAs and deeply analyze the function of soybean linRNAs gene.
本文被邀请作为Journal of AGRICULTURAL and FOOD CHEMISTRY期刊的封面文章
Professor Liu Shanshan and Professor Zhang Shuzhen of Northeast Agricultural University are the co-corresponding authors, and Associate Professor Song Bo and doctoral student Luo Tingting are the co-first authors.
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