文献类型： 外文期刊

作者： Song, Bo ¹ ; Luo, Tingting ¹ ; Fan, Yuanhang ¹ ; Li, Ming ³ ; Qiu, Zhendong ¹ ; Tian, Yusu ¹ ; Shang, Yuzhuo ¹ ; Ma, Chongxuan ¹ ; Liu, Chang ¹ ; Cao, Qingqian ¹ ; Peng, Yuhan ¹ ; Xu, Pengfei ¹ ; Krishnan, Hari B. ⁴ ; Wang, Zhenhui ⁶ ; Zhang, Shuzhen ¹ ; Liu, Shanshan ¹ ;

作者机构： 1.Northeast Agr Univ, Soybean Res Inst, Key Lab Soybean Biol, Chinese Educ Minist, Harbin 150030, Peoples R China

2.Harbin Normal Univ, Coll Life Sci & Technol, Key Lab Mol & Cytogenet, Harbin 150025, Heilongjiang, Peoples R China

3.Heilongjiang Acad Agr Sci, Keshan Branch, Qiqihar 161000, Peoples R China

4.ARS, Plant Genet Res Unit, USDA, Columbia, MO 65211 USA

5.Univ Missouri, Plant Sci Div, Columbia, MO 65201 USA

6.Jilin Agr Univ, Fac Agron, Changchun 130118, Peoples R China

关键词： soybean; lincRNA; loss of function; beta-conglycinin deficiency; Cas9-free line

期刊名称：JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY （ 2023影响因子：5.7； 五年影响因子：6.0 ）

ISSN： 0021-8561

年卷期： 2024 年 72 卷 26 期

页码：

收录情况： SCI

摘要： Soybean beta-conglycinin is a major allergen that adversely affects the nutritional properties of soybean. Soybean deficient in beta-conglycinin is associated with low allergenicity and high nutritional value. Long intergenic noncoding RNAs (lincRNAs) regulate gene expression and are considered important regulators of essential biological processes. Despite increasing knowledge of the functions of lincRNAs, relatively little is known about the effects of lincRNAs on the accumulation of soybean beta-conglycinin. The current study presents the identification of a lincRNA lincCG1 that was mapped to the intergenic noncoding region of the beta-conglycinin alpha-subunit locus. The full-length lincCG1 sequence was cloned and found to regulate the expression of soybean seed storage protein (SSP) genes via both cis- and trans-acting regulatory mechanisms. Loss-of-function lincCG1 mutations generated using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system led to the deficiency of the allergenic alpha '-, alpha-, and beta-subunits of soybean beta-conglycinin as well as higher content of proteins, sulfur-containing amino acids, and free arginine. The dominant null allele LincCG1, and consequently, the beta-conglycinin-deficient phenotype associated with the lincCG1-gene-edited line was stably inherited by the progenies in a Mendelian fashion. The dominant null allele LincCG1 may therefore be exploited for engineering/developing novel hypoallergenic soybean varieties. Furthermore, Cas9-free and beta-conglycinin-deficient homozygous mutant lines were obtained in the T1 generation. This study is the first to employ the CRISPR/Cas9 technology for editing a lincRNA gene associated with the soybean allergenic protein beta-conglycinin. Moreover, this study reveals that lincCG1 plays a crucial role in regulating the expression of the beta-conglycinin subunit gene cluster, besides highlighting the efficiency of employing the CRISPR/Cas9 system for modulating lincRNAs, and thereby regulating soybean seed components.