大豆脂肪酸脱氢酶GmFAD3C-1基因的克隆及功能分析  

Cloning and genetic transformation of soybean fatty acid dehydrogenase GmFAD3C-1 gene

作　　者：焦苏淇 周俊名 尚雨晴 王佳欣 张爱晶 何浩博 赵秋竹 李越 姚丹[1] JIAO Su-qi;ZHOU Jun-ming;SHANG Yu-qing;WANG Jia-xin;ZHANG Ai-jing;HE Hao-bo;ZHAO Qiu-zhu;LI Yue;YAO Dan(College of Life Sciences,Jilin Agricultural University,Changchun 130118,China)

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118

出　　处：《中国油料作物学报》2022年第5期1006-1017,共12页Chinese Journal of Oil Crop Sciences

基　　金：吉林省自然科学基金(20200201027JC);吉林省科技厅重点研发项目(20210202006NC)。

摘　　要：研究通过对大豆脂肪酸脱氢酶GmFAD3家族中4个关键酶基因序列进行比对,与对照JN18相比,大豆低亚麻酸突变体MT72中GmFAD3C-1基因在起始密码子后+966bp处存在一个碱基位点的缺失(G→•),产生移码突变,导致氨基酸序列上产生较大变化(该突变基因命名为gmfad3c-1)。构建GmFAD3C-1基因超表达及CRISPR/Cas9编辑载体,利用花粉管通道法获得T_(2)转化植株。脂肪酸脱氢酶酶活测定结果显示,超表达植株T_(2)籽粒中,酶活与对照相比上升47.62%~78.85%,编辑载体T_(2)籽粒中,酶活与对照相比下降25.67%~47.11%。脂肪酸相对含量测定的结果进一步表明,GmFAD3C-1基因的表达与植株亚麻酸含量密切相关。By comparing the sequences of four key enzyme genes in the soybean fatty acid dehydrogenase GmFAD3 family,the GmFAD3C-1 gene had a base site deletion(G)at+966bp in the low linolenic acid soybean mutant MT72 compared with the control JN18,producing a frameshift mutation resulting in large changes in the amino acid sequence(the mutant gene is named gmfad3c-1).The GmFAD3C-1 gene overexpression and its CRISPR/Cas9 editing vectors were constructed,and the T_(2) transformed plants were obtained using the pollen tube channel method.The fatty acid dehydrogenase enzyme activity assay showed that in the T_(2) grain of overexpressing plants,enzyme activity increased 47.62%-78.85% compared to control,and in the editing vector T_(2) generation,enzyme activity decreased 25.67%-47.11% compared to control.The results of the fatty acid relative content determination further indicate that the expression of the GmFAD3C-1 gene is closely related with plant linolenic acid content.

关 键 词：大豆 亚麻酸 突变体 脂肪酸脱氢酶 GmFAD3C-1基因 

分 类 号：S565.1[农业科学—作物学]