大豆脂肪酸脱氢酶GmFAD3C-1基因的克隆及功能分析

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T₂转化植株。脂肪酸脱氢酶酶活测定结果显示，超表达植株T₂籽粒中，酶活与对照相比上升47.62%~78.85%，编辑载体T₂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。

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

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 T₂ transformed plants were obtained using the pollen tube channel method. The fatty acid dehydrogenase enzyme activity assay showed that in the T₂ grain of overexpressing plants, enzyme activity increased 47.62%-78.85% compared to control, and in the editing vector T₂ 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.