拟南芥AtFAD6基因突变体的构建  

Construction of Arabidopsis AtFAD6 gene mutant

作　　者：徐雪珍 郑月萍 张夏婷 宋孜元 XU Xue-zhen;ZHENG Yue-ping;ZHANG Xia-ting;SONG Zi-yuan(School of Agricultural and Food Sciences,Zhejiang A&F University,Hangzhou 311300,China)

机构地区：[1]浙江农林大学农业与食品科学学院,浙江杭州311300

出　　处：《江苏农业学报》2021年第5期1125-1130,共6页Jiangsu Journal of Agricultural Sciences

基　　金：科技部转基因重大专项(2016ZX08010003-003);浙江省自然科学基金青年基金项目(Q21C020003)。

摘　　要：运用优化后的CRISPR/Cas9基因编辑载体,创建了2个不同的拟南芥脂肪酸去饱和酶6基因(AtFAD6)突变体,其AtFAD6基因的保守位点氨基酸序列均发生变化,同时终止密码子被提前引入,基因功能丧失。脂肪酸组分分析结果显示,这2种突变体的叶片中单不饱和脂肪酸16∶1和18∶1大量积累,多不饱和脂肪酸16∶3和18∶3含量则大幅下降,同时伴随着叶片发黄、地上部生物量显著降低、抽薹提前2~3 d的表型变化。多不饱和脂肪酸18∶3作为茉莉酸合成的前体物质,其含量的下降致使突变体中茉莉酸信号标记基因AtVSP1在叶片中的表达量有所降低,而茎中的表达量提高了40%以上。所获得的2个AtFAD6功能丧失型突变体为进一步研究脂类代谢与植物生长发育之间的关系提供了重要的遗传材料。The optimized CRISPR/Cas9 gene editing vector was used to create two distinct Arabidopsis AtFAD6 gene mutant.As the nucleotide sequence of the conserved amino acid residues was mutated,and the stop codon was introduced in advance,the fad6 gene function of these two mutants was completely lost.The results of fatty acid composition analysis showed that monounsaturated fatty acids 16∶1 and 18∶1 accumulated significantly while the content of polyunsaturated fatty acids 16∶3 and 18∶3 decreased significantly in the leaves of these two mutants.At the same time,phenotypes of the mutants were appeared,including yellowed leaves,significantly decreased above-ground biomass,and bolting 2-3 days earlier than wild-type.The paly unsaturated fatty acid 18∶3 was used as the precursor substance of jasmonic acid synthesis.The decrease of its content reduced the expression of the jasmonic acid signal marker gene AtVSP1 in the leaves,while the expression in stems increased by more than 40%.The two AtFAD6 loss-of-function mutants obtained in our study provide important genetic material for further research on the relationship between lipid metabolism and plant growth and development.

关 键 词：拟南芥 CRISPR/Cas9 脂肪酸去饱和酶(FAD) 基因突变体 

分 类 号：Q754[生物学—分子生物学]