续随子ElSAD2基因的克隆与功能分析  

Cloning and Functional Analysis of ElSAD2 Gene in Euphorbia lathyris

作　　者：王鑫雨 葛丽萍[1] 盛晓倩 牛听风 包鹏 李润植[2] WANG Xinyu;GE Liping;SHENG Xiaoqian;NIU Tingfeng;BAO Peng;LI Runzhi(College of Forestry,Shanxi Agricultural University,Taigu,Shanxi 030801,China;Institute of Molecular Agriculture and Bioenergy,Shanxi Agricultural University,Taigu,Shanxi 030801,China)

机构地区：[1]山西农业大学林学院,山西太谷030801 [2]山西农业大学分子农业与生物能源研究所,山西太谷030801

出　　处：《西北植物学报》2022年第4期549-557,共9页Acta Botanica Boreali-Occidentalia Sinica

基　　金：山西省自然科学基金(201901D111223)。

摘　　要：Δ^(9)-硬脂酰-ACP脱氢酶(SAD)是参与植物不饱和脂肪酸生物合成的关键酶。该研究从续随子(Euphorbia lathyris)种子转录组数据库中筛选得到续随子ElSAD2基因序列,对其序列表达特性进行分析,并鉴定ElSAD2基因的功能。结果显示:(1)续随子ElSAD2的cDNA全长1665 bp,ORF为1194 bp,编码397个氨基酸残基;系统进化分析显示ElSAD2蛋白与蓖麻(Ricinus communis)RcSAD1蛋白等亲缘关系较近。(2)ElSAD2在续随子各器官中均有表达,其中在花后30 d种子中表达量最高。(3)在BY4389缺陷型酵母中过表达ElSAD2,使缺陷酵母不饱和脂肪酸含量升高。(4)本氏烟草瞬时表达ElSAD2,使得烟草叶片总油脂和油酸含量分别提高2.46%和2.1%。研究发现,ElSAD2能催化单不饱和油酸的生物合成,可进一步应用于油料植物油脂产量和品质改良。Stearoyl-acyl carrier proteinΔ^(9) desaturase(SAD)is a key enzyme that determines the content of plant saturated fatty acids and unsaturated fatty acids.This study obtained the ElSAD2 gene sequence based on the transcriptome database and analyzed its sequence and expression characteristics,then we identified the function of the ElSAD2 gene.The results showed that:(1)the cDNA length of ElSAD2 was 1665 bp and ORF was 1194 bp,encoding 397 amino acid residues.Phylogenetic analysis showed that ElSAD2 protein was closely related to RcSAD1 protein in Ricinus communis.(2)ElSAD2 was expressed in all tissues,with the highest expression level in seeds of 30 days after anthesis.(3)Genetic transformation showed that overexpression of ElSAD2 could increase unsaturated fatty acid content in defective yeast BY4389.(4)The instantaneous expression of ElSAD2 in tobacco leaves increased the contents of total fatty acid and oleic acid by 2.46%and 2.1%,respectively.It was found that ElSAD2 could efficiently catalyze the biosynthesis of monounsaturated oleic acid,which could be further applied to the improvement of oil yield and quality.

关 键 词：Δ^(9)-硬脂酰-ACP脱氢酶 续随子 能源植物 油酸(18∶1Δ^(9)) 酵母功能互补实验 烟草瞬时表达 

分 类 号：Q785[生物学—分子生物学] Q786