甘蓝型油菜BnaLPAT4基因功能研究  

Function of BnaLPAT4 Gene in Brassica napus L.

作　　者：刘勇[1,2] 肖旦望 秦艺 陈静 陈婉瑜 熊兴华 LIU Yong;XIAO Danwang;QIN Yi;CHEN Jing;CHEN Wanyu;XIONG Xinghua(Hunan Branch of National Oil Improvement Center,Hunan Agricultural University,Changsha,Hunan 410128;College of Agronomy,Hunan Agricultural University/Key Laboratory of Rice Rape Disease Resistance Breeding of Hunan Province,Changsha,Hunan 410128)

机构地区：[1]湖南农业大学国家油料改良中心湖南分中心,湖南长沙410128 [2]湖南农业大学农学院/湖南省水稻油菜抗病育种重点实验室,湖南长沙410128

出　　处：《核农学报》2022年第8期1537-1545,共9页Journal of Nuclear Agricultural Sciences

基　　金：转基因重大专项(SQ2018ZD080068);国家重点研发计划项目(2017YFD0101703)。

摘　　要：为了验证油菜溶血磷脂酸酰基转移酶4(BnaLPAT4)在油脂合成中的作用,本试验通过同源克隆法从甘蓝型油菜湘油15号中获得2条全长分别为1143、1140 bp的BnaLPAT4基因,分别命名为BnaLPAT4-1和BnaLPAT4-2,构建pGAL::BnaLPAT4-1和pGAL::BnaLPAT4-2载体转化酵母INVSC,通过气相色谱法检测酵母中的脂肪酸组分,并以植物表达载体pBI121为骨架,构建pNapin::BnaLPAT4-1和pNapin::BnaLPAT4-2载体并通过农杆菌介导法转染拟南芥,获得转基因苗后检测拟南芥转基因种子脂肪酸组分。结果表明,酵母菌转化子与空转化子之间的脂肪酸组分发生显著变化,其中BnaLPAT4-1转化子饱和脂肪酸的含量达66.88%,是空转化子的2.61倍,酵母转化子生成了亚麻酸和花生酸;BnaLPAT4-2转化子中C16脂肪酸的含量较对照提高22.40个百分点。BnaLPAT4在拟南芥种子中特异表达,花生烯酸含量较野生哥伦比亚型提高4.7个百分点,且拟南芥转基因材料种子中的芥酸和花生三烯酸的含量为0。本研究结果为验证BnaLPAT4基因的生物学功能和油脂成分改良提供了理论参考。In order to verify the role of BnaLPAT4 in oil synthesis,two BnaLPAT4 genes with full length of 1143 bp and 1140 bp were obtained from Xiangyou 15 of Brassica napus by homologous cloning,named BnaLPAT4-1 and BnaLPAT4-2 respectively.pGAL::BnaLPAT4-1 and pGAL::BnaLPAT4-2 vectors were constructed to transform yeast INVSC,and the fatty acid components in transformed yeast were detected by gas chromatography.pNapin::BnaLPAT4-1 and pNapin::BnaLPAT4-2 vectors were constructed using plant expression vector pBI121 as the skeleton and transfected into Arabidopsis thaliana by agrobacterium-mediated method.Fatty acid components of Arabidopsis seeds were detected after transgenic seedlings were obtained.The results showed that linolenic acid and arachidonic acid were produced in the yeast transformers compared with the control,and the content of saturated fatty acid in BnaLPAT4-1 transformers was reached to 66.88%,1.61 times higher than that in the control.The content of C16 fatty acids in BnaLPAT4-2 transformants was increased by 22.40%and by nearly 70%compared with the control.BnaLPAT4 was specifically expressed in Arabidopsis seeds,and the content of arachidonic acid in BraLPAT4 transformauts was increased 4.7%and by 26%compared with the wild type,and the content of erucic acid and arachidonic acid in Arabidopsis transgenic seeds was 0.The results provide a theoretical reference for verifying the biological function of BnaLPAT4 gene and lipid composition improvement.

关 键 词：拟南芥 BnaLPAT4 酵母穿梭载体 脂肪酸分析 气相色谱-质谱法 

分 类 号：S565.4[农业科学—作物学]