过量表达棉花GDSL脂肪酶提高甘蓝型油菜油脂含量的研究  

Overexpression of a Cotton GDSL Lipase Increases the Oil Content of Brassica napus L.

作　　者：郝晓云[1] 蔡永智[1] 袁哈利 李榕[1] 王斐[1] 李鸿彬[1,2] 

机构地区：[1]石河子大学生命科学学院,石河子832003 [2]石河子大学农业生物技术重点实验室,石河子832003

出　　处：《中国粮油学报》2014年第6期63-68,73,共7页Journal of the Chinese Cereals and Oils Association

基　　金：国家自然科学基金(31260339);兵团种质资源创新专项(2012BB050);石河子大学杰出青年项目(2012ZRKXJQ03)

摘　　要：GDSL脂肪酶作为一种脂质水解酶具有广泛的底物特异性，在植物的油脂代谢、生长发育等众多生理过程中发挥着重要作用。利用RT—PCR技术从发育的陆地棉种子中克隆得到棉花的GDSL脂肪酶基因的cDNA，该基因开放读码框为1068bp，编码含有356个氨基酸的蛋白质。对棉花种子发育不同时期进行半定量PCR分析的结果表明，在种子发育15—20d时，GhGLIP基因的表达量较高。将该基因构建到真核表达载体pCAMBIA2300中，转入农杆菌GV3101后转化甘蓝型油菜野油19，通过筛选和鉴定获得转基因阳性油菜株系。转基因油菜的GDSL脂肪酶活力获得了显著提高，转基因油菜种子的油脂含量比野生型油菜提高了8．68％，提高比率为24．3％。GDSL lipase （GLIP）, a lipid hydrolase with broad substrate specificity, exists in plant tissues, playing important roles in plants＇lipid metabolism, growth and development and many other physiological processes. In the study, a cotton GDSL lipase named GhGLIP has been isolated from cotton ovules by RT -PCR method. The full open reading frame was 1 068 bp, containing a protein of 356 amino acids. Semi - quantitative PCR analysis of dif- ferent cotton ovule development periods showed that GhGLIP gene expressed higher values in 15 -20 d ovules. The plant over expression vector pCAMBIA2300 - GhGLIP was obtained then transformed into Brassica napus L. （wild -oil 19） by agrobacterium mediated method. Transgenic oilseed rape plants were obtained after screening and identification with a significant promotion of GDSL lipase enzyme activity ; the oil content of transgenic Brassiea napus seeds was significantly enhanced by 8.68% higher and a increase ratio of 24.3% than wide -type.

关 键 词：棉花种子 GDSL脂肪酶 油脂含量 甘蓝型油菜 

分 类 号：TS213.2[轻工技术与工程—粮食、油脂及植物蛋白工程]