禾本科植物富甲硫氨酸蛋白基因的克隆及其进化分析  

Cloning and Phylogenetic Analysis of Methionine-rich Storage Protein Genes in Gramineae Plants

作　　者：陈宏伟[1,2,3] 饶力群[1] 邱业先[1,2,3] 

机构地区：[1]湖南农业大学生物科学技术学院,湖南长沙410128 [2]苏州科技大学化学生物与材料工程学院,江苏苏州215009 [3]江西农业大学理学院,江西南昌330045

出　　处：《核农学报》2016年第12期2326-2335,共10页Journal of Nuclear Agricultural Sciences

基　　金：江西省新世纪学术与技术带头人项目(20030304);苏州市科技局计划项目(SYN201410);江苏高校优势学科建设工程项目

摘　　要：甲硫氨酸(Met)是豆科植物的第一限制性氨基酸。为了获取富Met贮藏蛋白基因,通过比对禾本科富Met贮藏蛋白基因序列来设计引物,以千金子、芦苇、稗、光头稗、东乡野生稻、茶陵野生稻和湖南野生稻等几种常见禾本科植物种子RNA为模板,经RT-PCR扩增获得c DNA片段,并进行序列比对、Motif预测和进化分析。测序结果显示,禾本科植物富Met贮藏蛋白基因序列存在保守程度不同的区域,在同一个属内的植物间,富Met贮藏蛋白基因的序列高度保守。Motif预测显示,富Met贮藏蛋白可能与植物抗病性等功能有关。进化分析表明,不同科的植物种子富Met贮藏蛋白基因存在较大差异,禾本科植物则较为保守,尤其是稻属中富Met贮藏蛋白基因高度保守,玉米中则存在不同类型的富Met贮藏蛋白。本研究丰富了富Met贮藏蛋白基因库,为豆科植物富Met基因工程提供了更多选择,为进一步理解富Met贮藏蛋白在植物发育代谢中的作用机理提供了参考。Methionine is the first limited amino acid in legume. To clone Met-rich storage protein gene,primers based on alignment of Met-rich storage protein genes were designed,and the c DNA of Met-rich storage protein genes were amplified by RT-PCR from several gramineae plants such as Leptochloa chinensis（ Linn.） Nees,Phragmites australis（ Cav.） Trin. ex Steud.,Echinochloa crus-galli（ Linn.） Beauv.,Echinochloa colona（ Linn.） Link,Oryza rufipogon Griff.（ including Dongxiang wild rice,Chaling wild rice and Hunan wild rice）,and then for sequence alignment,Motif prediction and evblutionary analysis. Sequence analysis showed that the sequences of Met-rich storage proteins in gramineae plants have regions with different conservativeness. Met-rich storage protein were highly conserved ameng the same genus. Motif prediction indicated that the function of the Met-rich storage protein may be related with some functions like plant disease resistance. Phylogenetic analysis showed that the Met-rich storage protein genes had great variation in different families,there were more conservative in gramineae,especially in Oryza and while existed as different types in maize. The study will help to enrich the pool of Met-rich storage protein genes,and offer more choice for genetic engineering of the Met-rich protein in legume,and serve as reference for further understanding the mechanism of Met-rich storage protein in plant development and metabolism.

关 键 词：富甲硫氨酸贮藏蛋白质 基因克隆 进化分析 

分 类 号：Q943.2[生物学—植物学]