苹果矮化中间砧“SH6”中GID2基因的克隆及生物信息学分析
2021-06-22王恩莹晁琳珂郝素晓卢艳芬姚允聪
王恩莹 晁琳珂 郝素晓 卢艳芬 姚允聪



摘要 探究GID2基因在调控植物生长发育方面的作用机制。从苹果矮化砧木“SH6”中克隆到GID2-1和GID2-2基因,运用MEGA7、SWISS-MODEL、DNAMAN 7.0等生物信息学工具分析其序列、蛋白质结构;通过qRT-PCR分析GID2-1和GID2-2基因在不同激素处理下的表达情况。序列分析表明,GID2-1基因的CDS序列长度为591 bp,GID2-2为597 bp,分别编码196、198个氨基酸的蛋白质;GID2基因具有F-box结构域。qRT-PCR分析表明,在GA处理下,表达量下调;相反,在PAC处理下其表达量上调。结果表明,GID2基因可能在苹果生长发育方面发挥重要作用。
关键词 SH6;GID2;生长发育;生物信息学;赤霉素
中图分类号 Q943.2 文献标识码 A 文章编号 0517-6611(2021)09-0104-05
doi:10.3969/j.issn.0517-6611.2021.09.026
Abstract In order to explore the mechanism of GID2 gene in regulating plant growth and development. The GID2-1 and GID2-2 genes from the apple dwarf rootstock “SH6” were cloned, and their sequence and protein structure were analyzed using bioinformatics tools such as MEGA7, SWISS-MODEL, DNAMAN7.0. The expression of GID2-1 and GID2-2 genes under different hormone treatments was analyzed by qRT-PCR. Sequence analysis showed that the CDS sequence of GID2-1 gene was 591 bp in length,GID2-2 was 597 bp, respectively encoding a protein of 196, 198 amino acids;GID2 gene had F-box domain. qRT-PCR analysis showed that the expression level was down-regulated under GA treatment;on the contrary, its expression level was up-regulated under PAC treatment. The results indicated that GID2 gene may play an important role in apple growth and development.
Key words SH6;GID2;Growth and development;Bioinformatics;Gibberellin
赤霉素(GA)是植物生長发育过程中不可或缺的一类重要植物激素,影响植物许多发育过程,包括种子萌发、茎伸长、花粉成熟和诱导开花等方面[1]。在20世纪60年代,赤霉素对植物株高的影响较大,在“绿色革命”期间,半矮化品种的发展表明了赤霉素响应信号途径的改变导致粮食产量的巨大增加[2]。随着赤霉素信号途径元件的克隆与表达分析,赤霉素信号通路的大部分成分已经在水稻和拟南芥中鉴定出来,主要包括GA受体GID1、生长抑制蛋白DELLAs和F-box蛋白(拟南芥为SLY1,水稻为GID2)[1],研究表明,DELLA蛋白对植物生长具有负调控作用,GA信号通过克服DELLA蛋白介导生长的抑制作用来促进植株高度[3],并提出了GA-GID1-DELLA复合物来感知GA信号从而调控植物生长的机制[4-6]。
赤霉素如何抑制DELLAs成为研究的一个热点,研究表明通过对功能缺失突变体gid2-1和sly1-10的GA不敏感矮化表型的分析,发现水稻中GID2蛋白……
