苹果砧木‘SH6’中RGLs基因的克隆及生物信息学分析

Cloning and bioinformatic analysis of RGLs in apple rootstock 'SH6'

【目的】从苹果砧木‘SH6’中分离出三个DELLA家族基因RGL1b,RGL2a,RGL2b,为探究苹果DELLA蛋白家族对植株生长发育的调控机理,对这三个基因进行了生物信息学分析。【方法】通过克隆技术从苹果砧木‘SH6’中分离得到RGL1b,RGL2a,RGL2b的编码序列,利用生物学工具分析其序列特征,采用实时荧光定量PCR方法分析其基因表达水平。【结果】序列分析表明,苹果砧木‘SH6’的3种RGLs序列具有较高相似性,均存在DELLA蛋白和GRAS家族的结构特征。但序列也存在部分差异,预测其编码的蛋白都具有亲水性能,RGL1b,RGL2a蛋白大部分定位在细胞质,而RGL2b主要分布在细胞核。用实时荧光定量PCR方法,检测了‘SH6’及其亲本‘国光’中这3种RGLs基因的相对表达水平,结果显示,RGL1b,RGL2a,RGL2b在‘SH6’中的表达量均明显高于‘国光’中的表达。【结论】苹果砧木‘SH6’中RGL1b,RGL2a,RGL2b基因可能在果树矮化中发挥重要作用。

【Objective】To explore the roles of the apple rootstock DELLA protein family on plant growth and development,we had identified three DELLA genes(RGL1b,RGL2a,RGL2b)from apple rootstock‘SH6’,and performed the bioinformatics analysis of these three genes.【Methods】The coding sequences of RGL1b,RGL2a,and RGL2b were obtained by gene cloning,and their sequence characteristics and expression levels were analyzed by a series of bioinformatic tools and real\|time PCR.【Results】The results showed that the sequences of these three RGLs proteins had a high similarity,especially at DELLA and GRAS Super\|family typical domains.All of the RGLs proteins are hydrophilic although there are some differences among their sequences.The RGL1b and RGL2a were predicted to be mainly located in the cytoplasm,while RGL2b mainly distributed in the nuclear.The relative expression levels of these genes were investigated by real\|time PCR.The results showed that the expression levels of all the RGL1b,RGL2a and RGL2b genes were higher in leaves of‘SH6’than in‘Ralls Janet’.【Conclusion】The RGL1b,RGL2a,and RGL2b in apple rootstock‘SH6’might play important roles in the dwarfing growth regulation of fruit trees.