薏苡ramosa2基因的克隆及表达分析  

Cloning and Expression Analysis of ramosa2 Gene in Coix lacryma-jobi L.

作　　者：蒙秋伊 杨玲玲 尚昆 李秀诗 朱加保[2] 王玉娇 付瑜华 Meng Qiuyi;Yang Lingling;Shang Kun;Li Xiushi;Zhu Jiabao;Wang Yujiao;Fu Yuhua(Institute of Subtropical Crops,Guizhou Academy of Agricultural Sciences,Xingyi,562400;Cotton Institute,Anhui Academy of Agricultural Sciences,Hefei,230031)

机构地区：[1]贵州省农业科学院贵州省亚热带作物研究所,兴义562400 [2]安徽省农业科学院棉花研究所,合肥230031

出　　处：《分子植物育种》2023年第19期6292-6299,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(32060471);贵州省农科院国基后补助项目([2021]58号),贵州省农科院院专项([2012]004号)共同资助;贵州省农业科学院青年科技基金项目([2020]06)。

摘　　要：ramosa基因属于LBD基因家族,在植物侧生器官原基的启动和形态形成中发挥重要作用。为了解薏苡ramosa基因的功能及表达特征,本研究采用RT-PCR和RACE技术克隆获得薏苡ramosa2(ClRA2)基因,并对其序列的生理生化特征及表达模式进行分析。ClRA2 cDNA序列全长1421 bp,ORF长813 bp,编码270个氨基酸,含有LOB结构域,位于26~124位置,属于LBD基因家族。ClRA2相对分子质量为49.96 kD,等电点为9.08,属于疏水性的不稳定蛋白。分子进化树分析发现ClRA2与高粱SbRA2亲缘关系最近。实时荧光定量PCR检测结果表明,ClRA2在根中的表达丰度最高,表达量远高于茎和花穗。本研究为解析ClRA2基因功能及其调控薏苡侧生器官发育机制提供了理论基础。ramosa gene belongs to the LBD gene family and plays an important role in the initiation and morphogenesis of plant lateral organs.In order to understand the function and expression characteristics of the Coix ramosa gene,ClRA2 gene was cloned by RT-PCR and RACE,and its physiological and biochemical characteristics and expression patterns were analyzed.The full length of ClRA2 cDNA sequence is 1421 bp,the length of ORF is 813 bp,encoding 270 amino acids,containing LOB domain,the protein domain is located in the position of 26~124 bp,and belonging to the LBD gene family.The molecular weight of ClRA2 was 49.96 kD and the isoelectric point was 9.08,which was a hydrophobic unstable protein.Molecular evolutionary tree analysis showed that ClRA2 was closely related to SbRA2 in conical Sorghum bicolor.The results of real-time quantitative PCR showed that the expression abundance of Cl RA2 was the highest in roots,and the expression level was much higher than that in stems and flower ears.This study provides a theoretical basis for elucidating the function of ClRA2 gene and its mechanism of regulating the development of Coix lacryma lateral organs.

关 键 词：薏苡(Coix lacryma-jobi L.) ramosa2 基因克隆 表达分析 

分 类 号：S519[农业科学—作物学]