楸树DELLA基因家族生信分析及CbuGRAS9的功能分析  

Bioinformatics Analysis of CbuDELLAs Gene Family and Functional Analysis of CbuGRAS9

作　　者：王珊珊[1] 王瑞[1] 樊二勤 付鹏跃 曲冠证[1] 王楠[2] WANG Shanshan;WANG Rui;FAN Erqin;FU Pengyue;QU Guanzheng;WANG Nan(State Key Laboratory of Tree Genetics and Breeding,Northeast Forestry University,Harbin 150040;Chinese Academy of Forestry,Beijing 100091)

机构地区：[1]林木遗传育种全国重点实验室,东北林业大学,哈尔滨150040 [2]中国林业科学研究院,北京100091

出　　处：《植物研究》2024年第1期139-151,共13页Bulletin of Botanical Research

基　　金：国家重点研发计划项目(2021YFD2200202)。

摘　　要：通过鉴定楸树(Catalpa bungei)DELLA家族基因并分析CbuGRAS9的基因功能,为楸树生殖调控性状的遗传改良提供理论依据。基于楸树基因组数据,鉴定并克隆了5个与拟南芥(Arabidopsis thaliana)同源的CbuDELLAs基因;利用ExPASy、SWISS-MODEL、Plant-mPloc、PlantCare等在线工具对CbuDELLAs蛋白进行等电点、蛋白结构、亚细胞定位及启动子顺式作用元件预测;以9-1(Catalpa bungei‘Luoqiu No.1’)和‘百日花’楸树(Catalpa‘Bairihua’)为材料,分析了CbuDELLAs基因的表达量差异,并通过异源转化拟南芥证实了CbuGRAS9的分子功能,利用酵母双杂交文库筛选了与CbuGRAS9互作的蛋白。结果表明:5个CbuDELLAs蛋白的氨基酸数目为455~588,蛋白相对分子质量为5.04~6.43 kDa,等电点为4.81~5.14;CbuDELLAs蛋白均含有DELLA和GRAS保守结构域,全部为亲水性蛋白。亚细胞定位预测结果显示CbuDELLAs蛋白均定位于细胞核中。启动子顺式作用元件分析发现,这5个DELLAs基因启动子区均含有参与赤霉素反应的顺式作用元件。qRT-PCR结果显示‘,百日花’楸中CbuDELLAs基因表达量显著高于对照9-1楸,其中CbuGRAS9为差异最显著的基因,CbuGRAS9转基因株系的开花时间被明显推迟。鉴定到与CbuGRAS9互作的蛋白质主要富集在核糖体、氨基酸合成、次级代谢、光合作用、TCA循环等代谢通路。To provide theoretical basis for genetic improvement of reproductive regulatory traits of Catalpa bungei,DELLAs family genes were identified and the function of CbuGRAS9 was analyzed.Based on the genomic data of Catalpa bungei,five CbuDELLAs genes homologous to Arabidopsis thaliana were identified and cloned.ExPASy,SWISS-MODEL,Plant-mPloc,PlantCare and other online tools were used to predict the isoelectric point,protein structure,sub-cellular localization and promoter cis-acting elements of CbuDELLAs protein.The expression differences of CbuDELLAs were analyzed by using Catalpa‘Bairihua’and Catalpa bungei‘Luoqiu No.1’as materials,and the molecular function of CbuGRAS9 was confirmed by heterologus transformation in Arabidopsis thaliana,and the proteins interacted with CbuGRAS9 were screened by yeast twohybrid library.The results showed that the amino acid number of the five CbuDELLAs proteins ranged from 455 to 588 aa,the relative molecular weight of the proteins ranged from 5.04 to 6.43 kDa,and the isoelectric point value ranged from 4.81 to 5.14.All CbuDELLAs proteins contained DELLA and GRAS conserved domains and are hydrophilic proteins.Sub-cellular localization prediction showed that CbuDELLAs protein was located in the nucleus.The analysis of promoter cis-acting elements showed that the five promoter regions of DELLAs all contained cis-acting elements involved in gibberellin reaction.T he results of qRT-PCR showed that the expression of CbuDELLAs in Catalpa‘Bairihua’were significantly higher than that in Catalpa bungei‘Luoqiu No.1’,and CbuGRAS9 was the most significantly gene,and the flowering time of CbuGRAS9 transgenic plants were significantly delayed.Proteins interacted with CbuGRAS9 were mainly concentrated in metabolic pathways such as ribosome,amino acid synthesis,secondary metabolism,photosynthesis and TCA cycle.

关 键 词：楸树 DELLAs 生物信息学分析 功能分析 互作蛋白 

分 类 号：Q949.777.9[生物学—植物学]