枸橼C-05小热激蛋白基因HSP23.3克隆与拟南芥遗传转化  

Cloning and Arabidopsis Genetic Transformation of HSP23.3 Gene in Citron C-05

作　　者：张凯[1] 李益 王萍 马先锋[1] 邓子牛[1] Zhang Kai;Li Yi;Wang Ping;Ma Xianfeng;Deng Ziniu(Engineering Research Center for Horticultural Crop Germplasm Creation and New Variety Breeding(Ministry of Education),National Center for Citrus Improvement Changsha,College of Horticulture,Hunan Agricultural University,Changsha,410128)

机构地区：[1]湖南农业大学园艺学院,园艺作物种质创新与新品种选育教育部工程研究中心,国家柑橘改良中心长沙分中心,长沙410128

出　　处：《分子植物育种》2023年第15期4981-4989,共9页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31720103915);湖南省自然科学基金项目(2020JJ4040);湖南创新型省份建设专项(2019RS1052)共同资助。

摘　　要：以枸橼C-05叶片为材料,对枸橼C-05中HSP23.3基因的ORF进行了克隆,并对其生物学信息进行了分析。结果显示:HSP23.3基因在枸橼C-05中的ORF为615 bp,编码204个氨基酸。枸橼C-05的HSP23.3蛋白属于跨膜亲水不稳定蛋白,预测定位于细胞质,二级结构由α-螺旋、β-转角、延伸链和无规则卷曲构成,其中,α-螺旋和无规则卷曲是其最主要的二级结构元件,且三级结构和二级结构预测结果高度相似。不同物种同源序列进化分析结果显示枸橼C-05的HSP23.3与漾濞槭的TXG74 151.1亲缘关系最近。含枸橼C-05种质HSP23.3基因的转基因拟南芥T2代株系接种丁香假单胞菌结果显示转基因拟南芥与野生型相比症状较轻且能抑制丁香假单胞菌的繁殖。本研究初步认为HSP23.3基因可能参与枸橼C-05抗溃疡病反应过程。The leaves of Citron C-05 were used as material.The HSP23.3 gene ORF was cloned and its biological information was analyzed.The results showed that the HSP23.3 gene ORF sequence was 615 bp,its encoding amino acids was 204.Biological information analysis results showed that the HSP23.3 protein is a transmembrane hydrophilic and unstable protein,which was predicted to be located in the cytoplasm.The secondary structure was composed ofα-helix,β-turn,extended strand and random coils.α-helix and random coil is the most important secondary structure element,and the prediction results were highly similar between tertiary structure and secondary structure.The results of evolutionary analysis of homologous sequences of different species showed that HSP23.3 of Citron C-05 was closely related to TXG74151.1 of Acer palmatum.The overexpression vector containing HSP23.3 gene of Citron C-05 germplasm was transferred into wild type Arabidopsis thaliana.The results showed that T2 Arabidopsis thaliana containing the HSP23.3 gene of Citron C-05 had milder symptoms than the Col_0 wild type and its inhibit the reproduction of Pst.DC3000 pathogen infection.This study preliminarily believes that the HSP23.3 may be involved in the anti citrus canker reaction process of Citron C-05.

关 键 词：柑橘 HSP23.3 基因克隆 超表达载体构建 遗传转化 

分 类 号：Q943.2[生物学—植物学] S666[农业科学—果树学]