棉花C_(3)HC_(4)型RING锌指蛋白基因GhSIZ1克隆及表达分析  

Cloning and Expression Analysis of a C_(3)HC_(4)-type RING Zinc finger Protein Gene GhSIZl in Gossypium hirsutum

作　　者：曾佳聪 王怡[2] 于月华[2] 倪志勇[1] Zeng Jiacong;Wang Yi;Yu Yuehua;Ni Zhiyong(College ofLife Sciences,Xinjiang Agricultural University,Urumqi,830052;College of Agronomy,Xinjiang Agricultural University,Urumqi,830052)

机构地区：[1]新疆农业大学生命科学学院,乌鲁木齐830052 [2]新疆农业大学农学院,乌鲁木齐830052

出　　处：《分子植物育种》2023年第16期5241-5246,共6页Molecular Plant Breeding

基　　金：自治区天山创新团队计划项目(2020D14002);国家自然科学基金项目(U1903203);棉花生物学国家重点实验室开放课题基金项目(CB2020A23);自治区重大科技专项计划项目(2021A02001-3)共同资助。

摘　　要：植物抗逆分子机制非常复杂,克隆并鉴定新的逆境相关基因,有助于解析植物抗逆机理。C_(3)HC_(4)型锌指蛋白在植物生长发育和抵抗非生物胁迫中发挥重要作用。本研究从陆地棉‘Y1169’中克隆了1个C_(3)HC_(4)型锌指蛋白的基因GhSIZ1(XM_041085878.1),并分析GhSIZ1对干旱、盐胁迫和脱落酸的转录响应。序列分析表明该基因cDNA全长3389 bp,含有1个2613 bp的开放阅读框,利用生物信息学分析了棉花GhSIZ1基因的理化性质及其结构功能域,结果显示棉花GhSIZ1基因编码870个氨基酸,为亲水性蛋白,预测分子量约为279.39 k D,理论等电点为4.86,脂肪指数为29.06。GhSIZ1中含有1个C_(3)HC_(4)型RING锌指结构域。系统进化树分析表明Gh SIZ1与达尔文棉(Gossypium darwinii)TYG_87958.1、海岛棉(Gossypium barbadense)KAB_2050370.1和夏威夷棉(Gossypium tomentosum)TYH_93561.1等植物的蛋白质相似性较高,与达尔文棉TYG_87958.1处于同一进化枝上,亲缘关系最近。亚细胞定位结果显示,GhSIZ1:GFP融合蛋白定位于细胞核中。实时荧光定量PCR分析表明在干旱、高盐和脱落酸处理后,GhSIZ1的表达量均先下降再上升,暗示棉花GhSIZ1基因与棉花抗逆有关。以上研究结果为进一步探究棉花抗逆机制提供了一定的参考。The molecular mechanism of plant stress resistance is very complex.Cloning and identifying new stress related genes is helpful to analyze the stress resistance mechanism of plants.This study cloned a C,HC4 zinc finger protein gene GhSIZ1(XM_041085878.1)from Gossypium hirsutum'Y1169'and analyzed the transcriptional response of GhSIZl to drought,salt stress and abscisic acid.Sequence analysis showed that the cDNA of the gene was 3389 bp,contained an open reading frame of 2613 bp.The physicochemical properties and structural functional domains of the GhSIZl gene were analyzed by bioinformatics.The results showed that it encoded 870 amino acids,which was hydrophilic protein,the predicted molecular weight was about 279.39 kD,the theoretical isoelectric point was 4.86,and the fat index was 29.06.GhSIZ1 contains a C,HC4 type RING zinc finger domain.Phylogenetic tree analysis showed that GhSIZ1 had high protein similarity with Gossypium darwini(TYG_87958.1),Gossypium barbadense(KAB_2050370.1),Gos sypium tomentosum(TYH_93561.1)and other plants.It had the highest protein similarity with Gossypium darwini.On the same clade,the genetic relationship is the closest.The subcel lular localization results found that the GhSIZ1:GFP fusion protein was localized in the nucleus.Real-time fluorescent quantitative PCR analysis showed that after drought,high salt and abscisic acid stress,the expression of Gh-SIZl first decreased and then increased,suggesting that the GhSIZl gene is related to cotton stress resistance.The above research results provide a certain reference for further exploring the resistance mechanism of cotton.

关 键 词：陆地棉 基因克隆 锌指蛋白 

分 类 号：S562[农业科学—作物学]