花生学报2024,Vol.53Issue(4) :28-36,47.DOI:10.14001/j.issn.1002-4093.2024.04.003

花生AhHSP90-B8基因克隆、表达分析及互作蛋白筛选

Cloning,Expression Analysis,and Interaction Protein Identification of AhHSP90-B8 Gene from Peanut

苑翠玲 曹凤岩 李春娟 牟艺菲 王奇 王倩倩 单世华 孙全喜
花生学报2024,Vol.53Issue(4) :28-36,47.DOI:10.14001/j.issn.1002-4093.2024.04.003

花生AhHSP90-B8基因克隆、表达分析及互作蛋白筛选

Cloning,Expression Analysis,and Interaction Protein Identification of AhHSP90-B8 Gene from Peanut

苑翠玲 1曹凤岩 1李春娟 1牟艺菲 1王奇 1王倩倩 1单世华 1孙全喜1
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作者信息

  • 1. 山东省花生研究所,山东青岛 266100
  • 折叠

摘要

HSP90基因在植物生长发育和抗病反应中发挥重要作用.本研究克隆到一个热激蛋白90基因(AhHSP90-B8),其CDS全长为2 100 bp,共编码699个氨基酸,蛋白质分子量为80.2 kDa,等电点pI为4.42,不存在跨膜结构,亚细胞定位预测该蛋白定位于内质网上,是亲水蛋白.启动子区域存在19个已知的与植物生长发育及抗逆等相关的顺式作用元件.qRT-RCR分析发现,AhHSP90-B8基因在花生不同组织中均有表达,其中在根及果针中的表达量相对较高.接种晚斑病病原菌后不同时期,其表达量存在显著差异.接菌后3 d基因表达量相对变化较小,其他时期(5、10、15、20 d)基因表达量均呈现较大幅度的上调,在处理15 d后表达量达到峰值.进一步筛选到SGT1等相互作用蛋白,并通过酵母双杂交验证其真实互作.这暗示AhHSP90-B8可能通过与SGT1形成蛋白复合体来调控对晚斑病的抗性.本研究为进一步解析AhHSP90-B8基因功能奠定了理论基础,同时为培育优异花生新品种提供了潜在的抗性基因资源.

Abstract

The Heat Shock Protein 90(HSP90)genes play important roles in plant growth,de-velopment,and disease resistance response.In this study,a HSP90 gene was cloned,its CDS is 2,100 base pairs(bp)in length,encoding 699 amino acids,with a protein molecular weight(MW)of 80.2 kDa and an isoelectric point(pI)of 4.42,and there was no transmembrane structure.Subcellular lo-calization prediction results showed that the protein was hydrophilic and mainly located in the endo-plasmic reticulum.Nineteen known cis-acting elements related to plant growth,development,and stress resistance were in the promoter region.qRT-PCR analysis revealed that the AhHSP90-B8 gene was expressed in different tissues of peanut,with a relatively higher expression level in roots and pegs.There were significant differences in the expression levels at different stages after late spot dis-ease pathogens infection.Except for the relatively minor change in gene expression at the third day(3 days)after infection,gene expression levels showed significant up-regulation at other stages(5,10,15,and 20 days),reaching a peak after 15 days after infection.Furthermore,SGT1 was identified as an in-teracting protein,and was further verified their true interaction relationship through yeast two-hybrid.These results suggested that AhHSP90-B8 may act in conjunction with SGT1 by forming a protein complex.This study lays a theoretical foundation for further elucidating the function of the AhHSP90-B8 gene and provided potential resistance gene resources for breeding excellent new peanut varieties.

关键词

花生/AhHSP90-B8/晚斑病/分子特征/表达分析/互作蛋白

Key words

peanut/AhHSP90-B8/late leaf spot disease/molecular characteristics/expression analysis/interaction protein

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出版年

2024
花生学报
山东省花生研究所

花生学报

CSTPCD北大核心
影响因子:0.577
ISSN:1002-4093
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