甜橙硫氧还蛋白基因<i>CsTRXh</i>1克隆与表达分析

王淘; 杨澄; 闫亚娜; 王雨潇; 李瑞民

doi:10.19303/j.issn.1008-0384.2022.007.008

甜橙硫氧还蛋白基因CsTRXh1克隆与表达分析

Cloning and Expression of a Thioredoxin Gene CsTRXh1 from Citrus sinensis

摘要

摘要:
目的通过甜橙硫氧还蛋白基因CsTRXh1克隆与表达分析，以期为解析甜橙硫氧还蛋白基因的功能及胁迫应答机制提供参考依据。
方法基于甜橙基因组数据使用同源克隆法克隆甜橙硫氧还蛋白基因，利用生物信息学分析CsTRXh1蛋白与其他物种同源蛋白的相似性、系统进化和理化性质，使用qRT-PCR方法分析CsTRXh1基因在健康甜橙树和感染黄龙病甜橙树叶片中的表达模式，通过瞬时转化烟草叶片分析CsTRXh1蛋白在植物细胞中的亚细胞定位。
结果获得1个甜橙硫氧还蛋白基因CsTRXh1，GenBank登录号为ON125405。蛋白质序列比对分析表明，CsTRXh1与其他植物来源的硫氧还蛋白具有较高的序列相似度，包含保守结构域Thioredoxin。聚类分析表明，CsTRXh1为h型硫氧还蛋白。表达分析结果表明，CsTRXh1基因在感染黄龙病甜橙叶片中上调表达。亚细胞定位分析表明，CsTRXh1定位于细胞质和细胞膜。
结论 CsTRXh1基因受黄龙病菌侵染诱导表达，推测CsTRXh1在黄龙病菌侵染柑橘过程中参与生理生化调控功能。

Abstract:
Objective A thioredoxin gene, CsTRXh1, was cloned from Citrus sinensis to determine the expression and subcellular localization for studying its function and role played in stress response.
Method CsTRXh1 from the homology sequence in the genome of C. sinensis was cloned. Homology, phylogenetic relationship, and physicochemical properties of the gene were analyzed, and expressions in healthy and Huanglongbing-infected trees detected. The transient expression of the gene in tobacco leaves was used to find its subcellular localization.
Result The successfully cloned CsTRXh1 was assigned with the GenBank accession ON125405. A multi-protein sequences alignment showed that it contained a conserved thioredoxin domain with a high similarity with other plant thioredoxin proteins. The phylogenetic analysis indicated it to be an h-type thioredoxin protein. CsTRXh1 was upregulated in the leaves of Huanglongbing-infected trees and localized in the cytoplasm and cytomembrane.
Conclusion The expression induced in the Huanglongbing-infected trees revealed a possible association of CsTRXh1 with the physiological and biochemical regulations of insect-disseminated Candidatus Liberibacter asiaticus that causes the destructive, incurable citrus greening disease.

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