茄科植物PMI基因进化研究及辣椒CaPMI 基因克隆和表达分析

Evolutionary study of PMI genes in Solanaceae plants and cloning and expression analysis of CaPMI genes in Capsicum annuum

摘要

[目的]本研究旨在鉴定茄科及辣椒磷酸甘露糖异构酶(PMI)基因,探究其进化过程,并系统解析辣椒 CaPMI 基因在非生物胁迫及激素处理下的表达调控网络。[方法]利用生物信息学与分子克隆技术解析 PMI 基因家族特征,分析转录组数据,阐明茄科 PMI 在器官发育与果实成熟中的表达特征;利用 RT-qPCR 技术检测 CaPMI 在多重胁迫下的表达模式。[结果]PMI 基因在进化过程中受到强纯化选择(K_a/K_s<1),形成 3 个独立分支,不同物种发生了特异性保留。在辣椒基因组中鉴定出 3 个 CaPMI 基因,茄科 PMI 基因启动子区中富集生长发育、逆境响应及激素调控相关元件,但数量与组合模式差异明显。茄科 PMI 基因具有组织表达特异性,其中 CaPMI1 高表达于各器官,CaPMI3 特异性富集于花器官。在辣椒 cDNA 文库中克隆获得 CaPMI1(1299bp/432aa)和 CaPMI2(1326bp/441aa)基因,两者所编码的蛋白均为酸性蛋白(pI4.83~4.91)。胁迫响应分析显示,CaPMI1 在低温、高温及赤霉素(GA₃)处理下被显著激活;CaPMI2 和 CaPMI3 则对低温、高温、干旱、GA₃和水杨酸(SA)胁迫快速响应。[结论]本研究揭示了茄科 PMI 基因在进化中的保守性与功能,辣椒 CaPMI 基因家族通过亚功能化形成了 CaPMI1、CaPMI2 和 CaPMI3 协作互补的胁迫响应模块,增强了辣椒对复杂环境压力的适应性,为辣椒抗逆种质创制提供了基因资源与理论支撑。

Abstract

[Objectives] This study aimed to identify the phosphomannose isomerase (PMI) gene in Solanaceae and Capsicum annuum, investigate its evolutionary processes, and systematically analyze the expression regulatory network of the CaPMI gene in C. annuum under abiotic stress and phytohormone treatments. [Methods] The characteristics of PMI gene family were analyzed by bioinformatics and molecular cloning techniques, and the transcriptome data were analyzed to clarify the expression characteristics of Solanaceae PMI in organ development and fruit maturation. The expression pattern of pepper under multiple stresses was detected by RT-qPCR. [Results] PMI gene was strongly purified and selected (K_a/K_s < 1) during the evolutionary process, forming three independent evolutionary branches, and specific preservation occurred in different species. Three CaPMI genes were identified in the pepper genome, and all the promoter regions of Solanaceae PMI genes were enriched with elements related to growth and development, stress response and hormone regulation, but there were significant differences in the number and combination patterns. The CaPMI1 (1299bp/432aa) and CaPMI2 (1326bp/441aa) genes were cloned from a pepper cDNA library, and their encoded proteins were acidic (pI 4.83-4.91). Stress response analysis revealed that CaPMI1 was significantly activated under cold, heat, and gibberellin (GA₃) treatments, whereas CaPMI2 and CaPMI3 responded rapidly to cold, heat, drought, GA₃, and salicylic acid (SA) stresses. [Conclusions] This study revealed the conservation and functional innovation of Solanaceae PMI genes in evolution, and the CaPMI gene family formed a stress-response module of CaPMI1, CaPMI2 and CaPMI3 through neo-functionalization, which enhanced the adaptability of pepper to complex environmental pressures and provided genetic resources and theoretical support for the creation of stress-resistant germplasm in pepper.

关键词

辣椒 / 磷酸甘露糖异构酶(PMI) / 进化关系 / 基因克隆 / 表达分析 / 非生物胁迫响应

Key words

pepper / phosphomannose isomerase(PMI) / phylogenetic evolution / gene cloning / expression analysis / abiotic stress

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基金资助

国家自然科学基金项目(31902021)