不同耐旱性二倍体马铃薯材料转录组分析及耐旱候选基因的初步挖掘

Transcriptome Analysis of Different Drought-tolerant Diploid Potato Materials and Preliminary Mining of Drought-tolerant Candidate Genes

摘要

受全球气候变化影响，土壤水分亏缺日益加剧，已成为马铃薯生产的主要限制因素之一。为探究缺水条件下，耐旱性不同的二倍体马铃薯材料对干旱胁迫响应的分子机制，挖掘耐旱相关基因，以耐旱材料‘A233’与干旱敏感材料‘A239’为试验材料，利用转录组测序技术对PEG-6000胁迫下耐旱性不同的马铃薯材料进行差异表达基因的分析，基于差异表达基因的GO功能注释和KEGG通路富集分析，结合ABA与Ca²⁺信号传导途径的关键节点基因筛选和基因差异表达水平变化，以及qRT-PCR技术从ABA与Ca²⁺信号传导通路中筛选到8个耐旱候选基因。对筛选获得的8个候选耐旱基因在敏感无性系‘A163’与耐旱无性系‘A90’中进行克隆与序列比对，结果表明，其编码区高度保守，未发生关键功能变异，表明两无性系的耐旱性差异并非由这些基因的编码序列差异直接导致。这些基因可能参与马铃薯干旱胁迫反应。该研究为揭示马铃薯干旱逆境信号转导机制及发掘关键耐旱基因资源提供了重要依据，为今后马铃薯耐旱分子育种相关研究提供一定的理论基础。

Abstract

Under the influence of global climate change, soil water deficit is increasing and has become one of the major limiting factors for potato production. In order to explore the molecular mechanism of drought stress response of diploid potato materials with different drought tolerance under water deficit conditions, and to mine drought tolerance related genes, using drought-tolerant material 'A233' and drought-sensitive material 'A239' as experimental materials, transcriptomic sequencing was utilized to assess differently expressed genes (DEGs) in potato clones exhibiting contrasting drought tolerance under PEG-6000-induced osmotic stress. Based on the GO functional annotation and KEGG pathway enrichment analysis of differentially expressed genes, combined with the screening of key node genes in the ABA and Ca²⁺ signaling pathways and the analysis of gene differential expression level changes, as well as the use of qRT-PCR technology, eight drought-tolerant candidate genes were selected from the ABA and Ca²⁺ signaling pathways. Cloning and sequence alignment of the eight candidate drought-tolerant genes identified in the screening process were conducted in the sensitive clone 'A163' and the drought-tolerant clone 'A90'. The results indicated that their coding regions exhibited high conservation and no critical functional mutations, suggesting that the difference in drought tolerance between the two clones was not directly caused by variations in the coding sequences of these genes. These genes may be involved in the potato's drought stress response. This study provides crucial evidence for elucidating the signal transduction mechanisms in potatoes under drought stress and for identifying key drought-tolerant gene resources. It lays a theoretical foundation for future research on molecular breeding for drought tolerance in potatoes.

关键词

马铃薯 / 二倍体 / 干旱胁迫 / 耐旱候选基因

Key words

potato / diploid / drought stress / drought-tolerant candidate gene

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基金资助

财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-09)