基于比较基因组学的柑橘耐黄龙病关键基因鉴定与功能解析

doi:10.13560/j.cnki.biotech.bull.1985.2025-0389

摘要/Abstract

摘要：

目的耐病基因的发掘是培育耐黄龙病（Huanglongbing, HLB）柑橘材料的重要基础，为解析柑橘与黄龙病菌互作的分子机制，筛选柑橘属（Citrus spp.）及近缘种植物中耐黄龙病的关键基因。方法基于比较基因组学方法，系统鉴定了耐病材料中的关键基因，并利用特异性基因簇鉴定及共有直系同源群的分析，结合gene ontology（GO）富集分析，转录组数据分析以及蛋白质-蛋白质互作网络分析，鉴定耐病材料中的核心差异表达基因。结果黄龙病菌诱导的差异表达的防御相关基因中多个HSP70基因表达模式受黄龙病菌侵染的显著调控，而过表达HSP70显著提高了柑橘对细菌性病害的抗病性。结论耐病材料中防御相关基因HSP70正调控柑橘对病原菌的抗病性。

关键词: 柑橘, 黄龙病, 比较基因组学, 耐病性, 热休克蛋白70

Abstract:

Objective The identification of disease-tolerance genes is essential for breeding Huanglongbing (HLB)-resistant citrus germplasm. In order to elucidating the molecular mechanisms underlying citrus-Candidatus Liberibacter asiaticus (CLas) interactions, key HLB-resistant genes in Citrus spp. and closely-related plants were screened. Method Key genes in dieease-resistant materials were systematically identified using comparative genomics. Core differentially expressed genes (DEGs) were characterized through resistance-specific gene cluster analysis, conserved orthologous group identification, Gene Ontology (GO) enrichment, transcriptome profiling, and protein-protein interaction network analysis. Result Multiple HSP70 genes demonstrated significantly modulated expression patterns upon CLas infection among defense-related DEGs induced by the pathogen. The overexpression of HSP70 substantially enhanced citrus resistance against bacterial pathogens. Conclusion The defense-associated gene HSP70 positively regulates disease resistance to pathogens in citrus.

Key words: citrus, Huanglongbing, comparative genomics, disease tolerance, heat shock protein 70

胡衍铵, 代鑫吕, 钟娇艳, 李瑞民, 黄桂艳. 基于比较基因组学的柑橘耐黄龙病关键基因鉴定与功能解析[J]. 生物技术通报, 2025, 41(11): 282-292.

HU Yan-an, DAI Xin-lyu, ZHONG Jiao-yan, LI Rui-min, HUANG Gui-yan. Comparative Genomics-based Identification and Functional Characterization of Key Genes Resisting Huanglongbing in Citrus[J]. Biotechnology Bulletin, 2025, 41(11): 282-292.

图/表 9

图1 11个物种基因家族聚类分析Proteins代表各物种非冗余蛋白序列，Clusters代表各物种直系同源群，Singletons代表各物种单拷贝基因簇

Fig. 1 Clustering analysis of gene families across 11 speciesProteins indicate the non-redundant protein sequences of each species. Clusters indicate the orthogroups (orthologous gene clusters) across species. Singletons indicate the single-copy gene clusters specific to each species

图2 物种间的共有直系同源群鉴定

Fig. 2 Identification of common orthogroups in different species

图3 耐黄龙病材料共有直系同源群蛋白GO富集分析A：澳大利亚指橙和红河橙；B：宜昌橙和红河橙；C：莽山野橘和宜昌橙

Fig. 3 GO enrichment analysis of proteins in common orthogroups from HLB-resistant speciesA: Citrus australasica and C. hongheensis; B: C. ichangensis and C. hongheensis; C: C. mangshanensis and C. ichangensis

图4 五个耐黄龙病物种共有直系同源群分析A：共有直系同源群分析；B：GO富集分析；C：跨膜结构域分析

Fig. 4 Common orthogroups analysis from five HLB-resistant speciesA: Common orthogroups analysis; B: GO enrichment analysis; C: transmembrane domain analysis

图5 基因家族扩张与收缩分析

Fig. 5 Analysis of gene family expansion and contraction

图6 澳大利亚指橙扩张基因家族GO富集分析

Fig. 6 GO enrichment analysis of expanded gene family in C. australasica

图7 澳大利亚指橙基因组中扩张基因家族相关基因的表达模式A：“防御反应”相关基因； B：“苯丙烷代谢过程”相关基因

Fig. 7 Expression patterns of genes related to the expanded gene family in C. australasicaA: “Defense response” related genes; B: “phenylpropanoid metabolic process” related genes

图8 上调表达HSP70蛋白互作网络构建

Fig. 8 Constructing protein-protein interaction network of up-regulated HSP70 proteins

图9 HSP70的亚细胞定位及抗病性分析A：HSP70的亚细胞定位，标尺=50 μm；B：过表达HSP70对溃疡病的抗病性分析。以t-test进行统计学显著性分析，***表示P<0.001

Fig. 9 Subcellular localization and disease resistance analysis of HSP70A: Subcellular localization of HSP70, scale bar=50 μm. B: Analysis of the disease resistance of overexpressed HSP70 to citrus canker. Used t-test for statistical significance analysis, *** denotes P<0.001

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