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甜椒中辣椒褪绿病毒诱导的过敏抗性反应的转录组分析

Transcriptome Analysis of Capsicum Chlorosis Virus-Induced Hypersensitive Resistance Response in Bell Capsicum.

作者信息

Widana Gamage Shirani M K, McGrath Desmond J, Persley Denis M, Dietzgen Ralf G

机构信息

Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, Queensland, Australia.

Queensland Department of Agriculture and Fisheries, AgriScience Queensland, Gatton, Queensland, Australia.

出版信息

PLoS One. 2016 Jul 11;11(7):e0159085. doi: 10.1371/journal.pone.0159085. eCollection 2016.

Abstract

BACKGROUND

Capsicum chlorosis virus (CaCV) is an emerging pathogen of capsicum, tomato and peanut crops in Australia and South-East Asia. Commercial capsicum cultivars with CaCV resistance are not yet available, but CaCV resistance identified in Capsicum chinense is being introgressed into commercial Bell capsicum. However, our knowledge of the molecular mechanisms leading to the resistance response to CaCV infection is limited. Therefore, transcriptome and expression profiling data provide an important resource to better understand CaCV resistance mechanisms.

METHODOLOGY/PRINCIPAL FINDINGS: We assembled capsicum transcriptomes and analysed gene expression using Illumina HiSeq platform combined with a tag-based digital gene expression system. Total RNA extracted from CaCV/mock inoculated CaCV resistant (R) and susceptible (S) capsicum at the time point when R line showed a strong hypersensitive response to CaCV infection was used in transcriptome assembly. Gene expression profiles of R and S capsicum in CaCV- and buffer-inoculated conditions were compared. None of the genes were differentially expressed (DE) between R and S cultivars when mock-inoculated, while 2484 genes were DE when inoculated with CaCV. Functional classification revealed that the most highly up-regulated DE genes in R capsicum included pathogenesis-related genes, cell death-associated genes, genes associated with hormone-mediated signalling pathways and genes encoding enzymes involved in synthesis of defense-related secondary metabolites. We selected 15 genes to confirm DE expression levels by real-time quantitative PCR.

CONCLUSION/SIGNIFICANCE: DE transcript profiling data provided comprehensive gene expression information to gain an understanding of the underlying CaCV resistance mechanisms. Further, we identified candidate CaCV resistance genes in the CaCV-resistant C. annuum x C. chinense breeding line. This knowledge will be useful in future for fine mapping of the CaCV resistance locus and potential genetic engineering of resistance into CaCV-susceptible crops.

摘要

背景

辣椒褪绿病毒(CaCV)是澳大利亚和东南亚辣椒、番茄及花生作物中一种新出现的病原体。目前尚无具有CaCV抗性的商业辣椒品种,但在辣椒中鉴定出的CaCV抗性正被导入商业甜椒中。然而,我们对导致对CaCV感染产生抗性反应的分子机制的了解有限。因此,转录组和表达谱数据为更好地理解CaCV抗性机制提供了重要资源。

方法/主要发现:我们使用Illumina HiSeq平台结合基于标签的数字基因表达系统组装了辣椒转录组并分析了基因表达。在R品系对CaCV感染表现出强烈过敏反应的时间点,从接种CaCV/模拟接种的CaCV抗性(R)和易感(S)辣椒中提取的总RNA用于转录组组装。比较了在接种CaCV和缓冲液条件下R和S辣椒的基因表达谱。模拟接种时,R和S品种之间没有差异表达基因,而接种CaCV时,有2484个基因差异表达。功能分类显示,R辣椒中上调最显著的差异表达基因包括病程相关基因、细胞死亡相关基因、与激素介导信号通路相关的基因以及编码参与防御相关次生代谢物合成的酶的基因。我们选择了15个基因通过实时定量PCR来确认差异表达水平。

结论/意义:差异转录谱数据提供了全面的基因表达信息,有助于了解潜在的CaCV抗性机制。此外,我们在CaCV抗性辣椒(C. annuum x C. chinense)育种系中鉴定出了候选CaCV抗性基因。这些知识未来将有助于对CaCV抗性位点进行精细定位,并将抗性潜在地基因工程导入CaCV易感作物中。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5a4d/4939944/325829433548/pone.0159085.g001.jpg

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