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对一种抗性花生基因型和一种感病花生基因型在响应由[病原体名称未给出]引起的叶锈病感染时进行的比较RNA测序分析。

Comparative RNA-Seq profiling of a resistant and susceptible peanut () genotypes in response to leaf rust infection caused by .

作者信息

Rathod Visha, Hamid Rasmieh, Tomar Rukam S, Patel Rushika, Padhiyar Shital, Kheni Jasminkumar, Thirumalaisamy P P, Munshi Nasreen S

机构信息

Institute of Science, Nirma University, Ahmedabad, Gujarat India.

Department of Biotechnology and Plant Breeding, Ferdowsi University of Mashhad, Mashhad, Iran.

出版信息

3 Biotech. 2020 Jun;10(6):284. doi: 10.1007/s13205-020-02270-w. Epub 2020 Jun 1.

Abstract

The goal of this study was to identify differentially expressed genes (DEGs) responsible for peanut plant () defence against (causative agent of rust disease). Genes were identified using a high-throughput RNA-sequencing strategy. In total, 86,380,930 reads were generated from RNA-Seq data of two peanut genotypes, JL-24 (susceptible), and GPBD-4 (resistant). Gene Ontology (GO) and KEGG analysis of DEGs revealed essential genes and their pathways responsible for defence response to . DEGs uniquely upregulated in resistant genotype included pathogenesis-related (PR) proteins, MLO such as protein, ethylene-responsive factor, thaumatin, and F-box, whereas, other genes down-regulated in susceptible genotype were Caffeate -methyltransferase, beta-glucosidase, and transcription factors (WRKY, bZIP, MYB). Moreover, various genes, such as Chitinase, Cytochrome P450, Glutathione S-transferase, and R genes such as NBS-LRR were highly up-regulated in the resistant genotype, indicating their involvement in the plant defence mechanism. RNA-Seq analysis data were validated by RT-qPCR using 15 primer sets derived from DEGs producing high correlation value (  = 0.82). A total of 4511 EST-SSRs were identified from the unigenes, which can be useful in evaluating genetic diversity among genotypes, QTL mapping, and plant variety improvement through marker-assisted breeding. These findings will help to understand the molecular defence mechanisms of the peanut plant in response to infection.

摘要

本研究的目的是鉴定与花生植株抵御锈病病原菌(锈病致病因子)相关的差异表达基因(DEG)。采用高通量RNA测序策略鉴定基因。从两个花生基因型JL-24(感病)和GPBD-4(抗病)的RNA-Seq数据中总共产生了86380930条读数。对DEG进行基因本体论(GO)和KEGG分析,揭示了对锈病病原菌防御反应的关键基因及其途径。在抗病基因型中独特上调的DEG包括病程相关(PR)蛋白、MLO类蛋白、乙烯响应因子、thaumatin和F-box,而在感病基因型中下调的其他基因有咖啡酸-O-甲基转移酶、β-葡萄糖苷酶和转录因子(WRKY、bZIP、MYB)。此外,几丁质酶、细胞色素P450、谷胱甘肽S-转移酶等多种基因以及NBS-LRR等R基因在抗病基因型中高度上调,表明它们参与了植物防御机制。利用来自DEG的15个引物组通过RT-qPCR验证了RNA-Seq分析数据,其产生了高相关值(r = 0.82)。从单基因中总共鉴定出4511个EST-SSR,这对于评估基因型间的遗传多样性、QTL定位以及通过标记辅助育种改良植物品种可能有用。这些发现将有助于了解花生植株对锈病病原菌感染的分子防御机制。

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