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葎草中的抗性基因:来自叶片转录组的计算机预测调查以及与辣椒疫霉菌接种挑战后的表达研究。

Resistance Genes in Piper colubrinum: In Silico Survey From Leaf Transcriptome and Expression Studies Upon Challenge Inoculation with Phytophthora capsici.

机构信息

ICAR-Indian Institute of Spices Research, Marikunnu P O, Kozhikode, 673012, India.

出版信息

Appl Biochem Biotechnol. 2018 Mar;184(3):987-1008. doi: 10.1007/s12010-017-2600-7. Epub 2017 Sep 20.

Abstract

The oomycetes, Phytophthora capsici, cause foot rot disease in black pepper. Piper colubrinum Link, a distant relative of cultivated black pepper, is highly resistant to this destructive pathogen. Identification of resistance (R) genes in P. colubrinum and the study of its expression profile during interaction with the pathogen can help in understanding the resistance mechanism involved. In the present study, 1289 R gene-related transcripts were mined from P. colubrinum transcriptome, clustered, and classified according to the conserved motifs and domains. Transcripts belonging to four major R gene classes were identified in P. colubrinum, but TIR-NBS-LRR-type R genes were absent. The relative expression of 12 selected R genes was studied using two virulent isolates of P. capsici, and these were found to be upregulated in the initial hours of plant pathogen interaction. The R genes studied were expressed even in aseptically maintained tissue-cultured plants and uninoculated greenhouse-grown plants at basal level suggesting that the plants are geared up with the R gene all the time and are under continuous surveillance for the pathogen and basal level of R gene expression do not require a pathogen trigger. ACT, ATUB, and EIF3E were identified as the most stable reference genes that can be used for real-time PCR study. The present study identified promising R genes in P. colubrinum which can be used in developing Phytophthora-resistant black pepper.

摘要

卵菌纲中的辣椒疫霉会引起黑胡椒根腐病,而胡椒的远亲菾菜(Piper colubrinum Link)对这种具有破坏性的病原体具有高度抗性。鉴定菾菜中的抗性(R)基因,并研究其与病原体相互作用过程中的表达谱,有助于理解相关的抗性机制。在本研究中,从菾菜转录组中挖掘了 1289 个与 R 基因相关的转录本,根据保守基序和结构域进行聚类和分类。在菾菜中鉴定出属于四大 R 基因类别的转录本,但不存在 TIR-NBS-LRR 型 R 基因。使用两种致病力强的辣椒疫霉菌分离株研究了 12 个选定的 R 基因的相对表达情况,发现这些基因在植物与病原体相互作用的最初几个小时内被上调表达。研究的 R 基因甚至在无菌维持的组织培养植物和未接种温室种植的植物中也以基础水平表达,这表明植物一直配备有 R 基因,并对病原体进行持续监测,而基础水平的 R 基因表达不需要病原体触发。ACT、ATUB 和 EIF3E 被鉴定为可用于实时 PCR 研究的最稳定的参考基因。本研究鉴定了菾菜中具有潜力的 R 基因,可用于开发抗辣椒疫霉的黑胡椒。

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