黑尾叶蝉NcPGRP基因克隆及表达模式分析

发布时间：2018-08-15 17:52

【摘要】：为明确黑尾叶蝉肽聚糖识别蛋白(Nephotettix cincticeps peptidoglycan recognition protein,Nc PGRP)的功能,利用RT-PCR克隆长度为552 bp的Nc PGRP开放阅读框,其编码蛋白大小为19.9 k D,无信号肽,含3个锌离子结合/酰胺酶催化位点,1个二氨基庚二酸型(Dap型)肽聚糖识别位点。系统发育分析表明,Nc PGRP与其他昆虫亲缘关系较远。利用显微注射技术对黑尾叶蝉进行病原细菌诱导,定量PCR结果显示Nc PGRP在受大肠杆菌及藤黄微球菌刺激后,其转录水平显著上调。组织分布研究表明,Nc PGRP的m RNA在各组织中均有表达,且头部表达量最高。黑尾叶蝉取食感染水稻普通矮缩病毒RDV的水稻后,Nc PGRP转录水平受抑制。本文研究表明,Nc PGRP是一种诱导型表达的PGRP,黑尾叶蝉感染RDV能显著抑制其转录,进而削弱黑尾叶蝉免疫能力,这可能有利于RDV与黑尾叶蝉共生。
[Abstract]:In order to clarify the function of peptidoglycan recognition protein (Nephotettix cincticeps peptidoglycan recognition protein Nc PGRP, an open reading frame of NC PGRP was cloned by RT-PCR, which encoded a protein size of 19.9kD and no signal peptide. There are three zinc-binding / acylaminase catalytic sites and one Dap type peptidoglycan recognition site. Phylogenetic analysis showed that Nc PGRP was closely related to other insects. The microinjection technique was used to induce the pathogenic bacteria of Cicada nigra. The results of quantitative PCR showed that the transcription level of NC PGRP was upregulated significantly after being stimulated by E. coli and Micrococcus luteus. The study of tissue distribution showed that m RNA of Nc PGRP was expressed in all tissues, and the highest expression was found in the head. The transcription level of NC PGRP in rice infected with rice common dwarf virus (RDV) was inhibited. The results showed that Nc PGRP was an inducible expression of PGRP.The infection of RDV could significantly inhibit its transcription and further weaken its immune ability, which might be beneficial to the symbiosis of RDV and Cicadas nigra.
【作者单位】：植物病虫害生物学国家重点实验室/中国农业科学院植物保护研究所;浙江大学昆虫科学研究所/农业部农业昆虫学重点实验室;
【基金】：国家“973计划”(2014CB138404)
【分类号】：S435.112

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