一个调控抗菌肽表达的小菜蛾肽聚糖识别蛋白（PxPGRP-SA）

doi:10.16380/j.kcxb.2016.05.002

摘要/Abstract

摘要： 【目的】肽聚糖识别蛋白（peptidoglycan recognition proteins, PGRPs）是昆虫免疫系统中一类重要的模式识别蛋白。本研究旨在阐明经苏云金芽孢杆菌Bacillus thuringiensis侵染后，小菜蛾 Plutella xylostella PGRP-SA基因 (命名为PxPGRP-SA)在体内的表达模式和对抗菌肽基因的表达调控。【方法】本研究利用实时荧光定量PCR (qRT-PCR)技术分析B. thuringiensis侵染小菜蛾幼虫后PxPGRP-SA的转录模式，通过RNAi技术结合抗血清封闭实验检测PxPGRP-SA对小菜蛾抗菌肽基因的表达调控作用。【结果】qRT-PCR检测表明，小菜蛾4龄幼虫在注射具有活性的B. thuringiensis 6 h后，PxPGRP-SA在脂肪体和血细胞中表达量迅速上升，其中脂肪体中的表达量在注射24 h后达到高峰，而在血细胞中的表达量在18 h后达到高峰。RNAi沉默小菜蛾4龄幼虫PxPGRP-SA的转录后，可显著降低小菜蛾脂肪体中cecropin, moricin-2, lysozyme和defensin 4个抗菌肽基因及Dorsal和Sp?tzle基因的mRNA转录水平；注射anti-PxPGRP-SA封闭小菜蛾体内PxPGRP-SA的活性后，也可降低小菜蛾脂肪体中4个抗菌肽基因的 mRNA转录水平；PxPGRP-SA转录沉默后，同时导致添食B. thuringiensis的小菜蛾幼虫的存活率明显降低。【结论】 PxPGRP-SA参与了小菜蛾体内抗菌肽cecropin, moricin-2, lysozyme和defensin基因的表达调控，并在免疫防御B. thuringiensis的侵染过程中起了重要的作用。

关键词: 小菜蛾, 肽聚糖识别蛋白, RNAi, 抗血清, 抗菌肽表达

Abstract: 【Aim】 Peptidoglycan recognition proteins (PGRPs) are one kind of important pattern recognition proteins in insect immune system. In this study, we aim to investigate the expression patterns of PGRP-SA from Plutella xylostella (designated as PxPGRP-SA) and its involvement in the regulation of the expression of antimicrobial peptide genes in P. xylostella after infection by Bacillus thuringiensis. 【Methods】 The transcription pattern of PxPGRP-SA in P. xylostella larvae infected by B. thuringiensis was analyzed by quantitative real-time reverse transcription PCR (qRT-PCR), and the regulation of the expression of antimicrobial peptide genes by PxPGRP-SA in P. xylostella was investigated by RNAi technology and antiserum blocking assay. 【Results】 qRT-PCR analysis revealed that the transcripts of PxPGRP-SA increased significantly at 6 h after injection of live B. thuringiensis into the fat body and hemocytes of the 4th instar larvae of P. xylostella, with the highest expression level at 24 h in fat body and 18 h in hemocytes, respectively. The transcript of PxPGRP-SA in the 4th instar larvae of P. xylostella decreased due to RNAi. Blocking down the transcript of PxPGRP-SA by RNAi also reduced the transcriptional levels of four antibacterial peptide genes including cecropin, moricin-2, lysozyme and defensin in the 4th instar larvae and the mRNA levels of Dorsal and Sp?tzle genes significantly. Blocking the activity of PxPGRP-SA by injection of anti-PxPGRP-SA serum into the 4th instar larvae of P. xylostella also decreased the transcription levels of antibacterial peptide genes in the fat body rapidly. Silence of the expression of PxPGRP-SA decreased the survival rate of P. xylostella larvae feeding live B. thuringiensis significantly. 【Conclusion】 PxPGRP-SA is involved in regulating the expression of downstream antibacterial peptide genes including cecropin, moricin-2, lysozyme and defensin in P. xylostella, and plays an important role in the immune defense of P. xylostella against B. thuringiensis infection.

Key words: Plutella xylostella, peptidoglycan recognition protein, RNAi, antiserum, antimicrobial peptides

郑志华, 许小霞, 余静, 高延富, 张玉清, 欧阳莉娜, 金丰良. 一个调控抗菌肽表达的小菜蛾肽聚糖识别蛋白（PxPGRP-SA）[J]. 昆虫学报, 2016, 59(5): 489-499.

ZHENG Zhi-Hua, XU Xiao-Xia, YU Jing, GAO Yan-Fu, ZHANG Yu-Qing, OUYANG Li-Na , JIN Feng-Liang. A peptidoglycan recognition protein (PxPGRP-SA) regulating the expression of antimicrobial peptides in Plutella xylostella (Lepidoptera: Plutellidae)[J]. Acta Entomologica Sinica, 2016, 59(5): 489-499.

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链接本文: http://www.insect.org.cn/CN/10.16380/j.kcxb.2016.05.002

http://www.insect.org.cn/CN/Y2016/V59/I5/489

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一个调控抗菌肽表达的小菜蛾肽聚糖识别蛋白（PxPGRP-SA）

A peptidoglycan recognition protein (PxPGRP-SA) regulating the expression of antimicrobial peptides in Plutella xylostella (Lepidoptera: Plutellidae)

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