›› 2017, Vol. 60 ›› Issue (5): 505-512.doi: 10.16380/j.kcxb.2017.05.002

• 研究论文 • 上一篇    下一篇


唐芬芬1, 2, 郑亚强1, 白兴荣2,*, 陈斌1,*   

  1. (1. 云南农业大学植物保护学院, 云南生物资源保护与利用国家重点实验室, 昆明 650201; 2. 云南省农业科学院蚕桑蜜蜂研究所, 云南蒙自 661101)
  • 出版日期:2017-05-20 发布日期:2017-05-20

Changes in the resistance to nucleopolyhedrovirus and the expression levels of the related antimicrobial peptide and heat shock protein genes in Bombyx mori exposed to short-time heat shock

TANG Fen-Fen1, 2, ZHENG Ya-Qiang1, BAI Xing-Rong2,*, CHEN Bin1,*   

  1. (1. State Key Laboratory for Conservation and Utilization of BioResources in Yunnan, College of Plant Protection, Yunnan Agricultural University, Kunming 650201, China; 2. Institute of Sericulture and Apiculture, Yunnan Academy of Agricultural Sciences, Mengzi, Yunnan 661101, China)
  • Online:2017-05-20 Published:2017-05-20

摘要: 【目的】探讨短时热激对家蚕Bombyx mori抵抗核型多角体病毒能力的影响,为深入研究家蚕热激响应的抗病毒免疫机制提供参考。【方法】家蚕5龄幼虫42℃热激15 min后,经口喂食家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus, BmNPV)(5.4×106 PIBs/头)后正常饲养,观察热激对家蚕感染病毒的影响;同时,利用实时荧光定量RT-PCR技术检测热激后7类抗菌肽主基因(BmcecD, Bmmor, Bmglve2, BmdefeB, Bmatta1, Bmenbo2和Bmlebo3)以及6种热激蛋白基因(Bmhsp90, Bmhsp70, Bmhsp40, Bmhsp19.9, Bmhsp21.4和Bmhsp25.4)在家蚕5龄幼虫中肠组织中的表达变化;利用MEGA 5.0软件对家蚕所有热激蛋白进行系统进化分析。【结果】与未进行热激处理相比,短时42℃热激能够明显提高感染BmNPV的家蚕5龄幼虫的存活率。实时荧光定量RT-PCR结果表明,与未热激处理相比,热激处理后感染BmNPV的家蚕5龄幼虫中肠组织中Bmglve2和Bmhsp25.4的表达量显著上升。另外,系统进化分析显示,小分子热激蛋白BmHSP25.4在进化上特殊,单独与大分子热激蛋白(BmHSP90, BmHSP70和 BmHSP40)聚为一类。【结论】短时热激能够提高家蚕对BmNPV的抗性,热激蛋白基因Bmhsp25.4和抗菌肽基因Bmglve2可能在家蚕热激响应的抗病毒免疫中发挥关联功能。

关键词: 家蚕, 热激, BmNPV, 抗菌肽, 热激蛋白, mRNA表达水平

Abstract: 【Aim】 This study aims to investigate the effect of short-time heat shock on the resistance of the silkworm, Bombyx mori to nucleopolyhedrovirus, so as to provide references for further study on the antiviral immune mechanism of the silkworm in response to heat shock. 【Methods】 The 5th instar larvae of B. mori were exposed to 42℃ for 15 min, and then fed with B. mori nucleopolyhedrovirus (BmNPV) (5.4×106 PIBs/individual). All the tested larvae were reared as usual. The transcription levels of seven major antimicrobial genes (BmcecD, Bmmor, Bmglve2, BmdefeB, Bmatta1, Bmenbo2 and Bmlebo3) and six heat shock protein genes (Bmhsp90, Bmhsp70, Bmhsp40, Bmhsp19.9, Bmhsp21.4 and Bmhsp25.4) in the midgut of the 5th instar larvae after heat-shock treatment were analyzed by quantitative real-time PCR. The amino acid sequences of all BmHSPs were also phylogenetically analyzed using MEGA 5.0. 【Results】 Short-time heat shock significantly increased the survival rate of the BmNPV-infected 5th instar larvae of B. mori, as compared with the nonheat shock group. Quantitative real-time PCR results showed that the expression levels of Bmglve2 and Bmhsp25.4 were significantly up-regulated in the midgut of the BmNPV-infected 5th instar larvae of B. mori exposed to heat shock in comparison with the non-heat shock groups. The phylogenetic analysis showed that BmHSP25.4 was special in evolution, and individually classified into the same cluster with large HSPs (BmHSP90, BmHSP70 and BmHSP40). 【Conclusion】 Short-time heat shock before BmNPV feeding can enhance the resistance of B. mori to BmNPV. Bmhsp25.4 and Bmglve2 may relatively function in the immunity induction of B. mori against BmNPV through heat shock response.

Key words:  Bombyx mori, heat shock, BmNPV, antimicrobial peptide, heat shock protein, mRNA expression level