Effects of BmNPV Infection on β‑N‑acetylglucosidase Activity and Transcriptional Level of Related Genes in Bombyx mori

doi:10.15933/j.cnki.1004-3268.2024.05.016

Abstract

Abstract: In order to further analyze the action mechanism of β‑N‑acetylgucosaminidase(GlcNAcases)during Bombyx mori nuclear polyhedrovirus(BmNPV)infection,the expression of GlcNAcases genes was detected by fluorescence quantitative PCR and the activity of GlcNAcases was determined using the enzyme activity assay kit in heamolymph,fat body and midgut of silkworm larvae after oral infection with BmNPV at 3,6,9,12 and 24 h.The results indicated that the GlcNAcases activity in hemolymph,fat body and midgut showed a trend of gradual increase and then sharp decrease,while the transcription levels of the four GlcNAcases coding genes were different in these tissues. In hemolymph,BmGlcNAcase1 and BmGlcNAcase2 were significantly up‑regulated at 3 h to 12 h after infection with BmNPV,while the expression levels of BmGlcNAcase3 and BmGlcNAcase4 showed no significant change compared with the control group(no infection).At 24 h,the expression of four GlcNAcases genes began to down‑regulate and the expression levels were significantly lower than those in the control group.After infection with BmNPV,the expression of BmGlcNAcase2 and BmGlcNAcase3 in fat body was first up‑regulated and then down‑regulated. BmGlcNAcase2 began to up‑regulate at 3 h and BmGlcNAcase3 began at 9 h.In addition,the expression levels of BmGlcNAcase1 and BmGlcNAcase4 did not change significantly.In midgut,BmGlcNAcase2 and BmGlcNAcase4 were gradually up‑regulated firstly,their expression levels were gradually increased and showed a peak,then sharply decreased and the expression was significantly inhibited. However,BmGlcNAcase1 and BmGlcNAcase3 were down‑regulated continuously and their expression levels gradually decreased after infection with BmNPV. In conclusion,β‑N‑acetylglucosidases encoded by BmGlcNAcase2 may play a certain immune defense function in the process of silkworm resisting BmNPV infection.

Key words: Bombyx mori nuclear polyhedrovirus, β?N?acetylglucosidases, Bombyx mori, Enzymatic activity, Gene expression

摘要： 为了深入解析β-N-乙酰葡萄糖苷酶（GlcNAcases）在病毒感染过程中的作用机制，将家蚕核型多角体病毒（BmNPV）经口感染5龄起家蚕，在感染后3、6、9、12、24 h提取家蚕血淋巴、脂肪体和中肠，采用荧光定量PCR方法检测GlcNAcases基因的表达水平，同时利用GlcNAcases测试盒测定GlcNAcases活性变化情况。结果显示，经口感染BmNPV后，家蚕血淋巴、脂肪体和中肠组织中GlcNAcases活性整体呈现先逐渐升高后急剧降低的趋势；而家蚕机体4个GlcNAcases编码基因的转录水平在不同组织中存在一定差异。在感染BmNPV后3~12 h，血淋巴中BmGlcNAcase1 和BmGlcNAcase2 的表达量均明显上调，BmGlcNAcase3 和BmGlcNAcase4 表达量与对照组（未感染）无差异；在感染后24 h，血淋巴中4个GlcNAcases 基因表达量均开始下调，且表达量明显低于对照组。脂肪体组织中BmGlcNAcase2 和BmGlcNAcase3 在感染BmNPV后其表达量呈现先上调后下调的趋势，其中BmGlcNAcase2 的表达量在感染BmNPV后3 h就开始上调，BmGlcNAcase3 的表达量在9 h才开始上调，BmGlcNAcase1 和BmGlcNAcase4的表达量无明显变化。感染BmNPV后，中肠BmGlcNAcase2 和BmGlcNAcase4 的表达量整体先逐渐上调，增高至一个峰值，随后急剧减弱，表达受到明显抑制；而BmGlcNAcase1 和BmGlcNAcase3 在感染BmNPV后其表达量呈现下调趋势，表达量逐渐降低。综上，BmGlcNAcase2编码的GlcNAcases在家蚕抵御BmNPV的侵染过程中发挥了一定的免疫防御功能。

关键词: 家蚕核型多角体病毒, β-N-乙酰葡萄糖苷酶, 家蚕, 酶活性, 基因表达

CLC Number:

S881.2

YANG Weike, ZHANG Lusi, FAN Yonghui, LIU Zenghu, TANG Fenfen. Effects of BmNPV Infection on β‑N‑acetylglucosidase Activity and Transcriptional Level of Related Genes in Bombyx mori[J]. Journal of Henan Agricultural Sciences, 2024, 53(5): 150-156.

杨伟克, 张露斯, 范永慧, 刘增虎, 唐芬芬. BmNPV 侵染对家蚕β-N-乙酰葡萄糖苷酶活性及其相关基因转录水平的影响[J]. 河南农业科学, 2024, 53(5): 150-156.

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