家蚕抗核型多角体病毒(BmNPV)相关基因的表达分析及其实用性分子标记的筛选

doi:10.3969/j.issn.1674-7968.2019.12.012

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摘要家蚕核型多角体病毒(Bombyx mori nucleopolyhedrovirus, BmNPV)是一种对家蚕具有高致病性的病毒,会导致蚕业生产上的严重经济损失。迄今已在家蚕中发现了几种对BmNPV有抑制作用的蛋白,但各基因的表达水平与家蚕不同遗传类型材料BmNPV抗性的相关程度仍不清晰。本研究以高抗材料(highly resistant parent, R)和高感材料(highly susceptible parent, S)组配成F₁、F₂及回交等不同遗传类型材料,利用添毒实验与荧光定量PCR技术,检测家蚕不同遗传类型材料在不同感染条件下对BmNPV的抵抗能力及其相应的5个抗性相关基因(家蚕脂肪酶Ⅰ基因(Bmlipase-1), 家蚕丝氨酸蛋白酶Ⅱ基因(serine protease Ⅱ, BmSP-2), 家蚕核糖体蛋白S3A基因(ribosomal protein S3A, BmS3A), 家蚕抑制蛋白酶Ⅱ基因(suppressor of profilin 2, BmSop2), 家蚕烟酰胺腺嘌呤二核苷酸磷酸氧化还原酶基因(nicotinamide adenine dinucleotide phosphate oxidoreductase, BmNOX)的相对表达量,分析其与抗性的关联性。结果表明：Bmlipase-1和BmSP-2在家蚕中肠特异表达,其他基因没有组织表达特异性;Bmlipase-1、BmSP-2、BmNOX和BmSop2四个基因在不同遗传类型材料中肠的相对表达量之间均存在明显差异,且抗性越强的遗传类型材料的相对表达量也越高,各基因的相对表达量与材料抗性呈正相关性。而BmS3A基因的表达差异在各遗传类型材料间没有呈现出一定的相关性;Bmlipase-1在不同遗传类型材料的相对表达量与其抗性水平的一致性最高,该基因的相对表达水平可以在家蚕育种过程中作为抗核型多角体病毒材料选择与鉴定的依据之一。

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关键词 ：家蚕核型多角体病毒(BmNPV), 抗性基因, 荧光定量PCR, 表达水平, 相关性

Abstract：Bombyx mori nucleopolyhedrovirus (BmNPV) is a highly pathogenic virus in the sericultural industry and often results in severe economic losses. Some proteins are found to show antiviral activity against BmNPV in previous studies, but little is known about the correlations between expression levels of these antiviral related genes and resistance abilities of different genetic materials against BmNPV. In this study, to explore the correlations, the highly resistant parent (R) was crossed with the highly susceptible parent (S) to generate different types of genetic materials such as F₁, F₂, and backcrosses. The resistance to BmNPV and the corresponding relative expression levels of 5 antiviral related genes including Bmlipase-1, serine protease Ⅱgene (BmSP-2), ribosomal protein S3A gene (BmS3A), suppressor of profilin 2 gene (BmSop2) and nicotinamide adenine dinucleotide phosphate oxidoreductase gene (BmNOX) in different genetic materials were detected by BmNPV attack test and fluorescent quantitative PCR (qRT-PCR) technology.The results showed that BmSP-2 and Bmlipase-1 specifically expressed in midgut, other genes showed no tissue specificity. The relative expression levels of 4 genes (Bmlipase-1, BmSP-2, BmNOX and BmSop2) were higher in midguts of different types of genetic materials with stronger viral resistance, which had significant differences in different types of genetic materials, being consistent with their resistance abilities. However, there was no correlation among the expression levels of BmS3A. The highest level of consistency between expression levels of these antiviral related genes and resistance abilities of different types of genetic materials against BmNPV had been observed in Bmlipase-1. It was concluded that relative expression levels of Bmlipase-1 could be one of the basis in selecting anti -BmNPV materials in silkworm breeding.

Key words： Bombyx mori nucleopolyhedrovirus (BmNPV) Resistance gene Fluorescent quantitative PCR Expression level Correlation

收稿日期: 2019-05-09

ZTFLH:

S881.2

基金资助:湖南省自科基金(No. 2017JJ2137)和现代农业产业技术体系建设专项(CARS-18)

通讯作者: * jwai718@sina.com

引用本文:

刘勇, 艾均文, 唐芸, 薛宏, 何行, 健郑颖. 家蚕抗核型多角体病毒(BmNPV)相关基因的表达分析及其实用性分子标记的筛选[J]. 农业生物技术学报, 2019, 27(12): 2207-2215.
LIU Yong AI, Jun-Wen, TANG Yun, XUE Hong, HE Xing-Jian, ZHENG Ying. Expression Analysis of Antiviral Genes Related to BmNPV in Silkworm (Bombyx mori) and Screening of Practical Molecular Markers. 农业生物技术学报, 2019, 27(12): 2207-2215.

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http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.12.012 或 http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I12/2207

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