分子连锁分析探讨家蚕高抗BmNPV品系的抗性遗传基础

doi:10.3864/j.issn.0578-1752.2017.01.017

Abstract

Abstract: 【Objective】Bombyx mori nucleopolyhedrovirus (BmNPV) can lead the B. mori nucleopolyhedrosis, which caused a huge loss in the sericulture industry. The objective of this study is to map the controlling genes and understand the genetic basis underlying the BmNPV resistance to serve the theory support for resistant variety breeding. 【Method】Backcross population BC₁F (using for linkage analysis) and BC₁M (using for mapping analysis) were derived from crosses between the highly resistant B. mori strain 99R and the sensitive strain Dazao-N. Concentration gradient of virus was used to treat the parents, and the number of infected individuals was recorded. Then by using SPSS17.0, the LD₅₀ for each parent was calculated. Based on appraisal of resistance performance of two parents, the virus adding concentration for BC₁population was determined and then the virus was fed to larva quantitatively at the start of 4th instar one by one. The infected individuals in the BC₁F were chosen as linkage analysis materials. By using the screened polymorphism markers covering all B. mori autosomes, linkage analysis was conducted and then the genotype data were analyzed by T test to get the linkage significance level of each polymorphism marker to show whether it linked with resistance. Thereafter, polymorphism markers will be enriched on the chromosomes which include the resistance linked polymorphism markers to map resistance locus on them. 【Result】Median lethal dose (LD₅₀) of 99R and Dazao-N is 2.92×10⁶and 9.78×10⁵ polyhedral bodies, respectively, and the dose 2.0×10⁶was chosen as the infection dosage of BC₁ backcross population, which is slightly higher than the average of two parents’ LD₅₀. Two independent linkage analyses were conducted successively in the autumn of 2014 and spring of 2015, which got a discrepant result that the linkage analysis result could not be repeated in the following confirmation; i.e., one marker, S2205, on Chr 22 was identified to link with resistance in the 1st experiment, while no linked marker was identified in the following one. Comparison with the previous reports on the linkage and mapping analysis of BmNPV resistance, it was found that the nonrepeatability was ubiquitous in the related researches in B. mori. Previously, a molecular marker (AY380833) was found to highly link with the resistant trait in NB and 871C, which are both the highly resistance strains to BmNPV. However, the linkage relationship was not detected in the linkage analysis population of 99R and 871C here.【Conclusion】the genetic complexity of resistance by molecular linkage analysis was further proved, including the variation among different resistant strains, as well the plausible multiple resistant loci in one single strain. At the same time, it was proposed that the BmNPV resistance may be a complex trait, as a prerequisite to be one type of qualitative-quantitative traits, the quantitative trait characteristic of BmNPV resistance is very obvious.

Key words: Bombyx mori, BmNPV, resistance, molecular markers, linkage analysis

GAO Rui, LI ChunLin, TONG XiaoLing, CAO MingYa, SHI MeiNing, XU AnYing, LU Cheng, DAI FangYin. Insight into Genetic Basis of Bombyx mori Resistant Strains with Resistance to BmNPV by Molecular Linkage Analysis[J].Scientia Agricultura Sinica, 2017, 50(1): 195-204.

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Insight into Genetic Basis of Bombyx mori Resistant Strains with Resistance to BmNPV by Molecular Linkage Analysis

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