关键词: 柞蚕, 柞蚕微孢子虫, GST, ApGSTo1, 抗逆性

Abstract: 【Aim】 Nosema pernyi can cause the tussah microparticle disease of Antheraea pernyi, threatening the conservation and production safety of A. pernyi germplasm resources, and be vertically transmitted through eggs. Therefore, it is necessary to search for immune genes in A. pernyi to deal with the N. pernyi infection, so as to provide assistance for the subsequent search and cultivation of new varieties resistant to tussah microparticle disease. 【Methods】 The transcriptome of A. pernyi eggs infected with N. pernyi was constructed, and the immune-related genes were identified through GO functional annotation and KEGG analysis. The glutathione S-transferase (GST) gene of A. pernyi ApGSTo1 was cloned by PCR and bioinformatically analyzed. RT-qPCR was used to detect the expression level of ApGSTo1 in the 0-10-day-old eggs of A. pernyi infected by N. pernyi.【Results】 The number of expressed genes detected in the transcriptome of A. pernyi eggs infected and uninfected with N. pernyi was 17 453, and we found 89 significantly differentially expressed genes. In GO functional annotation, the numbers of genes involved in cellular anatomical entity and catalytic activity were the highest, while in KEGG analysis, the numbers of genes involved in the three pathways of transportation and catabolism, signal transduction, and digestive system were the highest. Based on transcriptome data analysis of A. pernyi eggs infected by N. pernyi, ApGSTo1 was used as the target gene. Results of PCR cloning and phylogenetic analysis proved that ApGSTo1 belongs to the omega-class GST. The expression level of ApGSTo1 in the A. pernyi eggs infected by N. pernyi was higher than that of the normal control group, and the expression level of ApGSTo1 in the 2-day-old eggs of A. pernyi infected by N. pernyi was extremely significantly higher than that of the normal control group. 【Conclusion】 Transcriptome data of A. pernyi eggs infected by N. pernyi were obtained, and an immune-related gene ApGSTo1 was identified. It was preliminarily demonstrated from molecular biology and transcriptional levels that ApGSTo1 participates in the stress resistance to N. pernyi infection in A. pernyi eggs. It was also found that the key time for the stress resistance to N. pernyi infection in A. pernyi eggs is at 2 d after egg laying. These results provide a research foundation for future research on the functions of ApGSTo1 in stress resistance and redox.

Key words: Antheraea pernyi, Nosema pernyi, GST, ApGSTo1, resistance