昆虫学报 ›› 2022, Vol. 65 ›› Issue (9): 1075-1089.doi: 10.16380/j.kcxb.2022.09.001

• 研究论文 •    下一篇

西伯利亚蝗卵发育过程的比较转录组分析及滞育关联基因筛选

赵娜1, 吕雪峰2, 扈鸿霞1, 宋余1, 蒋思涵1, 季荣1, 叶小芳1,*   

  1. (1. 新疆师范大学生命科学学院, 中亚区域跨境有害生物联合控制国际研究中心, 新疆特殊环境物种保护与调控生物学实验室, 乌鲁木齐 830054; 2. 新疆畜牧科学院畜牧业质量标准研究所, 乌鲁木齐830000)
  • 出版日期:2022-09-20 发布日期:2022-10-16

Comparative transcriptome analysis and screening of diapause-associated genes during the egg development of Gomphocerus sibiricus(Orthoptera: Acrididae)

ZHAO Na1, LÜ Xue-Feng2, HU Hong-Xia1, SONG Yu1, JIANG Si-Han1, JI Rong1, YE Xiao-Fang1,*    

  1. (1. International Center for the Collaborative Management of Cross-border Pest in Central Asia, Xinjiang Key Laboratory of Special Species Conservation and Regulatory Biology, College of Life Science, Xinjiang Normal University, Urumqi 830054, China; 2. Institute of Animal Husbandry Quality Standards, Xinjiang Academy of Animal Sciences, Urmuqi 830000, China)
  • Online:2022-09-20 Published:2022-10-16

摘要: 【目的】通过筛选西伯利亚蝗Gomphocerus sibiricus卵滞育基因及代谢通路,初步明确西伯利亚蝗卵滞育分子机理。【方法】利用Illumina NovaSeq 6000测序平台对西伯利亚蝗早期发育(ES)、滞育(DS)和滞育后发育阶段(PS)的卵进行转录组测序;采用GO和KEGG富集分析并结合文献,预测滞育相关通路并聚类热图分析,筛选蝗卵滞育关联基因,并选取其中6个重要基因进行qRT-PCR验证。【结果】DS vs ES和PS vs DS两比较组分别富集到12 419和4 789个差异表达基因,且多上调表达;两比较组共表达差异基因为2 206个,主要与糖代谢、环境胁迫和生长发育相关。DS vs ES组富集最显著的GO条目为蛋白结合,PS vs DS组GO条目主要包含酶活性、细胞骨架构建及蛋白结合等过程。滞育关联基因主要集中在Wnt信号通路、胰岛素信号通路、细胞周期通路以及昆虫激素生物合成等通路。6个重要的滞育关联基因的表达量变化趋势与转录组数据结果基本一致。【结论】本研究初步明确了西伯利亚蝗卵滞育调控的重要代谢途径,并筛选出20个滞育关联基因,为后续深入研究该物种的适应机理奠定了基础。

关键词: 西伯利亚蝗, 卵, 转录组, 滞育关联基因, KEGG, qRT-PCR

Abstract: 【Aim】This study aims to preliminarily clarify the molecular mechanism of diapause in Gomphocerus sibiricus eggs through screening diapause genes and metabolic pathways of the eggs.【Methods】 Transcriptome sequencing was performed on G. sibiricus eggs at different developmental stages [early developmental stage (ES), diapause stage (DS), and post-diapause developmental stage (PS)], with the Illumina NovaSeq 6000 sequencing platform. The diapause-associated pathways of G. sibiricus eggs were predicted 
by GO and KEGG enrichment analysis, and analyzed by cluster heat map analysis combined with literature reports to screen diapause-associated genes. qRT-PCR was used to verify six important genes of the screened diapause-associated genes. 【Results】 In the DS vs ES and PS vs DS comparative groups, 12 419 and 4 789 differentially expressed genes (DEGs) were enriched, respectively, and most of them were up-regulated. A total of 2 206 DEGs of the two comparative groups were mainly related to glucose metabolism, environmental stress and growth and development. The most significant enrichment of GO items in the DS vs ES group was protein binding. The GO items in the PS vs DS group mainly included enzymatic activity, cytoskeleton construction and protein binding. Diapause-associated genes were mainly involved in Wnt signaling pathway, insulin signaling pathway, cell cycle pathway and insect hormone biosynthesis pathway. The expression trends of the six important diapause-associated genes were consistent with the transcriptome data. 【Conclusion】 In this study, important metabolic pathways that regulate diapause of G. sibiricus eggs were preliminarily identified, and a total of 20 diapause-associated genes were screened out, laying a foundation for further study on the adaptation mechanism of this species.

Key words: Gomphocerus sibiricus; eggs, transcriptome, diapause-associated genes, KEGG; qRT-PCR