Discovering genes responsible for silk synthesis in Bombyx mori by piggyBac-based random insertional mutagenesis

• 来源期刊: 中国昆虫科学（英文版）
• 关键词: aquaporin Bombyx mori development mutant piggyBac silk gland
• 年，卷（期）: 2019,（5）
• 页码范围: 821-830
• 页数: 10页
• 语种: 英文
• DOI: 10.1111/1744-7917.12595

摘要

Silkworm mutants are valuable resources for both transgenic breeding and gene discovery.PiggyBac-based random insertional mutagenesis has been widely used in gene functional studies.In order to discover genes involved in silk synthesis,a piggyBac-based random insertional library was constructed using Bombyx mori,and the mutants with abnormal cocoon were particularly screened.By this means,a "thin cocoon" mutant was identified.This mutant revealed thinner cocoon shell and shorter posterior silk gland (PSG) compared with the wild type.The messenger RNA (mRNA) levels of all the three fibroin genes,including Fib-H,Fib-L and P25,were significantly down-regulated in the PSG of mutants.Four piggyBac insertion sites were identified in Aquaporin (AQP),Longitudinals lacking protein-like (Lola),Glutamyl aminopeptidase-like (GluAP) and Loc 101744460.The mRNA levels of all the four genes were significantly altered in the silk gland of mutants.In particular,the mRNA amount of AQP,a gene responsible for the regulation of osmotic pressure,decreased dramatically immediately prior to the spinning stage in the anterior silk gland of mutants.The identification of the genes disrupted in the "thin cocoon" mutant in this study provided useful information for understanding silk production and transgenic breeding of silkworms in the future.