Jiangsu Key Laboratory of Sericultural Biology and Biotechnology, School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang, 212018, Jiangsu, China.
The Key Laboratory of Silkworm and Mulberry Genetic Improvement, Ministry of Agriculture, Sericultural Research Institute, Chinese Academy of Agricultural Science, Zhenjiang, 212018, Jiangsu, China.
PLoS One. 2022 Jul 7;17(7):e0270840. doi: 10.1371/journal.pone.0270840. eCollection 2022.
Bombyx mori is an important economic insect, its economic value mainly reflected in the silk yield. The major functional genes affecting the silk yield of B. mori have not been determined yet. Bombyx mori vacuolar protein sorting-associated protein 13d (BmVps13d) has been identified, but its function is not reported. In this study, BmVps13d protein shared 30.84% and 34.35% identity with that of in Drosophila melanogaster and Homo. sapiens, respectively. The expressions of BmVps13d were significantly higher in the midgut and silk gland of JS (high silk yield) than in that of L10 (low silk yield). An insertion of 9 bp nucleotides and two deficiencies of adenine ribonucleotides in the putative promoter region of BmVps13d gene in L10 resulted in the decline of promoter activity was confirmed using dual luciferase assay. Finally, the functions of BmVps13d in B. mori were studied using the CRISPR/Cas9 system, and the mutation of BmVps13d resulted in a 24.7% decline in weight of larvae, as well as a 27.1% (female) decline and a 11.8% (male) decline in the silk yield. This study provides a foundation for studying the molecular mechanism of silk yield and breeding the silkworm with high silk yield.
家蚕是一种重要的经济昆虫，其经济价值主要体现在产丝量上。影响家蚕丝产量的主要功能基因尚未确定。已鉴定出家蚕液泡分选相关蛋白 13d（BmVps13d），但它的功能尚未报道。在本研究中，BmVps13d 蛋白与果蝇和人类的同源性分别为 30.84%和 34.35%。在中肠和丝腺中，JS（产丝量高）的 BmVps13d 表达量明显高于 L10（产丝量低）。双荧光素酶检测证实，L10 中 BmVps13d 基因启动子区的 9 个核苷酸插入和两个腺嘌呤核糖核苷酸缺失导致启动子活性下降。最后，利用 CRISPR/Cas9 系统研究了 BmVps13d 在家蚕中的功能，BmVps13d 的突变导致幼虫体重下降 24.7%，雌蚕产丝量下降 27.1%，雄蚕产丝量下降 11.8%。本研究为研究产丝量的分子机制和培育高产丝量家蚕提供了基础。
