N-聚糖合成通路中内质网α-1,2甘露糖苷酶和α-甘露糖苷酶Ⅱ基因对家蚕胚胎滞育的调控作用  

Regulating Function of N⁃Glycan Biosynthesis Pathway Genes ER 1,2⁃Alpha⁃Mannosidase and Alpha⁃MannosidaseⅡon Embryonic Diapause in Bombyx mori

作　　者：阿依努尔·亚森 樊丙炎 陈艳花 宁阳威 王东岳 吴赛 朱娟 王梅仙[1,2] 唐顺明 沈兴家 Ayinuer Yasen;Fan Bingyan;Chen Yanhua;Ning Yangwei;Wang Dongyue;Wu Sai;Zhu Juan;Wang Meixian;Tang Shunming;Shen Xingjia(Jiangsu Key Laboratory of Sericulture and Animal Biotechnology,School of Biotechnology,Jiangsu University of Sci-ence and Technology,Zhenjiang Jiangsu 212100,China;Key Laboratory of Silkworm and Mulberry Genetic Improve-ment,Ministry of Agriculture and Rural Affairs,Sericultural Scientific Research Institute,Chinese Academy of Agricultural Sciences,Zhenjiang Jiangsu 212100,China)

机构地区：[1]江苏科技大学生物技术学院,江苏省蚕桑与畜牧生物技术重点实验室,江苏镇江212100 [2]中国农业科学院桑蚕研究中心,农业农村部蚕桑遗传改良重点实验室,江苏镇江212100

出　　处：《蚕业科学》2024年第2期106-115,共10页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(32072791,32102609);江苏省高校研究生科研创新项目(KYCX22⁃3761,SJCX22⁃1983)。

摘　　要：家蚕滞育是基因与环境协同作用的结果,其调控网络十分复杂,至今仍未完全阐明。为了解家蚕N-聚糖生物合成通路是否参与胚胎滞育调控,调查家蚕不同化性品系蛹期、胚胎发育早期以及滞育激素(DH)、蜕皮激素(20E)处理后BmN细胞中内质网α-1,2甘露糖苷酶基因(BmMAN1B)和α-甘露糖苷酶Ⅱ基因(BmMAN2)的转录水平。qRT-PCR结果显示:这2个基因在不同化性品系中的转录水平存在较大差异,蛹晚期的转录水平高于早期;分别用100 nmol/L DH和10 ng/mL 20E处理BmN细胞,BmMAN1B和BmMAN2的转录水平都极显著上调,提示2个基因与家蚕滞育密切相关。对BmN细胞和家蚕二化性品系秋丰165℃催青产非滞育卵组(QFLT)蛹期2 d雌蛹进行BmMAN1B和BmMAN2基因的RNA干扰,结果发现滞育相关基因BmDH、BmDHR-1、BmTRE2和BmSDH-2a等的转录水平发生了变化;当在个体水平进行BmMAN1B和BmMAN2基因的共同干扰时,382%的子代卵由非滞育转变为滞育。结果表明,BmMAN1B和BmMAN2在家蚕胚胎滞育中具有重要的调控作用。Diapause of the silkworm,Bombyx mori is the result of the synergy of genes and environment,and its regulatory network is very complicated and has not been fully elucidated yet.To better understand whether the N-glycan biosynthesis pathway of the silkworm is involved in the regulation of embryonic diapause,the transcription levels of BmMAN1B and BmMAN2 were analyzed in dif-ferent tissues of different voltinism silkworm strains during the pupal stage and early embryonic development stage,and in BmN cells after treating of diapause hormone(DH)or 20-hydroxyecdysone(20E).The qRT-PCR results showed that the transcription levels of the two genes varied greatly among different voltinism strains,with higher levels observed in the late pupal stage compared to that of the early pupal stage.When BmN cells were treated with 100 nmol/L DH and 10 ng/mL 20E respectively,the transcription levels of BmMAN1B and BmMAN2 genes were significantly upregulated,suggesting that both BmMAN1B and BmMAN2 were closely related to the diapause process of the silkworm.The changes of transcription levels of diapause-related genes in-cluding BmDH,BmDHR-1,BmTRE2 and BmSDH-2a were detected post RNA interference(RNAi)of BmMAN1B and BmMAN2 in BmN cells and in day-2 female pupae developed from bivoltine silkworm variety Qiufeng eggs incubated in 165℃(QFLT),respectively.RNAi targeting both of BmMAN1B and BmMAN2 in individuals resulted in a shift of 382%progeny eggs from non-diapause to diapause,indicating that BmMAN1B and BmMAN2 play important regulatory roles in the diapause of Bombyx mori embryos.

关 键 词：家蚕 滞育 N-糖基化 内质网α-1 2甘露糖苷酶 α-甘露糖苷酶Ⅱ 

分 类 号：S881.2[农业科学—特种经济动物饲养]