Diapause is a programmed developmental arrest process in insects.Diapause can occur at various stages of insect development and is frequently restricted to a specific developmental stage within a single species.In Bom...Diapause is a programmed developmental arrest process in insects.Diapause can occur at various stages of insect development and is frequently restricted to a specific developmental stage within a single species.In Bombyx mori,embryonic diapause is elicited by the diapause hormone(DH)and DH receptor(DHR)in diapause strains.Nevertheless,the regulatory mechanism through which BmDHR functions as a G protein-coupled receptor(GPCR),to exert other physiological functions in nondiapause silkworms,remains unclear.In this study,we found that BmDHR had 7 alternative splice isoforms.A knockout experiment confirmed that BmDHR mediated the transduction of diapause signals.Interestingly,the loss of BmDHR caused partial precocious metamorphosis and an embryo-lethal phenotype in nondiapause silkworms.An assessment of global transcriptional patterns revealed that BmDHR knockout affected physiological responses induced by manifold cellular processes,including the Toll/immune deficiency(Imd),Wnt,insulin-like growth factor,Hedgehog and P38/mitogen-activated protein kinase(MAPK)signaling pathways.This study expands our knowledge of the physiological roles for DHR in insect growth and development.展开更多
[Objectives]The present study was conducted to investigate the change rule ofβ-fructofuranosidase gene expression and its enzyme activity in the midgut of 5 th instar silkworm(Bombyx mori),in order to provide a refer...[Objectives]The present study was conducted to investigate the change rule ofβ-fructofuranosidase gene expression and its enzyme activity in the midgut of 5 th instar silkworm(Bombyx mori),in order to provide a reference for illustrating the enzymatic mechanism of usingβ-fructofuranosidase to absorb sucrose nutrition from mulberry leaves.[Methods]Real-time fluorescent quantitative PCR was applied to analyze the expression of BmSuc1 and BmSuc2 in midgut of 5 th-instar silkworm larvae,meanwhile the activities ofβ-fructofuranosidase was determined.[Results]BmSuc1 was expressed in the midgut of 5 th-instar silkworm larvae at different developmental stages.Its expression was upregulated at the beginning of the 5 th instar and during the peak feeding period,whereas BmSuc2 expression remained very low throughout the entire 5 th instar.The activity ofβ-fructofuranosidase was relatively high during the peak feeding period of 5 th-instar larvae,showing a trend of increasing first and then decreasing.[Conclusions]The expression pattern of the BmSuc1 gene and the changes inβ-fructofuranosidase activity were generally consistent with the physiological process of sugar nutrient absorption and utilization from mulberry leaves in 5 th-instar silkworms.It suggests that BmSuc1,as a sucrose hydrolase gene,plays a major role in the digestion and absorption of sucrose nutrients from mulberry leaves in the midgut tissue.展开更多
【目的】航天蚕Bombyx mori后代小茧突变体sc的幼虫发育缓慢,食桑量少,推测其生长发育相关通路基因受到基因突变的影响。前期研究表明,sc突变受一对位于家蚕第3连锁群的隐性基因控制,但其控制基因并未被鉴定。本研究拟通过比较分析sc与...【目的】航天蚕Bombyx mori后代小茧突变体sc的幼虫发育缓慢,食桑量少,推测其生长发育相关通路基因受到基因突变的影响。前期研究表明,sc突变受一对位于家蚕第3连锁群的隐性基因控制,但其控制基因并未被鉴定。本研究拟通过比较分析sc与航天蚕后代正常茧品系TG的转录组数据,为sc突变体相应基因的鉴定及分子机制的解析提供参考。【方法】分别取sc和TG 5龄第4天幼虫的头部和中肠组织进行转录组测序(RNA-seq),并通过比较转录组分析获得差异表达基因(differentially expressed genes,DEGs),对DEGs进行GO注释和KEGG富集分析。利用qRT-PCR验证随机选取的DEGs在sc和TG中的表达量。此外,利用qRT-PCR调查感兴趣基因在sc中的表达量。【结果】TG vs sc比较组头部检测到1528个DEGs,其中820个上调表达,708个下调表达;TG vs sc比较组中肠检测到1401个DEGs,其中683个上调表达,718个下调表达。GO分析表明头部和中肠DEGs在生物学过程中,大多数DEGs参与细胞过程、代谢过程、生物调节和刺激反应等;在分子功能中,大多数DEGs参与结合、催化活性、结构分子活性、转运蛋白活性和ATP依赖活性等。头部和中肠DEGs均涉及Hippo,Insulin和mTOR等与家蚕生长发育相关的信号通路。qRT-PCR分析显示,基因的表达趋势与转录组测序结果一致;与TG相比,sc中生长发育相关通路中BMSK0008105,BMSK0009907,BMSK0002689,BMSK0000286,BMSK0012340和BMSK00083629等关键基因差异表达。【结论】sc和TG中生长发育相关的信号通路关键基因的差异表达,通过影响生长发育过程中的能量代谢、器官发生和细胞生长、增殖及凋亡等生理过程,进而影响小茧突变体sc的体型发育。研究结果有助于阐明sc突变体形成的分子机制,并为家蚕体型调控研究积累实验数据。展开更多
基金supported by the National Natural Science Foundation of China(32170483 and 31970460)the Natural Science Foundation of Zhejiang Province(LR22C040001 and LQ23C040001)the Zhejiang Province Agricultural Technology Major Collaborative Promotion Project(2024ZDXT09).
文摘Diapause is a programmed developmental arrest process in insects.Diapause can occur at various stages of insect development and is frequently restricted to a specific developmental stage within a single species.In Bombyx mori,embryonic diapause is elicited by the diapause hormone(DH)and DH receptor(DHR)in diapause strains.Nevertheless,the regulatory mechanism through which BmDHR functions as a G protein-coupled receptor(GPCR),to exert other physiological functions in nondiapause silkworms,remains unclear.In this study,we found that BmDHR had 7 alternative splice isoforms.A knockout experiment confirmed that BmDHR mediated the transduction of diapause signals.Interestingly,the loss of BmDHR caused partial precocious metamorphosis and an embryo-lethal phenotype in nondiapause silkworms.An assessment of global transcriptional patterns revealed that BmDHR knockout affected physiological responses induced by manifold cellular processes,including the Toll/immune deficiency(Imd),Wnt,insulin-like growth factor,Hedgehog and P38/mitogen-activated protein kinase(MAPK)signaling pathways.This study expands our knowledge of the physiological roles for DHR in insect growth and development.
基金Supported by General Project of Yunnan Provincial Agricultural Basic Research Joint Special Project(202301BD070001-229)Yunnan Provincial Key R&D Program(202403AK140075)+1 种基金Modern Sericulture Industry Technology System of Yunan Province(KJTX-07)Honghe Comprehensive Test Station of National Sericulture Industry Technology System(CARS-18).
文摘[Objectives]The present study was conducted to investigate the change rule ofβ-fructofuranosidase gene expression and its enzyme activity in the midgut of 5 th instar silkworm(Bombyx mori),in order to provide a reference for illustrating the enzymatic mechanism of usingβ-fructofuranosidase to absorb sucrose nutrition from mulberry leaves.[Methods]Real-time fluorescent quantitative PCR was applied to analyze the expression of BmSuc1 and BmSuc2 in midgut of 5 th-instar silkworm larvae,meanwhile the activities ofβ-fructofuranosidase was determined.[Results]BmSuc1 was expressed in the midgut of 5 th-instar silkworm larvae at different developmental stages.Its expression was upregulated at the beginning of the 5 th instar and during the peak feeding period,whereas BmSuc2 expression remained very low throughout the entire 5 th instar.The activity ofβ-fructofuranosidase was relatively high during the peak feeding period of 5 th-instar larvae,showing a trend of increasing first and then decreasing.[Conclusions]The expression pattern of the BmSuc1 gene and the changes inβ-fructofuranosidase activity were generally consistent with the physiological process of sugar nutrient absorption and utilization from mulberry leaves in 5 th-instar silkworms.It suggests that BmSuc1,as a sucrose hydrolase gene,plays a major role in the digestion and absorption of sucrose nutrients from mulberry leaves in the midgut tissue.
文摘【目的】航天蚕Bombyx mori后代小茧突变体sc的幼虫发育缓慢,食桑量少,推测其生长发育相关通路基因受到基因突变的影响。前期研究表明,sc突变受一对位于家蚕第3连锁群的隐性基因控制,但其控制基因并未被鉴定。本研究拟通过比较分析sc与航天蚕后代正常茧品系TG的转录组数据,为sc突变体相应基因的鉴定及分子机制的解析提供参考。【方法】分别取sc和TG 5龄第4天幼虫的头部和中肠组织进行转录组测序(RNA-seq),并通过比较转录组分析获得差异表达基因(differentially expressed genes,DEGs),对DEGs进行GO注释和KEGG富集分析。利用qRT-PCR验证随机选取的DEGs在sc和TG中的表达量。此外,利用qRT-PCR调查感兴趣基因在sc中的表达量。【结果】TG vs sc比较组头部检测到1528个DEGs,其中820个上调表达,708个下调表达;TG vs sc比较组中肠检测到1401个DEGs,其中683个上调表达,718个下调表达。GO分析表明头部和中肠DEGs在生物学过程中,大多数DEGs参与细胞过程、代谢过程、生物调节和刺激反应等;在分子功能中,大多数DEGs参与结合、催化活性、结构分子活性、转运蛋白活性和ATP依赖活性等。头部和中肠DEGs均涉及Hippo,Insulin和mTOR等与家蚕生长发育相关的信号通路。qRT-PCR分析显示,基因的表达趋势与转录组测序结果一致;与TG相比,sc中生长发育相关通路中BMSK0008105,BMSK0009907,BMSK0002689,BMSK0000286,BMSK0012340和BMSK00083629等关键基因差异表达。【结论】sc和TG中生长发育相关的信号通路关键基因的差异表达,通过影响生长发育过程中的能量代谢、器官发生和细胞生长、增殖及凋亡等生理过程,进而影响小茧突变体sc的体型发育。研究结果有助于阐明sc突变体形成的分子机制,并为家蚕体型调控研究积累实验数据。
