20-hydroxyecdysone(20E)induced transcription factor E93 is important for larval–adult transition,which functions in programmed cell death of larval obsolete tissues,and the formation of adult new tissues.However,the ...20-hydroxyecdysone(20E)induced transcription factor E93 is important for larval–adult transition,which functions in programmed cell death of larval obsolete tissues,and the formation of adult new tissues.However,the apoptosis-related genes directly regulated by E93 are still ambiguous.In this study,an E93 mutation fly strain was obtained by clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9-mediated long exon deletion to investigate whether and how E93 induces apoptosis during larval tissues metamorphosis.The transcriptional profile of E93 was consistent with 3 RHG(rpr,hid,and grim)genes and the effector caspase gene drice,and all their expressions peaked at the initiation of apoptosis during the degradation of salivary glands.The transcription expression of 3 RHG genes decreased and apoptosis was blocked in E93 mutation salivary gland during metamorphosis.In contrast,E93 overexpression promoted the transcription of 3 RHG genes,and induced advanced apoptosis in the salivary gland.Moreover,E93 not only enhance the promoter activities of the 3 RHG genes in Drosophila Kc cells in vitro,but also in the salivary gland in vivo.Our results demonstrated that 20E induced E93 promotes the transcription of RHG genes to trigger apoptosis during obsolete tissues degradation at metamorphosis in Drosophila.展开更多
The brown planthopper(Nilaparvata lugens) is the main migratory pest in many rice growing areas in Asia. E78 is a member of the nuclear hormone receptor superfamily which plays an important role in egg development and...The brown planthopper(Nilaparvata lugens) is the main migratory pest in many rice growing areas in Asia. E78 is a member of the nuclear hormone receptor superfamily which plays an important role in egg development and maternal regulation of early embryogenesis. In this study, brown planthopper E78(NlE78) was cloned, and the predicted amino acid sequence showed that it contains two conserved domains: NR-LBD and DBD. qRT-PCR showed that the expression of NlE78 is high in the fifth instar nymphs and the ovaries of females. After downregulation of NlE78, the rate of moulting failure(33.2%) increased significantly, and ovarian development was delayed. However, when NlE78was downregulated together with NlE93, the emergence rate increased significantly(78.79%), and ovarian development was similar to that when NlE78 was downregulated but not delayed. A co-immunoprecipitation experiment showed that NlE78 interacts with NlE93, a crucial downstream transcription factor of the ecdysone signalling pathway. Cellular localization by immunofluorescence revealed that NlE78 and NlE93 are expressed in the nucleus. This study indicates that NlE78 regulates ovarian development and moulting, possibly through its interaction with NlE93. This study is of great significance for the development of new pesticides and control methods based on newly discovered targets.展开更多
Ecdysone-induced protein 93F(E93)plays important roles during the metamorphosis process in insects.In this study,a cDNA of the LmE93 gene was identified from the transcriptome of Locusta migratoria,which consists of t...Ecdysone-induced protein 93F(E93)plays important roles during the metamorphosis process in insects.In this study,a cDNA of the LmE93 gene was identified from the transcriptome of Locusta migratoria,which consists of the 3378-nucleotide open-reading frame(ORF)and encodes 1125 amino acids with helix–turn–helix(HTH)motifs.Reverse transcription quantitative polymerase chain reaction analysis revealed that LmE93 was highest expressed in ovary.The LmE93 expression level was markedly low from the 3rd to 4th instar nymphs,and greatly increased in 1-day-old 5th instar nymphs with a peak on middle nymphal days,then declined in the late nymphal days.Moreover,injected dsLmE93 into 4th and 5th instar nymphs greatly reduced LmE93 transcripts,respectively,and prevented the process of metamorphosis,causing supernumerary nymphal stages.Hematoxylin–eosin staining of the integument showed that the apolysis occurred in advance in 4th instar nymphs,and old cuticle degradation was decreased in dsLmE93-injected locusts of 5th instar nymphs.Smaller and no fully developed wings with reduced columns between the anterior and posterior regions were found in N6 and N7 supernumerary nymphs.In addition,the development of the ovary in dsLmE93-injected locusts was severely blocked,the yolk was almost not formed and there was no development of ovarioles.The results indicated that LmE93 play key roles in the metamorphosis,cuticle,wing and ovarian development of locusts.展开更多
基金the National Science Foundation of China(Grants No.32070491 to Kang Li)Laboratory of Lingnan Modern Agriculture Project(NT2021003 to Sheng Li)the National Science Foundation of Guangdong Province(Grants No.2022A1515010457 to Kang Li),English was polished by the Nature Publishing Group.
文摘20-hydroxyecdysone(20E)induced transcription factor E93 is important for larval–adult transition,which functions in programmed cell death of larval obsolete tissues,and the formation of adult new tissues.However,the apoptosis-related genes directly regulated by E93 are still ambiguous.In this study,an E93 mutation fly strain was obtained by clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein 9-mediated long exon deletion to investigate whether and how E93 induces apoptosis during larval tissues metamorphosis.The transcriptional profile of E93 was consistent with 3 RHG(rpr,hid,and grim)genes and the effector caspase gene drice,and all their expressions peaked at the initiation of apoptosis during the degradation of salivary glands.The transcription expression of 3 RHG genes decreased and apoptosis was blocked in E93 mutation salivary gland during metamorphosis.In contrast,E93 overexpression promoted the transcription of 3 RHG genes,and induced advanced apoptosis in the salivary gland.Moreover,E93 not only enhance the promoter activities of the 3 RHG genes in Drosophila Kc cells in vitro,but also in the salivary gland in vivo.Our results demonstrated that 20E induced E93 promotes the transcription of RHG genes to trigger apoptosis during obsolete tissues degradation at metamorphosis in Drosophila.
基金supported by the National Natural Science Foundation of China (32172390, 1672023 and 31741107)the Natural Science Foundation of Zhejiang Province, China (LZ20C140002)。
文摘The brown planthopper(Nilaparvata lugens) is the main migratory pest in many rice growing areas in Asia. E78 is a member of the nuclear hormone receptor superfamily which plays an important role in egg development and maternal regulation of early embryogenesis. In this study, brown planthopper E78(NlE78) was cloned, and the predicted amino acid sequence showed that it contains two conserved domains: NR-LBD and DBD. qRT-PCR showed that the expression of NlE78 is high in the fifth instar nymphs and the ovaries of females. After downregulation of NlE78, the rate of moulting failure(33.2%) increased significantly, and ovarian development was delayed. However, when NlE78was downregulated together with NlE93, the emergence rate increased significantly(78.79%), and ovarian development was similar to that when NlE78 was downregulated but not delayed. A co-immunoprecipitation experiment showed that NlE78 interacts with NlE93, a crucial downstream transcription factor of the ecdysone signalling pathway. Cellular localization by immunofluorescence revealed that NlE78 and NlE93 are expressed in the nucleus. This study indicates that NlE78 regulates ovarian development and moulting, possibly through its interaction with NlE93. This study is of great significance for the development of new pesticides and control methods based on newly discovered targets.
基金supported by the National Natural Science Foundation of China(grant no.32072419,31872010,and 31701794)Research Project Supported by Shanxi Scholarship Council of China(grant no.2020-017)+1 种基金Natural Science Foundation of Shanxi Province for The Excellent Youth(grant no.201901D211194)Scientific and Technological Innovation Programs of Higher Education Institutions in Shanxi(Grant No.2017104).
文摘Ecdysone-induced protein 93F(E93)plays important roles during the metamorphosis process in insects.In this study,a cDNA of the LmE93 gene was identified from the transcriptome of Locusta migratoria,which consists of the 3378-nucleotide open-reading frame(ORF)and encodes 1125 amino acids with helix–turn–helix(HTH)motifs.Reverse transcription quantitative polymerase chain reaction analysis revealed that LmE93 was highest expressed in ovary.The LmE93 expression level was markedly low from the 3rd to 4th instar nymphs,and greatly increased in 1-day-old 5th instar nymphs with a peak on middle nymphal days,then declined in the late nymphal days.Moreover,injected dsLmE93 into 4th and 5th instar nymphs greatly reduced LmE93 transcripts,respectively,and prevented the process of metamorphosis,causing supernumerary nymphal stages.Hematoxylin–eosin staining of the integument showed that the apolysis occurred in advance in 4th instar nymphs,and old cuticle degradation was decreased in dsLmE93-injected locusts of 5th instar nymphs.Smaller and no fully developed wings with reduced columns between the anterior and posterior regions were found in N6 and N7 supernumerary nymphs.In addition,the development of the ovary in dsLmE93-injected locusts was severely blocked,the yolk was almost not formed and there was no development of ovarioles.The results indicated that LmE93 play key roles in the metamorphosis,cuticle,wing and ovarian development of locusts.
