The ecdysone-induced transcription factor E93 in model insects plays multiple roles in the insect metamorphosis processes,such as remodeling larval tissues and determining adult tissue formation.The knockdown of E93in...The ecdysone-induced transcription factor E93 in model insects plays multiple roles in the insect metamorphosis processes,such as remodeling larval tissues and determining adult tissue formation.The knockdown of E93in insects leads to incomplete metamorphosis,suggesting that E93 is a potential target for pest control.In this study,the HaE93 gene in the cotton bollworm Helicoverpa armigera,a polyphagous pest of various commercial crops worldwide,was identified and found to have high expression in the egg,prepupal,and pupal stages.The injection of ds HaE93 induced about 60%mortality in H.armigera at the larval-pupal stage.About 30%survived but showed delayed pupation and abnormal wings,and the females developed reduced ovaries.Therefore,about 90%of the HaE93 knockdown individuals failed to reproduce before they died.The results of qRT-PCR showed that the expression levels of ecdysone primary-response genes,chitin synthesis-related genes,and wing and ovary development-related genes were reduced in HaE93 knockdown H.armigera.These results indicated that HaE93plays a critical role in larva-pupa-adult metamorphosis and the development of the cuticle,wing,and ovary in female H.armigera by regulating the expression of the associated genes.Bioassays of ds HaE93 administered by either oral delivery or injection showed similar knockdown results,which suggested that HaE93 can be used as a target gene for the RNAi control of the pest H.armigera.展开更多
Mating behavior is essential for sexual reproduction,and it is often modulated by key chemical cues.In many moth species,males find compatible mates through the reception of sex pheromones which are released by female...Mating behavior is essential for sexual reproduction,and it is often modulated by key chemical cues.In many moth species,males find compatible mates through the reception of sex pheromones which are released by females.Pheromone receptors(PRs)are key elements in sensing these chemical signals.Concurrently,male moths emit a complex blend of volatile compounds during courtship;however,the mechanisms for recognizing putative male pheromones remain poorly understood.Here,we employed gas chromatography coupled with electroantennographic detection and mass spectrometry to analyze the volatile compounds produced by males of the cotton bollworm,Helicoverpa armigera.Three candidate male sex pheromones were identified,with(Z)-7-dodecen-1-yl acetate(Z7-12:OAc)eliciting the most pronounced electrophysiological response in the male antenna.The olfactory receptor neuron(ORN)ORN-a in Type A trichoid sensilla was shown to respond to Z7-12:OAc by conducting single sensillum recording(SSR)assays.Additionally,we found that the OR13s from five Heliothinae species responded to Z7-12:OAc by using the Xenopus oocyte expression system and two-electrode voltage-clamp recording.Our findings identified a candidate for evaluation in future behavioral studies of the poorly understood chemosensory recognition mechanisms underlying male sex pheromones.If its relevance is supported by behavioral data,this knowledge may facilitate the design of novel olfactory regulators for effective pest control strategies involving mating disruption.展开更多
[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene ...[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene in cotton bollworm. [Method] Total RNA was extracted from midgut of cotton bollworm. The full length cDNA of Habeta5 gene was cloned by using rapid amplification of cDNA ends (RACE) technology, then sequence analysis was carried out. [Result] The full length cDNA sequence was successfully cloned and isolated, named as Habeta5. It was 947 bp in length, contained an ORF (843 bp) and encoded 280 amino acid residues, with the predicted mass of 30.87 kD and isoelectric point(pI) of 9.60. In the deduced amino acid sequence, a proteasome β5 subunit domain lies between 74th to 261st amino acid residues. It has more than 62% identity to other insects such as Drosophila melanogaster. The proteasome β5 subunit conservative regions were very similar with each other. Molecular evolution by Neighbor Joining method indicated that Habeta5 was homologous with other proteasome β5 subunit of species. [Conclusion] Sequence alignment shows that the cloned fragment is a proteasome β5 subunit gene (GenBank accession number: FJ358434).展开更多
[ Objective ] The paper was to confirm the effect of mating and black light treatment on cryptochromes mRNA expression of Hclicoverpa armigera. [ Method ] Quantitative real-time PCR （ SYBR Green） technique was appli...[ Objective ] The paper was to confirm the effect of mating and black light treatment on cryptochromes mRNA expression of Hclicoverpa armigera. [ Method ] Quantitative real-time PCR （ SYBR Green） technique was applied to detect the expression of cryptochromes gene （ cryl and cry2 ） of H. armigera under different conditions. Total RNA was extracted from the head of H. armigera, and carried out reverse transcription to synthesize cDNA after digested by DNase I. Specific primers were used to carry out quantitative real-time PCR on cryl, cry2 and EF-Ict gene, respectively. [ Result] The expression of cryl mRNA of H. ar- migera significantly decreased after exposure to black light for 2 h, the mRNA expression of cry2 was smaller than control, they had no significant difference with each other. Mating had significant effect on mRNA expression of cryl and cry2 of H. armigera, and the mRNA expression of cryl and cry2 of male and female adults showed decreasing trend with the prolongation of time after mating. [ Conclusion] The result had important significance for further studying on function of cry gene and the control of cotton bollwonn.展开更多
基金supported by the National Natural Science Foundation of China(32001912 and 32370525)the Henan Provincial Natural Science Foundation,China(232300420012)。
文摘The ecdysone-induced transcription factor E93 in model insects plays multiple roles in the insect metamorphosis processes,such as remodeling larval tissues and determining adult tissue formation.The knockdown of E93in insects leads to incomplete metamorphosis,suggesting that E93 is a potential target for pest control.In this study,the HaE93 gene in the cotton bollworm Helicoverpa armigera,a polyphagous pest of various commercial crops worldwide,was identified and found to have high expression in the egg,prepupal,and pupal stages.The injection of ds HaE93 induced about 60%mortality in H.armigera at the larval-pupal stage.About 30%survived but showed delayed pupation and abnormal wings,and the females developed reduced ovaries.Therefore,about 90%of the HaE93 knockdown individuals failed to reproduce before they died.The results of qRT-PCR showed that the expression levels of ecdysone primary-response genes,chitin synthesis-related genes,and wing and ovary development-related genes were reduced in HaE93 knockdown H.armigera.These results indicated that HaE93plays a critical role in larva-pupa-adult metamorphosis and the development of the cuticle,wing,and ovary in female H.armigera by regulating the expression of the associated genes.Bioassays of ds HaE93 administered by either oral delivery or injection showed similar knockdown results,which suggested that HaE93 can be used as a target gene for the RNAi control of the pest H.armigera.
基金funded by the National Natural Science Foundation of China(32272540 and 32072509)the National Key R&D Program of China(2022YFD1400800)the Innovation Program of Chinese Academy of Agricultural Sciences(CAAS-CSCB-202302)。
文摘Mating behavior is essential for sexual reproduction,and it is often modulated by key chemical cues.In many moth species,males find compatible mates through the reception of sex pheromones which are released by females.Pheromone receptors(PRs)are key elements in sensing these chemical signals.Concurrently,male moths emit a complex blend of volatile compounds during courtship;however,the mechanisms for recognizing putative male pheromones remain poorly understood.Here,we employed gas chromatography coupled with electroantennographic detection and mass spectrometry to analyze the volatile compounds produced by males of the cotton bollworm,Helicoverpa armigera.Three candidate male sex pheromones were identified,with(Z)-7-dodecen-1-yl acetate(Z7-12:OAc)eliciting the most pronounced electrophysiological response in the male antenna.The olfactory receptor neuron(ORN)ORN-a in Type A trichoid sensilla was shown to respond to Z7-12:OAc by conducting single sensillum recording(SSR)assays.Additionally,we found that the OR13s from five Heliothinae species responded to Z7-12:OAc by using the Xenopus oocyte expression system and two-electrode voltage-clamp recording.Our findings identified a candidate for evaluation in future behavioral studies of the poorly understood chemosensory recognition mechanisms underlying male sex pheromones.If its relevance is supported by behavioral data,this knowledge may facilitate the design of novel olfactory regulators for effective pest control strategies involving mating disruption.
基金Supported by National Basic Research Program of China(2005CB121005)~~
文摘[Objective] Using molecular biotechnology to clone the proteasome β5 gene from cotton bollworm (Helicoverpa armigera), this research aimed to provide basis for further research on the function of proteasome β5 gene in cotton bollworm. [Method] Total RNA was extracted from midgut of cotton bollworm. The full length cDNA of Habeta5 gene was cloned by using rapid amplification of cDNA ends (RACE) technology, then sequence analysis was carried out. [Result] The full length cDNA sequence was successfully cloned and isolated, named as Habeta5. It was 947 bp in length, contained an ORF (843 bp) and encoded 280 amino acid residues, with the predicted mass of 30.87 kD and isoelectric point(pI) of 9.60. In the deduced amino acid sequence, a proteasome β5 subunit domain lies between 74th to 261st amino acid residues. It has more than 62% identity to other insects such as Drosophila melanogaster. The proteasome β5 subunit conservative regions were very similar with each other. Molecular evolution by Neighbor Joining method indicated that Habeta5 was homologous with other proteasome β5 subunit of species. [Conclusion] Sequence alignment shows that the cloned fragment is a proteasome β5 subunit gene (GenBank accession number: FJ358434).
基金Supported by National"973"Project(No.2006CB102006)Special Research Project in Public Service Sectors(Agriculture)(201103012)~~
文摘[ Objective ] The paper was to confirm the effect of mating and black light treatment on cryptochromes mRNA expression of Hclicoverpa armigera. [ Method ] Quantitative real-time PCR （ SYBR Green） technique was applied to detect the expression of cryptochromes gene （ cryl and cry2 ） of H. armigera under different conditions. Total RNA was extracted from the head of H. armigera, and carried out reverse transcription to synthesize cDNA after digested by DNase I. Specific primers were used to carry out quantitative real-time PCR on cryl, cry2 and EF-Ict gene, respectively. [ Result] The expression of cryl mRNA of H. ar- migera significantly decreased after exposure to black light for 2 h, the mRNA expression of cry2 was smaller than control, they had no significant difference with each other. Mating had significant effect on mRNA expression of cryl and cry2 of H. armigera, and the mRNA expression of cryl and cry2 of male and female adults showed decreasing trend with the prolongation of time after mating. [ Conclusion] The result had important significance for further studying on function of cry gene and the control of cotton bollwonn.
