A cDNA encoding aminopeptidase N was cloned by degenerated PCR combined with RACE technique in this paper. The full-length of APN-Harm is 3 043 bp. Open reading frame is 2 856 bp in length, encoding 951 amino acid res...A cDNA encoding aminopeptidase N was cloned by degenerated PCR combined with RACE technique in this paper. The full-length of APN-Harm is 3 043 bp. Open reading frame is 2 856 bp in length, encoding 951 amino acid residues. Its predicted molecular weight and isoelectric point are 108. 3 kDa and 5.29, respectively. This deduced amino acid sequence shares some common structural features with aminopeptidase N from several moth species, including the consensus zinc-binding motif HEXXHX18E and the GAMEN motif common to gluzincin aminopeptidases. The first 20 amino acid residues at N-termini is hydrophobic transmembrane helix. The sequence of APN-Harm was deposited in GenBank and the accession number is AY181026.展开更多
Helicoverpa armigera H. is a polyphagous notorious pest of a number of economic crops including tomato. In tomato, it can reduce its yield as high as 70% due to fruit boring. Although some predators and parasitoids ha...Helicoverpa armigera H. is a polyphagous notorious pest of a number of economic crops including tomato. In tomato, it can reduce its yield as high as 70% due to fruit boring. Although some predators and parasitoids have been recorded in the field for their natural control, their manipulation to a successful crop management is still awaited. To overcome this pest, insecticides play a significant role in its effective crop management program globally. Tomato is an absolutely perishable commodity and needs the specified period required between the application, of conventional pesticides of organophosphate group, and consumption which can hardly be afforded. Therefore, nine insecticides from the new chemistries, which were relatively safer to human and environment, i.e. Coragen (chlorantraniliprole), steward (indoxacarb), Belt (flubendamide), Delegate (spintoram), Volium Flexy (chlorantraniliprole + thiamethaxim), Fipronil (grafter), Proclaim (emamectin benzoate), Pirate (chlofenapyr) and Lufenuron (lufenuron), were tested on tomato crop heavily infested by this pest in the farmers field in districts Lodhran and Bahawalpur (Pakistan) in the month of April 2014. The mortality was compared after 4, 7 and 10 days and treatments were compared with control as well with one another. The average maximum % mortality, i.e. 89.36 and 85.09 of the pest, was observed with Volium Flexy, and Delegate also worked well even after 7 and 10 days. Similarly, the results on the basis of damaged fruits and percent loss of yield pointed out that Chlorantraniliprole, Flubendiamide and Indoxacarb had resulted better as compared with others, although the difference was statistically non significant. These new chemistry pesticides are suggested to be used at tomato crop against Helicoverpa armigera which are best fitted in the IPM program for the control of the pest.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
Sublethal effects of chlorantraniliprole to Helicoverpa armigera （Hiibner） larvae were evaluated through exposure of third instar larvae to the insecticide incorporated into an artificial diet. When larvae were fed ...Sublethal effects of chlorantraniliprole to Helicoverpa armigera （Hiibner） larvae were evaluated through exposure of third instar larvae to the insecticide incorporated into an artificial diet. When larvae were fed on the insecticide-containing diet for 7 d, the LC10, LC20, LC40, and LC50values were 3.790, 7.977, 21.577, and 33.121 μg active ingredient L^-1, respectively. Chlorantraniliprole at sublethal concentrations significantly reduced the larval body mass, emergence ratio, adult longevity and egg hatching rate in both the parental and offspring generations. The pupation and copulation rate in the parental generation and the pupal mass in the offspring also strongly decreased. Reproduction was seriously disturbed in both the parental and offspring groups even when only one of the partners was exposed to chlorantraniliprole at larval stages. However, at the lowest concentration of exposure （LC^0）, offspring fecundity was strongly reduced only when both partners were exposed. The net reproductive rate （R0）, intrinsic rate of increase （rm）, and gross reproduction rate （GRR） at LC20, LC40 concentrations were significantly lower than that of the control. Post-exposure effects also included an extended larval developmental time and increased male proportion in both generations. The doubling time （Dt） at LC20, LC40 concentrations as well as gross reproduction rate （GRR） at LC10 concentration were also significantly increased. Chlorantraniliprole might have significant effects on H. armigera population dynamics even at sublethal concentrations on both parental and offspring generations.展开更多
The full-length sequence of the odorant binding protein 5 gene,HarmOBP5,was obtained from an antennae cDNA library of cotton bollworm,Helicoverpa armigera （Hübner）.The cDNA contains a 444 bp open reading frame,...The full-length sequence of the odorant binding protein 5 gene,HarmOBP5,was obtained from an antennae cDNA library of cotton bollworm,Helicoverpa armigera （Hübner）.The cDNA contains a 444 bp open reading frame,encoding a protein with 147 amino acids,namely HarmOBP5.HarmOBP5 was expressed in Escherichia coli and the recombinant protein was purified by affinity chromatography.SDS-PAGE and Western blot analysis demonstrated that the purified protein can be used for further investigation of its binding characteristics.Competitive binding assays with 113 odorant chemicals indicated that HarmOBP5 has strong affinity to some special plant volatiles,including （E）-β-farnesene,ethyl butyrate,ethyl heptanoate,and acetic acid 2-methylbutyl ester.Based on three-dimensional （3D） model of AaegOBP1 from Aedes aegypti,a 3D model of HarmOBP5 was predicted.The model revealed that some key binding residues in HarmOBP5 may play important roles in odorant perception of H.armigera.This study provides clues for better understanding physiological functions of OBPs in H.armigera and other insects.展开更多
The growth, development and consumption of successive three generations of cotton bollworm, Helicoverpa armigera （Htibner）, fed on cotton bolls grown under elevated CO2 （double-ambient vs. ambient） in open-top cha...The growth, development and consumption of successive three generations of cotton bollworm, Helicoverpa armigera （Htibner）, fed on cotton bolls grown under elevated CO2 （double-ambient vs. ambient） in open-top chambers were examined. Significant decreases in protein, total amino acid, water and nitrogen content and increases in free fatty acid were observed in cotton bolls. Changes in quality of cotton bolls affected the growth, development and food utilization of H. armigera. Significantly longer larval development duration in three successive generations and lower pupal weight of the second and third generations were observed in cotton bollworm fed on cotton bolls grown under elevated CO2. Significantly lower fecundity was also found in successive three generations of H. armigera fed on cotton bolls grown under elevated CO2. The consumption per larva occurred significant increase in successive three generations and frass per larva were also significantly increased during the second and third generations under elevated CO2. Significantly lower relative growth rate, efficiency of conversion of ingested food and significant higher relative consumption rate in successive three generations were observed in cotton bollworm fed on cotton bolls grown under elevated CO2. Significantly lower potential female fecundity, larval numbers and population consumption were found in the second and third generations of cotton bollworm fed on cotton bolls grown under elevated CO2. The integrative effect of higher larval mortality rate and lower adult fecundity resulted in significant decreases in potential population consumption in the latter two generations. The results show that elevated CO2 adversely affects cotton bolls quality, which indicates the potential population dynamics and potential population consumption of cotton bollworm will alleviate the harm to the plants in the future rising CO2 atmosphere.展开更多
The brush border membrane vesicles (BBMVs) in midgut of Helicoverpa armigera weresuccessfully separated, and most of the Aminopeptidase N (APN) activities in BBMV werepreserved. The 3-[(3-chlor-amidopropyl) dimethylam...The brush border membrane vesicles (BBMVs) in midgut of Helicoverpa armigera weresuccessfully separated, and most of the Aminopeptidase N (APN) activities in BBMV werepreserved. The 3-[(3-chlor-amidopropyl) dimethylammonio]-1-propane-sulphonate (CHAPS)can enhance the dissolution of BBMV, and phosphatidylinositol-specific phosopholipase C(PI-PLC) can cleave the APN from midgut membrane. The APN was primarily purified usinga Mono-Q column. The results of immunoblotting showed that the 120 and 170kDa proteinsin the BBMV could bind Cry1Ac, and 120kDa APN was a glycosylphosphalidylinositol(GPI)-anchored protein. Two Bt-resistant strains (Bt-P, Bt-M) were obtained after beingselected for more than five years in laboratory using Bt insecticides and Bt transgeniccotton incorporated into diet separately. The resistance of Bt-P and Bt-M were 1083.3and 48.7 times that of susceptible strain. The genes encoding APN1 in midgut ofsusceptible and resistant H.armigera were cloned by PCR and RACE techniques. Theinferred amino acid sequences of APN1 possessed the common character of APN family ininsects. In comparison with APN1 in susceptible strain, three nucleotide mutations wereobserved in the APN1 of Bt-M strain and resulted in two amino acid replace in theputative protein sequences, and eight nucleotide mutations were observed in Bt-P strainand resulted in five amino acid replace.展开更多
Objective:To evaluate the antifeedant,insecticidal and growth inhibition activities of Solanum pseudocapsicum(S.pseudocapsicum)seed extracts against Spodoptera litura(S.litura)and Helicotverpa armigera(H.armigera).Met...Objective:To evaluate the antifeedant,insecticidal and growth inhibition activities of Solanum pseudocapsicum(S.pseudocapsicum)seed extracts against Spodoptera litura(S.litura)and Helicotverpa armigera(H.armigera).Methods:Hcxane,diethyl cther,dichloromethanc and ethyl acetate seed extracts were prepared and tested for autifeedant,insecticidal and growth inhibitory activities against fourth instar larvae of&litura and H.armigera.Results:Ethyl acetate extract showed promising antifeedant and insecticidal activities against S.litura and H.armigera.Percentage of deformed larvae,pupae and adults were maximum in treatment of ethyl acetate extract.Percentage of successful adult emergence was deteriorated by seeds on extract treated larvae.Conclusions:Ethyl acetate extracts of S.pseudocapsicum,showed higher efficiency of antifeedant,insecticidal and growth inhibition activities.Hence,it can be used to controll agricultural insect pests,S.litura and H.armigera.展开更多
文摘A cDNA encoding aminopeptidase N was cloned by degenerated PCR combined with RACE technique in this paper. The full-length of APN-Harm is 3 043 bp. Open reading frame is 2 856 bp in length, encoding 951 amino acid residues. Its predicted molecular weight and isoelectric point are 108. 3 kDa and 5.29, respectively. This deduced amino acid sequence shares some common structural features with aminopeptidase N from several moth species, including the consensus zinc-binding motif HEXXHX18E and the GAMEN motif common to gluzincin aminopeptidases. The first 20 amino acid residues at N-termini is hydrophobic transmembrane helix. The sequence of APN-Harm was deposited in GenBank and the accession number is AY181026.
文摘Helicoverpa armigera H. is a polyphagous notorious pest of a number of economic crops including tomato. In tomato, it can reduce its yield as high as 70% due to fruit boring. Although some predators and parasitoids have been recorded in the field for their natural control, their manipulation to a successful crop management is still awaited. To overcome this pest, insecticides play a significant role in its effective crop management program globally. Tomato is an absolutely perishable commodity and needs the specified period required between the application, of conventional pesticides of organophosphate group, and consumption which can hardly be afforded. Therefore, nine insecticides from the new chemistries, which were relatively safer to human and environment, i.e. Coragen (chlorantraniliprole), steward (indoxacarb), Belt (flubendamide), Delegate (spintoram), Volium Flexy (chlorantraniliprole + thiamethaxim), Fipronil (grafter), Proclaim (emamectin benzoate), Pirate (chlofenapyr) and Lufenuron (lufenuron), were tested on tomato crop heavily infested by this pest in the farmers field in districts Lodhran and Bahawalpur (Pakistan) in the month of April 2014. The mortality was compared after 4, 7 and 10 days and treatments were compared with control as well with one another. The average maximum % mortality, i.e. 89.36 and 85.09 of the pest, was observed with Volium Flexy, and Delegate also worked well even after 7 and 10 days. Similarly, the results on the basis of damaged fruits and percent loss of yield pointed out that Chlorantraniliprole, Flubendiamide and Indoxacarb had resulted better as compared with others, although the difference was statistically non significant. These new chemistry pesticides are suggested to be used at tomato crop against Helicoverpa armigera which are best fitted in the IPM program for the control of the pest.
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.
基金supported by the National 863 Program of China(2012AA101502)the Special Fund for Agro-Scientific Research in the Public Interest, China (200903033)
文摘Sublethal effects of chlorantraniliprole to Helicoverpa armigera （Hiibner） larvae were evaluated through exposure of third instar larvae to the insecticide incorporated into an artificial diet. When larvae were fed on the insecticide-containing diet for 7 d, the LC10, LC20, LC40, and LC50values were 3.790, 7.977, 21.577, and 33.121 μg active ingredient L^-1, respectively. Chlorantraniliprole at sublethal concentrations significantly reduced the larval body mass, emergence ratio, adult longevity and egg hatching rate in both the parental and offspring generations. The pupation and copulation rate in the parental generation and the pupal mass in the offspring also strongly decreased. Reproduction was seriously disturbed in both the parental and offspring groups even when only one of the partners was exposed to chlorantraniliprole at larval stages. However, at the lowest concentration of exposure （LC^0）, offspring fecundity was strongly reduced only when both partners were exposed. The net reproductive rate （R0）, intrinsic rate of increase （rm）, and gross reproduction rate （GRR） at LC20, LC40 concentrations were significantly lower than that of the control. Post-exposure effects also included an extended larval developmental time and increased male proportion in both generations. The doubling time （Dt） at LC20, LC40 concentrations as well as gross reproduction rate （GRR） at LC10 concentration were also significantly increased. Chlorantraniliprole might have significant effects on H. armigera population dynamics even at sublethal concentrations on both parental and offspring generations.
基金supported by the National Basic Research Program of China(2012CB114104)the National Natural Science Foundation of China(30871640,31071694)+1 种基金the National High-Tech R&D Program of China(2008AA02Z307)the International Cooperation and Exchange Foundation of NSFC-RS of China(31111130203).
文摘The full-length sequence of the odorant binding protein 5 gene,HarmOBP5,was obtained from an antennae cDNA library of cotton bollworm,Helicoverpa armigera （Hübner）.The cDNA contains a 444 bp open reading frame,encoding a protein with 147 amino acids,namely HarmOBP5.HarmOBP5 was expressed in Escherichia coli and the recombinant protein was purified by affinity chromatography.SDS-PAGE and Western blot analysis demonstrated that the purified protein can be used for further investigation of its binding characteristics.Competitive binding assays with 113 odorant chemicals indicated that HarmOBP5 has strong affinity to some special plant volatiles,including （E）-β-farnesene,ethyl butyrate,ethyl heptanoate,and acetic acid 2-methylbutyl ester.Based on three-dimensional （3D） model of AaegOBP1 from Aedes aegypti,a 3D model of HarmOBP5 was predicted.The model revealed that some key binding residues in HarmOBP5 may play important roles in odorant perception of H.armigera.This study provides clues for better understanding physiological functions of OBPs in H.armigera and other insects.
基金Project supported by the National Basic Research Program(973)of China(No.2006CB102002)the Pilot Project of Knowledge Innovation Program of Chinese Academy of Sciences(No.KSCX2-YW-N-006)National Natural Science Foundation of China(No.30571253,30621003)
文摘The growth, development and consumption of successive three generations of cotton bollworm, Helicoverpa armigera （Htibner）, fed on cotton bolls grown under elevated CO2 （double-ambient vs. ambient） in open-top chambers were examined. Significant decreases in protein, total amino acid, water and nitrogen content and increases in free fatty acid were observed in cotton bolls. Changes in quality of cotton bolls affected the growth, development and food utilization of H. armigera. Significantly longer larval development duration in three successive generations and lower pupal weight of the second and third generations were observed in cotton bollworm fed on cotton bolls grown under elevated CO2. Significantly lower fecundity was also found in successive three generations of H. armigera fed on cotton bolls grown under elevated CO2. The consumption per larva occurred significant increase in successive three generations and frass per larva were also significantly increased during the second and third generations under elevated CO2. Significantly lower relative growth rate, efficiency of conversion of ingested food and significant higher relative consumption rate in successive three generations were observed in cotton bollworm fed on cotton bolls grown under elevated CO2. Significantly lower potential female fecundity, larval numbers and population consumption were found in the second and third generations of cotton bollworm fed on cotton bolls grown under elevated CO2. The integrative effect of higher larval mortality rate and lower adult fecundity resulted in significant decreases in potential population consumption in the latter two generations. The results show that elevated CO2 adversely affects cotton bolls quality, which indicates the potential population dynamics and potential population consumption of cotton bollworm will alleviate the harm to the plants in the future rising CO2 atmosphere.
基金Research was supported by the National Natural Science Fundation of China(30200182)973 Program of Ministry of Science and Techno1ogy,China(001CB109004).
文摘The brush border membrane vesicles (BBMVs) in midgut of Helicoverpa armigera weresuccessfully separated, and most of the Aminopeptidase N (APN) activities in BBMV werepreserved. The 3-[(3-chlor-amidopropyl) dimethylammonio]-1-propane-sulphonate (CHAPS)can enhance the dissolution of BBMV, and phosphatidylinositol-specific phosopholipase C(PI-PLC) can cleave the APN from midgut membrane. The APN was primarily purified usinga Mono-Q column. The results of immunoblotting showed that the 120 and 170kDa proteinsin the BBMV could bind Cry1Ac, and 120kDa APN was a glycosylphosphalidylinositol(GPI)-anchored protein. Two Bt-resistant strains (Bt-P, Bt-M) were obtained after beingselected for more than five years in laboratory using Bt insecticides and Bt transgeniccotton incorporated into diet separately. The resistance of Bt-P and Bt-M were 1083.3and 48.7 times that of susceptible strain. The genes encoding APN1 in midgut ofsusceptible and resistant H.armigera were cloned by PCR and RACE techniques. Theinferred amino acid sequences of APN1 possessed the common character of APN family ininsects. In comparison with APN1 in susceptible strain, three nucleotide mutations wereobserved in the APN1 of Bt-M strain and resulted in two amino acid replace in theputative protein sequences, and eight nucleotide mutations were observed in Bt-P strainand resulted in five amino acid replace.
基金Supported by University Grant Commission(UGC)X plan(Grant No.UGC/TN/GAC-2008)
文摘Objective:To evaluate the antifeedant,insecticidal and growth inhibition activities of Solanum pseudocapsicum(S.pseudocapsicum)seed extracts against Spodoptera litura(S.litura)and Helicotverpa armigera(H.armigera).Methods:Hcxane,diethyl cther,dichloromethanc and ethyl acetate seed extracts were prepared and tested for autifeedant,insecticidal and growth inhibitory activities against fourth instar larvae of&litura and H.armigera.Results:Ethyl acetate extract showed promising antifeedant and insecticidal activities against S.litura and H.armigera.Percentage of deformed larvae,pupae and adults were maximum in treatment of ethyl acetate extract.Percentage of successful adult emergence was deteriorated by seeds on extract treated larvae.Conclusions:Ethyl acetate extracts of S.pseudocapsicum,showed higher efficiency of antifeedant,insecticidal and growth inhibition activities.Hence,it can be used to controll agricultural insect pests,S.litura and H.armigera.
