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Determination of Resistance to Seven Insecticides in Plutella xylostella L. in Fields of Northern Hunan 被引量:1
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作者 王雅菲 魏润洁 +1 位作者 朱航 周小毛 《Agricultural Science & Technology》 CAS 2015年第3期553-555,606,共4页
The resistance of field populations of Plutella xylostella, from the three vegetable producing areas (Nianyuxu Town of Yueyang City, Canggang Town of Changde City and Shatou Town of Yiyang City) in northern Hunan, t... The resistance of field populations of Plutella xylostella, from the three vegetable producing areas (Nianyuxu Town of Yueyang City, Canggang Town of Changde City and Shatou Town of Yiyang City) in northern Hunan, to seven insecticides was determined using leaf dipping method in door. The results showed that Plutella xylostella showed an extremely high-level resistance to beta-cypermethrin (resistance ratio, RR=257.13), a high-level resistance to abamectin (RR=135.15) and indoxacarb (RR=103.08) and a moderate-level resistance to chlorfenapyr and emamectin benzoate. But Plutella xylostella is relatively sensitive to chlorantraniliprole and Bacillus thuringiensis (Bt). Therefore, the prevention of Plutella xylostella in northern Hunan should focus on the alternative use of chlorfenapyr, emamectin benzoate, chlorantraniliprole and Bacillus thuringiensis and avoid the use of beta-cypermethrin so as to delay the generation and development of resistance to insecticides in Plutella xylostella. 展开更多
关键词 plutella xylostella insecticide Drug resistance determination
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Characterization of carboxylesterase PxαE8 and its role in multi-insecticide resistance in Plutella xylostella(L.) 被引量:2
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作者 LI Ran SUN Xi +1 位作者 LIANG Pei GAO Xi-wu 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2022年第6期1713-1721,共9页
Carboxylesterase(CarE)was considered as important phase-I detoxifying enzymes which participated in detoxification of different types of insecticides.Up-regulation of CarE genes has been proved playing a major role in... Carboxylesterase(CarE)was considered as important phase-I detoxifying enzymes which participated in detoxification of different types of insecticides.Up-regulation of CarE genes has been proved playing a major role in insecticide resistance in many pest insects,but its involvement in resistance to insecticides in Plutella xylostella has been rarely reported.In this study,a CarE cDNA named PxαE8 was identified in P.xylostella,which has an open reading frame of 1599 nucleotides and putatively encodes 532 amino acids.The investigation of spatial expression profiles of PxαE8 revealed that it was expressed in all developmental stages,especially in larvae and adults.The body part/tissue-specific expression profiles showed that the PxαE8 mainly expressed in fat body,malpighian tubule and hemolymph of larvae.Further,the relative expression of PxαE8 in two multi-resistant field populations,Hainan(HN)and Guangdong(GD)populations,was found 24.4-and 15.5-fold higher than that in susceptible population,respectively.Knockdown of PxαE8 by RNA interference dramatically increased the mortalities of larvae of HN population treated with LC_(50) of beta-cypermethrin and phoxim by 25.3 and 18.3%,respectively.These results suggested that up-regulation of PxαE8 was involved in resistance to both beta-cypermethrin and phoxim in P.xylostella,which shed light on further understanding of molecular mechanisms of multi-insecticide-resistance in P.xylostella and other pest insects. 展开更多
关键词 plutella xylostella CARBOXYLESTERASE insecticide resistance BETA-CYPERMETHRIN PHOXIM
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CRISPR/Cas9-based functional characterization of Px ABCB1 reveals its roles in the resistance of Plutella xylostella(L.)to Cry1Ac,abamectin and emamectin benzoate 被引量:1
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作者 OUYANG Chun-zheng YE Fan +5 位作者 WU Qing-jun WANG Shao-li Neil CRICKMORE ZHOU Xu-guo GUO Zhao-jiang ZHANG You-jun 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2023年第10期3090-3102,共13页
The identification of functional midgut receptors for pesticidal proteins produced by Bacillus thuringiensis(Bt)is critical for deciphering the molecular mechanism of Bt resistance in insects.Reduced expression of the... The identification of functional midgut receptors for pesticidal proteins produced by Bacillus thuringiensis(Bt)is critical for deciphering the molecular mechanism of Bt resistance in insects.Reduced expression of the PxABCB1 gene was previously found to be associated with Cry1Ac resistance in the diamondback moth,Plutella xylostella(L.).To directly validate the potential receptor role of PxABCB1 and its contribution to Bt Cry1Ac toxicity in P.xylostella,we used CRISPR/Cas9 to generate a homozygous knockout ABCB1KO strain with a 5-bp deletion in exon 3 of its gene.The ABCB1KO strain exhibited a 63-fold resistance to Cry1Ac toxin compared to the parental DBM1Ac-S strain.Intriguingly,the ABCB1KO strain also exhibited significant increases in susceptibility to abamectin and emamectin benzoate.No changes in susceptibility to various other Bt Cry proteins or synthetic insecticides were observed.The knockout strain exhibited no significant fitness costs.Overall,our study indicates that PxABCB1 can protect the insect against avermectin insecticides on one hand,while on the other hand it facilitates the toxic effect of the Bt Cry1Ac toxin.The results of this study will help to inform integrated pest management approaches against this destructive pest. 展开更多
关键词 Bacillus thuringiensis plutella xylostella CRISPR/Cas9 ABCB1 bioinsecticide resistance
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Sex-Related Larval Susceptibility of Diamondback Moth, Plutella xylostella (Lepidoptera:Plutellidae) to some Reduced-Risk Insecticides
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作者 Hongyi Wei Henry Yemisi Fadamiro 《Journal of Agricultural Science and Technology(A)》 2013年第11期870-877,共8页
Five reduced-risk insecticides were compared for their toxicities to male and female third instar larvae of diamondback moth (DBM), Plutella xylostella L. (Lepidoptera:Plutellidae) obtained from a laboratory colo... Five reduced-risk insecticides were compared for their toxicities to male and female third instar larvae of diamondback moth (DBM), Plutella xylostella L. (Lepidoptera:Plutellidae) obtained from a laboratory colony. Leaf-dip bioassays were used to assess larval mortality at different insecticides rates (ranging from 0.01 mg AI/L to 100 mg A1/L) and exposure times (24, 48 and 72 h after treatment). Toxicity of the insecticides generally increased with rate and exposure time. At 72 h after treatment, median lethal concentrations (LCs0s) of methoxyfenozide, spinosad, novaluron, indoxacarb and Bacillus thuringiensis (Bt) against the male third instar larvae of P, xylostella were 0.0524, 0.1117, 0.9149, 1.1939 and 4.4983 mg AI/L, respectively, and were 0.1008, 0.2583, 1.0649, 0.2850 and 5.3053 mg AI/L against female third instar larvae, respectively. At 0.1% of the approximate recommended field rates, methoxyfenozide (60%-75%) and spinosad (53%-57%) were the most toxic, while Bt (30%-45% mortality) was the least toxic to male and female P. xylostella larvae. Spinosad was the fastest acting and the only insecticide that caused significant larval mortality (17%) at 24 h after exposure. Significant sexual differences were recorded in the susceptibility of P. xylostella larvae to two of the insecticides: spinosad and indoxacarb. Male larvae were two-fold more susceptible to spinosad but 10-fold more tolerant to indoxacarb than female larvae. 展开更多
关键词 DBM plutella xylostella insecticideS toxicity susceptibility.
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Biochemical Mechanism of Chlorantraniliprole Resistance in the Diamondback Moth, Plutella xylostella Linnaeus 被引量:14
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作者 HU Zhen-di FENG Xia +5 位作者 LIN Qing-sheng CHEN Huan-yu LI Zhen-yu YIN Fei LIANG Pei GAO Xi-wu 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2014年第11期2452-2459,共8页
The insecticide chlorantraniliprole exhibits good efifcacy and plays an important role in controlling the diamondback moth, Plutella xylostella Linnaeus. However, resistance to chlorantraniliprole has been observed re... The insecticide chlorantraniliprole exhibits good efifcacy and plays an important role in controlling the diamondback moth, Plutella xylostella Linnaeus. However, resistance to chlorantraniliprole has been observed recently in some ifeld populations. At present study, diamondback moths with resistance to chlorantraniliprole (resistant ratio (RR) was 82.18) for biochemical assays were selected. The assays were performed to determine potential resistance mechanisms. The results showed that the selected resistant moths (GDLZ-R) and susceptible moth could be synergized by known metabolic inhibitors such as piperonyl butoxide (PBO), triphenyl phosphate (TPP) and diethyl-maleate (DEM) at different levels (1.68-5.50-fold and 2.20-2.89-fold, respectively), and DEM showed the maximum synergism in both strains. In enzymes assays, a high level of glutathione-S-transferase (GST) was observed in the resistant moth, in contrast, moths that are susceptible to the insecticide had only 1/3 the GST activity of the resistant moths. The analysis of short-term exposure of chlorantraniliprole on biochemical response in the resistant strain also showed that GST activity was signiifcantly elevated after exposure to a sub-lethal concentration of chlorantraniliprole (about 1/3 LC50, 12 mg L-1) 12 and 24 h, respectively. The results show that there is a strong correlation between the enzyme activity and resistance, and GST is likely the main detoxiifcation mechanism responsible for resistance to chlorantraniliprole in P. xylostella L., cytochrome P450 monooxygenase (P450) and carboxy-lesterase (CarE) are involved in to some extent. 展开更多
关键词 plutella xylostella CHLORANTRANILIPROLE resistance biochemical mechanism
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Influence of seasonal migration on evolution of insecticide resistance in Plutella xylostella 被引量:2
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作者 Menglun Wang Bin Zhu +3 位作者 Lei Zhang Yutao Xiao Pei Liang Kongming Wu 《Insect Science》 SCIE CAS CSCD 2022年第2期496-504,共9页
The diamondback moth,Plutella xylostella(L.),is one of the most destructive migratory pest species of cruciferous vegetables worldwide and has developed resistance to most of the insecticides used for its control.The ... The diamondback moth,Plutella xylostella(L.),is one of the most destructive migratory pest species of cruciferous vegetables worldwide and has developed resistance to most of the insecticides used for its control.The migration regularity,migratory behavior,and relationship between flight and reproduction of P.xylostella have been widely reported.However,the effect of migration on insecticide resistance in this pest is still unclear.In this study,the effect of migration on P.xylostella resistance to seven insecticides was investigated using populations across the Bohai Sea that were collected in the early and late seasons during 2017–2019.The bioassay results showed that the early season populations of P.xylostella from south China possessed much higher resistance to insecticides because of intensive insecticide application;alternatively,the late season populations migrated from northeast China,where the insecticides were only used occasionally,showed much lower insecticide resistance.The genome re-sequencing results revealed that,among the eight mutations involved in insecticide resistance,the frequencies of two acetylcholinesterase mutations(A298S and G324A)responsible for organophosphorus insecticide resistance were significantly decreased in the late season populations.The results indicated that P.xylostella migration between tropical and temperate regions significantly delayed the development of insecticide resistance.These findings illustrated the effect of regional migration on the evolution of insecticide resistance in P.xylostella,and provided foundational information for further research on the relationship between migration and insecticide resistance development in other insects. 展开更多
关键词 Bohai Sea insecticide resistance MIGRATION plutella xylostella
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cDNA cloning and characterization of t he carboxylesterase pxCCE016b from the diamondback moth, Plutella xylostella L. 被引量:1
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作者 HU Zhen-di FENG Xia +5 位作者 LIN Qing-sheng CHEN Huan-yu LI Zhen-yu YIN Fei LIANG Pei GAO Xi-wu 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2016年第5期1059-1068,共10页
Carboxylesterase is a multifunctional superfamily and can be found in almost all living organisms. As the metabolic enzymes, carboxylesterases are involved in insecticides resistance in insects for long time. In our p... Carboxylesterase is a multifunctional superfamily and can be found in almost all living organisms. As the metabolic enzymes, carboxylesterases are involved in insecticides resistance in insects for long time. In our previous studies, the enhanced c arboxylesterase activities were found in the chlorantraniliprole resistance strain of diamondback moth(DBM). However, t he related enzyme gene of chlorantraniliprole resistance has not been clear in this strain. Here, a full-length c DNA of carboxylesterase pxCCE016 b was cloned and exogenously expressed in Escherichia coli at the first time, which contained a 1 693 bp open reading frame(ORF) and encoded a protein of 542 amino acids. Sequence analysis showed that this c DNA has a predicted mass of 61.56 k Da and a theoretical isoelectric point value of 5.78. The sequence of deduced amino acid possessed the classical structural features: a type-B carboxylesterase signature 2(EDCLYLNVYTK), a type-B carboxylesterase serine active site(FGGDPENITIFGESAG) and the catalytic triad(S er186, Glu316, and His444). The real-time quantitative PCR(q PCR) analysis showed that t he expression level of the p x CCE016 b was significantly higher in the chlorantraniliprole resistant strain than in the susceptible strain. Furthermore, pxCCE016 b was highly expressed in the midgut and epidermis of the DBM larvae. When the 3rd-instar larvae of resistant DBM were exposed to abamectin, alpha-cypermethrin, chlorantraniliprole, spinosad, c hlorfenapyr and indoxacarb insecticides, the up-regulated expression of pxCCE016 b was observed only in the group treated by chlorantraniliprole. In addition, recombinant vector p ET-pxCCE016 b was constructed with the most coding region(1 293 bp) and large number of soluble recombinant proteins(less than 48 k Da) were expressed successfully with prokaryotic cell. Western blot analysis showed that it was coded by pxCCE016 b. All the above findings provide important information for further f unctional study, although we are uncertainty whether the pxCCE016 b gene is actually i nvolved in chlorantraniliprole resistance. 展开更多
关键词 plutella xylostella CARBOXYLESTERASE CHLORANTRANILIPROLE insecticide resistance pxCCE016b
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Tebufenozide resistance is associated with sex-linked inheritance in Plutella xylostella 被引量:2
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作者 Guang-Chun Cao Zhao-Jun Han 《Insect Science》 SCIE CAS CSCD 2015年第2期235-242,共8页
The diamondback moth (DBM), Plutella xylostella (L.), is a major pest of cruciferous crops. Tebufenozide, a novel nonsteroidal ecdysone agonist, exhibits good efficacy and has played an increasingly important role... The diamondback moth (DBM), Plutella xylostella (L.), is a major pest of cruciferous crops. Tebufenozide, a novel nonsteroidal ecdysone agonist, exhibits good efficacy and has played an increasingly important role in the control of Lepidopteran pests in China. For its resistance management, the genetic basis of tebufenozide resistance was studied using a laboratory selected resistant strain of DBM (resistant ratio, RR = 268). A series of crosses with laboratory susceptible and resistant strains revealed that tebufenozide resistance in this pest was partially biased toward female heredity, with a large difference in RR for F1 (RR = 29) and rF1 progeny (RR = 147). The dominance calculated for these 2 cross progeny was -0.788 and 0.09, respectively. Further analysis showed that the susceptible male and female larvae were similar in theft sensitivity to tebufenozide, but the resistant female larvae showed significantly higher resistance than the resistant male larvae. The heredity of tebufenozide resistance in DBM might be linked with the W sex chromosome, which suggested that DBM has the ability to develop high levels of resistance to tebufenozide. This is the first report of sex-linked inheritance oftebufenozide resistance in P xylostella (L.). 展开更多
关键词 insecticide resistance plutella xylostella sex linkage TEBUFENOZIDE
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Seasonal changes of resistance to fenvalerate and cypermethrin in the larval parasitoid Cotesia plutellae (Hymenoptera: Braconidae)
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作者 GANG WU TADASHI MIYATA 《Insect Science》 SCIE CAS CSCD 2005年第5期351-357,共7页
The seasonal changes of insecticide resistance and stability in hymenopteran Cotesia plutellae, collected from Jianxin, Fuzhou-City, and Shangjie, Minhou-County, Fujian, China, were assessed by using a dry residual fi... The seasonal changes of insecticide resistance and stability in hymenopteran Cotesia plutellae, collected from Jianxin, Fuzhou-City, and Shangjie, Minhou-County, Fujian, China, were assessed by using a dry residual film method. The resistance to two insecticides in the field populations of C. plutellae was not stable under insecticide-free conditions in the insectarium. Compared with susceptible F11 progeny of C. plutellae in the insectarium, the resistance ratios (RR) in F0 parents were 18.4 for fenvalerate and 11.4 for cypermethrin based on LC50 at 9 hours, and 32.8 for fenvalerate and 28.5 for cypermethrin based on LC50 at 24 hours when the parasitoids were left in contact with the insecticides for 1 hour and mortalities were recorded at 9 and 24 hours, respectively. However, the RR in a field population of C. plutellae were 9.2 for fenvalerate and 12.7 for cypermethrin, if the parasitoids were left in contact with the insecticides for 24 hours. The resistances to the two pyrethroids in other field populations collected from Jianxin and Shangjie from November 2000 and July 2004 were also determined. Significant seasonal variations of resistance to the two insecticides in the field populations of C. plutellae were found. The RR were 3.0-18.4 for fenvalerate and 4.8-20.6 for cypermethrin in Jianxin populations from November 2000 to April 2002 based on LC50 at 9 h, and 2.3-13.6 for fenvalerate and 3.6-16.0 for cypermethrin in Shangjie populations from May 2002 to July 2004 based on LC50 at 24 hours. The resistance levels were high in spring and autumn and decreased sharply in summer. In addition, significant recovery from the knocked-down caused by the insecticides was found in the F0 and field populations of C. plutellae which were resistant to fenvalerate and cypermethrin if the parasitoids were left in contact with the pyrethroids for 1 hour. However, no recovery was found in susceptible F11 progeny. 展开更多
关键词 Cotesia plutellae insecticide resistance FENVALERATE CYPERMETHRIN seasonal dynamics
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Effects of abamectin selection on the genetic differentiation within Plutella xylostella (Lepidoptera: Plutellidae) based on amplified fragment length polymorphism
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作者 Xiao-Mao Zhou Qing-Jun Wu +2 位作者 You-Jun Zhang Lian-Yang Bai Xiong-Ying Huang 《Insect Science》 SCIE CAS CSCD 2010年第4期353-360,共8页
Diamondback moth, Plutella xylostella (L.) is one of the most serious insect pests for its remarkable ability to develop resistance to virtually every insecticide that has been used against it. In the present study,... Diamondback moth, Plutella xylostella (L.) is one of the most serious insect pests for its remarkable ability to develop resistance to virtually every insecticide that has been used against it. In the present study, amplified fragment length poly- morphism (AFLP) is used to study the genetic differentiation as well as the effects of abamectin-selection on population genetic differentiation for P. xylostella. A dendrogram was constructed from the matrix of genetic distances using the STATISTICA software (Version 4.5) and unweighted pair-group method with arithmetic averages (UPGMA). The data demonstrated that compared to the susceptible strain (ABM-s), the heterozy- gosity in the abamectin-resistant strain (ABM-r) decreased with the increased selection pressure and resistant level. When the resistance ratio was below 4.3, there was no sig- nificant differentiation at the genome DNA level. When the resistance ratio reached 5.8, a fairly significant differentiation began to appear, and when the resistance ratio reached and exceeded 8.1, there appeared a significant genetic differentiation. The results suggest that abamectin selection is associated with increased genetic polymorphism in P. xylostella. 展开更多
关键词 abamectin resistance AFLP genetic differentiation plutella xylostella
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小菜蛾(Plutella xylostella L.)抗药性的研究现状 被引量:7
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作者 张桂芝 赤国彤 +1 位作者 张金林 白凤红 《河北农业大学学报》 CAS CSCD 北大核心 2003年第z1期201-204,共4页
小菜蛾是十字花科蔬菜上最主要的害虫 ,也是目前产生抗药性最严重的害虫之一。本文就国内外小菜蛾在抗药性发展、抗性消长规律、抗性机理、抗性遗传。
关键词 小菜蛾 抗药性 蔬菜
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Toxic activity of a protein complex purified from Xenorhabdus nematophila HB310 to Plutella xylostella larvae 被引量:7
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作者 Qin-Ying Wang Zi-Yan Nangong +4 位作者 Jun Yang Ping Song Yi Wang Liwang Cui Long Cui 《Insect Science》 CAS CSCD 2012年第3期329-336,共8页
Xenorhabdus nematophila, a Gram-negative proteobacterium belonging to the family Enterobacteriaceae and associated symbiotically with soil entomopathogenic nematodes, Steinernema carpocapsae, is pathogenic to a wide r... Xenorhabdus nematophila, a Gram-negative proteobacterium belonging to the family Enterobacteriaceae and associated symbiotically with soil entomopathogenic nematodes, Steinernema carpocapsae, is pathogenic to a wide range of insects. A protein complex with insecticidal activity was isolated from the cells ofX. nematophila HB310 strain using methods of salting out and native polyacrylamide gel electrophoresis (PAGE). Seven polypeptides ranging 50~250 kDa were well separated from the protein complex (named Xnpt) by sodium dodecyl sulfate (SDS)-PAGE, five of which are identified as XptA2, xptC 1, XptB 1, GroEL and hypothetical protein by matrix-assisted laser desorption- time-of-flight mass spectrometry (MALDI-TOFMS). Xnpt showed high oral virulence to larvae of diamondback moth (DBM), Plutella xylostella L. (Lepidoptera, Plutellidae) as its median lethal concentration (LC50) against second and third instar larvae were 331.45 ng/mL and 553.59 ng/mL at 72 h, respectively. The histological analysis of Xnptfed DBM larvae showed extensive histopathological effects on the midgut. Biochemical analysis indicated that Xnpt markedly inhibited the activities of three important enzymes in the midgut. Overall, our data showed that the protein complex isolated from X. nematophila HB310 induced the antifeedant and death of insects by destroying midgut tissues and inhibiting midgut proteases activities. 展开更多
关键词 insecticidal toxin MIDGUT plutella xylostella proteases Xenorhabdus nematophila
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Management of diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae) by mating disruption 被引量:4
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作者 Qing-Jun Wu Shu-Fa Zhang +3 位作者 Jin-Liang Yao Bao-Yun Xu Shao-Li Wang You-Jun Zhang 《Insect Science》 CAS CSCD 2012年第6期643-648,共6页
Field trials were conducted in China in 2008 and 2009 to evaluate the efficacy of mating disruption (MD) on diamondback moth, Plutella xylostella, in cabbage, Brassica oleracea var. capitata. Effectiveness was posit... Field trials were conducted in China in 2008 and 2009 to evaluate the efficacy of mating disruption (MD) on diamondback moth, Plutella xylostella, in cabbage, Brassica oleracea var. capitata. Effectiveness was positively correlated with the MD dispenser density in the field. A density of 167 MD dispensers per ha produced an average population decrease of about 50% compared to the conventional-practice field. Significant fewer males were captured in pheromone-treated and conventional-practice fields than in the blank control field, but the difference was not significant between the pheromone-treated and conventional-practice fields. In addition, fewer eggs and larvae were observed in pheromone-treated fields. Our results suggest mating disruption coupled with minimal insecticidal supplements is a promising solution for resistance management and control of diamondback moth infestation. 展开更多
关键词 mating disruption plutella xylostella resistance management sexpheromone
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Insecticidal Toxicity of Spilanthol from <i>Spilanthes acmella</i>Murr. against <i>Plutella xylostella</i>L. 被引量:1
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作者 Anuradha Sharma Vishal Kumar +2 位作者 Rameshwar Singh Rattan Neeraj Kumar Bikram Singh 《American Journal of Plant Sciences》 2012年第11期1568-1572,共5页
The present study explored the Spilanthes acmella Murr. for insecticidal principle, a plant of high value. The seed extract showed insecticidal activity against Plutella xylostella. Further, bioassay guided isolation ... The present study explored the Spilanthes acmella Murr. for insecticidal principle, a plant of high value. The seed extract showed insecticidal activity against Plutella xylostella. Further, bioassay guided isolation of bioactive compounds resulted in insecticidal active molecule, which was identified with the help of ESI-MS and NMR. Highest activity of 95 - 100 percent was observed at low dose of 2 g/l with spilanthol, while 60 - 70 and 80 - 90 percent mortality at 5 g/l in crude seed extracts prepared in methanol and hexane after 48 hours exposure, respectively. LC50 of 1.49, 5.14, 5.04, 11.75 g/l was observed with spilanthol, crude seed extract of methanol, hexane, deltamethrin, respectively. The findings indicate the potential of S. acmella with potent insecticidal toxicity for the management of P. xylostella and other insects of agricultural importance. 展开更多
关键词 Spilanthes acmella INSECTICIDAL Activity plutella xylostella Spilanthol Alkamides Extracts TOXICITY
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变温条件下小菜蛾对不同温度效应杀虫剂敏感性的变化
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作者 安静杰 窦亚楠 +4 位作者 郭江龙 袁文龙 党志红 高占林 李耀发 《河北农业大学学报》 CAS CSCD 北大核心 2024年第4期70-75,共6页
为了明确变温环境条件对小菜蛾药剂敏感性的影响,本研究采用食料浸渍法比较了环境温度变化下四氯虫酰胺(正温度效应杀虫剂)和溴虫腈(无温度效应杀虫剂)处理对小菜蛾药剂敏感性的影响;模拟田间变温条件,研究这两种杀虫剂不同施药时间对... 为了明确变温环境条件对小菜蛾药剂敏感性的影响,本研究采用食料浸渍法比较了环境温度变化下四氯虫酰胺(正温度效应杀虫剂)和溴虫腈(无温度效应杀虫剂)处理对小菜蛾药剂敏感性的影响;模拟田间变温条件,研究这两种杀虫剂不同施药时间对小菜蛾毒力的影响。结果表明,四氯虫酰胺施药前,15、25、35℃预处理24 h,其35℃下温度系数分别为+28.53、+25.84和+18.77,均表现为显著的正温度效应,但温度系数随预处理温度上升而降低。溴虫腈在15℃预处理24 h,对小菜蛾表现为弱正温度效应,25℃和35℃下预处理均无温度效应。表明药剂处理前温度变化会影响杀虫剂的温度效应。在模拟田间变温的情况下,四氯虫酰胺经过高温预适应后于16:00(30℃)对小菜蛾的毒力最高(0.41 mg/L),显著高于清晨6:00(22℃)持续低温时的毒力(32.33 mg/L),毒力差异倍数高达78.85倍。一天中无温度效应杀虫剂溴虫腈的毒力受温度变化的影响较小。本研究为利用杀虫剂的温度效应指导田间高效防治小菜蛾提供了依据。 展开更多
关键词 温度系数 小菜蛾 杀虫剂 毒力 变温
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动物源化改型单链抗体的设计、表达及杀虫活性
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作者 仲建锋 高美静 +1 位作者 卢莉娜 张志勇 《农药学学报》 CAS CSCD 北大核心 2024年第6期1069-1079,共11页
抗独特型抗体制备技术是开发新型抗虫蛋白资源的一项创新策略。为设计构造以基因工程抗体为基础的杀虫蛋白资源,制备了对小菜蛾(Plutella xylostella)具有杀虫活性的猪、牛源改型单链抗体(single chain variable fragment,sc Fv)。通过N... 抗独特型抗体制备技术是开发新型抗虫蛋白资源的一项创新策略。为设计构造以基因工程抗体为基础的杀虫蛋白资源,制备了对小菜蛾(Plutella xylostella)具有杀虫活性的猪、牛源改型单链抗体(single chain variable fragment,sc Fv)。通过NCBI数据库搜索猪、牛源抗体重、轻链可变区的8个骨架区,在其中植入人源抗独特型sc Fv 3E1的6个互补决定区,并进行人工合成及替换成噬菌体展示表达载体p IT2;利用间接竞争ELISA对动物源化改型sc Fv进行分型鉴定;利用表面等离子共振(surface plasmon resonance,SPR)技术分析动物源化改型sc Fv与小菜蛾刷状缘膜囊泡(brush border membrane vesicles,BBMV)的结合能力;采用浸叶法测定动物源化改型sc Fv对小菜蛾的生物活性。通过替换表达载体获得了猪源swine-3E1-p IT2质粒和牛源bovine-3E1-p IT2质粒,结果显示:swine-p IT2和bovine-p IT2表达上清液对Cry1B与其多克隆抗体(p Ab)结合的抑制率分别为44.3%和43.0%,模拟抗原Cry1B的某些结构功能,发现两者均属于β型抗独特型sc Fv。SPR分析发现,swine-p IT2、bovine-p IT2与小菜蛾BBMV的结合能力分别为209.48和195.31 RU。生物活性测定结果显示,改造后的swinep IT2和bovine-p IT2对小菜蛾幼虫均具有杀虫活性,LD_(50)值分别为5.90×10^(7)和6.22×10^(7)CFU/m L,均低于亲本3E1的6.86×10^(7)CFU/m L;并且改造后2种材料的毒力回归方程的斜率分别为1.50和1.48,均高于3E1(1.23)。综上,动物源化swine-p IT2和bovine-p IT2改型sc Fv属β型抗独特型sc Fv,能够与小菜蛾的BBMV结合,且具有杀虫活性,为新型生物农药蛋白资源挖掘提供了新的思路。 展开更多
关键词 动物源化 BT毒素 改型抗体 杀虫活性 表面等离子共振 小菜蛾
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小菜蛾杀虫抗体的对靶设计及验证
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作者 谢雅晶 杨立应 +8 位作者 胡晓丹 徐重新 张霄 高美静 卢莉娜 仲建锋 朱庆 刘媛 刘贤金 《江苏农业学报》 CSCD 北大核心 2024年第7期1212-1219,共8页
本研究拟通过模拟Cry毒素创制新型杀虫蛋白质用于小菜蛾防治,主要利用三维结构模拟及分子对接技术,以前期获得的抗Cry1Ab抗体为模板设计2个基因工程抗体(GEAb)。其中,具有杀虫活性的GEAb-GGCC与小菜蛾中肠刷状边缘膜囊泡(BBMV)具有较高... 本研究拟通过模拟Cry毒素创制新型杀虫蛋白质用于小菜蛾防治,主要利用三维结构模拟及分子对接技术,以前期获得的抗Cry1Ab抗体为模板设计2个基因工程抗体(GEAb)。其中,具有杀虫活性的GEAb-GGCC与小菜蛾中肠刷状边缘膜囊泡(BBMV)具有较高结合活性,并与Cry1A和Cry1B具有重叠的BBMV结合位点。BBMV免疫沉淀分析鉴定结果显示,与GEAb-GGCC结合的中肠蛋白质包括氨肽酶N(APN)、V-ATP酶B亚基和polycalin。由于GEAb-GGCC缺乏Cry1A类蛋白质中负责成孔的α-螺旋结构,推测GEAb-GGCC或通过结合小菜蛾中肠受体激活下游信号通路,引起中肠损伤,导致虫体死亡。 展开更多
关键词 对靶设计 杀虫基因工程抗体 分子对接 小菜蛾 Cry毒素
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天维菌素B杀虫活性测定及防治小菜蛾复配配方筛选
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作者 陈滢冲 江雅琴 +5 位作者 陈静 李威汛 周梦诗 王继栋 陈杰 袁静 《农药学学报》 CAS CSCD 北大核心 2024年第4期765-772,共8页
天维菌素B是中国自主创制的新型大环素内酯类杀虫剂。为挖掘其市场潜力,延缓害虫抗药性的产生,本研究采用浸叶法测定了天维菌素B对小菜蛾Plutella xylostella、玉米螟Pyrausta nubilalis、美国白蛾Hyphantria cunea等10种害虫的活性;采... 天维菌素B是中国自主创制的新型大环素内酯类杀虫剂。为挖掘其市场潜力,延缓害虫抗药性的产生,本研究采用浸叶法测定了天维菌素B对小菜蛾Plutella xylostella、玉米螟Pyrausta nubilalis、美国白蛾Hyphantria cunea等10种害虫的活性;采用喷雾法测定了其对棉蚜Aphis gossypii、二斑叶螨Tetranychus urticae和美洲斑潜蝇Liriomyza sativae的活性;采用孙云沛共毒系数法评价了其与硫虫酰胺、虫螨腈和茚虫威复配对小菜蛾的增效作用,并选择最优配比进行了田间药效试验。室内活性测定结果表明:天维菌素B对小菜蛾、玉米螟和二斑叶螨具有优异的杀虫、杀螨活性,LC50值分别为0.115、0.043和0.012 mg/L,活性显著高于或与阿维菌素相当;对黏虫Mythimna separata、美国白蛾、稻纵卷叶螟Cnaphalocrocis medinalis、西花蓟马Frankliniella occidentalis和棉蚜也具有较好的活性,LC50值为0.337~9.784 mg/L。复配配方筛选结果显示:天维菌素B与硫虫酰胺、茚虫威的各组配比均无增效作用;而天维菌素B与虫螨腈在质量比为1:14时的共毒系数为178.59,增效作用明显。选择天维菌素B与虫螨腈质量比1:14复配混合物开展防治小菜蛾的田间药效试验,结果显示,该复配组合在有效成分用量75 g/hm2时,药后7 d和14 d的防效均可达80%以上。研究表明,天维菌素B对小菜蛾、二斑叶螨、棉蚜等多种害虫具有较好的活性,杀虫谱广,具有较好的市场开发前景;天维菌素B与虫螨腈按质量比1:14复配具有增效作用,可以用于防治田间小菜蛾,并有助于延缓其抗药性的产生。 展开更多
关键词 天维菌素B 农药复配 虫螨腈 杀虫活性 小菜蛾
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小菜蛾RyR调控蛋白FKBP基因克隆及其时空表达谱
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作者 张万里 葛天成 +4 位作者 陈琦椿 彭争科 肖勇 李振宇 尹飞 《南方农业学报》 CAS CSCD 北大核心 2024年第10期3023-3032,共10页
【目的】明确FK506结合蛋白(FKBP)基因在小菜蛾对氯虫苯甲酰胺抗性形成中的作用,筛选出起主要作用的关键FKBP基因,为阐明小菜蛾抗双酰胺类杀虫剂的分子机理提供新思路。【方法】以抗氯虫苯甲酰胺种群和敏感种群小菜蛾为研究对象,采用浸... 【目的】明确FK506结合蛋白(FKBP)基因在小菜蛾对氯虫苯甲酰胺抗性形成中的作用,筛选出起主要作用的关键FKBP基因,为阐明小菜蛾抗双酰胺类杀虫剂的分子机理提供新思路。【方法】以抗氯虫苯甲酰胺种群和敏感种群小菜蛾为研究对象,采用浸叶法测定不同种群小菜蛾对氯虫苯甲酰胺的敏感性差异;利用cDNA末端快速扩增(RACE)方法克隆3个FKBP基因(FKBP8、FKBP12和FKBP52)的全长序列,并通过GSDS 2.0、CDD和MEME在线分析软件分析其基因结构与蛋白保守结构域和Motif;采用实时荧光定量PCR分析抗性种群不同发育阶段和敏感、抗性种群4龄幼虫不同组织FKBP基因的时空表达特性。【结果】生物测定结果显示,抗性种群小菜蛾对氯虫苯甲酰胺的抗性倍数为122.67倍,属于高水平抗性。实时荧光定量PCR分析发现,抗性种群小菜蛾体内3个FKBP基因的相对表达量均显著高于敏感种群(P<0.05,下同),其中FKBP8基因的表达差异最显著,是敏感种群的968倍。通过RACE方法克隆得到FKBP52、FKBP8和FKBP12基因的全长序列,长度分别为1473、1525和649 bp,分别编码423、307和108个氨基酸残基,其蛋白均为亲水蛋白。FKBP52、FKBP8和FKBP12基因具有相同的基因结构,FKBP52和FKBP12蛋白具有相同的保守结构域FkpA,而FKBP8蛋白具有2个不同的保守结构域。抗性种群小菜蛾4龄幼虫体内3个FKBP基因表达量均高于蛹期,其中FKBP12和FKBP8基因在4龄幼虫中的表达量最高,显著高于其他龄期和蛹期。FKBP8基因在抗性种群小菜蛾中肠和表皮中的表达量显著高于敏感种群,分别是敏感种群的3.2和3.7倍。【结论】FKBP52、FKBP8和FKBP12基因在氯虫苯甲酰胺抗性种群小菜蛾中均过量表达,其中FKBP8基因在高抗氯虫苯甲酰胺种群小菜蛾的中肠高表达,表明其可能通过过量表达参与小菜蛾对氯虫苯甲酰胺的抗药性。 展开更多
关键词 FK506结合蛋白 FKBP基因 小菜蛾 氯虫苯甲酰胺 时空表达特性 抗药性
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含有伯酰胺和氰基的苯基噻唑衍生物的设计、合成及杀虫活性评价
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作者 杨帅 唐豪毅 +1 位作者 赵晨 徐汉虹 《华南农业大学学报》 CAS CSCD 北大核心 2024年第3期354-363,共10页
【目的】噻唑是一类具有广泛生物活性的五元杂环化合物,本文设计并合成一系列含有伯酰胺及氰基的苯基噻唑衍生物,进一步挖掘噻唑骨架在防治鳞翅目害虫领域的潜力。【方法】以不同取代基的苯甲酸为原料合成含有伯酰胺及氰基的噻唑衍生物... 【目的】噻唑是一类具有广泛生物活性的五元杂环化合物,本文设计并合成一系列含有伯酰胺及氰基的苯基噻唑衍生物,进一步挖掘噻唑骨架在防治鳞翅目害虫领域的潜力。【方法】以不同取代基的苯甲酸为原料合成含有伯酰胺及氰基的噻唑衍生物,并经1H NMR、13C NMR和ESI-MS确证结构。采用浸液法测定目标化合物对小菜蛾Plutella xylostella的生物活性,并通过密度泛函理论(Density functional theory,DFT)计算从分子轨道层面解释该骨架具有杀虫活性的原因和差异。【结果】经结构确证,合成了22个含有伯酰胺及氰基的噻唑衍生物,其中6个化合物(6d、7d、13a、13b、14a和14b)对小菜蛾表现出很好的杀虫活性(校正死亡率>50%),尤其是化合物6d和7d,对小菜蛾的LC50达到了23.94和30.37 mg/L。构效关系表明,苯环仅取代4−CF3且5−位噻唑未取代时,表现出最优的杀虫活性。DFT计算结果表明,4−三氟甲基苯基噻唑既传递电子又接受电子,且化合物6d比7d具有更小的HOMO−LUMO间隙(ΔE)。【结论】化合物6d可作为先导化合物进一步开发,本研究可为含有噻唑骨架杀虫剂的分子设计与优化提供有价值的线索。 展开更多
关键词 苯基噻唑 合成 杀虫活性 小菜蛾 密度泛函理论
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