In assessing an insect-resistant genetically engineered (IRGE) crop before its commercialization, researchers normally use so-called "Tier-1 assays" as the initial step to determine the effects of the crop on non-...In assessing an insect-resistant genetically engineered (IRGE) crop before its commercialization, researchers normally use so-called "Tier-1 assays" as the initial step to determine the effects of the crop on non-target organisms. In these tests, the insecticidal proteins (IPs) produced by the IRGEs are added to the diets of test organisms in the laboratory. Test organisms in such assays can be directly exposed to much higher concentrations of the test IPs than they would encounter in the field. The results of Tier-1 assays are thus more conservative than those generated in studies in which the organisms are exposed to the IPs by feeding on IRGE plant tissue or in the case of predators or parasites, by feeding on invertebrate prey or hosts that have fed on IRGE plant tissue. In this report, we consider three important factors that must be considered in Tier-1 assays: (i) methods for delivery of the IP to the test organisms; (ii) the need for and selection of compounds used as positive controls; and (iii) methods for monitoring the concentration, stability and bioactivity of the IP during the assay. We also analyze the existing data from Tier-1 assays regarding the toxicity of Bt Cry proteins to non-target arthropod species. The data indicate that the widely used Bt proteins have no direct toxicity to non-target organisms展开更多
The fall armyworm Spodoptera frugiperda,an invasive insect pest native to the Americas,has established populations throughout eastern China.The North China Plain一a key corn-producing area in East China with a unique ...The fall armyworm Spodoptera frugiperda,an invasive insect pest native to the Americas,has established populations throughout eastern China.The North China Plain一a key corn-producing area in East China with a unique topography一was invaded by fall armyworm in 2019 and is seriously threatened by this migratory pest.However,the spatiotemporal extent of the migratory movements of fall armyworm from the North China Plain remains poorly understood.Using an air transport-based trajectory modeling approach that incorporates flight behavior,we simulated the potential nocturnal migration trajectories of fall armyworm from the North China Plain based on historical meteorological data from June to October of 2015-2019,and examined the night-time atmospheric conditions associated with their possible flights.The emigration patterns showed monthly variation in the main landing area and common migration direction.The displacement of newly emerged moths from the North China Plain was concentrated in the Northeast China Plain(including Liaoning,Jilin and Heilongjiang provinces)before late summer,after which they were most likely to undertake return flights to the south(especially into Hubei,Anhui and Hunan provinces).This southwest-northeast aerial migration corridor follows the topography of East China and is affected by the East Asian monsoon.These topographic-atmospheric conditions have resulted in the North China Plain becoming a key stopover for fall armyworm populations engaging in multigenerational long-distance migration across East China.These findings contribute to our knowledge of fall army-worm migration and will aid in the implementation of management and control strategies against this highly migratory agricultural pest.展开更多
Helicoverpa armigera, cotton bollworm, is one of the most disastrous pests worldwide, threatening various food and economic crops. Functional genomic tools may provide efficient approaches for its management. The clus...Helicoverpa armigera, cotton bollworm, is one of the most disastrous pests worldwide, threatening various food and economic crops. Functional genomic tools may provide efficient approaches for its management. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, dependent on a single guide RNA (sgRNA), has been used to induce indels for targeted mutagenesis in cotton bollworm. However, genomic deletions may be more desirable to disrupt the function of noncoding genes or regulatory sequences. By injecting two sgRNAs with Cas9 protein targeting different exons, we obtained predictable genomic deletions of several hundred bases. We achieved this type of modification with different combinations of sgRNA pairs, including HaCad and HaABCC2. Our finding indicated that CRISPR/Cas9 can be used as an efficient tool to engineer genomes with chromosomal deletion in H. armigera.展开更多
Aphis gossypii Glover (Hemiptera: Aphididae) is a key pest in cotton crops, notably owing to its increasing resistance to commonly used pesticides. Such resistance prompts for the development of integrated pest man...Aphis gossypii Glover (Hemiptera: Aphididae) is a key pest in cotton crops, notably owing to its increasing resistance to commonly used pesticides. Such resistance prompts for the development of integrated pest management (IPM) programs that include novel pesticides being effective against the aphid. In the present study, we assessed lethal and sublethal effects of cycloxaprid, a novel chiral neonicotinoid pesticide developed in China, on A. gossypii. The lethal concentration at 50% (LCs0) value of cycloxaprid on A. gossypii was estimated, using the dipping method, at 7.73 mg/L. The impact of a sublethal concentration (LC10) and a lethal concentration (LC40) of cycloxaprid on A. gossypii population growth and feeding behavior (using electrical penetration graph technique [EPG]), and its transgenerational effect were further assessed. Adult longevity and fecundity significantly decreased after exposure to LEa0 or LC10 of cycloxaprid. Cycloxaprid with sublethal concentrations (especially LC40) had negative effects on phloem ingestion by A. gossypii. Additionally, the offspring of the adults exposed to LC40 of cycloxaprid had shorter nymphal development duration and adult longevity than the control, and those from LC10 and LC40 treatments had lower adult fecundity and net productive rate. We demonstrated that cycloxaprid is a pesticide showing both lethal and sublethal activities, and transgenerational effects on,4. gossypii; it may be useful for implementation in IPM programs against this aphid pest.展开更多
Brush border membrane vesicles (BBMV) isolated from insect midguts have been widely used to study CrylA binding proteins. Sample preparation is important in two- dimensional electrophoresis (2-DE), so to determine...Brush border membrane vesicles (BBMV) isolated from insect midguts have been widely used to study CrylA binding proteins. Sample preparation is important in two- dimensional electrophoresis (2-DE), so to determine a suitable BBMV preparation method in Helicoverpa armigera for 2-DE, we compared three published BBMV preparation methods mostly used in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS- PAGE). All methods yielded similar types and numbers of binding proteins, but in different quantities. The Abdul-Rauf and Ellar protocol was the best of the three, but had limitations. Sufficient protein quantity is important for research involving limited numbers of insects, such as studies of insect resistance to Bacillus thuringiensis in the field. Consequently, we integrated the three BBMV isolation methods into a single protocol that yielded high quantities of BBMV proteins from H. armigera larval midguts, which proved suitable for 2- DE analysis.展开更多
The flight activity of Spodoptera litura in tethered conditions is evaluated using a computer-mediated flight-mill in the laboratory. The 3-4-day-old moths fly longer and farther than those of other ages. Male and fem...The flight activity of Spodoptera litura in tethered conditions is evaluated using a computer-mediated flight-mill in the laboratory. The 3-4-day-old moths fly longer and farther than those of other ages. Male and female moths exhibit similar flight activity. Mating status does not influence the flight duration and distance of 2-day-old females. However, these two flight parameters with a 6-day-old mated female is significantly lower than that of tmmated ones. The optimum temperature for flight ranged from 16-24℃, whereas the optimum RH ranged from 60%--100%. During 72-h period, the total flight duration and distance of 1-day-old male and female moths were 19.6 h (± 5.8) and 83.3 km (± 28.4), and 24.0 h (± 7.0) and 105.4 km (± 37.4), respectively. These results indicate that S. litura has a great potential to undertake long-distance migratory flights.展开更多
The rice stem borer, Chilo suppressalis Walker is one of the most important insect pests on rice in Asia, north Africa and southern Europe. Transgenic Bt rice has been developed in the laboratory with good resistance ...The rice stem borer, Chilo suppressalis Walker is one of the most important insect pests on rice in Asia, north Africa and southern Europe. Transgenic Bt rice has been developed in the laboratory with good resistance to this pest and other Lepidopteran insects, which will provide a possible alternative tool for this pest control. The full-length cDNAs encoding an aminopeptidase N (CsAPN) and a cadherin (CsCad) were cloned from C. suppressalis. CsAPN showed common features of, and high identities to, other insect APNs in its deduced amino acid sequence. Although a full-length cDNA encoding cadherin-like protein has been reported in GenBank, the newly isolated cadherin here (CsCad) showed some differences in its amino acid sequence, especially at the 7th cadherin repeat region (CR7), which indicated the newly isolated CsCad might be another allele. CsAPN and CsCad were successfully expressed in insect Tn cells, and the blot analysis showed these two proteins could bind Bt toxin CrylAb. The results will provide valuable information for the studies of toxin mode of action and the possible toxin resistance mechanisms in this pest.展开更多
The objective of this study was to identify rice genes that are in response to the striped stem borer (SSB) (Chilo suppressalis Walker) feeding at the first to second larval stage. Using combined suppression subtr...The objective of this study was to identify rice genes that are in response to the striped stem borer (SSB) (Chilo suppressalis Walker) feeding at the first to second larval stage. Using combined suppression subtractive hybridization (SSH) and dot blot approaches, we analyzed the induced defense genes that took place during the first 72 h of infesting intact rice (Oryza sativa L.) plants in sheath tissues with SSB larvae. By sequencing the whole SSH library, 39 expressed sequence tags involved in disease stress, insect stress or other stress responses were identified to be up-regulated by SSB larvae feeding. Among these genes, rice allene oxide cyclase (AOC), terpene synthase (TPS) and four proteinase inhibitor (PI) genes were up-regulated by SSB larvae feeding. Real-time quantitative reverse transcription polymerase chain reaction analysis showed that four rice PI genes were already up-regulated at 6 h, and reached peaks between 6 h to 12 h. In addition, the transcription ofgene involving in jasmonate signaling pathway such as allene oxide cyclase (AOC) concerning rice early defense response to SSB feeding was activated after rice feeding by SSB for 2 h. Although the expression office terpene synthase (TPS) gene, involved in the biosynthesis ofmonoterpenes or diterpenes, was already up-regulated at 7 h, a significant increase in the expression was delayed until 12 h and reached its peak at 24 h. The present study identified six SSB-response genes and their expression patterns, which provides evidence and information to understand insect stress-response in plants.展开更多
文摘In assessing an insect-resistant genetically engineered (IRGE) crop before its commercialization, researchers normally use so-called "Tier-1 assays" as the initial step to determine the effects of the crop on non-target organisms. In these tests, the insecticidal proteins (IPs) produced by the IRGEs are added to the diets of test organisms in the laboratory. Test organisms in such assays can be directly exposed to much higher concentrations of the test IPs than they would encounter in the field. The results of Tier-1 assays are thus more conservative than those generated in studies in which the organisms are exposed to the IPs by feeding on IRGE plant tissue or in the case of predators or parasites, by feeding on invertebrate prey or hosts that have fed on IRGE plant tissue. In this report, we consider three important factors that must be considered in Tier-1 assays: (i) methods for delivery of the IP to the test organisms; (ii) the need for and selection of compounds used as positive controls; and (iii) methods for monitoring the concentration, stability and bioactivity of the IP during the assay. We also analyze the existing data from Tier-1 assays regarding the toxicity of Bt Cry proteins to non-target arthropod species. The data indicate that the widely used Bt proteins have no direct toxicity to non-target organisms
基金the National Natural Science Foundation of China(31901873 and 31727901)the China Postdoctoral Science Foundation(2019M660896)+1 种基金the Central Public-interest Scientific Institution Basal Research Fund,China(CAAS-ZDRW202007,Y2019YJ06)the National Key R&D Program of China(2019YFD0300105).
文摘The fall armyworm Spodoptera frugiperda,an invasive insect pest native to the Americas,has established populations throughout eastern China.The North China Plain一a key corn-producing area in East China with a unique topography一was invaded by fall armyworm in 2019 and is seriously threatened by this migratory pest.However,the spatiotemporal extent of the migratory movements of fall armyworm from the North China Plain remains poorly understood.Using an air transport-based trajectory modeling approach that incorporates flight behavior,we simulated the potential nocturnal migration trajectories of fall armyworm from the North China Plain based on historical meteorological data from June to October of 2015-2019,and examined the night-time atmospheric conditions associated with their possible flights.The emigration patterns showed monthly variation in the main landing area and common migration direction.The displacement of newly emerged moths from the North China Plain was concentrated in the Northeast China Plain(including Liaoning,Jilin and Heilongjiang provinces)before late summer,after which they were most likely to undertake return flights to the south(especially into Hubei,Anhui and Hunan provinces).This southwest-northeast aerial migration corridor follows the topography of East China and is affected by the East Asian monsoon.These topographic-atmospheric conditions have resulted in the North China Plain becoming a key stopover for fall armyworm populations engaging in multigenerational long-distance migration across East China.These findings contribute to our knowledge of fall army-worm migration and will aid in the implementation of management and control strategies against this highly migratory agricultural pest.
基金the National Natural Science Foundation of China (No.31601646)Dapeng New District Industry Development Funds (KY20160103)Shenzhen Science and Technology Project (JCYJ20170303154440838).
文摘Helicoverpa armigera, cotton bollworm, is one of the most disastrous pests worldwide, threatening various food and economic crops. Functional genomic tools may provide efficient approaches for its management. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system, dependent on a single guide RNA (sgRNA), has been used to induce indels for targeted mutagenesis in cotton bollworm. However, genomic deletions may be more desirable to disrupt the function of noncoding genes or regulatory sequences. By injecting two sgRNAs with Cas9 protein targeting different exons, we obtained predictable genomic deletions of several hundred bases. We achieved this type of modification with different combinations of sgRNA pairs, including HaCad and HaABCC2. Our finding indicated that CRISPR/Cas9 can be used as an efficient tool to engineer genomes with chromosomal deletion in H. armigera.
文摘Aphis gossypii Glover (Hemiptera: Aphididae) is a key pest in cotton crops, notably owing to its increasing resistance to commonly used pesticides. Such resistance prompts for the development of integrated pest management (IPM) programs that include novel pesticides being effective against the aphid. In the present study, we assessed lethal and sublethal effects of cycloxaprid, a novel chiral neonicotinoid pesticide developed in China, on A. gossypii. The lethal concentration at 50% (LCs0) value of cycloxaprid on A. gossypii was estimated, using the dipping method, at 7.73 mg/L. The impact of a sublethal concentration (LC10) and a lethal concentration (LC40) of cycloxaprid on A. gossypii population growth and feeding behavior (using electrical penetration graph technique [EPG]), and its transgenerational effect were further assessed. Adult longevity and fecundity significantly decreased after exposure to LEa0 or LC10 of cycloxaprid. Cycloxaprid with sublethal concentrations (especially LC40) had negative effects on phloem ingestion by A. gossypii. Additionally, the offspring of the adults exposed to LC40 of cycloxaprid had shorter nymphal development duration and adult longevity than the control, and those from LC10 and LC40 treatments had lower adult fecundity and net productive rate. We demonstrated that cycloxaprid is a pesticide showing both lethal and sublethal activities, and transgenerational effects on,4. gossypii; it may be useful for implementation in IPM programs against this aphid pest.
基金This paper was contributed to the International Symposium on Insect Midgut Biology, April 7-11, 2008, Guangzhou, China.Acknowledgments This research was supported by the National Natural Science Foundation of China (30771424) and National Basic Research Program of China (973-2007CB 109204). We thank Dr. Jun Zhao (University of West Virginia, USA) for reviewing this manuscript.
文摘Brush border membrane vesicles (BBMV) isolated from insect midguts have been widely used to study CrylA binding proteins. Sample preparation is important in two- dimensional electrophoresis (2-DE), so to determine a suitable BBMV preparation method in Helicoverpa armigera for 2-DE, we compared three published BBMV preparation methods mostly used in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS- PAGE). All methods yielded similar types and numbers of binding proteins, but in different quantities. The Abdul-Rauf and Ellar protocol was the best of the three, but had limitations. Sufficient protein quantity is important for research involving limited numbers of insects, such as studies of insect resistance to Bacillus thuringiensis in the field. Consequently, we integrated the three BBMV isolation methods into a single protocol that yielded high quantities of BBMV proteins from H. armigera larval midguts, which proved suitable for 2- DE analysis.
基金We thank Dr. Yong-Jun Zhang, Dr. Gui-Rong Wang and Dr. Ge-Mei Liang for help in this work, and Mrs. LanZhi Han, Guo-Ping Li, Hong-Hua Su, and Mrs. Ke-Jian Lin, Shu-Dong Luo for their help with the data analysis, and Dr. Gul Zamin Khan for reviewing this paper critically. This research was supported by financial assistance from the National Natural Science Foundation of China (30625028), Chinese Ministry of Science and Technology (2006CB 102007, 2006BAD08A07).
文摘The flight activity of Spodoptera litura in tethered conditions is evaluated using a computer-mediated flight-mill in the laboratory. The 3-4-day-old moths fly longer and farther than those of other ages. Male and female moths exhibit similar flight activity. Mating status does not influence the flight duration and distance of 2-day-old females. However, these two flight parameters with a 6-day-old mated female is significantly lower than that of tmmated ones. The optimum temperature for flight ranged from 16-24℃, whereas the optimum RH ranged from 60%--100%. During 72-h period, the total flight duration and distance of 1-day-old male and female moths were 19.6 h (± 5.8) and 83.3 km (± 28.4), and 24.0 h (± 7.0) and 105.4 km (± 37.4), respectively. These results indicate that S. litura has a great potential to undertake long-distance migratory flights.
文摘The rice stem borer, Chilo suppressalis Walker is one of the most important insect pests on rice in Asia, north Africa and southern Europe. Transgenic Bt rice has been developed in the laboratory with good resistance to this pest and other Lepidopteran insects, which will provide a possible alternative tool for this pest control. The full-length cDNAs encoding an aminopeptidase N (CsAPN) and a cadherin (CsCad) were cloned from C. suppressalis. CsAPN showed common features of, and high identities to, other insect APNs in its deduced amino acid sequence. Although a full-length cDNA encoding cadherin-like protein has been reported in GenBank, the newly isolated cadherin here (CsCad) showed some differences in its amino acid sequence, especially at the 7th cadherin repeat region (CR7), which indicated the newly isolated CsCad might be another allele. CsAPN and CsCad were successfully expressed in insect Tn cells, and the blot analysis showed these two proteins could bind Bt toxin CrylAb. The results will provide valuable information for the studies of toxin mode of action and the possible toxin resistance mechanisms in this pest.
基金Acknowledgments We greatly appreciate the grant support from National Natural Science Foundation of China (No. 30871640, No. 30330410), China national "973" Basic Research Program (No. 2007CB 109202), and Research Foundation of State Key Laboratory for Biology of Plant Diseases and Insect Pests (SKL2007SR01).
文摘The objective of this study was to identify rice genes that are in response to the striped stem borer (SSB) (Chilo suppressalis Walker) feeding at the first to second larval stage. Using combined suppression subtractive hybridization (SSH) and dot blot approaches, we analyzed the induced defense genes that took place during the first 72 h of infesting intact rice (Oryza sativa L.) plants in sheath tissues with SSB larvae. By sequencing the whole SSH library, 39 expressed sequence tags involved in disease stress, insect stress or other stress responses were identified to be up-regulated by SSB larvae feeding. Among these genes, rice allene oxide cyclase (AOC), terpene synthase (TPS) and four proteinase inhibitor (PI) genes were up-regulated by SSB larvae feeding. Real-time quantitative reverse transcription polymerase chain reaction analysis showed that four rice PI genes were already up-regulated at 6 h, and reached peaks between 6 h to 12 h. In addition, the transcription ofgene involving in jasmonate signaling pathway such as allene oxide cyclase (AOC) concerning rice early defense response to SSB feeding was activated after rice feeding by SSB for 2 h. Although the expression office terpene synthase (TPS) gene, involved in the biosynthesis ofmonoterpenes or diterpenes, was already up-regulated at 7 h, a significant increase in the expression was delayed until 12 h and reached its peak at 24 h. The present study identified six SSB-response genes and their expression patterns, which provides evidence and information to understand insect stress-response in plants.