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 effects of mepiquat chloride(DPC)on the Cry1Ac protein content in Bacillus thuringiensis(Bt)cotton boll shells under high temperature and drought stress were investigated to provide a theoretical reference for Bt ...The effects of mepiquat chloride(DPC)on the Cry1Ac protein content in Bacillus thuringiensis(Bt)cotton boll shells under high temperature and drought stress were investigated to provide a theoretical reference for Bt cotton breeding and high-yield and-efficiency cotton cultivation.This study was conducted using Bt cotton cultivar‘Sikang 3'during the 2020 and 2021 growing seasons at Yangzhou University Farm,Yangzhou,Jiangsu Province,China.Potted cotton plants were exposed to high temperature and drought stress,and sprayed with either 20 mg L^(-1)DPC or water(CK).Seven days after treatment,the Cry1Ac protein content,α-ketoglutarate content,pyruvic acid content,glutamate synthase activity,glutamic oxaloacetic transaminase activity,soluble protein content,and amino acid content were measured,and transcriptome sequencing was performed.DESeq was used for differential gene analysis.Under the DPC treatment,the Cry1Ac protein content increased by 4.7-11.9% compared to CK.Theα-ketoglutarate content,pyruvic acid content,glutamate synthase activity,glutamic oxaloacetic transaminase activity,soluble protein content,and amino acid content all increased.Transcriptome analysis revealed 7,542 upregulated genes and 10,449 downregulated genes for DPC vs.CK.Gene ontology(GO)and Kyoto Encyclopedia of Gene and Genomes(KEGG)analyses showed that the differentially expressed genes were mainly involved in biological processes,such as carbon and amino acid metabolism.For example,genes encoding 6-phosphofructokinase,pyruvate kinase,glutamic pyruvate transaminase,pyruvate dehydrogenase,citrate synthase,isocitrate dehydrogenase,2-oxoglutarate dehydrogenase,glutamate synthase,1-pyrroline-5-carboxylate dehydrogenase,glutamic oxaloacetic transaminase,amino-acid N-acetyltransferase,and acetylornithine deacetylase were all significantly upregulated.The DPC treatment increased pyruvate,α-ketoglutarate,and oxaloacetate by increasing the operational rate of the glycolytic pathway of the citric acid cycle.It also significantly upregulated the gen展开更多
Studies on the behavior of Helicoverpa armigera in relation to soybean cultivars expressing Bt proteins are fundamentally important for IPM. We determine the no-choice and feeding choice of H. armigera, the no-choice ...Studies on the behavior of Helicoverpa armigera in relation to soybean cultivars expressing Bt proteins are fundamentally important for IPM. We determine the no-choice and feeding choice of H. armigera, the no-choice and oviposition choice in different Bt and non-Bt soybeans cultivars. In the first step it was carried out the experiment to determine the no-choice and food choice in test for 12 Bt and non-Bt soybean cultivars. Simultaneously the consumption was determined. In a second step it was evaluated the no-choice and oviposition choice in different soybean cultivars. Helicoverpa armigera caterpillars showed neither attractiveness and preference for food nor attractiveness and oviposition preference for Bt and non-Bt soybean leaves when simultaneously contrasting the tested cultivars. This study showed that Bt toxin did not influence the oviposition preference, and H. armigera adults did not differentiate Bt and non-Bt soybean cultivars for oviposition indicating no behavioral preference. Third instar caterpillars showed no discrimination between Bt and non-Bt soybean leaves. Research in this area must focus on the possibility of widespread planting of genetically modified soybeans containing the Bt protein, which is associated with selection pressure and the behavior of pest species in relation to their hosts, as well as on the adequacy of management tactics that is able to prevent the loss of technology efficiency.展开更多
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supported by the National Natural Science Foundation of China(31901462)the Natural Science Foundation of the Jiangsu Higher Education Institutions,China(22KJA210005)+1 种基金the Priority Academic Program Development of Jiangsu Higher Education Institutions,China(PAPD)the Brand Professional Construction Program of Jiangsu Higher Education Institutions,China。
文摘The effects of mepiquat chloride(DPC)on the Cry1Ac protein content in Bacillus thuringiensis(Bt)cotton boll shells under high temperature and drought stress were investigated to provide a theoretical reference for Bt cotton breeding and high-yield and-efficiency cotton cultivation.This study was conducted using Bt cotton cultivar‘Sikang 3'during the 2020 and 2021 growing seasons at Yangzhou University Farm,Yangzhou,Jiangsu Province,China.Potted cotton plants were exposed to high temperature and drought stress,and sprayed with either 20 mg L^(-1)DPC or water(CK).Seven days after treatment,the Cry1Ac protein content,α-ketoglutarate content,pyruvic acid content,glutamate synthase activity,glutamic oxaloacetic transaminase activity,soluble protein content,and amino acid content were measured,and transcriptome sequencing was performed.DESeq was used for differential gene analysis.Under the DPC treatment,the Cry1Ac protein content increased by 4.7-11.9% compared to CK.Theα-ketoglutarate content,pyruvic acid content,glutamate synthase activity,glutamic oxaloacetic transaminase activity,soluble protein content,and amino acid content all increased.Transcriptome analysis revealed 7,542 upregulated genes and 10,449 downregulated genes for DPC vs.CK.Gene ontology(GO)and Kyoto Encyclopedia of Gene and Genomes(KEGG)analyses showed that the differentially expressed genes were mainly involved in biological processes,such as carbon and amino acid metabolism.For example,genes encoding 6-phosphofructokinase,pyruvate kinase,glutamic pyruvate transaminase,pyruvate dehydrogenase,citrate synthase,isocitrate dehydrogenase,2-oxoglutarate dehydrogenase,glutamate synthase,1-pyrroline-5-carboxylate dehydrogenase,glutamic oxaloacetic transaminase,amino-acid N-acetyltransferase,and acetylornithine deacetylase were all significantly upregulated.The DPC treatment increased pyruvate,α-ketoglutarate,and oxaloacetate by increasing the operational rate of the glycolytic pathway of the citric acid cycle.It also significantly upregulated the gen
文摘Studies on the behavior of Helicoverpa armigera in relation to soybean cultivars expressing Bt proteins are fundamentally important for IPM. We determine the no-choice and feeding choice of H. armigera, the no-choice and oviposition choice in different Bt and non-Bt soybeans cultivars. In the first step it was carried out the experiment to determine the no-choice and food choice in test for 12 Bt and non-Bt soybean cultivars. Simultaneously the consumption was determined. In a second step it was evaluated the no-choice and oviposition choice in different soybean cultivars. Helicoverpa armigera caterpillars showed neither attractiveness and preference for food nor attractiveness and oviposition preference for Bt and non-Bt soybean leaves when simultaneously contrasting the tested cultivars. This study showed that Bt toxin did not influence the oviposition preference, and H. armigera adults did not differentiate Bt and non-Bt soybean cultivars for oviposition indicating no behavioral preference. Third instar caterpillars showed no discrimination between Bt and non-Bt soybean leaves. Research in this area must focus on the possibility of widespread planting of genetically modified soybeans containing the Bt protein, which is associated with selection pressure and the behavior of pest species in relation to their hosts, as well as on the adequacy of management tactics that is able to prevent the loss of technology efficiency.
