Response Of Important Enzyme Genes To Defense Responses Of Kidney Bean And Broad Bean Plants In Frankliniella Occidentalis(Perganda)

Western flower thrips,Frankliniella occidentalis?Pergande?,is a worldwide invasive pest that causes serious damage to flowers and vegetables.Based on the previous gene research and the results of the transcriptome data measured in the laboratory,real-time PCR was used to determine expression levels of various enzymes in western flower thrips after they were shifted from kidney bean pods to the favorite kidney bean plants and the non-favorite broad bean plants.The expression levelsof detoxificationenzymes?cytochromeP450,glutathione-S-transferaseand carboxylesterase?genes,digestive enzymes?amylase,trypsin and tryptase?genes and protective enzymes?superoxide dismutase,catalase and peroxidase?genes in 2^nd instar nymphs and adults at F₁,F₂,F₂ and F₃ generations are determined.The results will lay the foundation for further exploration of the adaptability of the host and the interaction between plants and insects.Below are key research findings:1 Responses of detoxification enzyme genes in western flower thrips to kidney bean and broad beanIn this paper,nine of P450 genes,including 5 CYP4 genes?CYP4-1,CYP4-2,CYP4-3,CYP4-4 and CYP4-5?and 4 CYP6 genes?CYP6-1,CYP6-2,CYP6-3 and CYP6-4?,four of GSTs genes?CL992.contig6,CL992.contig14,CL1914.Contig2 and Unigene 18748?,and CL4503.Contig2 annotated as the CarE gene were subjected to primer design and assayed for expression after shifted to kidney bean and broad bean plants.The results showed that CYP4-1 and CYP4-2 were significantly up-regulated in3 generations and were the highest in F₁ generation,CYP4-3 and CYP6-4 were significantly up-regulated only observed in 2^nd instar nymphs at F₁after shift from the bean pods to kidney bean plants.CYP4-4 and CYP6-2 were significantly up-regulated only atF₃,and CYP4-5 and CYP6-3 were significantly down-regulated at 3 generations after shifted to kidney bean and broad bean plants,and which was lowest at F₂.CYP6-1 was significantly down-regulated only at F₂ generation.The expression levels of CYP4-1,CYP4-2 and CYP4-3 in adults were significantly down-regulated at F₂ and F₃,and CYP4-1 and CYP4-2 were lowest at F₂,while CYP4-3 was the lowest at F₃generation.CYP4-4 was only significantly up-regulated at F₁ generation;CYP4-5 and CYP6-4 were significantly up-regulated at F₁ generation after shifting to kidney bean plants,while their expression levels were significantly down-regulated at F₂ and F₃generation.CYP6-1,CYP6-2 and CYP6-3 were significantly down-regulated at 3generations,and there was no significant difference between F₂ and F₃,and was significantly lower than F₁ generation.The expression levels of CYP4-3,CYP6-1,CYP6-2 and CYP6-4 in 2^nd instar nymphs of the western flower thrips at F₃ generation were significantly higher than those of the adults after the control and conversion to kidney bean plants.There was no significant difference of the expression levels of CYP4-4,CYP4-5 and CYP6-3 between 2 stages at F₁ generation.The expression levels were significantly higher in 2 nd instar nymps than those in the adults at the control,F₂and F₃ generations.The expression level of CYP4-1 and CYP4-2 was significantly higher in adults than those in 2^nd instar nymphs when feeding kidney bean pod.After shifted,the expression of CYP4-1 in the 2^nd instar nymphs of each generation was significantly higher than that in adults,while the expression of CYP4-2 in 2^nd instar nymphs of F₂ generation was significantly higher than that in adults.After western flower thrips was shifted to broad bean plants,the expression levels of CYP4-1,CYP4-3,CYP6-3 and CYP6-4 in the 2^nd instar nymphs were significantly higher at F₂ generation than the control.The expression levels of CYP4-4 and CYP4-5were significantly down-regulated at 3 generations after shifted to broad bean plants,and there was no significant difference among 3 generations.The expression level of CYP4-2 was significantly higher at F₂ and F₃ than that in control.The expression level of CYP6-2 were significantly higher than the control in all 3 generations,and the expression levels of CYP4-2 and CYP6-2 were highest at F₂ generation.The expression level of CYP6-1 was also highest at F₂ generation,and was significantly higher than the control,and at F₁ and F₃ generations were significantly lower than the control.In adults,the expression levels of CYP4-1,CYP4-4 and CYP4-5 were significantly lowerat 3generations after shifted to broad bean plants,while CYP4-3,CYP6-2 and CYP6-4 were significantly higher than the control.The expression levels of CYP4-2 at F₂ and F₃generations were significantly higher than the controlThe expression levels of CYP6-3was significantly higher at F₁ and F₃ generations than the control.Tthe expression levels of CYP6-1 weren't significantly different in each generation.The expression levels of CYP4-3,CYP6-1 and CYP6-2 in the 2^nd instar nymphs in the control and broad bean plants were significantly higher than those in the adult,while CYP4-2 was the opposite,and other genes varied from generation to generation.The expression levels of detoxification enzymes genes in different plants were different.The expression levels of CYP4-2,CYP4-3,CYP6-2 and CYP6-3 in adults fed on broad bean plants were significantly higher than those in kindey bean plants.The differences of other genes between the two plants were different according to generations and insect stages.The results of GSTs gene after host shift showed that the expression of CL992.contig6 in each generation was significantly higher than that in the control,and the expression of CL992.contig14 was significantly higher at F₂ and F₃.The expression level of CL1914.Contig2 was significantly higher than that of the control,while its expression at F₂ was significantly higher than that of the control,while the expression at F₃ was significantly lower than that of the control.After the conversion to kidney bean plants,the expression level of Unigene18748 at all generations were significantly lower than the control,and there was no significant difference among generations.the expression of CL992.contig6 in adults was significantly higher at F₃than control.The expression of CL992.contig14 was only significantly lower at F₂,and the expression of CL1914.Contig2 were significantly higher than the control at F₁ and F₃ generation.The expression level of Unigene18748 was not significantly different between each generation and the control.After shifting to broad bean plants,the expression level of CL992.contig6 in 2ⁿdd instar nymphs was significantly down-regulated only at F₁,and which were down-regulated in the adult at F₁ and F₂ generations.The expression levels of CL992.contig14,CL1914.Contig2 and Unigene18748 were significantly down-regulated in all generations after shifted to broad bean plants in both 2^nd instar nymphs and adults,and there was no significant difference among generations.The GSTs gene responds differently to different hosts.The expression levels of the four genes between the two hosts were not significantly different except in adult at F₂,and the expression levels of other genes in thrips feeding kidney bean plants were significantly higher than those in thrips feeding broad bean plants in both 2^nd instar nymphs and adults.The host response of the CarE gene in western flower thrips showed that the expression of CL4503.Contig2 was significantly down-regulated after the conversion of the western flower thrips to kidney bean plants in 3 generations in2^nd instar nymphs and adults,and after the conversion to the broad bean plants,the expression of CL4503.Contig2 was significantly down-regulated.The expression levels of CL4503.Contig2 were significantly up-regulated in 2^nd instar nymphs and adults.The response of CL4503.Contig2 was significantly different among different host plants.The expression levels in 2^nd instar nymphs and adults in broad bean plants were significantly higher than those in kidney bean plants.2 Responses of protective enzyme genes in western flower thrips to kidney bean and broad beanThe responses of the protective enzyme gene of the western flower thrips showed that FoCAT was significantly down-regulated in the 2^nd instar nymphs and adults after shifting to the kidney bean plants.There was no significant difference among generations in 2^nd instar nymphs.As to the adult,the expression level FoCAT at F₂ was significantly lower than F₁ and F₃ generation.After conversion to the broad bean plants,the expression in 2^nd instar nymphs were significantly down-regulated in all generations,and was the lowest at F₁ generation.The expression level of FoCAT were significantly lower in the adult at F₁ and F₃ generations.The expression levels of FoCAT in the broad bean plants in 2^nd instar nymphs and adults were significantly higher than those in the kidney bean plants in all three generations.The response of SOD gene showed that the expression levels of Unigene3401 and Unigene15728 in 2^nd instar nymphs feeding kindey bean plants were significantly down-regulated,while its expression were opposite in feeding broad bean plants.In the adult,the expression levels of Unigene3401 and Unigene15728 were significantly up-regulated at F₁ and F₃,while the expression were significantly up-regulated and highest in feeding broad bean plants at F₂ generation.The expression levels of Unigene3401and Unigene15728 in 2^nd instar nymphs and adults in the broad bean plants were significantly higher than those in kidney bean plants.The response of POD gene showed that Unigene10079 in 2nd instar nymph was significantly up-regulated at F₂ and F₃,and CL4306.Contig1 was significantly up-regulated at F₃,while Unigene10079 in thrips feeding broad bean was significantly up-regulated in each generation.CL4306.Contig1 was significantly up-regulated at F₂ and F₃.As to adult,the expression of Unigene10079 and CL4306.Contig1 in thrips feeding broad bean were up-regulated at each generation,and the expressions of Unigene10079and CL4306.Contig1 were different in thrips feeding kidney bean plants.The expression of Unigene10079 in 2^nd instar nymphs and adults in broad bean plants was significantly higher than that in kidney bean plants except that its expression in 2nd instar nymph at F₃.The expression of CL4306.Contig1 in different plants was different at different stage.3 Responses of digestive enzyme genes in wesstern flower thrips to kidney bean and broad beanIn the digestive enzyme genes,the expression level of the amylase gene CL7175.Contig1 significantly decreased in 2^nd instar nymphs and adults of the western flower thrips,and which were significantly up-regulated in feeding broad bean plants.After switching from pods to kidney bean plants,the expression of trypsin gene CL10275.Contig2 in 2^nd instar nymphs was significantly down-regulated at all generations,while CL4520.Contig1 was only significantly down-regulated at F₁,There was no significant difference of CL10275.Contig2 in thrips feeding broad bean plants with controls at 3 generations.The expression level of CL4520.Contig1 was significantly up-regulated at F₂ and F₃.As to adult,the changes of expression levels of CL10275.Contig2 and CL4520.Contig1 were different after shifting to kidney bean plants.The expression level of CL4520.Contig1 were significantly up-regulated when feeding broad bean plants.The responses of tryptone trypsin gene CL2175.Contig1 and CL579.Contig4showed that the expression of CL2175.Contig1 was significantly down-regulated in the2^nd instar nymphs and adults after from the bean pod was transferred to the kidney bean plant.The expression level of CL2175.Contig1 in the 2^nd instar nymphs was not significantly different from the control at each generation.As to adult,The expression level of CL2175.Contig1 decreased at F₁ and F₂ generations,while they was significantly up-regulated at F₃ generation.After conversion to the broad bean plants,the expression levels of CL2175.Contig1 and CL579.Contig4 were significantly up-regulated in all generations both at the 2^nd instar nymphs and adults.The expression of CL10275.Contig2 in adult of thrips on different plants was not significantly different at F₂ generation.The expression of other digestive enzymes was significantly higher in broad bean plants than in common bean plants.