The Roles Of Heat Shock Transcription Factor(Hsf) And Heat Shock Protein Gene(Hsp70,Hsp21,sHsp21) On The High Temperature Tolerance Of Agasicles Hygrophila Selman&vogt(Coleoptera:Cheysomelidae)

Alternanthera philoxeroides is a serious perennial invasive weed.In present,It is the outbreak of hazard in many region of the south.Agasicles hygrophila,an effective specialized natural enemy insect of Al.philoxeroides,and Adopted by many countries,and achieved some success.In the subtropical area of China,the population dynamics of Ag.hygrophila is affected by the temperature and its thermal adaptability is poor.In some areas its population density declines sharply from August to September when the temperature is extremely high,which limits the control efficiency of A.philoxeroides in the summer.2009,however,we are released into the Ag.hygrophila in Hainan population,currently in Hainan it can happen all the year round,so what is to improve the ability of high temperature tolerance? Vis ible,the Ag.hygrophila has been hot adaptive differentiation in our country.The expression of heat shock protein has a close relation with the organism resistance,organism product heat shock protein can protect the body from damaged under high temperature.Therefore,in order to reveal the heat shock protein(Hsps)gene in Ag.hygrophila regulation mechanism in the high temperature tolerance,this study targeted on Hunan and Ha inan population as the research object,aim at the population density of Ag.hygrophila keeps high and this phenomenon occurs throughout the year in Hainan,around the heat shock protein gene in the relation between temperature stress adaptation the scientific problems,Using high-throughput sequencing and fluorescence quantitative PCR technology and method of research on cloned the Ag.hygrophila?s Hsp70,Hsp21,s Hsp21 and HSF gene and its biological characteristics,and studied its function under high temperature stress.To understand the adaptability of Ag.hygrophila to temperature and the climate warming effective bio-control of Al.philoxeroides has the vital significance.1.The clone of Hsp70,Hsp21,s Hsp21 and Hsf in Ag hygrophilaThis research obtained the complete c DNA sequences of Hsp70,Hsp21,s Hsp21 gene and partial Hsf sequence in Ag.hygrophila.Total length of 2053 bp of Hsp70 gene sequences,open reading frame(ORF)length of 1940 bp,the gene encoding 646 amino acids;Hsp21 gene sequence is 549 bp and 538 bp open reading frame,the gene encoding 180 amino acids;s Hsp21 gene sequence is 693 bp and open reading frame(ORF)length of 567 bp,the gene encoding 188 amino acids;And obtained the length of 790 bp of partial Hsf sequence.2.Phylogenetic analysis of Hsp70,Hsp21,s Hsp21 and Hsf genePhylogenetic tree showed that the Hsp70,Hsp21,s Hsp21 and Hsf gene is relatively conservative in evolution,the same of order can be list the same clade.The homology of the Hsp70,Hsp21,s Hsp21 gene in Lissorhoptrus oryzophilus,Monochamus alternates and Leptinotarsa decemlineata,which reached 70%~85% similar ity,and on the phylogenetic relationship.the Hsf sequence results are Blast comparison,the results showed according to sequence amplification by all Hsf is predicted.So,we inference this experiment the cloning of Hsp70,Hsp21,s Hsp21 and Hsf gene is Ag.hygrophila.3.The Hsp70,Hsp21,s Hsp21 and HSF gene m RNA expression analysis of Ag.hygrophila under the different temperature treatmentThis study analyzed the expression level of Hsp70,Hsp21,sHsp21 and Hsf gene under different temperature in two geographical populations of Ag.hygrophila using the RT-PCR technique.The results showed that gene expression quantity of Hsp70,Hsp21,s Hsp21 were basic significantly higher than that of control group of 30? under the 36? and 39? high temperature treatment.But For Hunan populations,gene expression quantity of Hsf were significantly lower than the control group of 30?;On the other hand,it was signif icantly higher than that of control group of 30?in Hainan population.Using RNAi to identify the function of the Hsp70,Hsp21,s Hsp21 and Hsf gene of two geographical populations under high temperature stress.We fed Ag.hygrophila with the ds RNA of Hsp70,Hsp21,s Hsp21 and Hsf gene,makes the purpose gene is part of the silence,Ag.hygrophila cannot effectively the perception of the outside high temperature stress,reduced of heat shock protein expression in cells.So,the heat tolerant ability of Ag.hygrophila significantly increased under high temperature condition that the death rate signif icantly increased.Research results can be further suggests that by feeding ds RNA method to suppress the target gene expression in Ag.hygrophila is feasible.And show that Hsp70,Hsp21,s Hsp21 and Hsf gene plays a key role in the high temperature adaptation of two geographic populations.