Identification Of Bmptp And Bmcyp333a2 Gene And Its Involvement In Fluoride-resistance

In this research,the fluoride-resistent strain T6 and the sensitive strain 734 were used to identify genes linked to the model insects in Lepidoptera silkworm resisttance to fluoride by the differential display technique.The fluoride caused a large number of deaths to the silkworm when mulberry leaves were polluted by fluoride.this could cause serious harm to the sericultural industry.the human and other living thing as also.Elucidating the mechanism of resistance to fluoride would provide a new direction for the fluoride prevention and treatment of fluorosis.The research process and the main conclusions are presented as the following:1.the fluoride-resistent strain T6 and the sensitive strain 734 were normal reared,then feeding 200mg/kg sodium fluoride in fifth instar.the statistical analysis shows the physiological and biochemical changes of silkworm.In the 200mg/kg concentration of sodium fluoride,T6 strains grow nomal but the 734 strains obvious showed the symptoms of fluorosis.the mortality of the fluoride-resistent strain T6 remarkable lower than the sensitive strain 734.Body weight growth index has notable difference between the two strains,it shows that T6 strain may have the Def gene.2.the different display technique-PCR(DD-PCR)was used to analyze difffe rentially expressed genes in fluoride-resistant silkworm Bombyx mori strain T6 and susceptible strain 734,and a differentially expressed gene,Phosphate transport protein(Bmptp),was obtained.the gene participate in Phosphate transport.RT-PCR analyzed that Bmptp had higher expression level in fat body of strain T6 than 734,which indicated that fluoride may influence the expression level of Bmptp cause the fail Synthesis of ATP,then silkworm lack of energy supply,causing poisoning symptoms.3.the Fluoride ion selective electrode method was used to measure the fluoride concentration in fat body of fluoride-resistant silkworm Bombyx mori strain T6 and susceptible strain 734,the result shows T6 strain could maintain the fluoride concentration below the safety threshold by fluoride metabolism,while the 734 strains was opposite,suggesting that the fat body may be involved in the metabolism of fluoride.4.another highly expressed gene,Bmcypcyp333a2,in resistant strain was obtained.RT-PCR results showed that Bmcypcyp333a2 had higher expresion level in strain T6 than 734,the gene expressed in all tissues and the fluoride ion gradually accumulated in tissues after adding fluoride in silkworm,indicating it may be related with fluoride-resistance.