Cloning And Expression Of Heat Shock Protein Gene(HSP70、HSP90) In Phyllonorycter Ringoniella(Matsumura)

Phyllonorycter ringoniella which undergo complete metamorphosis and has strong heat resistance, is a common pest in apple and several other fruit trees. In order to explore the possible physiological role that hsp90 and hsp70 play in Phyllonorycter ringoniella, the fragment of hsp90 and hsp70 gene was cloned and the biological information and expression patterns under different treatments were analyzed. The fragment was cloned using RACE-PCR technology. The expression patterns of hsp90 and hsp70 in different tissues,developmental stages and different heat treatments were analyzed using RT-qPCR.A 942bp-length hsp90 gene fragment was obtained, it was named Prhsp90 and its access number in GenBank was KP671600. And a 284bp-length hsp70 gene was obtained, it was named KR075714. The sequences analysis showed that hsp90 amino acid sequence share high homology with that from other insects, showing 98% and higher homology with hsp90 from Heliothis armigera(ADP37710.1) and Spodoptera litura(ADK55516.1); and hsp70 amino acid sequence also showed 87% and higher homology with other insects such as Carposina sasakii(AHN92449.1) and Plodia interpunctella(ACB55606.1).Expression analysis of PrHSP90 in different tissues indicated its expression level was significantly higher in the wing compared with the head, thorax, abdomen and foot. The PrHSP90 transcript can be detectable during all the developmental stages, but its expression level was dramatically higher in adults than that in larvae and pupae. Under heat treatments,the expression of PrHSP90 increased with the increased temperature. After heat treatments at35℃, 38℃, 41℃ and 44℃, its expression increased significantly, which was about 3~15times of that in control groups. The expression of hsp70 gene was significantly higher than hsp90, and the response in response to heat shock was more significant.The results suggest the expression of hsp90 and hsp70 is not only tissue-specified but also play key roles during development and heat resistance in Phyllonorycter ringoniella, our results also provide basis for further studies about the relationships between hsp90 or hsp70 and the development and heat resistance of this insect.