Molecular Cloning And Expression Of Cyp6Family Genes From Phyllonorycter Ringoniella

Phyllonorycter ringoniella（Lepidoptera: Gracilariidae）, endemic to east Asia, is acommon pest on the apple tree. For their research in ecological prevention and control, andrarely make the news and molecular aspects. Insect cytochrome P450enzymes （cytochromeP450monooxygenases, P450s）, a diverse class of enzymes found in virtually all insect tissues,fulfill many important tasks, ranging from the synthesis and degradation of ecdysteroids andjuvenile hormones to the metabolism of foreign toxical chemicals of natural or syntheticorigin. Thus, in this study, P450genes from P. ringoniella were cloned, their mRNAexpression of P450s was also detected. Especially the analysis of clone and expression ofP450genes, the main results are as follows:1. Three novel P450fragments and two novel P450genes which were1740bp and1940bp in length were isolated by PCR-RACE strategy. Their ORFs encoded544aa and546aa, respectively, and named phCYP6AN4and phCYP6ab by the International P450Nomenclature Committee.2. P. ringoniella was treated with cyhalothrin for24hours, according to phCYP6AN4and phCYP6ab genes expressed in P. ringoniella show that the quantity of inducers inducedeffect exists a certain degree of specificity, and one or more of the following compounds areeffective may not apply to other types of gene.3.phCYP6AN4and phCYP6ab genes in P. ringoniella mRNA expression in differenttissue volume detection, found two genes expressed in insect body have a certain amount ofdifferences, the most expression quantity of phCYP6AN4and phCYP6ab is foot、wing andfoot, respectively, the analysis concluded that the two P450gene for tissue specificity, mayplay a role in the development of insect physiology.4. The analysis of mRNA expression of phCYP6AN4and phCYP6ab in different instarsof P. ringoniella, the most expression quantity of phCYP6AN4and phCYP6ab is4th、5thinstars and4th instars, respectively, illustrate the similarities and differences in different ageexpression. Combining with the ecological control method, is commonly to applying pesticidecontrol in4th instars; Adjust measures to the expression of different genes, to have the bettereffect.