The Full Length Cloning Of Pieris Rapae Hsp90 Gene And Related Genes Effectiveness Exploration In Butterfly Phylogeny

Pieris rapae belongs to Lepidoptera,Pieridae,Pieris and it is distributed throughout the world and is widely distributed in the country.The larvae of Pieris rapae are known as Pieris rapae crucivora and it has many host plants which are mainly harmful to the important pests of Chinese cabbage,mustard,cabbage,radish and other cruciferous crops,which seriously affect the yield and quality of vegetables.HSP90 plays an important role in the life activities of insects.It is one of the most important molecular chaperones in insects and participates in important biological processes such as insect growth and development,diapause and signal transduction.In addition,HSP90 has a certain relationship with the insect resistance of insects,so the research on the related genes of the Pieris rapae hsp90 gene is helpful to prevent it.In this study,the full-length cDNA of Pieris rapae hsp90 gene was cloned by RT-PCR and SMARTer RACE method.The results were analyzed by various bioinformatics methods.In addition,we conducted the Phylogenetic analysis based on the amino acid sequence of insects HSP90.Besides,we also explored the reliability of phylogenetic relationships established by hsp90 and Pgi partial gene sequences.We amplified the sequences of hsp90 of 36 butterflies and the sequences of Pgi genes of 21 butterflies from various butterflies for the construction of individual phylogenetic trees,COI,16S,wg and EF-1? genes were constructed and identified by two tree methods.The conclusions are as follows:(1)The length of the Pieris rapae hsp90 cDNA was 2461 bp.The open reading frame(ORF)was 2157 bp,encoding 718 amino acids,the initiation codon was ATG,the stop codon was TAA.The homology of the Pieris rapae HSP90 with other insects is as high as 75%-95%.It belongs to the ATPase superfamily,which is a hydrophilic protein with unstable protein structure and a molecular weight of 81.9 kD.Homology analysis showed that the Pieris rapae hsp90 was closely related to the phylogenetic development of the Pieris canidia hsp90 gene and was able to classify the insects into their respective traditional taxonomic positions.(2)The topology of phylogenetic tree constructed by Pgi gene partial sequence and the phylogenetic tree constructed by the sequence of COI+16S+wg+EF-1? is not consistent and it proved that Pgi gene may not be used for Family and belowclassification order phylogenetic molecular marker.The topology of phylogenetic tree constructed by hsp90 gene partial sequence and the phylogenetic tree constructed by the sequence of COI+16S+wg+EF-1? is consistent and it proved that hsp90 gene can be used for Family and below classification order phylogenetic molecular marker.