Cloning And Expression Of Transient Receptor Potential Gene In Three Whiteflies And Their Function Identification Under High Temperature Stress

The temperature is an important factor affecting species population dynamics and diffusion distribution. One of important reasons of the success of invasion and outbreak for alien invasive species is the high resistance to the temperature. Temperature sensing system mediated by the transient receptor potential (TRP) is a key component affecting the insects’ perception of temperature. Studying the function of the key regulation factor in the temperature sensing mechanism assists to clear the temperature perception and provide theoretical basis for the temperature adaptive regulation mechanism. This study targeted on Bemisia tabaci MEAM1crytic specie, Bemisia tabaci MED crytic specie and Trialeurodes vaporariorum and focused on the clone of TRP gene in whitefly and the analysis of its bioinformatics characteristics. We also make function identification under high temperature stress. The main results and conclusions as follows:1. The clone of TRP gene in B. tabaci MEAM1, B. tabaci MED and T. vaporariorum.This research took the reverse transcription PCR and RACE technique. This is the first time obtained the complete cDNA sequences of TRP gene in B. tabaci MEAM1and B. tabaci MED and partial TRP sequence in T. vaporariorum. Bioinformatics analysis showed that the genes belong to the members of the family of transient receptor ion channels, named MEAM1BtTRP、MED BtTRP and Tv TRP, respectively.2. The TRP gene mRNA expression analysis of B. tabaci MEAMl and T. vaporariorum under temperature stress.This study analyzed the expression level of TRP gene under different temperature in B. tabaci MEAM1and T. vaporariorum using the qRT-PCR technique. The results showed that the mRNA expression level increased in both whiteflies under13℃temperature stress condition.In addition, under7℃stress condition, the mRNA expression level in B. tabaci ME AM1is higher than that of T. vaporariorum.3. Phylogenetic analysis of TRP gene.Phylogenetic tree showed that the TRP gene is relatively conservative in evolution, and the homology of the TRP gene in the three whiteflies is close to the TRP2gene in Acyrthosiphon pisum, which reached75%similarity and they also had the same amino acid with conservative domain structure. It was speculated that the TRPs obtained in this study were in the same family with the TRP2in A. pisum.4. Using RNAi to identify the function of the TRP gene of B. tabaci MEAM1under high temperature stress.We fed B. tabaci MEAM1with the dsRNA of ME AMI BtTRP gene and found that the death rate of B. tabaci MEAM1significantly increrased under high temperature, which suggested that the heat tolerant ability of B. tabaci MEAM1significantly increased under high temperature condition. This showed that MEAM1BtTRP plays a key role in the high temperature adaptation of B. tabaci MEAM1.