Effects of Extreme High Temperature on Trehalose Content and Trehalose Transporter Gene in Tetranychus truncatus
[Objective]The objective of this study is to clarify the effect of high temperature on the trehalose transporter gene of Tetranychus truncatus,and to provide a theoretical basis for the green control of pests.[Method]According to the transcriptome data of T.truncatus induced by high temperature,the CDS sequences and protein amino acid sequences of two trehalose transporter genes TtTret1-like and TtTret1 were obtained.Bioinformatics analysis of TtTret1-like and TtTret1 was carried out using analytical tools such as ExPASy,ProScale and MEGA.The expression characteristics of TtTret1-like and TtTret1 at different development stages and different high temperatures(38,42,46 and 50℃)were analyzed by qRT-PCR.The trehalose content of T.truncatus at different high temperatures was determined by microanalysis.RNA interference(RNAi)technique was used to silence TtTret1-like and TtTret1,and to explore their functions in coping with high temperature.[Result]Bioinformatics analysis showed that the full-length CDS of TtTret1-like and TtTret1 was 1 389 and 1 569 bp,respectively,encoding 463 and 523 amino acids.The predicted protein molecular weights were 50 189.03 and 57 358.10 Da,and the isoelectric points were 8.87 and 8.70,respectively.TtTret1-like and TtTret1 are both basic amino acids and hydrophobic amino acids,belonging to unstable proteins.The secondary structure is a spiral and coiled structure.Both trehalose transporters have conserved domains and 12 transmembrane domains of major facilitator superfamily(MFS).The amino acid sequence similarity and phylogenetic analysis showed that TtTret1-like and TtTret1 were highly consistent with the trehalose transporter sequences of other arachnids,especially with T.urticae.The expression level of TtTret1-like was higher in egg,larva and adult stages.TtTret1 was highly expressed in larva stage.With the increase of treatment temperature,the expression level of TtTret1-like increased,and reached the highest level at 50℃.However,with the increase of treatment temperature,the expression level of TtTret1 increased first and then decreased,reaching the maximum at 42℃.The trehalose content in T.truncatus increased significantly after high temperature stress.After silencing TtTret1-like and TtTret1 for 48 h,the trehalose content in T.truncatus increased,but the trehalose content in hemolymph decreased.The survival rates of T.truncatus were 11%and 46.67%after treated at 50℃for 2 h and then recovered for 96 h,which were significantly lower than those of the control group.[Conclusion]It is hypothesized that TtTret1-like and TtTret1 in T.truncatus play an important role in their responses to high temperature stress.
Tetranychus truncatushigh temperature stresstrehalosetrehalose transporterRNA interference(RNAi)
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