Cloning and expression profiling of the cathepsin genes CTSB-16D and CTSB-348 and their response to different environmental stresses in Myzus persicae(Hemiptera:Aphididae)
To explore the role of CTSB-16D and CTSB-348 of Myzus persicae(Sulzer)in response to high and low temperature and UV-B stresses,they were cloned by RT-PCR and their sequence characteristics were analyzed using bioinformatics approaches.The relative expression levels of the two genes in aphids at different developmental stages and in wingless adults exposed to high temperature,low temperature and UV-B stresses for different durations were detected by quantitative real-time PCR(RT-qPCR).The sequences of CTSB-16D and CTSB-348(GenBank acc.nos.MZ962352 and MZ962353)were 1 096 bp and 1 101 bp in length,with an open reading frame of 1 017 bp and 1 023 bp,encoding 338 and 340 amino acids,respectively.The relative molecular weight of the proteins are 38.14 kD and 37.52 kD,with an isoelectric point of 5.46 and 5.99,respectively.Phylogenetic analysis showed that both CTSB-16D and CTSB-348 had the closest relationship with CTSB of Acyrthosiphon pisum(XP_003246386.1,NP_001119608.1).RT-qPCR analysis showed that the two genes were expressed at different developmental stages,with the highest expression levels of CTSB-16D and CTSB-348 in the 1st-instar nymph and wingless adults,respectively.Low and high temperature and UV-B stresses obviously induced the expression of CTSB-16D and CTSB-348 genes,and the expression increasing first and then decreased with increasing exposure time.Under 4℃,the expression levels of CTSB-16D and CTSB-348 reached the peak after 90 min exposure,respectively,but under 36℃,their expression reached the peak after 30 min exposure.Under UV-B stress,the expression levels of CTSB-16D and CTSB-348 peaked after 60 min and 120 min exposure,respectively.The different expressions of CTSB-16D and CTSB-348 at different developmental stages and their different responses to temperature and UV-B stresses suggested that the two genes were involved in the growth and development of M.persicae in response to environmental stresses.
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