IDENTIFICATION AND CHARACTERIZATION OF TWO PHOSPHOLIPID HYDROPEROXIDE GLUTATHIONE PEROXIDASE GENES FROM THE MEDITERRANEAN SPECIES OF THE WHITEFLY Bemisia tabaci COMPLEX

Abstract

Phospholipid hydroperoxide glutathione peroxidases (PHGPXs) are essential enzymes of the cellular antioxidant defense system during insect-plant interactions. However, little attention has been devoted to the functional characterization of PHGXPs in the whitefly Bemisia tabaci. Here, we report the identification and characterization of two PHGPX genes, designated as BtQ-PHGPX1 and BtQ-PHGPX2 from the Mediterranean species of the B. tabaci complex. Sequence analysis indicated that the length of BtQ-PHGPX1 is of 942 bp with a 729 bp open-reading frame (ORF) encoding 242 amino acids, and BtQ-PHGPX2 is of 699 bp with a 567 bp ORF encoding 188 amino acids. Sequence alignment analysis showed that BtQ-PHGPX1 and BtQ-PHGPX2 shared high similarity with other known PHGPXs. The NVASXCGXT, FPCNQFXXQEPG, and IKWNFXKFLV surrounded the reactive cysteine, glutamine, and tryptophan residues, respectively. Recombinant BtQ-PHGPX1 and BtQ-PHGPX2 were overexpressed in Escherichia coli and purified. quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis with whiteflies of different development stages showed that the mRNA levels of BtQ-PHGPX2 were significantly higher in larvae than in other stages. The mRNA levels of BtQ-PHGPX2 were significantly higher than BtQ-PHGPX1 during all the developmental stages. The mRNA levels of BtQ-PHGPX1 and BtQ-PHGPX2 in female adults were relatively higher than in male adults. The expression of BtQ-PHGPX1 and BtQ-PHGPX2 was induced by the insecticide imidacloprid. These results suggest that BtQ-PHGPX1 and BtQ-PHGPX2 may participate in detoxification of oxidative hazards in B. tabaci.

Evolutionary and functional characterization of oxidative stress proteins in dendroctonus ponderosae hopkins (curculionidae: scolytinae)

“….3 -Alignments for glutathione peroxidase sequences from Dendroctonus ponderosae (DP), Drosophila melanogaster (DM), Bombyx mori (BM), Bicyclus anynana (BA), Tribolium castaneum (TC), Apis mellifera (AM), Pseudomyrmex gracilis (PG), Anopheles gambiae (AGa), and Anoplophora glabripennis (AG). The conserved glutathione peroxidase domains (A, B, and C respectively) are highlighted in hollow black boxes(Jiu et al 2015).Phylogenetic analysis of insect glutathione peroxidases showed that when both were present, the GPx1 and GPx2 sequences formed a species-specific cluster (Figure 2.4). An exception was the Asian longhorn beetle where a species-specific clade did not form.…”