Identification and characterization of two phospholipid hydroperoxide glutathione peroxidase genes from the Mediterranean species of the whitefly Bemisia tabaci complex.

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.

Cloning, expression and characterization of mitochondrial manganese superoxide dismutase from the Whitefly, Bemisia tabaci.

A mitochondrial manganese superoxide dismutase from an invasive species of the whitefly Bemisia tabaci complex (Bt-mMnSOD) was cloned and analyzed. The full length cDNA of Bt-mMnSOD is 1210 bp with a 675 bp open reading frame, corresponding to 224 amino acids, which include 25 residues of the mitochondrial targeting sequence. Compared with various vertebrate and invertebrate animals, the MnSOD signature (DVWEHAYY) and four conserved amino acids for manganese binding (H54, H102, D186 and H190) were observed in Bt-mMnSOD. Recombinant Bt-mMnSOD was overexpressed in Escherichia coli, and the enzymatic activity of purified mMnSOD was assayed under various temperatures. Quantitative real-time PCR analysis with whiteflies of different development stages showed that the mRNA levels of Bt-mMnSOD were significantly higher in the 4th instar than in other stages. In addition, the in vivo activities of MnSOD in the whitefly were measured under various conditions, including exposure to low (4 °C) and high (40 °C) temperatures, transfer from a favorable to an unfavorable host plant (from cotton to tobacco) and treatment with pesticides. Our results indicate that the whitefly MnSOD plays an important role in cellular stress responses and anti-oxidative processes and that it might contribute to the successful worldwide distribution of the invasive whitefly.

Molecular characterization and oxidative stress response of an intracellular Cu/Zn superoxide dismutase (CuZnSOD) of the whitefly, Bemisia tabaci.

Superoxide dismutases (SODs) are important for the survival of insects under environmental and biological stresses; however, little attention has been devoted to the functional characterization of SODs in whitefly. In this study, an intracellular copper/zinc superoxide dismutase of whitefly (Bemisia tabaci) (Bt-CuZnSOD) was cloned. Sequence analysis indicated that the full length cDNA of Bt-CuZnSOD is of 907 bp with a 471 bp open reading frame encoding 157 amino acids. The deduced amino acid sequence shares common consensus patterns with the CuZnSODs of various vertebrate and invertebrate animals. Phylogenetic analysis revealed that Bt-CuZnSOD is grouped together with intracellular CuZnSODs. Bt-CuZnSOD was then over-expressed in E. coli and purified using GST purification system. The enzymatic activity of purified Bt-CuZnSOD was assayed under various temperatures. When whiteflies were exposed to low (4°C) and high (40°C) temperatures, the in vivo activity of Bt-CuZnSOD was significantly increased. Furthermore, we measured the activities of several antioxidant enzymes, including SOD, catalase and peroxidase, in the whitefly after transferring the whitefly from cotton to tobacco (an unfavorable host plant). We found that the activity of SOD increased rapidly on tobacco plant. Taken together, these results suggest that the Bt-CuZnSOD plays a major role in protecting the whitefly against various stress conditions.

Robust Majorana edge modes with low frequency multiple time periodic driving.

Floquet Majorana edge modes capture the topological features of periodically driven p-wave superconductors. We present a Kitaev chain with multiple time periodic driving terms. Our results demonstrate how multiple driving will affect Floquet bands in frequency space, leading to more robust Floquet Majorana edge modes against driving frequency ω in comparison with the single driving scenario. Meanwhile, we have proposed how to predict Majorana edge modes via the Zak phase of Floquet bands. Besides, in contrast to the cases with single driving term, where the constant phase can be gauged out by properly choosing the initial time, we have shown the relative phase between multiple driving can not be gauged out and will play a dominant role in deciding topological phase transitions. For the sake of completeness, we also investigate the high frequency limit. Analytical results on effective Hamiltonian can be obtained via Magnus expansion and relative phase induced topological transitions can be shown explicitly.

Cloning of a putative extracellular Cu/Zn superoxide dismutase and functional differences of superoxide dismutases in invasive and indigenous whiteflies.

Superoxide dismutases (SODs) are a group of important antioxidant defense enzymes. In this study, a putative extracellular Cu/Zn superoxide dismutase (ecCuZnSOD) complementary DNA was cloned and characterized from the whitefly, Bemisia tabaci. Quantitative polymerase chain reaction analysis showed that the expression level of BtecCuZnSOD was more than 10-fold higher in the invasive Middle East Asia Minor 1 (MEAM1) than in the native Asia II 3 species of the B. tabaci species complex. After exposure to low temperature (4 °C), the expression of Bt-ecCuZnSOD gene was significantly up-regulated in MEAM1 but not in Asia II 3. Furthermore, the expression level of B. tabaci intracellular CuZnSOD (Bt-icCuZnSOD), Bt-ecCuZnSOD and mitochondrial MnSOD (Bt-mMnSOD) was compared after transferring MEAM1 and Asia II 3 whiteflies from favorable (cotton) to unfavorable host plants (tobacco). On cotton, both CuZnSOD genes were expressed at a higher level in MEAM1 compared with Asia II 3. Interestingly, after transferring onto tobacco, the expression of Bt-ecCuZnSOD was significantly induced in Asia II 3 but not in MEAM1. On the other hand, while Bt-mMnSOD was expressed equally in both species on cotton, Bt-mMnSOD messenger RNA was up-regulated in MEAM1 on tobacco. Consistently, enzymatic activity assays of CuZnSOD and MnSOD demonstrated that CuZnSOD might play an important protective role against oxidative stress in Asia II 3, whereas MnSOD activation was critical for MEAM1 whiteflies during host adaptation. Taken together, our results suggest that the successful invasion of MEAM1 is correlated with its constitutive high activity of CuZnSOD and inducible expression of MnSOD under stress conditions.