摘要: Thermal stress usually leads to excessive production of reactive oxygen species (ROS) in all aerobic organisms. Catalases (CAT) are the key antioxidant enzymes, which act as the first line of defense against ROS in the antioxidant pathway. The highly invasive and widely distributed whitefly Bemisia tabaci MED damages plants by feeding as well as by transmitting many plant viruses. Previous studies have shown that strong adaptability to high temperature helps explain the spread of MED around the world. However, the mechanism underlying high temperature adaptation of this pest is not well understood. In this study, 6 CAT genes were identified from the MED genome and transcriptome dataset, among which BtCAT1, BtCAT2, and BtCAT3 were found to be highly expressed in adults. The expression of BtCAT1, BtCAT2, or BtCAT3 increased with induction temperature and induction time. The MED was exposed with mean high temperature (30 degrees C or 35 degrees C) and a short-term extremely high temperature (39 degrees C or 41 degrees C) after the silencing of BtCAT1, BtCAT2, or BtCAT3 to significantly increased ROS levels by at least 0.5 times and significantly decreased survival rate and fecundity of MED adults. The ROS level in the treated specimens gradually returned to a normal level after 24 h at 25 degrees C, but the survival rate still declined significantly. Taken together, our results demonstrate that CAT could help B. tabaci adapt to long-term mean high temperatures and short-term extremely high temperatures by eliminating excessive ROS.

摘要: Field monitoring revealed that the infection ratio of the bacterial symbiont Cardinium in the whitefly (Bemisia tabaci MED) was relatively low in northern China. However, the role of this symbiont and the symbiont-whitefly-host plant interaction mechanism are poorly understood. We investigated the influence of Cardinium on the competitiveness of the host whitefly and the physiological interaction between the host plants and host whiteflies. Cardinium-infected whiteflies were displaced by uninfected whiteflies after 5 generations, which showed that Cardinium infection reduced whitefly competitiveness. The defense response genes of cotton significantly decreased under infestation by infected whiteflies compared to uninfected whiteflies. The expression of detoxification metabolism genes, especially the uridine 5MODIFIER LETTER PRIME-diphospho-glucuronyltransferase and P450 genes, in infected whiteflies significantly decreased. These results demonstrated that Cardinium could inhibit the defense response of the host plant and decrease the detoxification metabolism ability of the host whitefly. The reduced competitiveness of infected whiteflies may be associated with the inhibition of the whitefly detoxification metabolism by Cardinium, resulting in the reduced performance of infected whiteflies. However, Cardinium infection can suppress plant defenses, which may benefit both infected and uninfected whiteflies when they coexist. This research illustrates the symbiont-whitefly-host plant interaction mechanism and the population dynamics of the whitefly.

摘要: Fatty acid synthase (FAS) is a multifunctional enzyme that plays an important role in the formation of fatty acids. The fatty acids take part in many processes, such as cell signaling and energy metabolism, and in insects they are important in both cuticular hydrocarbon (CHC) formation and reproduction. Here we characterized the sequence structure and function of an FAS from the small brown planthopper (SBPH), Laodelphax striatellus. The full-length open reading frame (ORF) sequence of LsFAS1 was 7122 bp, encoding a predicted protein of 2373 amino acid residues. There were 7 functional domains in the LsFAS1 protein sequence. Gene expression screening by real-time quantitative polymerase chain reaction (RT-qPCR) showed that LsFAS1 was expressed in all developmental stages. Relative expression was highest at the 4th-instar and female adult stages. Among different tissues, the expression level of LsFAS1 in the ovary was the highest. Phylogenetic analysis showed that LsFAS1 clustered in a clade with 2 FASs from Nilaparvata lugens. Furthermore, these 3 FASs are related to cockroach BgFAS and locust LmFAS. After RNA interference-mediated knock-down, most treated insects died at eclosion. In addition, the lifespan of dsFAS1-treated female adults was shorter than that of the dsGFP-injected control, and offspring production decreased. Also, the expression of vitellogenin (Vg) and vitellogenin receptor (VgR) genes decreased. Virgin females dissected at days 2 and 4 post-eclosion showed many matured oocytes in planthoppers treated with dsGFP but not with dsFAS1. These data highlight the importance of LsFAS1 in SBPH, including a role in reproduction.

摘要: Evolution of resistance to Cry proteins in multiple pest insects has been threatening the sustainable use of Bacillus thuringiensis (Bt)-transgenic crops. Better understanding about the mechanism of resistance to Cry proteins in insects is needed. Our preliminary study reported that the transcription of HaABCC3 was significantly decreased in a near-isogenic line (LFC2) of a Cry1Ac-resistant strain (LF60) of the global pest Helicoverpa armigera. However, the causality between HaABCC3 downregulation and resistance to Cry1Ac remains to be verified, and the regulatory mechanism underlying the HaABCC3 downregulation is still unclear. In this study, our data showed that both HaABCC3 and HaABCC3 downregulation were genetically linked to resistance to Cry1Ac in LF60. However, no InDels were observed in the coding sequence of HaABCC3 from LF60. Furthermore, F-1 offspring from the cross of LF60 and a HaABCC2/3-knockout mutant exhibited moderate resistance to Cry1Ac toxin; this indicated that the high resistance to Cry1Ac toxin in LF60 may have resulted from multiple genetic factors, including HaABCC2 mis-splicing and HaABCC3 downregulation. Results from luciferase reporter assays showed that promoter activity of HaABCC3 in LF60 was significantly lower than that in the susceptible strain, which indicated that HaABCC3 downregulation was likely mediated by promoter variation. Consistently, multiple variations of the GATA- or FoxA-binding sites in the promoter region of HaABCC3 were identified. Collectively, all results in this study suggested that the downregulation of HaABCC3 observed in the H. armigera LF60 strain, which is resistant to Cry1Ac, may be mediated by a cis-regulatory mechanism.

摘要: Tomato spotted wilt orthotospovirus (TSWV) causes substantial economic losses to vegetables and other crops. TSWV is mainly transmitted by thrips in a persistent and proliferative manner, and its most efficient vector is the western flower thrips, Frankliniella occidentalis (Pergande). In moving from the thrips midgut to the salivary glands in preparation for transmission, the virions must overcome multiple barriers. Although several proteins that interact with TSWV in thrips have been characterized, we hypothesized that additional thrips proteins interact with TSWV and facilitate its transmission. In the current study, 67 F. occidentalis proteins that interact with G(N) (a structural glycoprotein) were identified using a split-ubiquitin membrane-based yeast 2-hybrid (MbY2H) system. Three proteins, apolipoprotein-D (ApoD), orai-2-like (Orai), and obstructor-E-like isoform X2 (Obst), were selected for further study based on their high abundance and interaction strength; their interactions with G(N) were confirmed by MbY2H, yeast beta-galactosidase and luciferase complementation assays. The relative expressions of ApoD and Orai were significantly down-regulated but that of Obst was significantly up-regulated in viruliferous thrips. When interfering with Obst in larval stage, the TSWV acquisition rate in 3 independent experiments was significantly decreased by 26%, 40%, and 35%, respectively. In addition, when Obst was silenced in adults, the virus titer was significantly decreased, and the TSWV transmission rate decreased from 66.7% to 31.9% using the leaf disk method and from 86.67% to 43.33% using the living plant method. However, the TSWV acquisition and transmission rates were not affected by interference with the ApoD or Orai gene. The results indicate that Obst may play an important role in TSWV acquisition and transmission in Frankliniella occidentalis.