Systematic analysis of a wasp parasitism arsenal

Parasitoid wasps are among the most diverse insects on earth with many species causing major mortality in host populations. Parasitoids introduce a variety of factors into hosts to promote parasitism, including symbiotic viruses, venom, teratocytes and wasp larvae. Polydnavirus‐carrying wasps use viruses to globally suppress host immunity and prevent rejection of developing parasites. Although prior results provide detailed insights into the genes viruses deliver to hosts, little is known about other products. RNAseq and proteomics were used to characterize the proteins secreted by venom glands, teratocytes and larvae from Microplitis demolitor, which carries M. demolitor bracovirus (MdBV). These data revealed that venom glands and teratocytes secrete large amounts of a small number of products relative to ovaries and larvae. Venom and teratocyte products exhibited almost no overlap with one another or MdBV genes, which suggested that M. demolitor effector molecules are functionally partitioned according to their source. This finding was well illustrated in the case of MdBV and teratocytes. Many viral proteins have immunosuppressive functions that include disruption of antimicrobial peptide production, yet this study showed that teratocytes express high levels of the antimicrobial peptide hymenoptaecin, which likely compensates for MdBV‐mediated immunosuppression. A second key finding was the prevalence of duplications among genes encoding venom and teratocyte molecules. Several of these gene families share similarities with proteins from other species, while also showing specificity of expression in venom glands or teratocytes. Overall, these results provide the first comprehensive analysis of the proteins a polydnavirus‐carrying wasp introduces into its host.     

Effects of the endoparasitoid Cotesia chilonis (Hymenoptera: Braconidae) parasitism,venom, and calyx fluid on cellular and humoral immunity of its host Chilo suppressalis (Lepidoptera: Crambidae) larvae

The larval endoparasitoid Cotesia chilonis injects venom and bracoviruses into its host Chilo suppressalis during oviposition. Here we study the effects of the polydnavirus (PDV)-carrying endoparasitoid C. chilonis (Hymenoptera: Braconidae) parasitism, venom and calyx fluid on host cellular and humoral immunity, specifically hemocyte composition, cellular spreading, encapsulation and melanization. Total hemocyte counts (THCs) were higher in parasitized larvae than in unparasitized larvae in the late stages following parasitization. While both plasmatocyte and granulocyte fractions and hemocyte mortality did not differ between parasitized and unparasitized hosts, in vitro spreading behavior of hemocytes was inhibited significantly by parasitism throughout the course of parasitoid development. C. chilonis parasitism suppressed the encapsulation response and melanization in the early stages. Venom alone did not alter cellular immune responses, including effects on THCs, mortality, hemocyte composition, cell spreading and encapsulation, but venom did inhibit humoral immunity by reducing melanization within 6 h after injection. In contrast to venom, calyx fluid had a significant effect on cell spreading, encapsulation and melanization from 6 h after injection. Dose–response injection studies indicated the effects of venom and calyx fluid synergized, showing a stronger and more persistent reduction in immune system responses than the effect of either injected alone.     

花椒窄吉丁触角转录组及嗅觉相关基因分析

【目的】建立花椒窄吉丁Agrilus zanthoxylumi成虫触角转录组数据库,挖掘嗅觉相关基因,为今后研究其触角的化学感受机制及生物防控提供理论支撑。【方法】采用高通量测序平台IlluminaNovaSeq 6000对花椒窄吉丁雌雄成虫触角进行转录组测序,用Trinity软件对获得的高质量reads进行序列拼接与组装;使用BLAST软件将触角转录组数据比对NR, NT, Swiss-Prot, GO, KEGG, BLASTX,eggNOG, Pfam, TmHMM, SignalP, KO, Map, BLASTP和RNAMMER公共数据库;基于初步筛选到的花椒窄吉丁候选气味结合蛋白(odorant binding protein, OBP)和化学感受蛋白(chemosensory protein, CSP)以及其他鞘翅目昆虫的同源蛋白的核苷酸序列,利用MEGA软件进行系统进化分析。运用RPKM (reads perkilobase per million mapped reads)值对嗅觉相关基因表达量进行分析。【结果】花椒窄吉丁雌雄成虫触角转录组测序共获得36 209条基因和90 982条转录本,其N₅₀分别为2 103和2 523 bp,组装完整性较高。注释到NR数据库的基因最多(41.62%),其中与赤拟谷盗Tribolium castaneum相似基因所占比例最高(19%)。在GO数据库中比对到11 614个基因,按功能分为细胞组分、分子功能与生物学进程三大类57个分支,其中分子功能大类中的结合(70.57%)与催化活性(45.51%)相关基因占比最多;KEGG代谢途径分析表明,7 427条基因参与了5类代谢通路,其中涉及信号转导的基因最多,为815条;筛选到7个候选OBP基因和5个具有全长开放阅读框的CSP基因,其编码蛋白均具有化学感受蛋白家族的典型特征。系统进化分析表明,花椒窄吉丁OBPs和CSPs分别与苹果小吉丁A. mali的OBPs和CSPs氨基酸序列一致性最高。RPKM值表明,嗅觉相关基因AzanOBP1和AzanOBP2在雌成虫触角中不表达,在雄成虫触角中微量表达;AzanOBP3在雄成虫触角中高丰度表达。【结论】首次获得了花椒窄吉丁成虫触角转录组数据,筛选到了花椒窄吉丁OBP, CSP, Or, IR和SNMP等的嗅觉相关基因。推测触角中高丰度表达的OBPs对雄成虫识别同类异性释放的信息素或寄主植物释放的挥发物起关键作用。研究结果可为花椒窄吉丁化学感受基因功能分析及嗅觉感受机制研究奠定分子基础。     

3种赤眼蜂对甜菜夜蛾和小菜蛾卵的寄生功能反应

为评估稻螟赤眼蜂Trichogramma japonicum Ashmead、螟黄赤眼蜂Trichogramma chilonis Ishii和玉米螟赤眼蜂Trichogramma ostriniae Pang et Chen对甜菜夜蛾Spodoptera exigua Hübner卵和小菜蛾Plutella xylostella Linnaeus卵的寄生潜能,本研究在室内条件下测试了3种赤眼蜂对不同密度甜菜夜蛾卵和小菜蛾卵的寄生率、寄生功能反应和搜寻效应。结果表明,稻螟赤眼蜂、螟黄赤眼蜂和玉米螟赤眼蜂对甜菜夜蛾卵和小菜蛾卵均有一定的寄生效应。螟黄赤眼蜂对甜菜夜蛾卵的寄生率、瞬间攻击率、寄生效能和搜寻效应均高于稻螟赤眼蜂和玉米螟赤眼蜂;3种赤眼蜂对小菜蛾卵的寄生率、瞬间攻击率、寄生效能和搜寻效应则表现为：稻螟赤眼蜂>螟黄赤眼蜂>玉米螟赤眼蜂。3种赤眼蜂对两种寄主的寄生功能反应均符合HollingⅡ型模型方程,表现为寄生量均随寄主卵密度的增加而升高,达到一定水平后趋于平稳。赤眼蜂的搜寻效应随寄主卵密度的增加而降低。综合分析可知,螟黄赤眼蜂对于甜菜夜蛾的控害能力优于稻螟赤眼蜂螟和玉米螟赤眼蜂,而稻螟赤眼蜂比螟黄赤眼蜂和玉米螟赤眼蜂更适合用于防控小菜蛾。     

A novel Meloidogyne graminicola effector,MgMO237, interacts with multiple host defence‐related proteins to manipulate plant basal immunity and promote parasitism

Plant‐parasitic nematodes can secrete effector proteins into the host tissue to facilitate their parasitism. In this study, we report a novel effector protein, MgMO237, from Meloidogyne graminicola, which is exclusively expressed within the dorsal oesophageal gland cell and markedly up‐regulated in parasitic third‐/fourth‐stage juveniles of M. graminicola. Transient expression of MgMO237 in protoplasts from rice roots showed that MgMO237 was localized in the cytoplasm and nucleus of the host cells. Rice plants overexpressing MgMO237 showed an increased susceptibility to M. graminicola. In contrast, rice plants expressing RNA interference vectors targeting MgMO237 showed an increased resistance to M. graminicola. In addition, yeast two‐hybrid and co‐immunoprecipitation assays showed that MgMO237 interacted specifically with three rice endogenous proteins, i.e. 1,3‐β‐glucan synthase component (OsGSC), cysteine‐rich repeat secretory protein 55 (OsCRRSP55) and pathogenesis‐related BetvI family protein (OsBetvI), which are all related to host defences. Moreover, MgMO237 can suppress host defence responses, including the expression of host defence‐related genes, cell wall callose deposition and the burst of reactive oxygen species. These results demonstrate that the effector MgMO237 probably promotes the parasitism of M. graminicola by interacting with multiple host defence‐related proteins and suppressing plant basal immunity in the later parasitic stages of nematodes.     

Superparasitism behavior and host discrimination of Campoletis chlorideae (Ichneumonidae: Hymenoptera) toward Mythimna separata (Noctuidae: Lepidoptera)

The oriental army worm, Mythimna separata Walker (Lepidoptera: Noctuidae), was adopted as a host to keep Campoletis chlorideae Uchida (Hymenoptera: Ichneumonidae) populations in the laboratory, because it can be mass reared. However, wasp cocoon production in this system via single parasitism was not satisfactory. To improve the rearing efficiency of C. chlorideae on M. separata, we studied the effects of superparasitism on the offspring production of C. chlorideae, as well as the host discrimination ability of the adult wasps in the laboratory. The results showed that, compared with single parasitism, both double and quintuple parasitism significantly increased C. chlorideae cocoon production in M. separata without a significant effect on emergence, weight, or sex ratio of adult wasps at the P < 0.05 level. Host selection experiments suggested that C. chlorideae preferred to lay eggs on unparasitized hosts rather than freshly parasitized hosts but could distinguish neither the 24-h postparasitization hosts from the unparasitized hosts nor the freshly self-parasitized hosts from the conspecifically parasitized hosts. No-choice behavior tests indicated that C. chlorideae took significantly longer time to accept the freshly parasitized hosts containing more than two eggs compared with the unparasitized hosts. In the end, superparasitism behavior and its application in the mass rearing of this endoparasitoid are discussed.