Parasitoid Serpins Evolve Novel Functions to Manipulate Host Homeostasis.

Parasitoids introduce various virulence factors when parasitism occurs, and some taxa generate teratocytes to manipulate the host immune system and metabolic homeostasis for the survival and development of their progeny. Host-parasitoid interactions are extremely diverse and complex, yet the evolutionary dynamics are still poorly understood. A category of serpin genes, named CvT-serpins, was discovered to be specifically expressed and secreted by the teratocytes of Cotesia vestalis, an endoparasitoid of the diamondback moth Plutella xylostella. Genomic and phylogenetic analysis indicated that the C. vestalis serpin genes are duplicated and most of them are clustered into 1 monophyletic clade. Intense positive selection was detected at the residues around the P1-P1' cleavage sites of the Cv-serpin reactive center loop domain. Functional analyses revealed that, in addition to the conserved function of melanization inhibition (CvT-serpins 1, 16, 18, and 21), CvT-serpins exhibited novel functions, i.e. bacteriostasis (CvT-serpins 3 and 5) and nutrient metabolism regulation (CvT-serpins 8 and 10). When the host-parasitoid system is challenged with foreign bacteria, CvT-serpins act as an immune regulator to reprogram the host immune system through sustained inhibition of host melanization while simultaneously functioning as immune effectors to compensate for this suppression. In addition, we provided evidence that CvT-serpin8 and 10 participate in the regulation of host trehalose and lipid levels by affecting genes involved in these metabolic pathways. These findings illustrate an exquisite tactic by which parasitoids win out in the parasite-host evolutionary arms race by manipulating host immune and nutrition homeostasis via adaptive gene evolution and neofunctionalization.

Comprehensive analysis of nanoplastic effects on growth phenotype, nanoplastic accumulation, oxidative stress response, gene expression, and metabolite accumulation in multiple strawberry cultivars.

Nanoplastics (NPs) have emerged as a novel environmental threat due to their potential impacts on both animals and plants. Currently, research on the ecotoxicity of NPs has mainly focused on marine aquatic organisms and freshwater algae, with very limited investigations conducted on horticultural plants. This study examined the effects of varying concentrations (0, 1, 10, 50 mg·L-1) of polystyrene NPs (PS-NPs) on strawberry growth. The findings revealed that low concentrations of PS-NPs stimulated strawberry growth, whereas high concentrations impeded it. Notably, diverse strawberry cultivars displayed considerable differences in their sensitivity to PS-NP exposure. Laser scanning confocal microscopy confirmed the absorption of PS-NPs by strawberry roots, with variations in PS-NP accumulation observed across different cultivars. Comparative transcriptomics analysis suggested that the differential expression of genes responsible for calcium ion transport played a significant role in the observed intervarietal differences in PS-NP accumulation among strawberry cultivars. Furthermore, distinct variations in endogenous oxidative responses were observed in different strawberry cultivars under PS-NP treatment. Further analysis indicated that the down-regulation of peroxidase (POD) gene expression and terpenoid compounds accumulation were responsible for heightened endogenous oxidative stress observed in certain strawberry cultivars under PS-NP treatment. Transcriptomic and metabolomic analyses were performed on six strawberry cultivars to investigate their response to PS-NPs in terms of endogenous gene expression and metabolite accumulation. The results identified one commonly up-regulated gene (wall-associated receptor kinase-like) and sixteen commonly down-regulated genes associated with lipid metabolism and carbohydrate metabolism. In addition, a significant reduction in fatty acid metabolite accumulation was observed in the six strawberry cultivars under PS-NP treatment. These findings have significant implications for understanding the effects of NPs on strawberry growth, metabolism, and antioxidant responses, as well as identifying marker genes for monitoring and evaluating the impact of NP pollution on strawberry.

The complete mitochondrial genome of Nasutitermes tiantongensis Zhou et Xu, 1993 (Isoptera: Termitidae: Nasutitermitinae).

Nasutitermes tiantongensis belongs to Nasutitermitinae and its mitochondrial genome was determined in this study. It consisted of 13 PCGs, 22 tRNAs, 2 rRNAs, and an A + T-rich control region, and its length was 15824 bp. The phylogenetic analysis suggested that the genus Nasutitermes was not monophyletic, and N. tiantongensis formed a sister group with Bultitermes laticephalus. The mitochondrial genome of N. tiantongensis provides a resource for evolutionary analysis within Nasutitermitinae.

A teratocyte-specific serpin from the endoparasitoid wasp Cotesia vestalis inhibits the prophenoloxidase-activating system of its host Plutella xylostella.

Many endoparasitoids adopt several parasitic factors, such as venom, polydnavirus and teratocytes, to suppress the immune response of their associated hosts including melanization for successful parasitism. A teratocyte-specific expressed serpin gene, designated as CvT-serpin6, was identified from the parasitoid Cotesia vestalis. The immunoblot result suggested that CvT-serpin6 was secreted into extracellular space. qPCR results showed that CvT-serpin6 was mainly transcribed at later stages of parasitism, and the transcriptional abundance of CvT-serpin6 in teratocytes was significantly increased in response to the challenge of bacteria. Inhibitory assay indicated that recombinant CvT-serpin6 (rCvT-serpin6) could inhibit the activation of Plutella xylostella prophenoloxidase and ultimately resulted in the inhibition of melanization in P. xylostella haemolymph. Furthermore, we confirmed that rCvT-serpin6 could form SDS-stable complexes with activated PxPAP1 and PxPAP3 in a dose-dependent manner. Altogether, our results further shed insight into the molecular mechanisms that teratocytes involved in controlling host immune response.

Genome-wide characterization of the SHORT INTER-NODES/STYLISH and Shi-Related Sequence family in Gossypium hirsutum and functional identification of GhSRS21 under salt stress.

Saline stress is a significant factor that caused crop growth inhibition and yield decline. SHORT INTERNODES/STYLISH (SHI/STY) and SHI-RELATED SEQUENCE (SRS) transcription factors are specific to plants and share a conserved RING-like zinc-finger domain (CX2CX7CX4CX2C2X6C). However, the functions of SHI/STY and SRS genes in cotton responses to salt stress remain unclear. In this study, 26 GhSRSs were identified in Gossypium hirsutum, which further divided into three subgroups. Phylogenetic analysis of 88 SRSs from8 plant species revealed independent evolutionary pattern in some of SRSs derived from monocots. Conserved domain and subcellular location predication of GhSRSs suggested all of them only contained the conserved RING-like zinc-finger domain (DUF702) domain and belonged to nucleus-localized transcription factors except for the GhSRS22. Furthermore, synteny analysis showed structural variation on chromosomes during the process of cotton polyploidization. Subsequently, expression patterns of GhSRS family members in response to salt and drought stress were analyzed in G. hirsutum and identified a salt stress-inducible gene GhSRS21. The GhSRS21 was proved to localize in the nuclear and silencing it in G. hirsutum increased the cotton resistance to salt using the virus-induced gene silencing (VIGS) system. Finally, our transcriptomic data revealed that GhSRS21 negatively controlled cotton salt tolerance by regulating the balance between ROS production and scavenging. These results will increase our understanding of the SRS gene family in cotton and provide the candidate resistant gene for cotton breeding.

A serpin (CvT-serpin15) of teratocytes contributes to microbial-resistance in Plutella xylostella during Cotesia vestalis parasitism.

BACKGROUND: Parasitic wasps are an important group of entomophagous insects for pest control. As parasitic wasps often lay eggs on or into their associated hosts, parasitoids evolve to utilize several factors including venom, polydnavirus (PDV) to alter host physiology for successful parasitism. Some taxa of endoparasitoids produce teratocytes, which are a type of cell that is released into host insects when wasp eggs hatch. Teratocytes display multifunction in parasitism such as host nutritional exploration, immune and developmental regulation, by secreting plenty of proteins into host hemocoel. RESULTS: A serpin (CvT-serpin15) secreted by teratocytes was characterized. QPCR results showed the expressional level of CvT-serpin15 was upregulated following bacterial challenges. Enzyme activity experiment indicated the recombinant CvT-serpin15 protein could interfere with the growth of Gram-positive bacteria Staphylococcus aureus. The survival rate assay demonstrated CvT-serpin15 increased survival rate of Plutella xylostella infected by S. aureus. CONCLUSION: CvT-serpin15 secreted by teratocytes would boost the host immune system when pathogens invade host hemocoel during parasitism, and ultimately protect the development of wasp larva from bacterial infection. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

The genomes of two parasitic wasps that parasitize the diamondback moth.

BACKGROUND: Parasitic insects are well-known biological control agents for arthropod pests worldwide. They are capable of regulating their host's physiology, development and behaviour. However, many of the molecular mechanisms involved in host-parasitoid interaction remain unknown. RESULTS: We sequenced the genomes of two parasitic wasps (Cotesia vestalis, and Diadromus collaris) that parasitize the diamondback moth Plutella xylostella using Illumina and Pacbio sequencing platforms. Genome assembly using SOAPdenovo produced a 178 Mb draft genome for C. vestalis and a 399 Mb draft genome for D. collaris. A total set that contained 11,278 and 15,328 protein-coding genes for C. vestalis and D. collaris, respectively, were predicted using evidence (homology-based and transcriptome-based) and de novo prediction methodology. Phylogenetic analysis showed that the braconid C. vestalis and the ichneumonid D. collaris diverged approximately 124 million years ago. These two wasps exhibit gene gains and losses that in some cases reflect their shared life history as parasitic wasps and in other cases are unique to particular species. Gene families with functions in development, nutrient acquisition from hosts, and metabolism have expanded in each wasp species, while genes required for biosynthesis of some amino acids and steroids have been lost, since these nutrients can be directly obtained from the host. Both wasp species encode a relative higher number of neprilysins (NEPs) thus far reported in arthropod genomes while several genes encoding immune-related proteins and detoxification enzymes were lost in both wasp genomes. CONCLUSIONS: We present the annotated genome sequence of two parasitic wasps C. vestalis and D. collaris, which parasitize a common host, the diamondback moth, P. xylostella. These data will provide a fundamental source for studying the mechanism of host control and will be used in parasitoid comparative genomics to study the origin and diversification of the parasitic lifestyle.

A trypsin inhibitor-like protein secreted by Cotesia vestalis teratocytes inhibits hemolymph prophenoloxidase activation of Plutella xylostella.

To establish successful infections, endoparasitoid wasps must develop strategies to evade immune responses of the host. Here, we identified and characterized a teratocytes-expressed gene encoding a trypsin inhibitor-like protein containing a cysteine-rich domain from Cotesia vestalis, CvT-TIL. CvT-TIL had a high expression level during the later developmental stage of teratocytes and was secreted into host hemolymph. Further experiments showed CvT-TIL strongly suppressed the prophenoloxidase activation of host hemolymph in a dose-dependent manner by interacting with PxPAP3 of PO cascade. Our results not only provide evidence for an inhibition between CvT-TIL gene and the host's melanization activity, but also expand our knowledge about the mechanisms by which parasitoids regulate humoral immunity of the host.

Parasitic insect-derived miRNAs modulate host development.

Parasitic wasps produce several factors including venom, polydnaviruses (PDVs) and specialized wasp cells named teratocytes that benefit the survival of offspring by altering the physiology of hosts. However, the underlying molecular mechanisms for the alterations remain unclear. Here we find that the teratocytes of Cotesia vestalis, an endoparasitoid of the diamondback moth Plutella xylostella, and its associated bracovirus (CvBV) can produce miRNAs and deliver the products into the host via different ways. Certain miRNAs in the parasitized host are mainly produced by teratocytes, while the expression level of miRNAs encoded by CvBV can be 100-fold greater in parasitized hosts than non-parasitized ones. We further show that one teratocyte-produced miRNA (Cve-miR-281-3p) and one CvBV-produced miRNA (Cve-miR-novel22-5p-1) arrest host growth by modulating expression of the host ecdysone receptor (EcR). Altogether, our results show the first evidence of cross-species regulation by miRNAs in animal parasitism and their possible function in the alteration of host physiology during parasitism.

Cotesia vestalis teratocytes express a diversity of genes and exhibit novel immune functions in parasitism.

Some endoparasitoid wasps lay eggs that produce cells called teratocytes. In this study, we sequenced and analyzed the transcriptome of teratocytes from the solitary endoparasitoid Cotesia vestalis (Braconidae), which parasitizes larval stage Plutella xylostella (Plutellidae). Results identified many teratocyte transcripts with potential functions in affecting host immune defenses, growth or metabolism. Characterization of teratocyte-secreted venom-like protein 8 (TSVP-8) indicated it inhibits melanization of host hemolymph in vitro, while two predicted anti-microbial peptides (CvT-def 1 and 3) inhibited the growth of bacteria. Results also showed the parasitized hosts lacking teratocytes experienced higher mortality after immune challenge by pathogens than hosts with teratocytes. Taken together, these findings indicate that C. vestalis teratocytes secrete products that alter host immune functions while also producing anti-microbial peptides with functions that help protect the host from infection by other organisms.