Multifaceted interactions between host ESCRT-III and budded virus-related proteins involved in entry and egress of the baculovirus Autographa californica multiple nucleopolyhedrovirus.

The endosomal sorting complex required for transport (ESCRT) is a conserved protein machine mediating membrane remodeling and scission. In the context of viral infection, different components of the ESCRT-III complex, which serve as the core machinery to catalyze membrane fission, are involved in diverse viruses' entry, replication, and/or budding. However, the interplay between ESCRT-III and viral factors in the virus life cycle, especially for that of large enveloped DNA viruses, is largely unknown. Recently, the ESCRT-III components Vps2B, Vps20, Vps24, Snf7, Vps46, and Vps60 were determined for entry and/or egress of the baculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV). Here, we identified the final three ESCRT-III components Chm7, Ist1, and Vps2A of Spodoptera frugiperda. Overexpression of the dominant-negative forms of these proteins or RNAi downregulation of their transcripts significantly reduced infectious budded viruses (BVs) production of AcMNPV. Quantitative PCR together with confocal and transmission electron microscopy analysis revealed that these proteins were required for internalization and trafficking of BV during entry and egress of nucleocapsids. In infected Sf9 cells, nine ESCRT-III components were distributed on the nuclear envelope and plasma membrane, and except for Chm7, the other components were also localized to the intranuclear ring zone. Y2H and BiFC analysis revealed that 42 out of 64 BV-related proteins including 35 BV structural proteins and 7 non-BV structural proteins interacted with single or multiple ESCRT-III components. By further mapping the interactome of 64 BV-related proteins, we established the interaction networks of ESCRT-III and the viral protein complexes involved in BV entry and egress.IMPORTANCEFrom archaea to eukaryotes, the endosomal sorting complex required for transport (ESCRT)-III complex is hijacked by many enveloped and nonenveloped DNA or RNA viruses for efficient replication. However, the mechanism of ESCRT-III recruitment, especially for that of large enveloped DNA viruses, remains elusive. Recently, we found the ESCRT-III components Vps2B, Vps20, Vps24, Snf7, Vps46, and Vps60 are necessary for the entry and/or egress of budded viruses (BVs) of Autographa californica multiple nucleopolyhedrovirus. Here, we demonstrated that the other three ESCRT-III components Chm7, Ist1, and Vps2A play similar roles in BV infection. By determining the subcellular localization of ESCRT-III components in infected cells and mapping the interaction of nine ESCRT-III components and 64 BV-related proteins, we built the interaction networks of ESCRT-III and the viral protein complexes involved in BV entry and egress. These studies provide a fundamental basis for understanding the mechanism of the ESCRT-mediated membrane remodeling for replication of baculoviruses.

Early monitoring of parasitism by Aphidiinae parasitoids on the grain aphid Sitobion miscanthi in wheat fields using DNA barcoding.

BACKGROUND: Sitobion miscanthi is a major wheat pest at the grain-filling stage found in China. Identifying parasitoid species and understanding parasitism rates are keys to controlling the aphids via natural enemies in the wheat field. RESULTS: In the present study, a method based on DNA barcoding for early determination of the community composition of Aphidiinae parasitoids and parasitism on the aphid was developed. The proposed method detected Aphidius gifuensis as the predominant parasite, with parasitism rates of 40.1 ± 2.8% in 2019 and 65.7 ± 3.7% in 2022, and found that the rate varied significantly among different wheat varieties. COI primers efficiently amplified the Aphidiinae parasitoids COI fragments and amplified the aphid COI fragments derived from parasitized (mummified) S. miscanthi. Thus, the COI barcode is not sufficiently specific to unambiguously detect immature parasitoids inside their S. miscanthi hosts. However, it can be used to detect the DNA extracted from mummified aphids. In contrast, the 16S and LWRh primers effectively amplified and identified the parasitoids in parasitized aphids. The 16S primer was reliable even in the early stages of parasitism (24 h) and for DNA samples stored at -20 °C for 5 days. The three barcodes from COI, 16S, and LWRh genes could not clearly distinguish a few certain Aphidiinae species owing to relatively low intraspecific and interspecific diversity. CONCLUSION: The morphological features remain indispensable when identifying Aphidiinae species. Nonetheless, the COI and 16S primers could be used in combination for monitoring the parasitism rates on S. miscanthi in wheat fields. © 2022 Society of Chemical Industry.

Effects of Crop Resistance on the Tritrophic Interactions between Wheat Lines, Schizaphis graminum (Hemitera: Aphididae), and Propylaea japonica (Coleoptera: Coccinellidae).

Crop resistance and biological control are both considered efficient and environmentally friendly methods of sustainable pest control. In this study, we aimed at investigating the direct influence of four wheat lines with varying resistance level on the life-history traits of the greenbug, Schizaphis graminum, and the mediational effect on the functional response of a predatory ladybird, Propylaea japonica, under laboratory conditions. Results showed that the aphid fitness was the lowest for aphids that had been feeding on wheat line '98-10-19' for one year. These aphids had the longest development time, and least adult mass, minimal mean relative growth rate, and lowest reproductive fitness. In contrast, the aphids that fed on wheat line '98-10-30' were the fittest, with the shortest development time and highest levels of reproductive fitness. The predatory activities of the ladybeetle, especially the adult male significantly decreased following the consumption of aphids belonging to the '98-10-19'-acclimated population. However, there were no significant differences in predatory efficiency (net attack frequency) among the four aphid acclimated populations. Our results showed that the wheat line '98-10-19' has a relative higher resistance to S. graminum than the other three wheat lines, which could further decrease the amount of prey available for consumption. However, the ecological effect of the resistance of '98-10-19' to S. graminum posed no negative influence on the biocontrol potential of P. japonica to these aphids, as their predatory efficiency increases at the fourth instar larvae phase.

Efficient entry of budded virions of Autographa californica multiple nucleopolyhedrovirus into Spodoptera frugiperda cells is dependent on dynamin, Rab5, and Rab11.

Autographa californica multiple nucleopolyhedrovirus (AcMNPV), a member of the Alphabaculovirus genus of the family Baculoviridae, is an enveloped double-stranded DNA virus. Budded virions (BVs) of AcMNPV enter host cells via clathrin-mediated endocytosis. However, the route of functional intracellular trafficking of AcMNPV BVs during entry is not well established. In the current study, we found that entering BVs were colocalized mainly with cellular Rab5 and Rab11. Expression of dominant-negative (DN) Rab5 and Rab11 or RNAi-mediated down regulation of these two cellular transcripts significantly reduced BVs entry into but not egress from Spodoptera frugiperda cells (Sf9), whereas similar treatments for Rab4 and Rab7 had no apparent effect on virus infection. Combined with data from RNAi knockdowns of dynamin, and dynasore inhibition assays, our results support a model in which AcMNPV BVs enter permissive host cells by clathrin-mediated endocytosis, followed by de-envelopment of BVs predominantly within early and maturing endosomes rather than within late endosomes. Additionally, Rab11 suppression studies suggest the Rab11-dependent recycling endosomal pathway is involved in virion entry.