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In cloud-native environments, containers are often deployed within lightweight virtual machines (VMs) to ensure strong security isolation and privacy protection. With the growing demand for customized cloud services, third-party vendors are turning to infrastructure-as-a-service (IaaS) cloud providers to build their own cloud-native platforms, necessitating the need to run a VM or a guest that hosts containers inside another VM instance leased from an IaaS cloud. State-of-the-art nested virtualization in the x86 architecture relies heavily on the host hypervisor to expose hardware virtualization support to the guest hypervisor, not only complicating cloud management but also raising concerns about an increased attack surface at the host hypervisor. This paper presents the design and implementation of PVM, a high-performance guest hypervisor for KVM that is transparent to the host hypervisor and assumes no hardware virtualization support. PVM leverages two key designs: 1) a minimal shared memory region between the guest and guest hypervisor to facilitate state transition between different privilege levels and 2) an efficient shadow page table design to reduce the cost of memory virtualization. PVM has been adopted by a major IaaS cloud provider for hosting tens of thousands of secure containers on a daily basis. Our experiments demonstrate that PVM significantly outperforms current nested virtualization in KVM for memory virtualization, particularly for concurrent workloads, while maintaining comparable performance in CPU and I/O virtualization.more » « less
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Introduction Nosema is a diverse genus of unicellular microsporidian parasites of insects and other arthropods.Nosema muscidifuracis infects parasitoid wasp species ofMuscidifurax zaraptor andM. raptor (Hymenoptera: Pteromalidae), causing ~50% reduction in longevity and ~90% reduction in fecundity.Methods and Results Here, we report the first assembly of the
N. muscidifuracis genome (14,397,169 bp in 28 contigs) of high continuity (contig N50 544.3 Kb) and completeness (BUSCO score 97.0%). A total of 2,782 protein-coding genes were annotated, with 66.2% of the genes having two copies and 24.0% of genes having three copies. These duplicated genes are highly similar, with a sequence identity of 99.3%. The complex pattern suggests extensive gene duplications and rearrangements across the genome. We annotated 57 rDNA loci, which are highly GC-rich (37%) in a GC-poor genome (25% genome average).Nosema -specific qPCR primer sets were designed based on 18S rDNA annotation as a diagnostic tool to determine its titer in host samples. We discovered highNosema titers inNosema -curedM. raptor andM. zaraptor using heat treatment in 2017 and 2019, suggesting that the remedy did not completely eliminate theNosema infection. Cytogenetic analyses revealed heavy infections ofN. muscidifuracis within the ovaries ofM. raptor andM. zaraptor , consistent with the titer determined by qPCR and suggesting a heritable component of infection and per ovum vertical transmission.Discussion The parasitoids-
Nosema system is laboratory tractable and, therefore, can serve as a model to inform future genome manipulations ofNosema -host system for investigations of Nosemosis. -
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Abstract Effective cellular signaling relies on precise spatial localization and dynamic interactions among proteins in specific subcellular compartments or niches, such as cell-to-cell contact sites and junctions. In plants, endogenous and pathogenic proteins gained the ability to target plasmodesmata, membrane-lined cytoplasmic connections, through evolution to regulate or exploit cellular signaling across cell wall boundaries. For example, the receptor-like membrane protein PLASMODESMATA-LOCATED PROTEIN 5 (PDLP5), a potent regulator of plasmodesmal permeability, generates feed-forward or feed-back signals important for plant immunity and root development. However, the molecular features that determine the plasmodesmal association of PDLP5 or other proteins remain largely unknown, and no protein motifs have been identified as plasmodesmal targeting signals. Here, we developed an approach combining custom-built machine-learning algorithms and targeted mutagenesis to examine PDLP5 in Arabidopsis thaliana and Nicotiana benthamiana. We report that PDLP5 and its closely related proteins carry unconventional targeting signals consisting of short stretches of amino acids. PDLP5 contains 2 divergent, tandemly arranged signals, either of which is sufficient for localization and biological function in regulating viral movement through plasmodesmata. Notably, plasmodesmal targeting signals exhibit little sequence conservation but are located similarly proximal to the membrane. These features appear to be a common theme in plasmodesmal targeting.
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