Transcriptome Analysis Reveals Functional Diversity in Salivary Glands of Plant Virus Vector, Graminella nigrifrons

Insect salivary glands play an important role for host feeding, specifically by secreting salivary proteins for digestion and potentially modulating host defenses. Compared to other hemipterans, the significance of salivary glands is less studied in the black-faced leafhopper, Graminella nigrifrons, a crop pest that vectors several agronomically important plant viruses. To identify functionally important genes in the salivary glands of the black-faced leafhopper, we compared transcriptomes between adult salivary glands (SG) and the remaining carcasses. We identified 14,297 salivary gland-enriched transcripts and 195 predicted secretory peptides (i.e., with a signal peptide and extracellular localization characteristics). Overall, the SG transcriptome included functions such as ‘oxidoreduction’, ‘membrane transport’, and ‘ATP-binding’, which might be important for the fundamental physiology of this tissue. We further evaluated transcripts with potential contributions in host feeding using RT-qPCR. Two SG-enriched transcripts (log2 fold change > 5), GnP19 and GnE63 (a putative calcium binding protein), were significantly upregulated in maize-fed adults relative to starved adults, validating their importance in feeding. The SG-enriched transcripts of the black-faced leafhopper could play a potential role for interacting with maize and could be targets of interest for further functional studies and improve pest control and disease transmission.

Perilla Mosaic Virus Is a Highly Divergent Emaravirus Transmitted by Shevtchenkella sp. (Acari: Eriophyidae)

Shiso (Perilla frutescens var. crispa) is widely grown as an important vegetable or herb crop in Japan. Beginning around the year 2000, occurrences of severe mosaic symptoms on shiso were documented and gradually spread across Kochi Prefecture, one of four major shiso production areas in Japan. Next generation sequencing and cloning indicated the presence of a previously unknown virus related to the members of the genus Emaravirus, for which we proposed the name Perilla mosaic virus (PerMV). The genome of PerMV consists of 10 RNA segments, each encoding a single protein in the negative-sense orientation. Of these proteins, P1, P2, P3a, P3b, P4, and P5 show amino acid sequence similarities with those of known emaraviruses, whereas no similarities were found in P6a, P6b, P6c, and P7. Characteristics of the RNA segments as well as phylogenetic analysis of P1 to P4 indicate that PerMV is a distinct and highly divergent emaravirus. Electron microscopy observations and protein analyses corresponded to presence of an emaravirus. Transmission experiments demonstrated that an eriophyid mite, Shevtchenkella sp. (family Eriophyidae), transmits PerMV with a minimum 30-min acquisition access period. Only plants belonging to the genus Perilla tested positive for PerMV, and the plant−virus−vector interactions were evaluated. The nucleotide sequences reported here are available in the DDBJ/ENA/GenBank databases under accession numbers LC496090 to LC496099.

Diagnostic of begomoviruses in complex infections –a case study

Complexes of viruses inducing a syndrome in plants strongly hinder the identification of the causal agent of a disease. The Sida micrantha mosaic disease is associated with a complex of begomoviruses. For more than twenty years, two DNAs A (DNA A2 and DNA A3) belonging to this complex could neither be detected nor isolated from Sida micrantha Schr. plants, although one of them (DNA A2) now appears to be the major component of the complex. A random unintended Bemisia tabaci-mediated transmission of begomoviruses from several Sida species – including S. micrantha – to experimental Malva parviflora plants resulted in the serendipitous finding of these new DNAs A. Simultaneously, a number of other begomoviruses infecting Sida plants from several Latin American countries were transmitted to M. parviflora plants and the convergence of them resulted in natural pseudorecombinants. Pseudorecombination, however, took place exclusively between heterologous genomic components that shared identical binding sites for the replication-associated protein AC1. This case study constitutes an exceptional opportunity for the analysis of plant–virus– vector interactions in a quasi-natural environment. In addition to that, the methodology described here may be used to isolate and characterize different begomoviruses inducing a syndrome during mix infections.

Discovery of Known and Novel Viral Genomes in Soybean Aphid by Deep Sequencing

The phytobiome includes not just cellular microorganisms, but also all viruses associated with plants: the virome. Plants and aphids exchange viruses regularly and efficiently; thus, the plant virome is tightly linked with the aphid virome. Yet, little is known about aphid viromes, particularly that of the soybean aphid (Aphis glycines), one of the most economically important pest insects of soybean. To sample the soybean aphid virome, and to seek new viruses as potential biological control agents, we employed next-generation sequencing. Genomes isolated from viruses in soybean aphids collected at four sites revealed many viruses, and six complete or nearly complete genomes were assembled. Most abundant were the picornavirus-like dicistroviruses Aphid lethal paralysis virus and Rhopalosiphum padi virus. We also sequenced the genome of a new dicistrovirus, Big Sioux River virus, fragments of which had been found previously in honey bee. Genome sequences that represent two entirely new virus families were obtained. These include an abundant tetravirus-like virus and a virus distantly related to cileviruses of plants and negeviruses of insects. Surprisingly, Cotton leafroll dwarf virus, a member of the genus Polerovirus, was found in soybean aphids from China, suggesting that the soybean aphid may be a vector of this virus. This virus had not been reported previously in China or in soybean. This study provides a peek into the rapidly expanding, largely unexplored world of insect viromes that will provide valuable knowledge for future understanding of plant−virus−vector interactions. [Formula: see text] Copyright © 2017 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .