Project description:The control of insect borne disease is recognized as one of the major agricultural, animal and human health challenges of today. Viruses in the family Luteoviridae are phloem-limited, plant viruses that are vectored by aphids in a circulative manner. They are responsible for a wide-range of economically important disease in almost all staple food crops. In order to be transmitted, these viruses require species-specific passage of the pathogen across several membrane barriers within the insect. After the pathogen is ingested from the sap of an infected plant, the virus moves across and through the aphid gut into the hemoceol (insect blood) where it circulates until it reaches and enters the main or accessory salivary glands. From here, the pathogen is injected into a new plant host when the aphid feeds. The identification of insect proteins that interact with circulative plant viruses is technically challenging and a major goal for the plant vector biology field. Such information is critical to develop novel control strategies that block virus transmission by insects. In this study, we used affinity purification-high-resolution mass spectrometry (AP-MS) to rapidly capture and identify aphid proteins in complex with Potato leafroll virus (PLRV), a luteovirid transmitted by the green peach aphid (Myzus persicae), directly from viruliferous aphid tissue.
Project description:70mer probes were designed to detect plant viruses infection in genus level. This microarray platform is able to detect 169 plant virus species of 13 virus genera.
Project description:Small RNAs play essential regulatory roles in genome stability, development and stress responses in most eukaryotes. Plants encode DICER-LIKE (DCL) RNaseIII enzymes, including DCL1, which produces miRNAs, and DCL2, DCL3 and DCL4, which produce diverse size classes of siRNA. Plants also encode RNASE THREE-LIKE (RTL) enzymes that lack DCL-specific domains and whose function is largely unknown. Small RNA sequencing in plants over-expressing RTL1 or RTL2 or lacking RTL2 revealed that RTL1 over-expression inhibits the accumulation of all types of small RNAs produced by DCL2, DCL3 and DCL4, indicating that RTL1 is a general suppressor of plant siRNA pathways. By contrast, RTL2 plays minor, if any, role in the small RNA repertoire. In vivo and in vitro assays revealed that RTL1 prevents siRNA production by degrading dsRNA before they are processed by DCL2, DCL3 and DCL4. The substrate of RTL1 cleavage is likely long perfect (or near-perfect) dsRNA, consistent with the RTL1-insensitivity of miRNAs, which derive from short imperfect dsRNA. RTL1 is naturally expressed only weakly in roots, but virus infection strongly induces its expression in leaves, suggesting that RTL1 induction is a general strategy used by viruses to counteract the siRNA-based plant antiviral defense. Accordingly, transgenic plants over-expressing RTL1 are more sensitive to TYMV infection than wild-type plants, likely because RTL1 prevents the production of antiviral siRNAs. However, TCV, TVCV and CMV, which encode stronger suppressors of RNA silencing (VSR) than TYMV, are insensitive to RTL1 over-expression. Indeed, TCV VSR inhibits RTL1 activity, suggesting that inducing RTL1 expression and dampening RTL1 activity is a dual strategy used by viruses to establish a successful infection. These results reveal another level of complexity in the evolutionary battle between viruses and plant defenses.