Capturing pathogenic plant waterborne viruses: A novel tool for agricultural diagnostics using three model viruses
Abstract
With continued growth of world population, a greater demand of natural resources is generated. Water, a natural resource used directly or indirectly by all is an essential resource capable of being contaminated with waterborne plant pathogens. For sustainable agriculture, growers take advantage of non-favorable cropping areas by pumping water from nearby lakes, rivers, wells, aquifers, or even runoff. These various water resources could be potential reservoirs for waterborne phytopathogens and act as a microbiome leading to their introduction in cropping systems. When considering large bodies of water and water dynamics, including dilution factors and volume, it becomes challenging to detect and identify potential pathogens, especially plant viruses, which are in very low numbers. Current plant pathogen detection tools are used retroactively by sampling various tissues or soils following disease symptoms. With highly virulent pathogens this can lead to unacceptable losses. There is a need to develop a system to monitor water sources for the presence of waterborne plant viruses to prevent accidental or intentional introduction from irrigation sources. The objective of this project is to develop a preemptive detection system that will readily sample water for plant waterborne viruses, develop laboratory protocols to process environmental water samples, and to establish biological significance of waterborne viruses. An inexpensive, scalable and robust water sampling device made from polyvinyl chloride (PVC) was developed. A protocol for elution, precipitation and RNA extraction of waterborne viruses from water samples was established. Plant based multiplex primers were adapted for nucleic acid based detection of three viral model viruses include Pepino mosaic virus (PepMV), Tomato bushy stunt virus (TBSV), and Pepper mild mottle virus (PMMoV) belonging to the genus Potexvirus, Tombusvirus, and Tobamovirus, respectively. All three viruses were recovered from water volumes up to 5 gal. Hydroponics test demonstrated movement of PepMV, TBSV, and PMMoV from inoculated plants to water and into healthy plants where they established diseases. Monitoring of microbial loads in agricultural irrigation systems, and other water sources, is essential for effective surveillance and disease prevention for plant and animal health.
Collections
- OSU Dissertations [11222]