The field of biosecurity has greatly benefited from the widespread adoption of high-throughput sequencing technologies, for its ability to deeply query plant and animal samples for pathogens for which no tests exist. However, the bioinformatics analysis tools designed for rapid analysis of these sequencing datasets are not developed with this application in mind, limiting the ability of diagnosticians to standardise their workflows using published tool kits. We sought to assess previously published bioinformatic tools for their ability to identify plant- and animal-infecting viruses while distinguishing from the host genetic material. We discovered that many of the current generation of virus-detection pipelines are not adequate for this task, being outperformed by more generic classification tools. We created synthetic MinION and HiSeq libraries simulating plant and animal infections of economically important viruses and assessed a series of tools for their suitability for rapid and accurate detection of infection, and further tested the top performing tools against the VIROMOCK Challenge dataset to ensure that our findings were reproducible when compared with international standards. Our work demonstrated that several methods provide sensitive and specific detection of agriculturally important viruses in a timely manner and provides a key piece of ground truthing for method development in this space.
To protect New Zealand’s unique ecosystems and primary industries, imported plant materials must be constantly monitored at the border for high-threat pathogens. Techniques adopted for this purpose must be robust, accurate, rapid, and sufficiently agile to respond to new and emerging threats. Polymerase chain reaction (PCR), especially real-time PCR, remains an essential diagnostic tool but it is now being complemented by high-throughput sequencing using both Oxford Nanopore and Illumina technologies, allowing unbiased screening of whole populations. The demand for and value of Point-of-Use (PoU) technologies, which allow for in situ screening, are also increasing. Isothermal PoU molecular diagnostics based on recombinase polymerase amplification (RPA) and loop-mediated amplification (LAMP) do not require expensive equipment and can reach PCR-comparable levels of sensitivity. Recent advances in PoU technologies offer opportunities for increased specificity, accuracy, and sensitivities which makes them suitable for wider utilization by frontline or border staff. National and international activities and initiatives are adopted to improve both the plant virus biosecurity infrastructure and the integration, development, and harmonization of new virus diagnostic technologies.
This collaborative work by over 180 researchers from 40+ countries addresses the challenges posed by “phantom agents”—putative pathogenic agents named in literature without supporting data on their existence. Those agents remain on regulatory lists, creating barriers in trade and plant certification. Historically identified based solely on symptoms, these agents lack isolates or sequence data, making reliable detection or risk assessment impossible. After reviewing over 120 such agents across 10 key plant genera, we recommend their removal from regulatory lists and call for revised standards aligned with modern diagnostics. This effort seeks to streamline germplasm exchange, benefiting global agriculture by removing the constraints imposed by phantoms.
More than 200 plant viruses and many of their invertebrate vectors have invaded New Zealand (NZ) in the last two centuries. All of these invaders are associated with introduced agricultural, horticultural and/or weed species. At least 16 of the viruses have invaded the native flora, including some rare and critically endangered species. Patterns are emerging: aphid transmitted viruses which are able to infect species from a number of families are prominent. For example, cucumber mosaic virus infects native species from seven families. There are also examples of viruses with more restricted host ranges invading individual families, particularly the native grasses. The yellow dwarf viruses have escaped from cereals and pasture into native grasses. Some of the species are also native to Australia and the Pacific Islands and the review and its literature should be of interest to those working further afield. Prospects for controlling or mitigating the effects of the viruses in agricultural systems have limited application in the native flora but they are relevant to propagation and rescue strategies. Biosecurity measures are not only the first line of defence against threats to agriculture but also to the conservation estate. The protection of native floras from virus invasion is another justification for the control measures and legislative procedures already in place to protect agriculture. High throughput sequencing to detect all viruses at the border, and genetic engineering and RNAi technologies to mitigate the effects of invasions are promising developments for the protection of the native flora.
Peonies (Paeonia lactiflora) are beautiful ornamental plants, popular in the South Island and lower North Island of New Zealand (NZ). From 1998 to the present, NZ peonies’ breeders have been introd...