Seed lots of tomato and capsicum (Solanum lycopersicon and Capsicum annuum, respectively) are required to be free of quarantine pests before their entry to Australia is permitted. Testing of samples from 118 larger seed lots in the period 2019–2021 revealed that 31 (26.3%) carried one or more of four Tobamovirus species, including tomato mottle mosaic virus (ToMMV), which is a quarantine pest for Australia. Testing of samples from a further 659 smaller seed lots showed that 123 (18.7%) carried a total of five Tobamovirus species, including ToMMV and tomato brown rugose fruit virus (ToBRFV), which is also a quarantine pest for Australia. Estimated prevalence of contamination by tobamoviruses ranged from 0.388% to 0.004% in contaminated larger seed lots. Analyses of these data allow us to estimate probabilities of detection of contamination under different regulatory settings.

The Tobamovirus, Tomato mottle mosaic virus (ToMMV), was first reported in Mexico in 2013. The virus is thought to pose a serious risk to capsicum (Capsicum annuum) and tomato (Solanum lycopersicum) crops as it is may break resistance. In May 2019 a shipment of imported capsicum seeds was submitted for testing on-shore and using a one-step RT-PCR which detects Solanaceous tobamoviruses, an amplicon of 811 base pairs (bp) was detected and direct sequencing of this amplicon indicated that it had 98% - 99% nucleotide (nt) identity to the same region in ToMMV isolates. A cDNA library was generated using the Illumina TruSeq Stranded Total RNA and sequenced using Illumina HiSeq 3000 technology, bioinformatic analysis confirmed the arrangement of a 6398 nt genome which was 98% - 99% nt identity with the type strain of ToMMV. This is the first report of ToMMV in Capsicum annuum seed.

Rapid and reliable detection tools are essential for disease surveillance and outbreak management, and genomic data is essential to determining pathogen origin and monitoring of transmission pathways. Low virus copy number and poor RNA quality can present challenges for genomic sequencing of plant viruses, but this can be overcome by enrichment of target nucleic acid. A targeted whole genome sequencing (TWG-Seq) approach for the detection of cucumber green mottle mosaic virus (CGMMV) has been developed where overlapping amplicons generated using two multiplex RT-PCR assays are then sequenced using the Oxford Nanopore MinION. Near complete coding region sequences were assembled with ≥100× coverage for infected leaf tissue dilution samples with RT-qPCR cycle quantification (Cq) values from 11.8 to 38 and in seed dilution samples with Cq values 13.8 to 27. Consensus sequences assembled using this approach showed greater than 99% nucleotide similarity when compared to genomes produced using metagenomic sequencing. CGMMV could be confidently detected in historical seed isolates with degraded RNA. Whilst limited access to, and costs associated with second-generation sequencing platforms can influence diagnostic outputs, the portable Nanopore technology offers an affordable high throughput sequencing alternative when combined with TWG-Seq for low copy or degraded samples.

Cucumber green mottle mosaic virus (CGMMV) is a Tobamovirus of economic importance affecting cucurbit crops and Asian cucurbit vegetables. CGMMV was detected in the Northern Territory (NT) in September 2014, the first record for Australia, with 26 properties confirmed as of May 2016. Research was undertaken to determine virus longevity in soils in the NT and investigate the use of disinfectants to remove viable CGMMV from the soil. An in-field trial at 12 months post-quarantine at four properties, and bioassays from collected soils indicate that CGMMV remained viable in at least two of the properties 12 months after plant hosts were removed from the ground. The infectivity of CGMMV from soil was also investigated in two trials with 140 watermelon seeds and 70 watermelon plants sown into CGMMV infested soils with or without the application of the disinfectants Virkon^TM (2%) and Bleach (1%). Watermelons grown in soil, not treated with the Virkon^TM or Bleach, showed CGMMV infection rates of 4% and 2.5% respectively. When Virkon^TM or Bleach was applied, no positive CGMMV detections were observed in the watermelons. This research highlights the importance of proper management of infested properties and the need for on-farm biosecurity to manage CGMMV.

Grapevine Pinot gris virus (GPGV; genus Trichovirus in the family Betaflexiviridae) was detected in Australia in 2016, but its impact on the production of nursery material and fruit in Australia is still currently unknown. This study investigated the prevalence and genetic diversity of GPGV in Australia. GPGV was detected by reverse transcription-polymerase chain reaction (RT-PCR) in a range of rootstock, table and wine grape varieties from New South Wales, South Australia, and Victoria, with 473/2171 (21.8%) samples found to be infected. Genomes of 32 Australian GPGV isolates were sequenced and many of the isolates shared high nucleotide homology. Phylogenetic and haplotype analyses demonstrated that there were four distinct clades amongst the 32 Australian GPGV isolates and that there were likely to have been at least five separate introductions of the virus into Australia. Recombination and haplotype analysis indicate the emergence of new GPGV strains after introduction into Australia. When compared with 168 overseas GPGV isolates, the analyses suggest that the most likely origin of Australian GPGV isolates is from Europe. There was no correlation between specific GPGV genotypes and symptoms such as leaf mottling, leaf deformation, and shoot stunting, which were observed in some vineyards, and the virus was frequently found in symptomless grapevines.

Cucumber green mottle mosaic virus (CGMMV) is a Tobamovirus of economic importance affecting cucurbit crops and Asian cucurbit vegetables. Non-host crops of CGMMV, including capsicum (Capsicum annum), sweetcorn (Zea mays), and okra (Abelmoschus esculentus), were tested for their susceptibility to the virus, with field and glasshouse trials undertaken. After 12 weeks post-sowing, the crops were tested for the presence of CGMMV, and in all cases, no CGMMV was detected. Commonly found within the growing regions of cucurbits and melons worldwide are weeds, such as black nightshade (Solanum nigrum), wild gooseberry (Physalis minima), pigweed (Portulaca oleracea), and Amaranth species. Several weeds/grasses were tested for their ability to become infected with CGMMV by inoculating weeds directly with CGMMV and routinely testing over a period of eight weeks. Amaranthus viridis was found to be susceptible, with 50% of the weeds becoming infected with CGMMV. To further analyse this, six Amaranth samples were used as inoculum on four watermelon seedlings per sample and tested after eight weeks. CGMMV was detected in three of six watermelon bulk samples, indicating that A. viridis is a potential host/reservoir for CGMMV. Further research into the relationship between CGMMV and weed hosts is required. This research also highlights the importance of proper weed management to effectively manage CGMMV.