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• W4382449102 abstract "Grapevine rubodvirus 2 (syn. Grapevine Muscat rose virus; GMRV, genus Rubodvirus, family Phenuiviridae) is a negative-strand RNA virus with a tri-segmented genome (Diaz-Lara et al., 2019). It has only been described in a symptomless grapevine (Vitis vinifera cv. Muscat Rose) from the USDA National Clonal Germplasm Repository, which originated from Argentina (Diaz-Lara et al., 2019). In December 2021, shoots with mottling and deformation symptoms on leaves were collected from a grapevine cv. Malbec, at a vineyard in the Sunraysia region of Victoria, Australia. Total RNA was extracted from 0.3 g leaf tissue (MacKenzie et al., 1997), libraries were prepared using a TruSeq® Stranded Total RNA Library Prep Plant with Ribo-Zero Plant kit (Illumina, USA) and high throughput sequencing was done using a NovaSeq platform (Illumina, USA) with 2 × 150 bp paired-end reads. Sequence reads were trimmed using Trim Galore! Version 0.6.5 and assembled into contigs using SPADES Version 3.13.0 (Bankevich et al., 2012). BLASTn analysis identified a total of 46 viral contigs, of which 36 contigs aligned most closely with three RNA genome segments of GMRV, one contig matched with Grapevine Pinot gris virus (GPGV) and nine contigs matched with Grapevine rupestris stem pitting-associated virus. Reference mapping using Bowtie2 Version 2.3.4.2 (Langmead & Salzberg, 2012) illustrated 100% completeness of the Australian GMRV genomic regions and RNA1, RNA2 and RNA3 had 97.6%, 99.3% and 98.6% nucleotide identity, respectively, to the three published GMRV RNA segments (GenBank Accession Nos. MK728654.1, MK728655.1 and MK728656.1). The presence of RNA 1 and RNA 3 in the sample was confirmed using two published GMRV-specific endpoint RT-PCR assays (Diaz-Lara et al., 2019). A primer pair (GMRV-MP-Forward 5′-TGGGTGGCTATCATCACTGT-3′; GMRV-MP-Reverse 5′-CATCTCTTGCACCTTTCGGC-3′) was developed in this study to amplify a 620 bp nucleotide region of the movement protein gene (MP) and confirmed the presence of RNA2. Direct bi-directional Sanger sequencing of amplicons also confirmed the detection of RNA1, RNA 2 and RNA3 by RT-PCR. The Australian GMRV isolate was also detected using RT-PCR and Sanger sequencing in a previously uninfected Cabernet Franc grapevine that was grafted with the infected Malbec material, further confirming its presence in the sample and graft transmissibility. Additional samples were collected from six Malbec grapevines at a second vineyard in Sunraysia, which had no visible symptoms and were grown from Malbec clones imported from Argentina. Amplicons of the expected size were observed in one sample after testing with each of the three RT-PCR assays. To our knowledge, this is the first report of GMRV in Australia and the sequences for RNA1, RNA2 and RNA3 are available in GenBank (OQ418650, OQ418651 and OQ418652, respectively). There are no known vectors of GMRV but the virus is transmitted through vegetative propagation. Therefore, use of planting material in which the virus hasn't been detected is advisable to minimise risk. The correlation between the Australian GMRV isolate and symptoms in diseased Malbec requires further investigation because of the presence of other viruses, particularly GPGV, that might also be associated with the symptoms observed. However, detection of both GMRV and GPGV in the asymptomatic cv. Malbec sample suggests that neither virus are the sole cause of the disease. Further surveillance is required to determine the distribution of GMRV and its association with disease in Australia. This research was funded by La Trobe University through their provision of a La Trobe Full Fee Research Scholarship and La Trobe University Graduate Research Scholarship, and a PhD research scholarship by Wine Australia. We would like to acknowledge Agriculture Victoria Research as this research was conducted using the facilities of Agriculture Victoria." @default.
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• W4382449102 date "2023-04-01" @default.
• W4382449102 modified "2023-09-26" @default.
• W4382449102 title "First report of <i>Grapevine rubodvirus 2</i> infecting <i>Vitis vinifera</i> cv. Malbec in Australia" @default.
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• W4382449102 doi "https://doi.org/10.1002/ndr2.12172" @default.
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