Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2023 Oct 17;15(10):2102.
doi: 10.3390/v15102102.

A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries

Mizuho Nita  1   2 , Taylor Jones  1 , Diana McHenry  1 , Elizabeth Bush  1 , Charlotte Oliver  1 , Akira Kawaguchi  3 , Akiko Nita  1 , Miyuki Katori  2
Affiliations

A NitroPure Nitrocellulose Membrane-Based Grapevine Virus Sampling Kit: Development and Deployment to Survey Japanese Vineyards and Nurseries

Mizuho Nita et al. Viruses. .

Abstract

We developed a NitroPure Nitrocellulose (NPN) membrane-based method for sampling and storing grapevine sap for grapevine virus detection. We devised an efficient nucleic acid extraction method for the NPN membrane, resulting in 100% amplification success for grapevine leafroll-associated virus 2 (GLRaV2) and 3 (GLRaV3), grapevine rupestris stem pitting-associated virus (GRSPaV), grapevine virus A, grapevine virus B, and grapevine red blotch virus (GRBV). This method also allowed the storage of recoverable nucleic acid for 18 months at room temperature. We created a sampling kit to survey GLRaV2, GLRaV3, and GRBV in Japanese vineyards. We tested the kits in the field in 2018 and then conducted mail-in surveys in 2020-2021. The results showed a substantial prevalence of GLRaV3, with 48.5% of 132 sampled vines being positive. On the other hand, only 3% of samples tested positive for GLRaV2 and none for GRBV.

Keywords: GLRaV2; GLRaV3; GRBV; NitroPure nitrocellulose; grapevine viruses.

PubMed Disclaimer

Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the study’s design, in the collection, analyses, or interpretation of data, in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Distribution of GLRaV2 and GLRaV3 among sampled prefectures. The total number of samples per prefecture is shown as a number within the prefecture border, and a darker color indicates more samples per prefecture. Pie charts represent the percentage of virus-positive and negative samples: blue = GLRaV3, red = GLRaV2, and light orange = negative.
Figure 2
Figure 2
Grouping of GLRaV3 isolates in this study based on the Tamura–Nei neighbor-joining tree with bootstrap values estimated from 1000 replicates (bootstrap values < 80% are not shown).
See this image and copyright information in PMC

Similar articles

See all similar articles

References

    1. Martelli G.P. Directory of virus and virus-like diseases of the grapevine and their agents. J. Plant Pathol. 2014;96:1–136. doi: 10.4454/JPP.V96I1SUP. - DOI
    1. Meng B., Martelli G.P., Golino D.A., Fuchs M. Grapevine Viruses: Molecular Biology, Diagnostics and Management. Springer; Berlin/Heidelberg, Germany: 2017. Grapevine Viruses: Molecular Biology, Diagnostics and Management; pp. 1–698. - DOI
    1. EFSA Panel on Plant Health (PLH) Bragard C., Dehnen-Schmutz K., Gonthier P., Jacques M., Miret J.A.J., Justesen A.F., MacLeod A., Magnusson C.S., Milonas P., et al. List of non-EU viruses and viroids of Cydonia Mill., Fragaria L., Malus Mill., Prunus L., Pyrus L., Ribes L., Rubus L. and Vitis L. EFSA J. 2019;17:e05501. doi: 10.2903/j.efsa.2019.5501. - DOI - PMC - PubMed
    1. Naidu R.A., Rowhani A., Fuchs M.F., Golino D., Martelli G.P. Grapevine Leafroll: A Complex Viral Disease Affecting a High-Value Fruit Crop. Plant Dis. 2014;98:1172–1185. doi: 10.1094/PDIS-08-13-0880-FE. - DOI - PubMed
    1. Wallis C.M., Sudarshana M.R. Effects of Grapevine red blotch-associated virus (GRBaV) infection on foliar metabolism of grapevines. Can. J. Plant Pathol. 2016;38:358–366. doi: 10.1080/07060661.2016.1227374. - DOI

Publication types

Supplementary concepts

Grants and funding

This research was funded by Japan Grants-in-Aid for Scientific Research (aka Kakenhi), grant number 19K06049.

LinkOut - more resources