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Elimination of Grapevine fleck virus from infected grapevines ‘Kyoho’ through meristem-tip culture of dormant buds
J Plant Biotechnol 2017;44:401-408
Published online December 31, 2017
© 2017 The Korean Society for Plant Biotechnology.

Mi Young Kim・Kang Hee Cho・Jae An Chun・Seo Jun Park・Se Hee Kim・Han Chan Lee

Fruit Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Wanju 55365, Korea
Correspondence to: H. C. Lee (⊠)
Fruit Research Division, National Institute of Horticultural and Herbal Science, Rural Development Administration, Wanju 55365, Korea
e-mail: l0hc0811@korea.kr
Received October 6, 2017; Revised November 7, 2017; Accepted November 7, 2017.
cc This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Herein, we report the meristem-tip culture from dormant buds of grape ‘Kyoho’ single-infected with Grapevine fleck virus (GFkV), which is phloem-limited and transmitted by graft inoculation. We produced GFkV-free shoots without thermo- or chemotherapy using meristem-tip explants approximately 0.3 mm (73 explants) and 0.8 mm long (five explants) including shoot apical meristem, 2–5 leaf primordia, and 1–4 uncommitted primordia from dormant buds of the infected woody cuttings (stored at 4°C). Explants were cultured on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 3.0 mg/L benzyladenine (BA) and 0.1 mg/L indole-3-butyric acid (IBA). After 16 weeks of culture, shoot (10-mm long) regeneration frequency achieved from 0.3-mm explants was 4.1% and that obtained from 0.8-mm explants was 40.0%. Virus-free efficiency (expressed as the percentage of RT-PCR negative shoots regenerated) from 0.3- and 0.8-mm explants was 100% and 50%, respectively. Following in vitro multiplication, RT-PCR assays revealed identical results to assays of the first regenerated shoots. Our new methodological approach could be applied for eliminating other viruses in grapevines, as well as for producing virus-free plants in many other deciduous tree species, including fruit trees.
Keywords : Tissue culture, Meristem excision, Grapevine, Virus detection, RT-PCR


December 2017, 44 (4)
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