Details

Autor(en) / Beteiligte

• Bi, Wen‐Lu
• Hao, Xin‐Yi
• Cui, Zhen‐Hua
• Pathirana, Ranjith
• Volk, Gayle M.
• Wang, Qiao‐Chun

Titel

Shoot tip cryotherapy for efficient eradication of grapevine leafroll‐associated virus‐3 from diseased grapevine in vitro plants

Ist Teil von

• Annals of applied biology, 2018-11, Vol.173 (3), p.261-270

Ort / Verlag

Oxford, UK: Blackwell Publishing Ltd

Erscheinungsjahr

2018

Link zum Volltext

Quelle

Alma/SFX Local Collection

Beschreibungen/Notizen

• We describe a droplet‐vitrification cryotherapy method for the eradication of grapevine leafroll‐associated virus‐3 (GLRaV‐3) from diseased in vitro shoots of Vitis plants. The procedure involved pre‐culture of 1.0‐mm shoot tips containing five to six leaf primordia (LPs) for 3 days with a pre‐culture medium containing 0.3 M sucrose, 0.16 mM glutathione and 0.14 mM ascorbic acid, treatment of the pre‐cultured shoot tips for 20 min at room temperature with a loading solution composed of 2 M glycerol and 0.4 M sucrose and exposure to plant vitrification solution 2 (PVS2), prior to freezing in liquid nitrogen of dehydrated shoot tips contained in 2.5‐μL PVS2 droplets on aluminium foil strips. Virus localisation showed GLRaV‐3 was not present in apical dome (AD) and LPs 1–4, but it was detected in the basal shoot tip region, approximately 0.5 mm from the AD, as well as in LP 5 and more mature tissues. Histological observations identify that only freezing in liquid nitrogen results in the death of all cells in areas of shoot tips harbouring virus, whereas PVS2 treatment does not. Thus, freezing in liquid nitrogen is a necessary step that eradicates GLRaV‐3. This cryotherapy procedure produced shoot regrowth levels that ranged from 43% to 59%, and all plants recovered after cryotherapy were free of GLRaV‐3 in two wine, one table and one rootstock cultivars. Thus, this procedure can be considered to be efficient and wildly applicable for eradication of GLRaV‐3 from Vitis spp. A shoot tip cryotherapy was described for efficient eradication of grapevine leafroll‐associated virus‐3. Mechanism for eradication of GLRaV‐3 by shoot tip cryotherapy was provided.