Fast and Effective Thermotherapy Treatment for <i>In Vitro</i> Virus Eradication in Apple and Pear Trees
- 1 Department of Plant Physiology, University of Santiago de Compostela, Santiago de Compostela, Spain
- 2 Center of Agrarian Research of Mabegondo INGACAL-CIAM, Abegondo, Spain
- 3 Department of Plant Physiology, University of Santiago de Compostela, Santiago de Compostela, Spain
Abstract
Heat therapy followed by the isolation and in vitro culture of apical meristems is a suitable procedure for virus eradication. However, the period of heat treatment is usually long (28 - 50 days) and the yield of viable plants free of viruses after treatment is often low (<50%). Here, we describe an alternative method to obtain virus-free plants. We used traditional Galician cultivars, six apple trees and two pear trees, infected with Apple chlorotic leaf spot virus (ACLSV) and Apple mosaic virus (ApMV). We combined heat therapy of in vitro shoots using a temperature gradient from 25 ℃ to 40 ℃ increasing 1 ℃ per day for a shorter period of time (18 days) with the posterior isolation and culture of apical meristems. All DNA samples analyzed, obtained from plants developed from meristems, were 100% free of ApMV and almost 90% free of ACLSV. With this in vitro procedure combined we obtained a good yield of tested plants free of viruses. Our method is fast and effective and it could be also useful to eradicate these and other viruses in other fruit trees.
- Lister, R.M. (1970) Apple Chlorotic Leaf Spot Virus C.M.I/A.A.B. Description of Plant Viruses No.30. Purdue University, Lafayette.
- Mink, G.I. (1992) Ilarvirus Vectors. In: Harris, K.F., Ed., Advances in Disease Vector Research, Springer, New York, 261-281. https://doi.org/10.1007/978-1-4612-2910-0_8
- Gella, R. and Errea, P. (1998) Application of In vitro Therapy for Ilarvirus Elimination in Three Prunus Species. Journal of Phytopathology, 146,445-449. https://doi.org/10.1111/j.1439-0434.1998.tb04779.x
- Fulton, R.W. (1972) Apple Mosaic Virus CMI/AAB Descriptions of Plant Viruses, No. 83. Association of Applied Biologists, Wellesbourne.
- Wang, L.P., Hong, N., Wang, G.P., Xu, W.X., Michelutti, R. and Wang, A.M. (2010) Distribution of Apple Stem Grooving Virus and Apple Chlorotic Leaf Spot Virus in Infected In Vitro Pear Shoots. Crop Protection, 29, 1447-1451. https://doi.org/10.1016/j.cropro.2010.08.003
- Ji, Z., Zhao, X., Duan, H., Hu, T., Wang, S., Wang, Y. and Cao, K. (2013) Multiplex RT-PCR Detection and Distribution of Four Apple Viruses in China. Acta Virologica, 57, 435-441. https://doi.org/10.4149/av_2013_04_435
- Panattoni, A., Luvisi, A. and Triolo, E. (2013) Review. Elimination of Viruses in Plants: Twenty Years of Progress. Spanish Journal of Agricultural Research, 11, 173-188. https://doi.org/10.5424/sjar/2013111-3201
- Paprstein, F., Sedlak, J., Polak, J., Svobodova, L., Hassan, M. and Bryxiova, M. (2008) Results of In Vitro Thermotherapy of Apple Cultivars. Plant Cell, Tissue and Organ Culture, 94, 347-352. https://doi.org/10.1007/s11240-008-9342-8
- Mink, G.I., Wample, R. and Howell, W.E. (1998) Heat Treatment of Perennial Plants to Eliminate Phytoplasmas, Viruses, and Viroids While Maintaining Plant Survival. In: Hadidi, A., Ed., Plant Virus Disease Control, Springer, St. Paul.
- Cieslińska, M. (2002) Elimination of Apple Chlorotic Leaf Spot Virus (ACLSV) from Pear by In Vitro Thermotherapy and Chemotherapy. ISHS Acta Horticulturae 596: VIII International Symposium on Pear, 481-484. https://doi.org/10.17660/ActaHortic.2002.596.80
- Tan, R., Wang, L., Hong, N. and Wang, G. (2010) Enhanced Efficiency of Virus Eradication Following Thermotherapy of Shoot-Tip Cultures of Pear. Plant Cell, Tissue and Organ Culture, 101, 229-235. https://doi.org/10.1007/s11240-010-9681-0
- Manganaris, G.A., Economou, A.S., Boubourakas, I.N. and Katis, N.I. (2003) Elimination of PPV and PNRSV through Thermotherapy and Meristem-Tip Culture in Nectarine. Plant Cell Reports, 22,195-200. https://doi.org/10.1007/s00299-003-0681-y