Obtaining seed-borne virus free mahaleb (Prunus mahaleb) genotypes through ‎meristem tip culture and in vitro thermotherapy

Document Type : Full Paper

Authors

1 Former M.Sc. Student, Islamic Azad University, Garmsar, Iran‎

2 Associate Professor, Agricultural Research, Extension and Education Organisation (AREEO), Horticultural Research Institute, ‎Temperate Fruit Research Center, Karaj, Iran‎

3 Associate Professor, AREEO, Seed and Plant Registration Organization, Karaj, Iran

4 Associate Professor, AREEO, Agriciltural Biotechnology Research Institute, Karaj, Iran

Abstract

Mahaleb (Prunus mahaleb) plant is one of the most important sources of cherry clonal and seed base rootstock production in Iran. Obtaining seed-borne viruse-free (Prune dwarf virus, Plum pox virus, Prunus necrotic ringspot virus) sources for selected mahaleb rootstocks (NB5176, NBVP1, NBVP2), For this purpose, the viral infection status of genotypes was initially studied in orchard using ELISA serological method. Then the meristem tips were cultured in vitro and 6 months later, transferred to MS medium containing 0.5mg/l BAP and 0.1 mg/l NAA, DKW contacting 0.5 mg/l BAP and WMP contacting 0.3 mg/l pectin. After the initial proliferation stage, the plantlets were heat-treated and then health status of the remained plantlets was studied using RT-PCR. Then the healthy plants were rooted and transferred to pots. Based on ELISA result, some genotypes were probably infected to PNRSV and PDV. The highest shoot tip establishment was recorded as 42.5% by NBPV1. No genotype was proliferated in MS and WPM, so only DKW was apropriate for proliferation. For plantlet rooting, 1/2DKW and modified QL containing 1.5 mg/l IBA were appropriate. NBVP2 was the best in all rooting media. Most of plantlets from three genotypes toleratedin-vitro thermotherapy condition and were free of PNRSV, PDV, PPV based on RT-PCR result. The healthy plants were transferred to pots and could be cultured in an isolated orchard to obtain healthy mahaleb seeds.

Keywords


  1. Albertini, A. & De Salvador, F.R. (1991). Giligio L Informatore Agrario. Supplemento Portinnesti Frutticoli, XLVII36, 13-18.
  2. Barba, M. (1992). Comparison of different method to produce virus free stone fruits, Acta Horticulturae, 309, 385-392.
  3. Boari, A., Boscia, B., Di Terlizzi, B. & Savino, V. (1998). Study on seed transmission of prune dwarf virus (PDV) in Prunus mahaleb L.. Advances in Horticultural Sciences, 12, 89-92.
  4. Bouzari, N. (2011). Collection and evaluation of local Cerasus germplasm to obtain rootstock and variety. Final Repot, Seed and Plant Improvement Institute, Karaj, Iran. (in Farsi)
  5. Boxus, Ph. (1984). Assainissement des arbres fruitiers et du fraisier par culture de meristemes. Parasitica, 40, 139-155.
  6. Bright, J. & Marte, S. (2004). Cherry growing in NSW. Agfact H5.1.2. Second edition.
  7. Cieślińska, M. (2007). Applicaton of thermo- and in vitro for eliminating some viruses infecting Prunus sp. fruit trees. Plant Research, 55, 117-124.
  8. Dardi, G., Vito, G. & Standardi, A. (1996). in vitro mass propagation of eleven Prunus mahaleb ecotypes. Acta Horticulturae, 410, 477-483.
  9. Deogratias, J.M., Dosba, F. & Lutz, A. (1989). Eradication of prune dwarf virus, prunus necrotic ringspot virus and apple chlorotic leaf spot virus in sweet cherries by a combination of chemtherapy, thermotherapy and in vitro culture. Canadian Journal of Plant pathology, 11, 337-342
  10. Driver, J.A. & Kuniyuki, A.M. (1984). in vitro propagation of Paradox walnut rootstock. Journal ofHorticultural Sciences, 19, 507-509.
  11. Fallah, T., Nasrolah-Neghad, S., Shahsavand, M. & Ebadi, A. (2008). Detection of PNRSV in Golestan stone fruits using DAS-ELISA and RT-PCR. (Page 517), Proceedings of 18th Iranian Plant Protection Congress (in Farsi).
  12. Fotopoulos, S. & Sotiropoulos, T.E. (2005). in vitro rooting of PR204/84 rootstock (Prunus persica x P.amygdalus) as influenced by mineral concentration of the culture medium and exposure to darkness for a period. Agronomy Research, 3, 3-8. (in Farsi)
  13. Ganji Moghadam, A., Bolandi, A.R. & Anahid, S. (2008). Proliferation of 4 dwarfing mahaleb genotypes. Pajouhesh-va-Sazandegi, 79, 55-61. (in Farsi)
  14. Gella, R. & Errea, P. (1998). Application of in vitro therapy for Ilarvirus elimination in tress Prunus species. Phytopathology, 146, 445-449.
  15. George, E.F. (1996). Plant Propagation by Tissue Culture: in Practice, Part II. 2nd edition. Exegetics Limited Press, London.
  16. Giorgio, V. & Standardi, A. (1993). Growth and production of two sweet cherry cultivars grafted on 60 ecotypes of Prunus mahaleb. Acta Horticulturae, 410, 471-476.
  17. Goudarzi, R., Majidi, I., Talaee, A. & Mostafavi , M. (1997). Micropropagation of cherry rootstock (Prunus avium CV F12/1) by shoot tip culture. Iranian Journal of Agricultural Sciences, 28, 133-143
  18. Hosseini, A.R.D., Moghadam, E.G., Khorasani, S.K. &, Bihamta, M.R. (2011). Effects of growth regulators on micropropagation of some mahaleb dwarf genotypes. Archives of Applied Science Research, 3, 118-125.
  19. Houang, S.C. & Millikan, D.F. (1980). in vitro micrografting of apple shoot-tips. Hortsciences, 15, 741-743.
  20. Hrotkó, K., Sebok, I., Magyar, L. & Gyeviki, M. (2009). Selection and evaluation of Prunus mahaleb L. clonal rootstocks. Kertgazdasag Horticulture, 41, 57-65.
  21. Jafarpour, B. & Khayat, M. (2013).  Detection PPV in Khorasan Razavi stone fruits.  Plant Protection, 27, 132-135.
  22. Leblay, C., Chevreau, E. & Robin, L.M. (1991). Adventitious shoot regeneration from in vitro leaves of several pear cultivars (Pyrus communis L.). Plant Cell, Tissue and Organ Culture, 25, 99-105
  23. Lenz, F., Baumann, G. & Kornkamhaeng, P. (1983). High temptrature treatment of Prunus avium L. 'F12-1' for virus elimination. Phytopathol. Z., 106, 373-375.
  24. Lioyd, G., Mc Cown, B. (1981). Commercially feasible micro propagation of mountain laurel, Kalmia latifolia, by the use of shoot tip culture. Proceedings of the International Plant Propagators Society, 30, 421-427.
  25. Mahdavian, M., Bouzari, N. & Abdollahi, H. (2010). Effect of culture medium and growth regulators on proliferation and rooting of mahaleb genotypes. Seed and Plant Journal, 26, 15-26. (in Farsi)
  26. Manganaris, G.A., Economou, A.S., Boubourakas, I. & Katis, N. (2003). Production of virus-free plant propagation material from infected nectarine trees. Acta Horticulturae, 616, 501-505
  27. Menzel, W., Jelkmann, W. & Maiss, E. (2002). Detection of four apple viruses by multiplex RT-PCR assays with coamplification of plant mRNA as internal control. Journal of Virological Methods, 99, 81-2.
  28. Milusheva, S., Gercheva, P., Bozhkova, V &, Kamenova, I. (2008). Experiments on transmission of plum pox virus through prunus seeds. Journal of Plant Pathology 90, S1.23
  29. Mohammadi, A., Goharzad, F. & Zaghi, A. (2012). Contribution of PPV with plum fall in Mazandaran province. Pest and Disease 80, 97-9. (in Farsdi)
  30. Moini, A. & Izadpanah, K.M. (2000). Serological identification of PNRSV and PPV in Dasht-e-Moghan. In: Proceedings of the 14th Plant Protection Congress of Iran, Isfahan, Iran. (Page 338). (in Frasi)
  31. Mozaffarian, V. (2004). Trees and shrubs of Iran. PP1003, Farhang-e-Moaser Publ. (in Farsi)
  32. Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiology, 15, 437-497.
  33. Myrta, A., Di Terlizzi, B., Savino, V. & Martelli, G.P. (2003). Virus diseases affecting the Mediterranean stone fruit industry: a decade of surveys. In: Myrta A., Di Terlizzi B., Savino V.(eds.). Virus and virus-like diseases of stone fruits, with particular reference to the Mediterranean region. Options Mediterranne, Series B: Studies and Research, 45, 15-23.
  34. Naderpour, M. & Sadeghi, L. (2015). Investigation of stone fruit orchards in Western Azarbaijan and Khorasan Razavi provinces to introduce healthier orchards for stone fruit propagation industries. Final Report 47168, Center for Agriculture Information and Scientific Documentation, AREEO, Ministry of Agriculture, Iran. (in Farsi)
  35. Naderpour, M. & Shahbazi, R. (2015). Investigation of pome fruit orchards in Western Azarbijan and Khorasan Razavi provinces to introduce healthier orchards for stone fruit propagation industries. Final Report 47282, Center for Agriculture Information and Scientific Documentation, AREEO, Ministry of Agriculture, Iran. (in Farsi)
  36. Perry, R.L. (1987). Cherry Rootstocks. In: Rom RC, Carlson RF (eds.) Rootstocks for Fruit Crops. (pages 217-264), John Wiley and Sons, New York.
  37. Rakhshandehroo, F., Zamanizadeh, H.R., Modarresi, A. & Hajmansoor, S. (2006). Occurrence of Prunus necrotic ringspot virus and Arabis mosaic virus on rose in Iran. Plant Disease, 90, 975-981. (in Farsi)
  38. Sabaghian, S. (2014). Investigation on PNRSV in weeds of rose plantings. In: Proceedings of 21st Iranian Plant Protection Congress. (Page 212).
  39. Sansavini, S. & Lougi, S. (1996). Performance of the sweet cherry cultivar Van on new clonal rootstocks. Acta Horticulturae, 410, 363-372.
  40. Saponari, M., Bottalico, G. & Savino, V. (1999). in vitro propagation of Prunus mahaleb and its sanitation from prune dwarf virus. Advances in Horticultural Sciences 13, 56-60.
  41. Sipahioglu, H.M., Usta, M., Polat, B. & Ocak, M. (2005). Distribution of Prunus necrotic ringspot (PNRSV) and Apple chlorotic leaf spot viruses (ACLSV) in prunus leaves. Tarim Bilimleri Dergisi, 11, 86-90.
  42. Snir, I. & Stein, A. (1985). in vitro selection and elimination of prunus necrotic ring spot virus in sweet cherry (Prunus avium). Riv Ortiflorofruit It, 69, 191-194.
  43. Stein, A., Spiegel, S., Faingersh, G. & Levy, S. (1991). Responses of micropropagated peach cultivars to thermotherapy for elimination of Prunus necrotic ring spot virus. Annals of Applied Biology, 104, 267-276.
  44. Tatari, M. & Mousavi, A. (2013). Optimization of in vitro culture of Tetra, Nemaguard and GF677 rootstocks. Iranian Journal of Agricultural Breeding, 15, 103-115. (in Farsi)
  45. Usta, M., Sipagolou, H.M. & Polat, B. (2005). Comparison of DAS-ELISA and RT-PCR methods for the detection of Prunus necrotic ringspot virus (PNRSV). Journal of Agricultural Sciences, 15, 153-158.
  46. Webester, A.D. & Looney, N.E. (1996). Cherries: Crop Physiology, Production and Uses. 513 pages, CABI Publ.