Molecular assessment of thermotherapy and meristem culture efficiency on some ‎virus’s eradication from important commercial pear cultivars (Pyrus communis L.)‎

Document Type : Full Paper

Authors

1 Former Ph.D. Student and Associate Professor, Department of Horticultural Science, Faculty of Agriculture, University of Tabriz, ‎Tabriz, Iran

2 Associate Professor, Department of Horticultural Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran

3 Associate Professor, Department of Agriculture Biotechnology Research Institute of Iran (ABRII), Karaj, Iran‎

Abstract

This study was aimed to eradicate three viruses, Apple chlorotic leaf spot virus (ACLSV),Apple stem grooving virus (ASGV) and Apple stem pitting virus (ASPV) from seven pear cultivars including Abate Fetel, Beiruti, Dargazi, Coscia, Louise Bonne, Mellina, Spadona. Experiments were performed by evaluation of the effectiveness of thermotherapy duration including 0, 7, 14 and 21 days at 38 °C and meristem culturing in size of less than 0.2 mm on virus eradication rates of explants. At first mother samples were tested for ACLSV, ASGV and ASPV by and RT-PCR and all samples were infected by all three viruses, except Abate Fetel and Beiruti samples, that were free of ASP virus. Thermotherapy and meristem culturing were performed in vitro. Explants from thermotherapy and meristem culturing were tested by RT-PCR for all three viruses and the results showed that eradication rates of ACLSV, ASGV and ASPV, respectively 78.46, 26.63 and 35.5 percent were different in the different pear cultivars. The highest rate of virus eradication was related to Coscia and the lowest to Spadona cultivars. There was a direct relationship between increasing the duration of thermotherapy and virus elimination of explants, but this increased period of time up to 21 days reduced the growth and multiplication and even destroyed the explants. Therefore, 14 days of thermotherapy was the most effective treatment for elimination of ASGV, ASPV and ACLSV, respectively 61.4, 100 and 45.5 percent, from pear explants. At the end of the experiment, samples that were diagnosed virus free using RT-PCR, were proliferated, rooted and transferred to greenhouse condition for acclimation stage.

Keywords

References

  1. Abdollahi, H. (2010). Pear: Botany, Cultivars and Rootstocks. Iranian Agricultural Ministry Publications, Tehran, Iran. 210pp.
  2. Adams, A.N., Guise, C.M. & Crossley, S. J. (1999). Plum pox virus detection in dormant plum trees by PCR and ELISA. Plant Pathology, 48, 240- 244.
  3. Adams, M.J., Antoniw, J.F., Bar-Joseph, M., Brunt, A.A., Candresse, T., Foster, G.D., Martelli, G.P., Milne, R.G. & Fauquet, C.M. (2004). The new plant virus family Flexiviridae and assessment of molecular criteria for species demarcation. Archives of Virology, 149, 1045-1060.
  4. Callaway, A., Giesman-Cookmeyer, D., Gillock, E.T., Sit, T. L. & Lommel, S.A. (2001). The multifunctional capsid proteins of plant RNA viruses. Annual Review of Phytopathology, 39, 419-460.
  5. Campbell, A.I. (1967). The effect of some pear viruses on the growth and compatibility of a number of Pyrus species and near relatives. Journal of Horticultural Science, 42(2), 133-138.
  6. Cieslinska, M. (2002). Elimination of apple chlorotic leaf spot virus (ACLSV) from pear by in vitro thermotherapy and chemotherapy. Acta Horticulturae, 596, 481-484.
  7. Cooper, V.C. & Walkey, D.G.A. (1978). Thermal inactivation of cherry leaf roll virus in tissue cultures of Nicotiana rustica rose from seeds and meristem tips. Annals of Applied Biology, 88, 273-278.
  8. Deng, X.Y., Hong, N., Hu, H.J. & Wang, G.P. (2004). Detection of latent viruses in Pyrus pyrifolia by IC-RT-PCR and TC-RT-PCR. Journal of Fruit Science, 21, 569-572.
  9. Dziedzic, E. (2008). Elimination of Prunus necrotic ring spot virus (PNRSV) from plum ‘Earliblue’ shoots through thermotherapy in vitro. Journal of Fruit and Ornamental Plant Research, 16, 101-109.
  10. Food and Agriculture Organization. (2016). FAOSTAT. Retrieved May 1, 2012, from http://www.fao.org/statistics/en.
  11. Gambino, G., Bondaz, J. & Gribaudo, I. (2006). Detection and elimination of viruses in callus, somatic embryos and regenerated plantlets of grapevine. European Journal of Plant Pathology, 114, 397-404.
  12. Grimova, L., Winkowska, L., Zíka, L. & Rysanek, P. (2016). Distribution of viruses in old commercial and abandoned orchards and wild apple trees. Journal of Plant Pathology, 98, 549-554.
  13. Hadidi, A. & Barba, M. (2011). Economic impact of pome and stone fruit viruses and viroids. Virus and Virus Like Diseases of Pome and Stone Fruits, 1(8), 1-7.
  14. Hosokawa, M. (2008). Leaf primordia-free shoot apical meristem culture: a new method for production of viroid-free Plants. The Japanese Society for Horticultural Science, 77, 341-349.
  15. Hu, G.J., Hong, N., Wang, L.P., Hu, H.J. & Wang, G.P. (2012). Efficacy of virus elimination from in vitro cultured sand pear (Pyrus pyrifolia) by chemotherapy combined with thermotherapy. Crop Protection, 37, 20-25.
  16. Hu, G., Dong, Y., Zhang, Z., Fan, X., Ren, F. & Zhou, J. (2015). Virus elimination from in vitro apple by thermotherapy combined with chemotherapy. Plant Cell, Tissue and Organ Culture, 121, 435-443.
  17. Kazemi, N., Zaree, N.F., Habashi, A.A. & Asadi, W. (2019). Molecular assessment of chemotherapy and meristem culture efficiency for production of seven cultivars of virus-free Pear (Pyrus communis L.). Journal of crops improvement, 21(1): 107-118. (In Farsi)
  18. Knapp, E., Hanzer, V., Weiss, H., da Caˆmara Machado, A., da Clmara Machado, A., Weiss, B., Wang, Q., Katinger, H. & Laimer da Clmara Machado, M. (1995). New aspects of virus elimination in fruit trees. Acta Horticulturae, 386, 409-418.
  19. Komorowska, B., Malinowski, T. & Michalczuk, L. (2010). Evaluation of several RT-PCR primer pairs for the detection of Apple stem pitting virus. Journal of Virological Methods, 168, 242-247.
  20. Koubouris, G.C., Maliogka, V.I., Efthimiou, K., Katis, N.I. & Vasilakakis, M.D. (2007). Elimination of Plum pox virus through in vitro thermotherapy and shoot tip culture compared to conventional heat treatment in apricot cultivar Bebecou. Journal of General Plant Pathology, 73, 370-373.
  21. Magome, H., Yoshikawa, N., Takahashi, T., Ito, T. & Miyakawa, T. (1997). Molecular variability of the genomes of capilloviruses from apple, Japanese pear, European pear, and citrus trees. Phytopathology, 87, 389-396.
  22. Magome, H., Yoshikawa, N. & Takahashi, T. (1999). Single-strand conformation polymorphism analysis of apple stem grooving capillovirus sequence variants. Phytopathology, 89, 136-140.
  23. Manganaris, G.A., Economou, A.S., Boubourakas, I.N. & Katis, N.I. (2003). Elimination of PPV and PNRSV through thermotherapy and meristem-tip culture in nectarine. Plant Cell Reports, 22, 195-200.
  24. Mathioudakis, M.M., Maliogka, V.I., Dovas, C.I., Paunovi´c, S. & Katis, N.I. (2008). Reliable RT-PCR detection of Apple stem pitting virus in pome fruits and its association with quince fruit deformation disease. Plant Pathology, 58, 228-236.
  25. Masoomi-Aladizgeh, F., Jabbari, L., Khayam Nekouei R. & Aalami A. (2016). A simple and rapid system for DNA and RNA isolation from diverse plants using handmade kit. Nature, Protocol Exchange site.
  26. Meijneke, C.A.R., Van Oosten, H.J., & Peerboom, H. (1973). Growth, yield, and fruit quality of virus-infected and virus-free Golden Delicious apple trees. Acta Horticulturae, 44, 209-212.
  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, 81-92.
  28. Murashige, T. & Skoog, F. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15, 473-497.
  29. Panattoni, A., Luvisi, A. & Triolo, E. (2013). Elimination of viruses in plants: twenty years of progress. Spanish Journal of Agricultural Research, 11, 173-188.
  30. Paprstein, F., Sedlak, J., Polak, J., Svobodova, L., Hassan, M. & Bryxiova, M. (2008). Results of in vitro thermotherapy of apple cultivars. Plant Cell, Tissue Organ Culture, 94, 347-352.
  31. Paprstein, F., Sedlak, J., Svobodova, L., Polak, J. & Gadiou, S. (2013). Results of in vitro chemotherapy of apple cv. Fragrance. Horticultural Science (Prague), 40, 186-190.
  32. Posnette, A.F., Cropley, R. & Ellemberger, C. (1963). The effect of virus infection on the growth and crop of apple, pear and plum trees. Phytopathologia Mediterranea, 2, 158-161.
  33. Prokhnevsky, A.I., Peremyslov, V.V., Napuli, A.J. & Dolja, V.V. (2002). Interaction between long-distance transport factor and Hsp70- related movement protein of Beet yellows virus. Virology, 76, 11003-11011.
  34. Quoirin, M. & Lepoivre, P.H. (1977). Improved media for in vitro culture of Prunus sp. In: Symposium on Tissue Culture for Horticultural Purposes, Sept., Gent, Belgium, 78, pp. 437-442. 
  35. Rana, T., Chandel, l.V., Kumar, Y., Ram, R., Hallan, V. & Zaidi, A.A., (2010). Molecular variability analyses of Apple chlorotic leaf spot virus capsid protein. BioScience, 35, 605-615.
  36. Retheesh, S.T. & Bhat, A.I., (2010). Simultaneous elimination of Cucumber mosaic virus and Cymbidium mosaic virus infecting Vanilla planifolia through meristem culture. Crop Protection, 29, 1214-1217.
  37. Sareila, O., Hohkuri, M., Wahlroos, T. & Susi, P. (2004). Role of viral movement and coat proteins and RNA in phloem-dependent movement and phloem unloading of tobamoviruses. Phytopathology, 152, 622-629.
  38. Sharma, S., Singh, B., Rani, G., Zaidi, A.A., Hallan, V., Nagpal, A. & Virk, G.S. (2007). Production of Indian citrus ringspot virus-free plants of Kinnow employing chemotherapy coupled with shoot tip grafting. Journal of Central European Agriculture, 1, 1-8.
  39. Shim, H.K., Min, Y.J., Hong, S.Y., Kwon, M.S., Kim, H.R., Choi, Y.M., Lee, S.C. & Yang, J.M. (2004). Nucleotide sequences of a Korean isolate of Apple stem grooving virus associated with black necrotic leaf spot disease on pear (Pyrus pyrifolia). Molecular Cell Biology, 18, 192-199.
  40. Sun, Q., Sun, H. & Bell, R.L. (2009). Effect of polyvinyl alcohol on in vitro rooting capacity of shoots in pear clones (Pyrus communis L.) of different ploidy. Plant Cell, Tissue Organ Culture, 99, 299-304.
  41. Tan, R.R., Wang, L.P., Hong, N. & Wang, G.P. (2010). Enhanced efficiency of virus eradication following thermotherapy of shoot-tip cultures of pear. Plant Cell, Tissue Organ Culture, 101, 229-235.
  42. Tatineni, S., Afunian, M.R., Hilf, M.E., Gowda, S., Dawson, W.O. & Garnsey, S.M. (2009). Molecular characterization of Citrus tatter leaf virus historically associated with Meyer lemon trees: complete genome sequence and development of biologically active in vitro transcripts. American Phytopathological Society, 99, 423-431.
  43. Valero, M., Ibanez, A. & Morte, A. (2003). Effects of high vine yard temperatures on the Grapevine leaf roll associated virus elimination from Vitis vinifera L. cv. Napoleon tissue cultures. Scientia Horticulturae, 97, 289-296. 
  44. Verma, N., Ram, R. & Zaidi, A.A. (2005). In vitro production of Prunus necrotic ringspot virus-free begonias through chemo- and thermotherapy. Scientia Horticulturae, 103, 239-247.
  45. Wang, L.P., Wang, G.P., Hong, N., Tan, R.R., Deng, X.Y. & Zhang, H. (2006). Effect of thermotherapy on elimination of Apple stem grooving virus and Apple chlorotic leaf spot virus for in vitro cultured pear shoot tips. Horticultural Science, 41, 729-732.
  46. Wang, Q.C. & Valkonen, J.P.T. (2008). Elimination of two viruses which interact synergistically from sweet potato by shoot tip culture and cryotherapy. Journal of Virological Methods, 154, 135-145.
  47. Wang, L.P., Hong, N., Matic, S., Myrta, A., Song, Y.S., Michelutti, R. & Wang, G.P. (2011). Pome fruit viruses at the Canadian clonal genebank and molecular characterization of apple chlorotic leaf spot virus isolates. Scientia Horticulturae, 130, 665-671.
  48. Yaegashi, H., Isogai, M., Tajima, H., Sano, T. & Yoshikawa, N. (2007). Combinations of two amino acids (Ala40 and Phe75 or Ser40 and Tyr75) in the coat protein of Apple chlorotic leaf spot virus are crucial for infectivity. Journal of General Virology, 88, 2611-2618.
Iranian Journal of Horticultural Science
Volume 50, Issue 4
March 2020
Pages 983-992

Files

History

  • Receive Date: 10 October 2018
  • Revise Date: 25 December 2018
  • Accept Date: 05 January 2019

Share

How to cite

Statistics