Australasian Plant Pathology

, Volume 48, Issue 2, pp 115–118 | Cite as

Elimination of Apple stem pitting virus from in vitro-cultured pear by an antiviral agent combined with thermotherapy

  • G. J. Hu
  • N. Hong
  • G. P. WangEmail author
Research Note


Six in vitro pear cultivars, Wonhwang, Xuehua, Conference, Stankimson, Starcrimson and Red Bastlett were infected by Apple stem pitting virus (ASPV) and were treated by combination of 25 μg/ml ribavirin and temperature 35 °C. Results showed that ribavirin could enhance the proliferation of Xuehua, Conference, Stankimson and Starcrimson and death plants were found in Xuehua and Red Bastlett and the total survival rate of the six cultivars was 93.5%. After treatment, the regenerated plants were detected by reverse transcription-polymerase chain reaction (RT-PCR). The results showed that ASPV in all regenerated plants could not be detected and the total elimination rate of the six cultivars was 92.3%. The combination showed high efficacy on elimination of ASPV.


Pear In vitro culture Apple stem pitting virus (ASPV) Chemotherapy Thermotherapy Virus elimination 



This work is supported by the China Agriculture Research System CARS-29-10.

Supplementary material

13313_2018_606_MOESM1_ESM.docx (17 kb)
ESM 1 (DOCX 16 kb)


  1. Astruc N, Marcos JF, Macquaire G, Candresse T, Pallfis V (1996) Studies on the diagnosis of Hop stunt viroid in fruit trees: identification of new hosts and application of a nucleic acid extraction procedure based on non-organic solvents. Eur J Plant Pathol 102:837–846CrossRefGoogle Scholar
  2. Brunt AA, Crabtree K, Dallwitz MJ, Gibbs AJ, Watson L, Zurcher EJ (1996) Apple stem pitting virus. Plant viruses online: descriptions and lists from the VIDE database. CAB International, LondonGoogle Scholar
  3. Cieślińska M (2007) Application of thermo- and chemotherapy In vitro for eliminating some viruses infecting Prunus sp. fruit trees. J Fruit Ornam Plant Res 15:117–124Google Scholar
  4. Cieślińska M, Zawadzka B (1999) Preliminary results of investigation on elimination of viruses from apple, pear and raspberry using thermotherapy and chemotherapy in vitro. Phytopatholgy 17:41–48Google Scholar
  5. Desvignes JC, Boyé R (1988) Different diseases caused by the Chlorotic leaf spot virus on the fruit trees. Acta Hortic 235:31–38Google Scholar
  6. Hauptmanová A, Polák J (2011) The elimination of Plum pox virus in plum cv. Bluefree and apricot cv. Hanita by chemotherapy of in vitro cultures. Hort Sci (Prague) 38:49–53CrossRefGoogle Scholar
  7. Hu GJ, Hong N, Wang LP, Hu HJ, Wang GP (2012) Efficacy of virus elimination from in vitro cultured sand pear (Pyrus pyrifolia) by chemotherapy combined with thermotherapy. Crop Prot 37:20–25CrossRefGoogle Scholar
  8. Hu GJ, Dong YF, Zhang ZP, Fan XD, Ren F, Zhou J (2015) Virus elimination from in vitro apple by thermotherapy combined with chemotherapy. Plant Cell Tissue Organ Cult 121:435–443CrossRefGoogle Scholar
  9. Hu GJ, Dong YF, Zhang ZP, Fan XD, Ren FX, Li ZN, Zhang SN (2018) Elimination of grapevine rupestris stem pitting-associated virus from vitis vinifera ‘kyoho’ by an antiviral agent combined with shoot tip culture. Sci Hortic 229:99–106CrossRefGoogle Scholar
  10. Jelkmann W, Paunovic S (2011) Apple stem pitting virus. In: Hadidi A, Barba M, Candresse T, Jelkmann W (eds) Virus and virus-like diseases of pome and stone fruits. APS Press, St. Paul, pp 35–40CrossRefGoogle Scholar
  11. Koubouris G, Maliogka V, Efthimiou K, Katis N, Vasilakakis M (2007) Elimination of Plum pox virus through in vitro thermotherapy and shoot tip culture compared to conventional heat treatment in apricot cultivar Bebecou. J Gen Plant Pathol 73:370–373CrossRefGoogle Scholar
  12. Lerch B (1987) On the inhibition of plant virus multiplication by ribavirin. Antivir Res 7:257–270CrossRefPubMedGoogle Scholar
  13. Pasquini G, Faggioli F, Pilotti M, Lumia V, Barba M, Hadidi A (1998) Characterization of Apple chlorotic leaf spot virus isolates from Italy. Acta Hortic 472:195–202CrossRefGoogle Scholar
  14. Paunovic S, Ruzic D, Vujovic T, Milenkovic S, Ievremovic D (2007) In vitro production of Plum pox virus-free plums by chemotherapy with ribavirin. Biotechnol Biotechnol Equip 21:417–421CrossRefGoogle Scholar
  15. Quecini V, Lopes ML, Pacheco FTH, Ongarelli MDG (2006) Ribavirin, a guanosin analogue mammalian antiviral agent, impairs Tomato spotted wilt virus multiplication in tobacco cell cultures. Arch Phytopathol Plant Protect 41:1–13CrossRefGoogle Scholar
  16. Schwarz K, Jelkmann W (1998) Detection and characterization of European Apple stem pitting virus isolates of apple and pear by PCR and partial sequence analysis. Acta Hortic 472:75–85CrossRefGoogle Scholar
  17. Smith RA (1980) Mechanisms of action of ribavirin. In: Smith RA, Kirkpatrick W (eds) Ribavirin a Broad Spectrum Antiviral Agent. Academic Press, New York, pp 99–118Google Scholar
  18. Stouffer RF (1989) Apple stem pitting. In: Fridlund PR (ed) Virus and virus-like diseases of pome fruits and simulating non-infectious disorders. Washington State University, Pullman, pp 138–144Google Scholar
  19. Tan RR, Wang LP, Hong N, Wang GP (2010) Enhanced efficiency of virus eradication following thermotherapy of shoot-tip cultures of pear. Plant Cell Tissue Organ Cult 101:229–235CrossRefGoogle Scholar
  20. Tiziano C, Folwell RJ, Wandschneider P, Eastwell KC, Howell WE (2003) Economic implications of a virus prevention program in deciduous tree fruits in the US. Crop Prot 22:1149–1156CrossRefGoogle Scholar
  21. Valero M, Ibáñez A, Morte A (2003) Effects of high vineyard temperatures on the Grapevine leafroll associated virus elimination from Vitis vinifera L. cv. Napoleon tissue cultures. Sci Hortic 97:289–296CrossRefGoogle Scholar
  22. Verma N, Ram R, Zaidi AA (2005) In vitro production of Prunus necrotic ringspot virus-free begonias through chemo-and thermotherapy. Sci Hortic 103:239–247CrossRefGoogle Scholar
  23. Wang LP, Wang GP, Hong N, Tang RR, Deng XY, 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. Hortscience 41:729–732CrossRefGoogle Scholar
  24. Wang QC, Cuellar WJ, Rajamäki M, Hirata Y, Valkonen JPT (2008) Combined thermotherapy and cryotherapy for efficient virus eradication: relation of virus distribution, subcellular changes, cell survival and viral RNA degradation in shoot tips. Mol Plant Pathol 8:1–14Google Scholar
  25. Welland CM, Cantos M, Troncoso A, Perez-Camacho F (2004) Regeneration of virus-free plants by in vitro chemotherapy of GFLV (Grapevine fanleaf virus) infected explants of Vitis vinifera L. cv. ‘Zalema’. Acta Hortic (ISHS) 652:463–466CrossRefGoogle Scholar
  26. Yanase H (1983) Back transmission of Apple stem grooving virus to apple seedlings and induction of symptoms of apple topworking disease in Mitsuba Kaido (Malus sieboldii) and Kobano Zumi (Malus sieboldii var. arborescens) rootstocks. Acta Hortic 130:117–122CrossRefGoogle Scholar

Copyright information

© Australasian Plant Pathology Society Inc. 2018
corrected publication December/2018

Authors and Affiliations

  1. 1.National Key Laboratory of Agromicrobiology, College of Plant Science and TechnologyHuazhong Agricultural UniversityWuhanPeople’s Republic of China
  2. 2.National Center for Eliminating Viruses from Deciduous Fruit Tree, Research Institute of PomologyChinese Academy of Agricultural SciencesXingchengPeople’s Republic of China

Personalised recommendations