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Elimination of PPV and PNRSV through thermotherapy and meristem-tip culture in nectarine

  • Cell Biology and Morphogenesis
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Abstract

The plum pox virus (PPV) and prunus necrotic ringspot virus (PNRSV) cause serious disease problems in stone-fruit trees. In this work, the possibility of obtaining plant material free from these viruses through thermotherapy and meristem-tip culture from infected nectarine shoots (Prunus persica var. nectarina Max, cv. 'Arm King') was studied. In addition, the detection of these viruses in in vitro cultures and young acclimatized plantlets with double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) and multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) was studied. Meristem-tip explants (0.8–1.3 mm) derived from sprouted buds of winter wood and spring shoots from field grown plants had a 2–5% regeneration response. However, application of thermotherapy to potted nectarine trees (3 weeks at a maximum temperature of 35°C) facilitated excision of longer meristem tips (1.3–2.0 mm) that resulted in a significantly higher regeneration response (38%) in woody plant medium (WPM) without plant growth regulators. Such explants formed multiple shoots with the addition of 8 μM benzylaminopurine and 0.8 μM indoleacetic acid. When they were tested for the presence of PPV and PNRSV, 86% and 81% were found to be virus-free as detected by DAS-ELISA and multiplex RT-PCR, respectively. Individual shoots excised from virus-free cultures readily rooted in vitro (half-strength WPM plus 2 μM indolebutyric acid) and grew to plantlets. The combination of an efficient protocol for virus elimination and the establishment of highly sensitive diagnostics resulted in the production of nectarine plants free from PPV and PNRSV.

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Abbreviations

cDNA :

Complementary DNA

DAS-ELISA :

Double antibody sandwich-ELISA

DSMZ :

Deutsche Sammlung von Mikroorganismen und Zelkulturen

ELISA :

Enzyme-linked immunosorbent assay

IC-RT-PCR :

Immunocapture-RT-PCT

IgG :

Immunoglobulin G

OD :

Optical density

PCR :

Polymerase chain reaction

PNRSV :

Prunus necrotic ringspot virus

PPV :

Plum pox virus

RT-PCR :

Reverse transcriptase-PCR

PVP-40 :

Polyvinylpyrrolidone-40

WPM :

Woody plant medium

References

  • Adams AN (1978) The detection of plum pox virus in Prunus spp. species by enzyme-linked immunosorbent assay (ELISA). Ann Appl Biol 90:215–221

    Google Scholar 

  • Adams AN, Guise CM, Crossley SJ (1999) Plum pox virus detection in dormant plum trees by PCR and ELISA. Plant Pathol 48:240–244

    Article  CAS  Google Scholar 

  • Ayabe M, Sumi S (2001) A novel and efficient tissue culture method—"stem-disc dome culture"—for producing virus-free garlic (Allium sativum L.). Plant Cell Rep 20:503–507

    Article  CAS  Google Scholar 

  • Brown GR, Kwiatkowski S, Martin MW, Thomas PE (1988) Eradication of PVS from potato clones through excision of meristems from in vitro heat-treated shoot tips. Am Potato J 65:633–638

    Google Scholar 

  • Candresse T, Macquaire G, Lanne M, Bousalem M, Quiot-Douine L, Quiot JB, Dunez J (1995) Analysis of plum pox virus variability and development of strain-specific assays. Acta Hortic 386:357–369

    CAS  Google Scholar 

  • Crosslin JM, Hammond R, Hammerschlag FA (1992) Detection of prunus necrotic ringspot virus serotypes in herbaceous and Prunus hosts with a complementary RNA probe. Plant Dis 76:1132–1136

    CAS  Google Scholar 

  • Dovas IC, Hatziloucas E, Salomon R, Barg E, Shiboleth Y, Katis NI (2001) Comparison of methods for virus detection in Allium spp. J Phytopathol 149:731–737

    Article  CAS  Google Scholar 

  • Escalettes V, Dosba F, Lansac M, Eyquard JP (1988) Genetic resistance to plum pox potyvirus in peaches. Acta Hortic 465:689–698

    Google Scholar 

  • Greber RS, Klose MJ, Milne JR, Teakle DS (1991) Transmission of prunus necrotic ringspot using plum pollen and thrips. Ann Appl Biol 118:580–593

    Google Scholar 

  • Hammerschlag FA, Scorza R (1991) Field performance of micropropagated own-rooted peach trees. J Am Soc Hortic Sci 116:1089–1091

    Google Scholar 

  • Hammerschlag FA, Bauchan GR, Scorza R (1987) Factors influencing in vitro multiplication and rooting of peach cultivars. Plant Cell Tissue Organ Cult 8:235–242

    CAS  Google Scholar 

  • Helguera PR, Taborda R, Docampo DM, Ducasse DA (2001) Immunocapture reverse transcription-polymerase chain reaction combined with nested PCR greatly increases the detection of prunus necrotic ring spot virus in the peach. J Virol Methods 95:93–100

    Article  CAS  PubMed  Google Scholar 

  • Heuss K, Liu Q, Hammerschlag FA, Hammond RW (1999) A cRNA probe detects prunus necrotic ringspot virus in three peach cultivars after micrografting and in peach shoots following long-term culture at 4°C. HortScience 34:346–347

    Google Scholar 

  • Heuss-La Rosa K, Hammond R, Crosslin JM, Hazel C, Hammerschlag FA (1995) Monitoring prunus necrotic ringspot virus infection by hybridization with a cRNA probe after in vitro micrografting. J Am Soc Hortic Sci 120:928–931

    Google Scholar 

  • Herrera G, Sepulvida P, Madariaga M (1998) Survey of sharka disease (plum pox virus) on stone fruit trees in Chile. Acta Hortic 472:393–399

    Google Scholar 

  • Kartha KK, Gamborg OL (1975) Elimination of cassava mosaic disease by meristem culture. Phytopathology 65:826–828

    Google Scholar 

  • Lloyd G, McCown B (1981) Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Comb Proc Intl Plant Prop Soc 30:421–427

    Google Scholar 

  • Navarro L (1988) Application of shoot-tip grafting in vitro to woody species. Acta Hortic 227:43–55

    Google Scholar 

  • Pusey PL, Yadava UL (1991) Influence of prunus necrotic ringspot virus on growth, productivity, and longevity of peach trees. Plant Dis 75:847–851

    Google Scholar 

  • Rosner A, Maslenin L, Spiegel S (1998) Differentiation among isolates of prunus necrotic ringspot virus by transcript conformation polymorphism. J Virol Methods 74:109–115

    Article  CAS  PubMed  Google Scholar 

  • Rowhani A, Maningas MA, Lile LS, Daubert SD, Golino DA (1995) Development of a detection system for viruses of woody plants based on PCR analysis of immobilized virions. Phytopathology 85:347–352

    CAS  Google Scholar 

  • Scott SW, Barnett OW, Burrows PM (1989) Incidence of prunus necrotic ringspot virus in selected peach orchards of South California. Plant Dis 73:913–916

    Google Scholar 

  • Shu W, Timon B (1993) Preliminary study on the methods of getting virus-free peach plantlets in vitro. Acta Hortic 374:191–194

    Google Scholar 

  • Skirvin RM, Kouider M, Joung A, Korban SS (1986) The tissue culture of apple Malus domestica Borkh. In: Bajaj YPS (ed) Trees. Biotechnology in agriculture and forestry, vol 1. Springer, Berlin Heidelberg New York, pp 183–197

  • Spiegel S, Scott SW, Bowman-Vance V, Tam Y, Galiak-Parov NN, Rosner A (1996) Improved detection of prunus necrotic ringspot virus by the polymerase chain reaction. Eur J Plant Pathol 102:681–685

    CAS  Google Scholar 

  • Uyemoto JK, Asai WK, Luhn F (1992) Ilarviruses: evidence for rapid spread and effects on vegetative growth and fruit yields of peach trees. Plant Dis 76:71–74

    Google Scholar 

  • Varveri C, Bem F (1995) Viruses of stone and pome fruit mother-tree plantations in Greece. Acta Hortic 386:431–438

    Google Scholar 

  • Varveri C, Candresse T, Cugusi M, Ravelonandro M, Dunez J (1988) Use of 32P labelled transcribed RNA probe for dot hybridization detection of plum pox virus. Phytopathology 78:1280–1283

    CAS  Google Scholar 

  • Walkey DGA (1980) Production of virus-free plants by tissue culture. In: Ingram DS, Helgeson IP (eds) Tissue culture methods for plant pathologists. Blackwell, Oxford, pp 109–117

  • Wetzel T, Candresse T, Ravelonandro M, Dunez J (1991) A polymerase chain reaction assay adapted to plum pox potyvirus detection. J Virol Methods 33:355–365

    CAS  PubMed  Google Scholar 

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Acknowledgements

The authors would like to thank the Pomology Institute of Naoussa (NAGREF) for the supply of plant material and Dr. T. Syros for technical assistance. GAM thanks the Greek Scholarship Foundation for financial support.

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Authors and Affiliations

  1. School of Agriculture, Aristotle University, P.O. Box 281, 541 24, Thessaloniki, Greece

    G. A. Manganaris, A. S. Economou, I. N. Boubourakas & N. I. Katis

Authors
  1. G. A. Manganaris
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  2. A. S. Economou
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  3. I. N. Boubourakas
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  4. N. I. Katis
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Corresponding author

Correspondence to A. S. Economou.

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Communicated by G.C. Phillips

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Manganaris, G.A., Economou, A.S., Boubourakas, I.N. et al. Elimination of PPV and PNRSV through thermotherapy and meristem-tip culture in nectarine. Plant Cell Rep 22, 195–200 (2003). https://doi.org/10.1007/s00299-003-0681-y

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  • DOI: https://doi.org/10.1007/s00299-003-0681-y

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