Micropropagation of Pear (Pyrus sp.)

  • Barbara M. Reed
  • Jeanine DeNoma
  • Sugae Wada
  • Joseph Postman
Part of the Methods in Molecular Biology book series (MIMB, volume 994)


Elements of micropropagation include establishment of shoot tip cultures, proliferation, rooting, and acclimatization of the resulting plantlets. The wide genetic variation in Pyrus makes micropropagation challenging for many genotypes. Initiation of shoots is most successful from forced dormant shoots or from scions grafted onto seedling rootstocks to impose juvenility. Clean shoots are recovered after testing for contaminants at the initiation stage on ½ strength Murashige and Skoog 1962 medium (MS), at pH 6.9 for 1 week or by streaking on nutrient agar. Although pear species and cultivars are cultured on several well-known media, MS is the most commonly used. Our studies showed that multiplication and growth of shoots are best on Pear Medium with higher concentrations of calcium chloride, potassium phosphate, and magnesium sulfate than MS medium and 4.4 μM N6 benzyladenine. Pear shoots are often recalcitrant to rooting; however, a 5 s dip in 10 mM indole-3-butyric acid or naphthalene acetic acid before planting on basal medium without plant growth regulators is effective for many genotypes. Pear shoots store well at 1–4°C, and can hold for as long as 4 years without reculture. Cryopreservation protocols are available for long-term storage of pear shoot tips. Acclimation of in vitro-rooted or micrografted shoots in a mist bed follows standard procedures.

Key words

Micrografting Micropropagation Mineral nutrition Multiplication Pyrus Shoot culture Rooting 


  1. 1.
    Zhao H, Gu N (1990) Pear. In: Chen Z, Evans DA, Sharp WR, Ammirato PV, Sondahl MR (eds) Handbook of plant cell culture, vol 6. McGraw-Hill, New York, pp 264–277Google Scholar
  2. 2.
    Singha S (1986) Pear (Pyrus communis). In: Bajaj YPS (ed) Biotechnology in agriculture and forestry, vol 1. Springer, Heidelberg, pp 198–206Google Scholar
  3. 3.
    Bell RL, Reed BM (2002) In vitro tissue culture of pear: advances in techniques for micropropagation and germplasm preservation. Acta Hortic 596:412–418Google Scholar
  4. 4.
    Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  5. 5.
    Quoirin M, Lepoivre P (1977) Improved media for in vitro culture of Prunus. Acta Hort 78:437–442Google Scholar
  6. 6.
    Driver JA, Kuniyuki AH (1984) In vitro propagation of Paradox walnut rootstock. HortScience 19(4):507–509Google Scholar
  7. 7.
    Lloyd G, McCown B (1980) Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture. Comb Proceed Int Plant Prop Soc 30:421–427Google Scholar
  8. 8.
    Bell RL, Srinivasan C, Lomberk D (2009) Effect of nutrient media on axillary shoot proliferation and preconditioning for adventitious shoot regeneration of pears. In Vitro Cell Dev Biol Plant 45:708–714CrossRefGoogle Scholar
  9. 9.
    Reed BM, Wada S, DeNoma J, Evans TJ, Niedz RP (2010) Improving in vitro mineral nutrition for diverse pear germplasm. In Vitro Cell Dev Biol 46:S64Google Scholar
  10. 10.
    Yeo DY, Reed BM (1995) Micropropagation of three Pyrus rootstocks. HortScience 30(3):620–623Google Scholar
  11. 11.
    Zimmerman RH, Bhardwaj SV, Fordham IM (1995) Use of starch-gelled medium for tissue culture of some fruit crops. Plant Cell Tiss Organ Cult 43:207–213Google Scholar
  12. 12.
    Rodriguez R, Diaz-Sala C, Cuozzo L, Ancora G (1991) Pear in vitro propagation using a double-phase culture system. HortScience 26(1):62–64Google Scholar
  13. 13.
    Moriguchi T, Kozaki I, Yamaki S, Sanada T (1990) Low temperature storage of pear shoots in vitro. Bull Fruit Tree Res Stn 17:11–18Google Scholar
  14. 14.
    Wanas WH, Callow JA, Withers LA (1986) Growth limitations for the conservation of pear genotypes. In: Withers LA, Alderson PG (eds) Plant Tissue Culture and Its Agricultural Applications. Butterworths, London, pp 285–290Google Scholar
  15. 15.
    Lambardi M, Roncasaglia R, Previati A, De Carlo A, Dradi G, Da Re F, Calamai L (2006) In vitro slow growth storage of fruit rootstocks inside gas-tight or gas-permeable containers. Acta Hort 725:483–488Google Scholar
  16. 16.
    Al Maarri K, Arnaud Y, Miginiac E (1994) Micropropagation of Pyrus communis cultivar `Passe Crassane’ seedlings and cultivar `Williams’: factors affecting root formation in vitro and ex vitro. Sci Hortic 58:207–214CrossRefGoogle Scholar
  17. 17.
    Reed BM, Paynter CL, DeNoma J, Chang Y (1998) Techniques for medium-and long-term storage of Pyrus L. genetic resources. Plant Gen Resources Newslett 115:1–4Google Scholar
  18. 18.
    Reed BM (2008) Plant cryopreservation: a practical guide. Springer Science and Business Media, New YorkCrossRefGoogle Scholar
  19. 19.
    Reed BM, DeNoma J, Luo J, Chang Y, Towill L (1997) Cryopreserved storage of a world pear collection. In Vitro Cell Dev Biol 33:51AGoogle Scholar
  20. 20.
    Reed BM (1995) Screening Pyrus germplasm for in vitro rooting response. HortScience 30:1292–1294Google Scholar
  21. 21.
    Dolcet-Sanjuan R, Mok DWS, Mok MC (1992) Characterization and in vitro selection for iron efficiency in Pyrus and Cydonia. In Vitro Cell Dev Biol 28P:25–29Google Scholar
  22. 22.
    Sakai A, Kobayashi S, Oiyama I (1991) Survival by vitrification of nucellar cells of navel orange (Citrus sinensis var. brasiliensis Tanaka) cooled to −196°C. J Plant Physiol 137:465–470CrossRefGoogle Scholar
  23. 23.
    Reed BM, DeNoma J, Luo J, Chang Y, Towill L (1998) Cryopreservation and long-term storage of pear germplasm. In Vitro Cell Dev Biol Plant 34:256–260CrossRefGoogle Scholar
  24. 24.
    Reed BM (1990) Survival of in vitro-grown apical meristems of Pyrus following cryopreservation. HortScience 25(1):111–113Google Scholar
  25. 25.
    Navarro L, Roistacher CN, Murashige T (1975) Improvement of shoot-tip grafting in vitro for virus-free citrus. J Am Soc Hortic Sci 100(5):471–479Google Scholar
  26. 26.
    Postman JD, Hadidi A (1995) Elimination of apple scar skin viroid from pears by in vitro thermo-therapy and apical meristem culture. Acta Hort 386:536–543Google Scholar

Copyright information

© Springer Science+Business Media New York 2012

Authors and Affiliations

  • Barbara M. Reed
    • 1
  • Jeanine DeNoma
    • 1
  • Sugae Wada
    • 2
  • Joseph Postman
    • 1
  1. 1.US Department of AgricultureAgricultural Research Service, National Clonal Germplasm RepositoryCorvallisUSA
  2. 2.Department of HorticultureOregon State UniversityCorvallisUSA

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