Abstract
The improvement of perennial fruit trees through traditional breedingmethods is a long-term effort because of their long generation time. Theproduction of haploid and doubled haploid plants should offer newpossibilities for genetic studies and breeding work. In this study, haploidclones of pear were treated in vitro by oryzalin for chromosomedoubling; the level of ploidy was assessed by flow cytometry. Oryzalinappeared to be an efficient agent for chromosome doubling, the optimalconcentrations range from 200 to 300 μM. For homozygosityassessment, analyses of isozyme markers were carried out, together withmicrosatellite markers PCR-amplified with primers initially developed forapple. The use of isozyme markers confirmed homozygosity of all thedoubled haploid clones except for one. The microsatellite markers can beused earlier than isozymes for checking homozygosity during theprogramme of haploid and doubled haploid clones production. Truedoubled haploid clones of pear were obtained in less than one year andtheir acclimatisation in greenhouse has already started.
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Bouvier, L., 1993. Haploïdie chez le pommier (Malus ×domestica Borkh.) et le poirier (Pyrus communis L.). Thèse de l'université de Paris VI, 138 pp.
Bouvier, L., F.R. Fillon & Y. Lespinasse, 1994. Oryzalin as an effi-cient agent for chromosome doubling of haploid apple shoots in vitro. Plant Breed 113: 343–346.
Bouvier, L., Y.Z. Zhang & Y. Lespinasse, 1993. Two methods of haploidization on pear, Pyrus communis L.: greenhouse seedling selection and in situ parthenogenesis induced by irradiated pollen. Theor Appl Genet 87: 229–232.
Chevreau, E., S. Leuliette & M. Gallet, 1997. Inheritance and linkage of isozyme loci in pear (Pyrus communis L.). Theor Appl Genet 94: 498–506.
Chevreau, E., A.G. Manganaris & M. Gallet, 1999. Isozyme segregation in five apple progenies and potential use for map construction. Theor Appl Genet 98: 329–336.
Cipriani, G., G. Lot, W.G. Huang, M.T. Marazzo, E. Peterlunger & R. Testolini, 1999. AC/GT and AG/CT microsatellite repeats in peach [Prunus persica (L) Batsch]: isolation, characterisation and cross-species amplification in Prunus. Theor Appl Genet 99: 65–72.
Doyle, J.J. & J.L. Doyle, 1990. Isolation of plant DNA from fresh tissue. Focus 12: 13–15.
Gianfranceschi, L., N. Seglias, R. Tarchini, M. Komjanc & C. Gessler, 1998. Simple sequence repeats for the genetic analysis of apple. Theor Appl Genet 96: 1069–1076.
Huang, W.G., G. Cipriani, M. Morgante & R. Testolini, 1998. Microsatellite DNA in Actinidia chinensis: isolation, characterisation and homology in related species. Theor Appl Genet 97: 1269–1278.
Lespinasse, Y., L. Bouvier, M. Djulbic & E. Chevreau, 1999. Haploidy in apple and pear. Acta Hort 484: 49–54.
Morejohn, L., T.E. Burau, J. Bajer, A.S. Bajer & D.E. Fosket, 1987. Oryzalin, a dinitraniline herbicide, binds to plant tubulin and inhibits microtubule polymerisation in vitro. Planta 172: 252–264.
Morgante, M. & A.M. Olivieri, 1993. PCR-amplified microsatellites as markers in plant genetics. Plant J 3(1): 175–182.
Peakall, R., S. Gilmore, W. Keys, M. Morgante & A. Rafalski, 1998. Cross species amplification of soybean (Glycine max) simplesequence-repeats (SSRs) within the genus and other legume genera. Mol Biol Evol 15: 1275–1287.
Quoirin, M., P. Lepoivre & P. Boxus, 1977. Un premier bilan de dix années de recherche sur les cultures de méristèmes et la multiplication in vitro de fruitiers ligneux. C R Rech Stn Fruit maraîchères, Gembloux, pp. 93–117.
Ramulu, K.S., H.A. Verhoeven & P. Dijkhuis, 1991. Mitotic blocking, micronucleation and chromosome doubling by oryzalin, amiprophosmethyl and colchicine in potato. Protoplasma 160: 65–71.
Verdoodt, L., A. Van Haute, I.J. Goderis, K. De Witte, J. Keulemans & W. Broothaerts, 1998. Use of the multi-allelic self-incompatibility gene in apple to assess homozygosity in shoots obtained through haploid induction. Theor Appl Genet 96: 294–300.
Vinatzer, B.A., H.B. Zhang & S. Sansavini, 1998. Construction and characterisation of a bacterial artificial chromosome library in apple. Theor Appl Genet 97: 1183–1190.
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Bouvier, L., Guérif, P., Djulbic, M. et al. Chromosome doubling of pear haploid plants and homozygosity assessment using isozyme and microsatellite markers. Euphytica 123, 255–262 (2002). https://doi.org/10.1023/A:1014998019674
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DOI: https://doi.org/10.1023/A:1014998019674