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‘Milam’ (a purported hybrid of Citrus jambhiri Lush) + ‘Femminello’ lemon (Citrus limon L. Burm. f.) allotetraploid somatic hybrids were used as pollen parents in interploid crosses with diploid ‘Femminello’ lemon to achieve mal secco tolerance in different populations of seedless triploid lemon types with good fruit quality. A total of 137 plantlets were obtained and subjected to screening experiments, in order to distinguish zygotic embryos from nucellars. Here we report on and discuss the results obtained with three techniques: flow cytometry, isozyme analysis and ISSR-PCR (the inter-simple sequence repeats-polymerase chain reaction). ISSR-PCR resulted to be a very efficient and reliable technique for the identification of zygotic plantlets.
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p.703-708,fig.
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- Istituto di Ricerca per la Genetica degli Agrumi - C.N.R. Viale delle Scienze n.11, 90128-Palermo, Italy
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Bibliografia
- 1. Tusa, N., Grosser, J. W., Gmitter, F. G. Jr and Louzada, E. S. Production of tetraploid somatic hybrid breeding parents for use in lemon cultivar improvement. Hort. Sci. 27 (1992) 445-447.
- 2. Starrantino, A. and Reforgiato Recupero, G. Citrus obtained in vitro from 2x females X 4x males. Proc. Intl. Citrus Symp. 1 (1981) 31-32.
- 3. Murashige, T. and Tucker, D. P. H. Growth factor requirements of citrus tissue culture. Proc. 1st Intl. Citrus Symp. 3 (1969) 1155-1161.
- 4. Dolezel, J. Binarova, P. and Lucretti, S. Analysis of nuclear DNA content in plant cells by flow cytometry. Biol Plant 31 (1989) 113-120.
- 5. Torres, A. J., Soost, R. K. and Diedenhofen, U. Leaf isozymes as genetic markers in Citrus. Amer. J. Bot. 65 (1978) 869-881.
- 6. Scora, R. W. and Torrisi, S. Relation of taxonomic, climatic and tissue maturity factors to the essential oil constituents in leaves and fruits in the Aurantioideae. Proc. Am. Soc. Hort. Sci. 88 (1966) 262-271.
- 7. Cardy, B. J., Stuber, C. W. and Goodman, M. M. Techniques for starch gel electrophoresis of enzymes from maize (Zea maize L.). Institute of Statistics, Mimeograph. Series no.1317. North Carolina State Univ., Raleigh (1981).
- 8. Vallejos, C. E. Enzyme activity staining In: Isozymes in Plant Genetics and Breeding, part A, (Tanskley, S. D., Orton, T. J. Eds), Elsevier: Amsterdam. (1983) 469-516.
- 9. Scarano, M.-T., Abbate, L., Ferrante, S., Lucretti, S. and Tusa, N. ISSR- PCR technique: a useful method for characterizing new allotetraploid somatic hybrids of mandarin (Citrus deliciosa Tenore). Plant. Cell Rep. 20 (2002) 1162-1166.
- 10. Fang, D. Q. and Roose, M. L. Identification of closely related citrus cultivars with inter-simple sequence repeat markers. Theor. Appl. Genet. 95 (1997) 408-417.
- 11. Zietkiewicz, E., Rafalski, A. and Labuda, D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20 (1994) 176-183.
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Bibliografia
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