, Volume 150, Issue 1–2, pp 271–280 | Cite as

Interspecific Hybridization in the Genus Lens Applying in Vitro Embryo Rescue



Four different hybridization experiments were carried out to obtain interspecific hybrids with Spanish cultivars of lentil (Lens culinaris M.). In hybridization experiments I and II, undertaken with only pollination and pollination with the addition of gibberellic acid after fertilization, respectively, no lentil hybrids were recovered. A single interspecific hybrid with L. odemensis was obtained in experiment III using the embryo rescue protocol of Cohen et al. (1984), in this case, a crossing efficiency of 0.11% and a rescue efficiency of 2.5% were obtained. Hybridization experiment IV used a specific embryo rescue protocol developed in this study. In this experiment, ovule-embryos of 18 DAP were cultured on MS salts with 1% sucrose and 1 μ M IAA + 0.8 μ M KN; after two weeks, embryos were released from the ovular integuments and cultured on the same medium for another two weeks in upright position. Afterwards, the embryos were transferred to test tubes containing the same medium and one month later plantlets were obtained. Using the above protocol, out of a total of 1707 pollinations, 6 interspecific hybrids with L. odemensis, 2 with L. nigricans and one with L. ervoides were recovered, yielding on average a crossing efficiency of 0.53% and an average rescue efficiency of 8.26%. Taking into consideration only the interspecific crossing blocks in which hybrids were recovered, the crossing efficiency with L. odemensis was 9%, while with both L. nigircans and L. ervoides the crossing efficiencies were 3%. Rescue efficiencies based on hybrids recovered per number of ovules cultured ranged between 50–100%.


embryo and ovule culture interspecific hybridization legume Lens lentil hybrids 



days after pollination


gibberellic acid


indole-3-acetic acid




polymerase chain reaction


random amplified polymorphic DNA




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  1. Abbo, S. & G. Ladizinsky, 1991. Anatomical aspects of hybrid embryo abortion in the genus Lens L. Botanical gazette 152: 316–320.CrossRefGoogle Scholar
  2. Abbo, S. & G. Ladizinsky, 1994. Genetical aspects of hybrid embryo abortion in the genus Lens L. Heredity 72: 193–200.Google Scholar
  3. Ahmad, M., A.G. Fautrier, D.L. McNeil, D.J. Burritt & G.D. Hill, 1995. Attempts to overcome postfertilization barrier in interspecific crosses of the genus Lens. Plant Breeding 114: 558–560.CrossRefGoogle Scholar
  4. Alvarez, M.T., P. García & M. Pérez de la Vega, 1997. RAPD polymorphism in Spanish lentil landraces and cultivars. J Genet & Breed 51: 91–96.Google Scholar
  5. Cohen, D., G. Ladizinsky, M. Ziv & F.J. Muehlbauer, 1984. Rescue of interspecific Lens hybrids by means of embryo culture. Plant Cell Tiss Org Cult 3: 343–347.CrossRefGoogle Scholar
  6. Dellaporta, S.L., J. Wood & J.B. Hicks, 1983. A plant DNA minipreparation: Version II. Plant Mol Biol Rep 1: 19–21.Google Scholar
  7. Duran, Y., R. Fratini, P. García & M. Pérez de la Vega, 2004. An intersubspecific genetic map of Lens. Theor Appl Genet 108: 1265–1273.PubMedCrossRefGoogle Scholar
  8. Ferguson, M., 2000. Lens spp: Conserved resources, priorities for collection and future prospects. In: R. Knight (Ed.), Linking Research and Marketing Opportunities for Pulses in the 21st Century, Current Plant Science and Biotecnology in Agriculture, Vol. 34, pp. 613–620. Kluwer Academic Publishers, Dordrecht.Google Scholar
  9. Ford, R., E.C.K. Pang & P.W.J. Taylor, 1997. Diversity analysis and species identification in Lens using PCR generated markers. Euphytica 96: 247–255.CrossRefGoogle Scholar
  10. Fratini, R. & M.L. Ruiz, 2002. Comparative study of different cytokinins in the induction of morphogenesis in lentil (Lens culinaris Medik.). In Vitro Cell Dev-Pl 38: 46–51.CrossRefGoogle Scholar
  11. Fratini, R. & M.L. Ruiz, 2003. A rooting procedure for lentil (Lens culinaris Medik.) and other hypogeous legumes (pea, chickpea and Lathyrus) based on explant polarity. Plant Cell Rep 21: 726–732.PubMedGoogle Scholar
  12. Fratini, R., M.L. Ruiz & M. Pérez de la Vega, 2004. Intra-specific and inter-sub-specific crossing in lentil (Lens culinaris Medik.). Can J Plant Sci 84: 981–986.Google Scholar
  13. Harlan, J.R. & J.M.J. de Wet, 1977. Pathways of genetic transfer from Tripsacum to Zea Mays. Proceedings of the National Academy of Science, USA 74: 3494–3497.CrossRefADSGoogle Scholar
  14. Ladizinsky, G., 1979. The origin of lentil and its wild genepool. Euphytica 28: 179–187.CrossRefGoogle Scholar
  15. Ladizinsky, G., D. Braun, D. Goshen & F.J. Muehlbauer, 1984. The biological species of the genus Lens L. [Lens nigricans]. Botanical gazette 145: 253–261.CrossRefGoogle Scholar
  16. Ladizinsky, G., D. Cohen & F.J. Muehlbauer, 1985. Hybridization in the genus Lens by means of embryo culture. Theor Appl Genet 70: 97–101.CrossRefGoogle Scholar
  17. Lazaridou, T.B., D.G. Roupakias & A.S. Economou, 1993. Embryo rescue in Vicia faba and Vicia narbonensis. Plant Cell Tiss Org Cult 33: 297–301.CrossRefGoogle Scholar
  18. Malik, K.A. & P.K. Saxena, 1992. Thidiazuron induces high frequency shoot regeneration in intact seedlings of pea (Pisum sativum), chickpea (Cicier arietinum) and lentil (Lens culinaris). Aust J Plant Physiol 19: 731–740.CrossRefGoogle Scholar
  19. Mejia-Jimenez, A., C. Muñoz, H.J. Jacobsen, W.M. Roca & S.P. Singh, 1994. Interspecific hybridization between common and tepary beans: Increased hybrid embryo growth, fertility, and efficiency through recurrent and congruity backcrossing. Theor Appl Genet 88: 324–331.Google Scholar
  20. McCoy, T.J. & L.Y. Smith, 1986. Interspecific hybridization of perennial Medicago species using ovule-embryo culture. Theor Appl Genet 71: 772–783.CrossRefGoogle Scholar
  21. Muehlbauer, F.J., N.F. Weeden & D.L. Hoffman, 1989. Inhertitance and linkage relationships of morphological and isozyme loci in lentil (Lens Miller). J Hered 80: 298–303.Google Scholar
  22. Muehlbauer, F.J., N.F. Weeden & D.L. Hoffman, 1989. Inhertitance and linkage relationships of morphological and isozyme loci in lentil (Lens Miller). J Hered 80: 298–303.Google Scholar
  23. Muench, D.G., A.E. Slinkard & G.J. Scoles, 1991. Determination of genetic variation and taxonomy in lentil (Lens Miller) species by chloroplast DNA polymorphism. Euphytica 56: 213–218.CrossRefGoogle Scholar
  24. Murashige, T. & F. Skoog, 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497.CrossRefGoogle Scholar
  25. Ocampo, B., C. Conicella & J.P. Moss, 2000. Wide crossing: Opportunities and progress. In: R. Knight (Ed.), Linking Research and Marketing Opportunities for Pulses in the 21st Century, Current Plant Science and Biotecnology in Agriculture, Vol. 34, pp. 411-419. Kluwer Academic Publishers, Dordrecht.Google Scholar
  26. Pickersgill, B., 1993. Interspecific hybridization by sexuals means. In: M.D. Hayward, N.O.Bosemark, I. Romagosa (Eds), Plant Breeding: Principles and prospects, pp. 63–78. Chapman & Hill, London.Google Scholar
  27. Pitarelli, G.W. & J.R. Stavely, 1975. Direct hybridisation of Nicotiana rependa , N. tabacum. J Hered 66: 281–284.Google Scholar
  28. Polanco, M.C. & M.L. Ruiz, 1997. Effect of benzylaminopurine on in vitro and in vivo root development in lentil (Lens culinaris Medik.). Plant Cell Rep 17: 22–26.CrossRefGoogle Scholar
  29. Polanco, M.C. & M.L. Ruiz, 2001. Factors that affect plant regeneration from in vitro culture of immature seeds in four lentil (Lens culinaris Medik.) cultivars. Plant Cell Tiss Org Cult 66: 133–139.CrossRefGoogle Scholar
  30. Saghai-Maroof, M.A., K.M. Soliman, R.A. Jorgensen & R.W. Allard, 1984. Ribosomal DNA spacer length polymorphism in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proc Natl Acad Sci USA 81: 8014–8018.PubMedCrossRefADSGoogle Scholar
  31. Vaillancourt, R.E. & A.E. Slinkard, 1992. Inhertitance of new genetic markers in lentil (Lens Miller). Euphytica 64: 227–236.CrossRefGoogle Scholar
  32. Vandenberg, A. & A.E. Slinkard, 1989. Inhertitance of four new quantitative genes in lentil. J Hered 80: 320–322.Google Scholar
  33. Williams, E.G., 1987. Interspecific hybridization in pasture legumes. In: J. Janick (Ed.), Plant Breeding Reviews No 5, pp. 237–305. AVI Publishing Co., Westport.Google Scholar
  34. Williams, E.G., G. Maheswaran, J.F. Hutchinson, 1987. Embryo and ovule culture in crop improvement. In: Janick J. (ed.), Plant Breeding Reviews No 5, pp. 181–236. AVI Publishing Co, Westport.Google Scholar
  35. Wilson, V.E., 1972. Morphology and technique for crossing Lens esculenta Moench. Crop Sci 12: 231–232.CrossRefGoogle Scholar
  36. Wilson, V.E. & A.G. Law, 1972. Natural crossing in Lens esculenta Moench. J Am Soc Hortic Sci 97: 142–143.Google Scholar
  37. Ye, G., D.L. McNeil, A.J. Conner & G.D. Hill, 2000. Improved protocol for the multiplication of lentil hybrids without genetic change by culturing single node explants. SABRAO Journal of Breeding and Genetics 32: 13–21.Google Scholar
  38. Ye, G., D.L. McNeil, A.J. Conner & G.D. Hill, 2002. Multiple shoot formation in lentil (Lens culinaris) seeds. New Zealand Journal of Crop and Horticultural Science 30: 1–8.MATHCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  1. 1.Área de Genética, Facultad de Ciencias Biológicas y AmbientalesUniversidad de LeónLeónSpain

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