Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 127, Issue 3, pp 585–590 | Cite as

In vitro embryo culture to shorten the breeding cycle in lentil (Lens culinaris Medik)

  • Carolina BermejoEmail author
  • Ileana Gatti
  • Enrique Cointry
Original Article


Breeding in lentil involves hybridization followed by different selection methods and requires 10 years to obtain a cultivar, as only one field generation per year can be produced. To shorten the breeding time it is essential to use biotechnological methods such as in vitro embryo culture combined with SSD method since only one seed is enough to produce the next generation. An efficient in vitro–in vivo system was developed. The best time to extract immature embryos and the best culture medium to obtain their complete development were analyzed. Embryos of Pardina, B1181 (microsperma type), B1051 and A1145 (macrosperma type) were collected at 15, 18, 21, and 24 days after pollination (DAP) and cultured on MS medium with five different concentrations of 6-benzylaminopurine (BAP) (0–0.025–0.05–0.1–0.25 mg L−1). An ANOVA test among genotypes, media, DAP and their interactions was performed. Genotypes, DAP and its interaction showed significant effects on the percentage of shoot production (F = 61.8; F = 79.3; F = 8.5; p < 0.01) and germination (F = 70.7; F = 69.8; F = 3.9; p < 0.01). Medium effect was only significant for germination (F = 8.7; p < 0.01). The microsperma genotypes gave higher percentages of shoot production (>80 %) and germination (>70 %). Although in vitro culture efficiency increased with DAP, 18 DAP was selected due to its high percentages of germination (13–70 %). The medium without BAP was the most suitable for embryo complete development (41–87 %). All plants obtained were morphologically normal and fertile. Using this approach, four generations per year were obtained allowing a rapid development of RILs.


Lentil In vitro culture Immature seeds Short generation cycles 





Days after pollination


Murashige and Skoog


Recombinant inbred lines


Single seed descent



Financial support for this research work was provided by the Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT).


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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  1. 1.Instituto de Investigaciones en Ciencias Agrarias de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas (IICAR-CONICET), Facultad de Ciencias AgrariasUniversidad Nacional de Rosario (UNR)ZavallaArgentina
  2. 2.CIUNR, Consejo de Investigadores de la Universidad Nacional de RosarioRosarioArgentina
  3. 3.Cátedra de Mejoramiento Vegetal y Producción de Semillas, Facultad de Ciencias AgrariasUNRZavallaArgentina

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