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ISSN 0864-4551 (Print)

2016, Number 2

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Biotecnol Apl 2016; 33 (2)

Phaseolus vulgaris L. regeneration systems and their application for Agrobacterium-mediated genetic transformation

Collado R, García L, Bermúdez I, Veitía N, Torres D, Romero C, Martirena A, Ocaña B, Rojas LE, Angenon G

Language: English
References: 17
Page: 2521-2523
PDF size: 433.31 Kb.


ABSTRACT

Procedures for regeneration and genetic transformation of Phaseolus vulgaris L. were established. Pre-culture of explants in liquid medium with TDZ, which has not been used for common bean regeneration before, was a critical step for multiple bud induction. The optimization of several parameters involved in DNA-transfer combined with efficient direct organogenesis allowed to regenerate plants from tissue transformed with Agrobacterium tumefaciens. Somatic embryo formation on cotyledons from immature seeds of P. vulgaris was achieved. For this species, it was the first time that somatic embryos reaching the cotyledonary stage were obtained by these procedures, and a protocol was established for the in vitro regeneration of P. vulgaris via indirect organogenesis. The use of cotyledonary nodes with one or two cotyledons attached, which have not been used as initial explants for common bean regeneration before, was an important factor for the successful formation of morphogenetic green nodular compact calli. This last was successfully used as target explants to obtain transformed plants, based on the indirect organogenesis regeneration pathway. The protocol established for the CIAP7247F variety was shown reproducible in other four commercial cultivars. These results provide for the first time ever a protocol for the genetic transformation of common bean via A. tumefaciens, which can be used for cultivar breeding. This research granted the 2015 Award of the Cuban National Academy of Sciences.


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