Use of the selective agents Geneticina G-418 and Higromicina B for genetic transformation into Phaseolus vulgaris variety CIAP 72-47
Journal Title: Biotecnología Vegetal - Year 2007, Vol 7, Issue 4
Abstract
Leguminous are a very important source of protein in the diet of human and animal. Phaseolus gender is the most used in the nutrition among them. Most of the systems for genetic transformation described up to the moment require selecting the plants that contain the transgen introduced by means of a selection marker. The selection of the system requires determining first their appropriate concentrations and inhibitory effect in the growth of the explants. Therefore, the objective of the present work was to determine the minimal inhibitory concentration of the antibiotics Geneticin G-418. Also the Hygromicin B that inhibited the growth of the explants of Phaseolus vulgaris var. CIAP 72-47. Stem segments of 5-6 mm with the cotyledonal nude obtained from mature seeds were used as plant material. Culture medium for buds induction with different concentrations of Geneticin G-418 (0, 20, 30, 40, 50 and 60 mg.l -1 ) and Hygromicin B (0, 10, 15, 20 and 25 mg.l -1 ) was used for the selection. Evaluations were carried out after eight weeks. Explants number with multiple buds, as well as number of necrotic explants were determined. Antibiotic Geneticin G-418 inhibited development of multiple buds. Explants were totally necrosed (100%) starting from concentration of 50 mg.l -1 . Formation of buds in the stem segment was not achieved.This differed statistically from the rest of treatments. According to results these one was selected as the minimal inhibitory concentration. Development of multiple buds was reached at a concentration superior to 20 mg.l -1 of antibiotic Hygromicin B. Explants turned necrosed and regeneration of multiple buds was not possible. That is why it was selected to be used in the experiments of genetic transformation of Phaseolus vulgaris variety CIAP 72-47 as minimal inhibitory concentration of this selective agent .
Authors and Affiliations
I. Bermúdez-Caraballoso , R. Collado , L. R. García , N. Veitía , D. Torres , C. Romero , G. Angenon
Keywords
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