Seed and embryo development in pod culture of Phaseolus vulgaris and P. vulgaris x P. acutifolius Public Deposited

http://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/gm80hz059

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  • Phaseolus vulgaris, the common bean, is susceptible to many bacterial, fungal and viral diseases. However, resistance to several diseases has been identified in related Phaseolus species, P. coccineus, P. acutifolius and P. lunatus. Thus, interspecific hybridization could result in transfer of beneficial characteristics. One of the problems encountered in interspecific hybridization of Phaseolus resides in the failure of the hybrid embryos to reach maturity. Embryo rescue techniques are required to recover interspecific hybrid plants. In vitro culturing of immature pods represents an alternative to embryo rescue. In order to develop appropriate pod culture methodology, immature selfed pods of P. vulgaris cv. Great Northern were cultured at three stages of development (containing embryos of 0.5, 0.6-1.0 and 1.5-2.5 mm in length respectively). Seed weights and embryo lengths increased significantly for all three classes of pods when cultured upright, supported by glass wool, in tubes containing a liquid medium. One or two embryos per pod, usually located at the middle positions, could develop further. The same pod culture procedure promoted precocious germination of cotyledonary embryos of P. vulgaris Great Northern x P. acutifolius P.I.321637, without advancing the embryos to further developmental stages. These findings suggest that the developmental arrest of the interspecific hybrid embryos in vivo is due to intrinsic developmental controls. Since growth regulators may be involved in the retardation of interspecific hybrid embryo growth, seed and embryo growth were measured after a pod culture period in the presence of 10⁻⁶M of N⁶-benzyladenine (BAP), gibberellic acid (GA₃) or abscisic acid. The growth regulators did not affect seed weight and embryo length in cultured P. vulgaris pods. However, the length of embryos in the largest class of P. vulgaris Great Northern x P. acutifolius P.I.321637 pods were significantly lower in the presence of BAP and GA₃ as compared to the control without growth regulators. Thus, at the concentration tested, growth regulators were not beneficial for further embryo development but BAP and GA₃ seemed to depress root elongation of the germinating embryos of the interspecific hybrid. Although mature hybrid seed can not be generated in pod culture, hybrid plantlets can be obtained after a brief phase of culturing the germinating embryos. The germinating embryos from pod culture offer some advantage over embryos dissected at an earlier stage, since the presence of roots enhances nutrient uptake and hardening. Further studies leading to identification of the factors that cause developmental arrest of interspecific hybrid embryos may aid in devising the optimal pod culture medium for further growth of hybrid seeds and embryos.
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