Abstract
Induction of prolific shoot formation in Phaseolus vulgaris L. cv. Kinghorn Wax was achieved by germinating mature seeds and growing seedlings on a medium supplemented with 10 μM thidiazuron (TDZ), a substituted phenylurea, or 80 μM N6-benzylaminopurine (BAP). Culture for 7 d in the presence of 10 μM TDZ was sufficient to induce maximal shoot formation, whereas a continuous presence of BAP was required for the induction and development of shoots. The differentiation of adventitious shoots occurred within four weeks of seed culture, from tissues in the regions of axillary buds on the cotyledonary node and also areas surrounding the shoot apex of the intact seedling. The number of shoots regenerated from intact seedlings was significantly higher than that obtained with expiants. Regenerated shoots developed into flowering plants. Similar results were obtained in six other bean cultivars.
Similar content being viewed by others
Abbreviations
- BAP:
-
N6-benzylaminopurine
- MS:
-
Murashige and Skoog (1962) medium
- TDZ:
-
N-phenyl-N 1-(1,2,3 thiadiazol-yl)urea (thidiazuron)
References
Allavena, A. (1984) Beans (Phaseolus). In: Handbook of plant cell culture, pp. 137–168, Sharp, W.R., Evans, D.A., Ammirato, P.V., Yamada, Y., eds. Macmillan, New York
Arndt, F., Rusch, R., Stillfried, H.V. (1976) SN 49537, a new cotton defoliant. Plant Physiol. 57, S-99
Capelle, S.C., Mok, D.W.S., Kirchner, S.C., Mok, M.C. (1983) Effects of thidiazuron on cytokinin autonomy and the metabolism of N6-(2-isopentenyl)[8-14C]adenosine in callus tissue of Phaseolus lunatus L. Plant Physiol. 73, 796–802
Crocomo, O.J., Sharp, W.R., Peters, J.C. (1976) Plantlet morphogenesis and the control of callus growth and root induction of Phaseolus vulgaris with addition of bean seed extract. Z. Pflanzenphysiol. 78, 458–460
Fiola, J.A., Hassan, M.A., Swartz, H.J., Bors, R.H., McNicols, R. (1990) Effect of thidiazuron, light fluence rates and kanamycin on in vitro shoot organogenesis from excised Rubus cotyledons and leaves. Plant Cell Tiss. Org. Cult. 20, 223–228
Franklin, C.I., Trieu, T.N., Gonzales, R.A., Dixon, R.A. (1991) Plant regeneration from seedling expiants of green bean (Phaseolus vulgaris L.) via organogenesis. Plant Cell Tiss. Org. Cult. 24, 199–206
Gamborg, O.L., Miller, R.A., Ojima, K. (1968) Nutrient requirements of suspension cultures of soybean root cells. J. Exp. Res. 50, 151–158
Hammatt, N., Ghose, T.K., Davey, M.R. (1986) Regeneration in legumes. In: Cell culture and somatic cell genetics of plants, vol. 3, pp. 66–95, Vasil, I.K., ed. Academic Press, New York
Johansen, D.A. (1940) Plant microtechnique. McGraw-Hill, New York
Kartha, K.K., Pahl, K., Leung, N.L., Morginski, L.A. (1981) Plant regeneration from meristems of grain legumes: soybean, cow-peas, peanut, chickpea and bean. Can. J. Bot. 59, 1671–1679
Lazzeri, P.A., Hildebrandt, D.F., Collins, G.B. (1987) Soybean somatic embryogenesis: effect of nutritional, physical and chemical factors. Plant Cell Tiss. Org. Cult. 10, 209–220
Leon, P., Planckaert, F., Walbot, V. (1991) Transient gene expression in protoplasts of Phaseolus vulgaris isolated from a cell suspension culture. Plant Physiol. 95, 968–972
Malik, K.A., Saxena, P.K. (1991) Regeneration in Phaseolus vulgaris L. Promotive role of N6-benzylaminopurine in cultures from juvenile leaves. Planta 184, 148–150
Martins, I.S., Sondahl, M.R. (1984a) Axillary bud development from nodal cultures of bean seedlings (Phaseolus vulgaris L.). Turrialba 34, 157–161
Martins, I.S., Sondahl, M.R. (1984b) Early stages of somatic embryo differentiation from callus cells of bean (Phaseolus vulgaris L.) grown in liquid medium. J. Plant Physiol. 117, 97–103
McClean, P., Grafton, K.F. (1989) Regeneration of dry bean (Phaseolus vulgaris L.) via organogenesis. Plant Sci. 60, 117–122
Mok, M.C., Mok, D.W.S., Armstrong, D.J., Shudo, K., Isogai, Y., Okanoto, T. (1982) Cytokinin activity of N-phenyl-N′-1,2,3- thidiazol-5ylurea (thidiazuron). Phytochemistry 21, 1509–1511
Morginski, L.A., Kartha, K.K. (1985) Tissue culture of legumes for crop improvement. Plant Breed. Rev. 2, 215–264
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497
Thomas, J.C., Katterman, F.R. (1986) Cytokinin activity induced by thidiazuron. Plant Physiol. 81, 681–683
Author information
Authors and Affiliations
Additional information
To whom correspondence should be addressed
This research was supported by operating grants from the Natural Sciences and Engineering Research Council of Canada and the University Research Board Grant Programs of the University of Guelph to P.K.S. We thank Drs. Jean Gerrath and R. Rastogi for helpful discussions. Technical assistance from Sangeeta Saxena is gratefully acknowledged.
Rights and permissions
About this article
Cite this article
Malik, K.A., Saxena, P.K. Regeneration in Phaseolus vulgaris L.: High-frequency induction of direct shoot formation in intact seedlings by N6-benzylaminopurine and thidiazuron. Planta 186, 384–389 (1992). https://doi.org/10.1007/BF00195319
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00195319