Abstract
In-vitro methods have been developed to regenerate clumps of multiple shoots and somatic embryos at high frequency from shoot tips of aseptically-grown seedlings as well as from shoot apices of precociously-germinated immature zygotic embryos of corn (Zea mays L.). About 500 shoots were produced from a shoot tip after eight weeks of culture (primary culture and one subculture of four weeks) in darkness on Murashige and Skoog basal medium (MS) supplemented with 500 mg/L casein hydrolysate (CH) and 9 μM N6-benzyladenine (BA). In this medium, shoots formed in shoot tips as tightly packed “multiple shoot clumps” (MSC), which were composed of some axillary shoots and many adventitious shoots. When the shoot tips were cultured on MS medium containing 500 mg/L CH, 9 μM BA and 2.25 μM 2,4-dichlorophenoxyacetic acid (2,4-D), most of the shoots in the clumps were adventitious in origin. Similar shoot tips cultured on MS medium containing 500 mg/L CH, 4.5 μM BA and 2.25 μM 2,4-D regenerated many somatic embryos within eight weeks of culture. Somatic embryos were produced either directly from the shoot apical meristems or from calli derived from the shoots apices. Both the MSC and the embryos produced normal shoots on MS medium containing 2.25 μM BA and 1.8 μM indole-3-butyric acid (IBA). These shoots were rooted on MS medium containing 3.6 μM IBA, and fertile corn plants were grown in the greenhouse. The sweet-corn genotype, Honey N Pearl, was used for the experiments described above, but shoot-tip cultures from all of 19 other corn genotypes tested also formed MSC on MS medium containing 500 mg/L CH and 9 μM BA.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- BA:
-
N6-benzyladenine
- CH:
-
casein hydrolysate
- CSM:
-
corn-shoot multiplication medium
- 2,4-D:
-
2,4-dichlorophenoxyacetic acid
- IBA:
-
indole-3-butyric acid
- MS:
-
Murashige and Skoog (basal medium)
- MSC:
-
multiple shoot clump(s)
References
Bhaskaran, S., Smith, R.H. (1990) Regeneration in cereal tissue culture: A Review. Crop Sci. 30, 1328–1336
Conger, B.V., Novak, F.J., Afza, R., Erdelsky, K. (1987) Somatic embryogenesis from cultured leaf segments of Zea mays. Plant Cell Rep. 6, 345–347
Dale, P.J. (1980) Embryoids from cultured immature embryos of Lolium multiflorum. Z. Pflanzenphysiol. 100, 73–77
Duncan, D.R., Williams, M.E., Zehr, B.E., Widholm, J.M. (1985) The production of callus capable of regeneration from immature embryos of numerous Zea mays genotypes. Planta 165, 322–332
Esau, K. (1977) Anatomy of seed plants. John Wiley & Sons, New York
Fromm, M.E., Morrish, F., Armstrong, C., Williams, R., Thomas, J., Klein, T.M. (1990) Inheritance and expression of chimeric genes in the progeny of transgenic maize plants. BioTechnology 8, 833–839
Gordon-Kamm, W.J., Spencer, T.M., Manganno M.L., Adams, T.R., Daines, R.J., Willets, N.G., Rice, T.B., Mackey, C.J., Krueger, R.W., Kaush, A.P., Lamaux, P.G. (1990) Transformation of maize cells and regeneration of fertile transgenic plants. Plant Cell 2, 603–608
Green, C.E., Phillips, R.L. (1975) Plant regeneration from tissue cultures of maize. Crop Sci. 15, 417–421
He, D.G. Yang, Y.M., Bertram, J., Scott, K.J. (1990) The histological development of the regenerative tissue derived from immature embryos of wheat (Triticum aestivum L.). Plant Sci. 68, 103–111
Hu, C.Y., Wang, P.J. (1983) Meristem, shoot tip, and bud culture. In: Handbook of plant tissue culture, vol. 1, pp. 177–227. Evans, D.A., Sharp, W.R., Ammirato, P.V., Yamada, Y., eds. Macmillan, New York
Hodges, T.K., Kamo, K.K., Imbrie, C.W., Becwar, M.R. (1986) Genotype specificity of somatic embryogenesis and regeneration in maize. BioTechnology 4, 219–223
Jones, T.J., Rost, T. (1989) The developmental anatomy and ultrastructure of somatic embryos from rice (Oryza sativa L.) scutellum epithelial cells. Bot. Gaz. 150, 41–49
Klomparens, K.S., Flegler, S., Hooper, G.R. (1986) Procedures for transmission and scanning electron microscopy for biological and medical sciences — A laboratory manual, Ladd Research Industries, Burlington, Vt., USA
Maheswaran, G., Williams, E.G. (1985) Origin and development of somatic embryos formed directly on immature embryos of Trifolium repens in vitro. Ann. Bot. 56, 619–630
McDaniel, C.N., Poethig, R.S. (1988) Cell-lineage patterns in the shoot apical meristem of the germinating maize embryo. Planta 175, 13–22
Murashige, T., Skoog, F. (1962) A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 15, 473–497
Potrykus, I. (1990) Gene transfer to plants: assessment and perspectives. Physiol. Plant. 79, 125–134
Raghavan, V. (1986) Embryogenesis in angiosperms — A developmental and experimental study. Cambridge University Press, Cambridge, UK
Rhodes, C.R., Green, C.E., Phillips, R.L. (1986) Factors affecting tissue culture initiation from maize tassels. Plant Sci. 46, 225–232
Rhodes, C.A., Pierce, D.A., Mettler, L.J., Mascarenhas, D., Detmer, J.J. (1988) Genetically transformed maize plant from protoplasts. Science 240, 204–207
Springer, W.D., Green, C.E., Kohn, K.A. (1979) A histological examination of tissue culture initiation from immature embryos of maize. Protoplasma 101, 269–281
Stolarz, A., Macewicz, K.J., Lorz, H. (1991) Direct somatic embryogenesis and plant regeneration from leaf explants of Nicotiana tabacum L.. J. Plant Physiol. 37, 347–357
Vasil, I.K. (1987) Developing cell and tissue culture systems for the improvement of cereal and grass crops. J. Plant Physiol. 128, 193–218
Vasil, I.K. (1990) The realities and challenges of plant biotechnology. BioTechnology 6, 296–300
Vasil, V., Lu, C., Vasil, I.K. (1985) Histology of somatic embryogenesis in cultured immature embryos of maize (Zea mays L.). Protoplasma 127, 1–8
Wang, A. (1987) Callus and plant regeneration from maize mature embryos, Plant Cell Rep. 6, 360–362
Wernicke, W., Milkovits, L. (1986) The regeneration potential of wheat shoot meristem in the presence and absence of 2,4-dichlorophenoxyacetic acid. Protoplasma 131, 131–141
Wernicke, W., Potrykus, I., Thomas, E. (1982) Morphogenesis from cultured leaf tissue of Sorghum bicolor—the morphogenetic pathways. Protoplasma 111, 53–62
Author information
Authors and Affiliations
Additional information
Heng Zhong is a Rockefeller Foundation Fellow on leave from the Institute of Botany, Academia Sinica, Beijing, P.R. China. This work was supported by a grant from the MidWest Plant Biotechnology Consortium and U.S.-A.I.D. grant No. DAN-4197-A-00-1126-00 to M.B. Sticklen. Thanks are due to Illinois Foundation Seeds, Minnesota Crop Improvement Association, and Michigan Agricultural Experiment Station, for the gift of corn seeds used in this research. Critical review of this manuscript by Indra K. Vasil, University of Florida, Gainesville; Wesley P. Hackett, University of Minnesota, St. Paul; Scott Poethig, University of Pennsylvania, Philadelphia; and Mark G. Bolyard, Michigan State University is gratefully acknowledged. The authors would also like to thank the Services of Center for Electron Optics, Michigan State University, for the electromicroscopic work as related to this publication.
Rights and permissions
About this article
Cite this article
Zhong, H., Srinivasan, C. & Sticklen, M.B. In-vitro morphogenesis of corn (Zea mays L.). Planta 187, 483–489 (1992). https://doi.org/10.1007/BF00199966
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00199966