Effect of nurse cultures on the production of macro-calli and fertile plants from maize embryogenic suspension culture protoplasts |
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Authors: | W L Petersen S Sulc C L Armstrong |
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Institution: | (1) Monsanto Agricultural Company, 700 Chesterfield Village Parkway, 63198 St. Louis, MO., USA;(2) Agrigenetics Company, 5649 East Buckeye Rd., 53716 Madison, WI., USA |
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Abstract: | Fertile plants have been obtained from maize (Zea mays L.) embryogenic suspension culture protoplasts. Friable, embryogenic callus initiated from an immature embryo from a cross involving the genotypes A188, B73, and Black Mexican sweetcorn was used to establish a rapidly growing embryogenic suspension culture. After nine months in culture, high yields of viable protoplasts (30×106/ gram fresh weight) were obtained following a 1.5 hour enzymatic digestion. Protoplasts cultured with feeder cells divided and formed embryogenic callus, from which male and female fertile plants were regenerated. Protoplast-derived R1 plants were self-pollinated and immature R2 embryos isolated for callus initiation. Female fertile plants have also been produced from protoplasts isolated from an R2-derived suspension culture. Significant interactions between protoplast and feeder-cell lines were observed.Abbreviations BC
backcross
- BMS
Black Mexican Sweetcorn
- 2,4-D
2,4-dichlorophenoxyacetic acid
- PWS
protoplast wash solution (0.2 M mannitol, 80 mM CaCl2)
- FDA
fluorescein diacetate
- ABA
abscisic acid |
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