ISSN:
1615-6102
Keywords:
Cold-treated buds
;
Donor plant flowering
;
Embryogenic pollen
;
Mineral-sucrose medium
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Summary Consistent high-frequency embryo formation, up to 30% of the cultured pollen, was possible when embryogenic pollen were selectively isolated from the gametophytic pollen on centrifugation, at 12–15 °C, in Percoll diluted by a low level of sucrose (4%). Higher level of sucrose in separation medium was inhibitory for embryogenesis. The embryogenic pollen were to be from the buds, petal length 2.4 ± 0.1 cm, of plants in early stage of flowering and induced to flower at 15 °C. The pollen from buds of plants in late stage of flowering gave a variable response. Prior to isolation of pollen, these buds were to be given an additional cold treatment at 10°C for 10 days. There was no response on mere water-sucrose but the nutrient requirements of the pollen for embryogenesis were very simple, filtersterilized mineral-sucrose (2%) medium at pH 6.8. Incubation of the cultures at 24°C than at 28°C resulted in high-frequency embryogenesis. Initiation of embryo formation, at high-frequency, was possible at low level of iron (2×10−5M Fe-EDTA) than customarily employed (10−4M) but for further development of embryos higher levels of iron were required. Pollen embryogenesis was also possible on either nitrate or ammonium as sole source of nitrogen. Increased level of sucrose (4%), growth regulators (cytokinins, gibberellin), monochromatic light (red or blue) and incubation in dark did not improve the response. An appreciable frequency of embryogenesis, up to 8% of the pollen, was also possible by taking pollen from buds of plants flowering at 24°C provided the buds were given a cold treatment at 10 °C for 10 days, prior to isolation of pollen.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/BF01279833
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