--V. R. Bommineni and E. Banasikowaska
Mature plants from cultured shoot apical meristems through "jiffy pot" technique: We reported the successful recovery of plantlets through shoot apical meristem culture (MNL 63: 87-88 and 64: 79-79; Plant Cell Tissue Organ Culture, 19: 225-234, 1989). The in vitro culture technology imposes stress upon the developing plantlets. We have developed a "jiffy pot" technique to minimize some of the stress-related conditions in the transfer sequence of excised meristem to plantlet, and at the same time to enable developing plantlets to be shipped in large numbers from a laboratory to a remote nursery.
The jiffy pots can be obtained from a garden/plant nursery store. First, the jiffy pots were soaked in water (in a tray) until the pots expand completely. The pots were then placed in Magenta(TM) plant cell culture vessels (3" x 3" x 4") (Sigma Chemical Company, USA) and the vessels autoclaved.
Murashige and Skoog's basic nutrient agar medium (MNL 63: 87-88; Plant Cell Tissue Organ Culture 19: 225-234, 1989) was used to support the explant apical meristems. One and one-half cm diameter agar blocks were removed and pressed into the autoclaved jiffy pot (top centre) under sterile conditions. The meristems were excised and placed on the agar blocks as described in Table 1. Two different methods were followed to compare the recovery of plantlets with the existing method (control, Table 1).
Recovery of fertile plants through in vitro culture of tassel meristems in Magenta plant cell culture vessels: We reported a summary of the recovery of plantlets from cultured tassel meristems in 125 ml conical flasks (Maydica 34: 263-275, 1989) derived from Oh43 and K21 genotypes. However, the number of plantlets recovered and the number of plantlets matured through that culture method were very low. To improve the success rate and increase the scale of the effort, Magenta plant cell culture vessels were used in the place of 125 ml conical flasks. As reported 40 ml of Murashige and Skoog's nutrient medium was placed into the vessels and the vessels were autoclaved.
The initial length of tassel meristems ranges from 1.0 to 1.5 cm; the tassel meristems were dissected by removing the sheath leaves, the dissecting meristems were placed into the Magenta vessels and incubated at 26+2 C for 25+ days. The plantlets which emerged after 25+ days of culture were allowed to grow further in the vessel until they were ready to be transferred to soil pots in the glasshouse.
A summary of the recovery of plantlets from cultured tassel meristems in both flasks and vessels is included in Table 2. A higher percentage of plantlets (19%) were recovered through tassel meristems cultured in Magenta vessels comparing to meristems cultured in 125 conical flasks (5%).
Table 1. Plants from cultured immature apical meristems (a C R b pl).
2. Recovery of plants from cultured tassel meristems (Oh43).
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