In vivo labelling of excised tassel florets from greenhouse-grown material and material grown in tissue culture

Individual florets or spikelets (S60) were excised from tassels grown in tissue culture (see Polowick MCGNL 55:116, 1981) or from greenhouse-grown material, and labelled with 35S-methionine for 2, 3 or 4 hours. The florets from the greenhouse material had immature pollen in the larger anthers and quartets in the smaller anthers. Florets from the cultured material contained anthers with either pollen mother cells or quartets. Results show that incorporation of label increased by a factor of approximately 6, from the 2 to 4 hr labelling period, for both the greenhouse material and the cultured material. Total incorporation after 4 hrs of labelling was higher in the cultured material than in the greenhouse material. Average values for each were 12,000 ± 3,000 counts/ug protein for the greenhouse material and 32,000 ± 22,000 counts/ug protein for the cultured material. Comparison of the one dimensional SDS PAGE patterns for extracted proteins from greenhouse grown material indicates similar patterns for the same stage/tissue labelled over the three time periods. The cultured material showed a difference in banding pattern from the greenhouse material, and a comparison of banding patterns from different developmental stages of the greenhouse tissue also showed differences.

M. J. Dunlop, R. I. Greyson, D. B. Walden and B. G. Atkinson

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