An attempt was made to transfer two sorts of DNA into maize via pollen grains. Controls for both pollen quality and DNA behaviour during the transformation experiments were included. When genomic DNA was used, no transformants were observed among the 1805 seeds screened. With plasmid DNA (harbouring the gene expressing kanamycin resistance in plant cells), three plants with kanamycin resistance were observed among the 1723 seeds screened, although no molecular evidence of transformation was obtained. Experiments indicated that under the conditions used, DNA was being degraded by both pollen and stigma nucleases. Consequently, we attempted to determine protocols which would inhibit these nuclease activities in order to preserve DNA integrity during transformation experiments, thus allowing fertilization. We found that a classic germination medium supplemented by 300 or 600 mM KNO3, or 20% PEG1550 satisfied all these conditions.
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Brewbaker and Kwack medium
Brewbaker and Kwack medium containing 15% (w/v) sucrose
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Roeckel, P., Heizmann, P., Dubois, M. et al. Attempts to transform Zea mays via pollen grains. Sexual Plant Reprod 1, 156–163 (1988). https://doi.org/10.1007/BF00193746
- Pollen and stigma nucleases