Flow cytometric probing of chromatin condensation in maize diploid nuclei

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Abstract
SUMMARY: Nuclei were isolated from stems, mesocotyls, nodal roots and root tips of two maize hybrids. The fluorochrome, propidium iodide (PI), was used to stain the nuclei that were passed through a flow cytometer‐cell sorter. Significant differences in fluorescence intensities were observed among the diploid nuclei from the different plant organs. These observations, in conjunction with previously reported DAPI fluorescence data, reflect the state of chromatin condensation in diploid nuclei isolated from meristematic and differentiating tissues. Nuclei from root meristematic tissues showed no or very little chromatin condensation. Mesocotyls and nodal roots, 1‐wk‐old, appeared to contain chromatin in a more diffuse state owing to completed differentiation which allows more DNA to be accessible to PI. Mesocotyls, 2‐wk‐old owing to senescence, contained degraded chromatin in which DNA was more accessible to PI because it lacked associated proteins. The diploid nuclei from stems had the highest amount of chromatin condensation. The results show the usefulness of flow cytometry for analyzing chromatin condensation in plant nuclei. The study also shows the importance of selecting a particular type of fluorochrome in the analysis of nuclear chromatin.