The Use of Stomatal Chloroplast Number for Rapid Determination of Ploidy Level in Maize

The usefulness of using the chloroplast number in epidermal guard cells as an indirect ploidy indicator was evaluated on seed‐grown and tissue culture‐derived maize plants. For seed‐grown plants, two maize genotypes (B89 and R75) which had both diploid and tetraploid seeds available were used as experimental materials. The ploidy levels of seed‐grown plants were confirmed by flow cytometric analysis of nuclear suspensions from these plants. For regenerated plants, haploid and diploid levels were examined and the ploidy levels of these plants were determined by chromosome counts of cells from root tips. A positive relationship between the chloroplast number and ploidy level was observed for both seed‐grown and regenerated plants. The stomatal guard cells of tetraploid plants had nearly double the number of chloroplasts as the diploid plants. Similar results were found from the regenerated plants. The differences in the mean chloroplast number between diploid and tetraploid seed‐grown plants and between haploid and diploid regenerated plants were highly significant. The results of this study demonstrate that counting chloroplasts in guard cells can be an efficient means of screening a large number of plants for ploidy levels. In addition, this study suggests the possibilities of using this method for detecting contaminated seed lots by different ploidy seed and for distinguishing plants of different genotypes.