Transmission Electron Microscopic Study of Maize Pachytene Chromosome 6

Abstract

Cytoplasm-free chromosomes are frequently obtained in meiotic chromosome spreads prepared from mildly-fixed maize microsporocytes. These chromosomes are suitable for detailed structural analysis using a published electron microscopic technique. In the electron micrograph, the knobs and heterochromatin regions that have been used for karyotype analyses in the light microscope are clearly visible. Therefore, the electron microscopic map can be easily aligned with the traditional cytological map. In addition to these prominent structural features, numerous electron-dense bands also are observed. To determine whether the bands can be used as markers for the identification of each chromosomal subregion, the banding pattern of chromosome 6 is analyzed. Chromosome 6 is frequently associated with the nucleolus and can be easily recognized. We observed that at the zygotene stage in prophase I, electron-dense regions are detected on each homolog of the synapsing chromosome. During synapsis, the electron-dense regions on both homologs are brought into register to form more conspicuous bands. At the early pachytene stage, the banding pattern is stable and reproducible. Chromosome 6 contains eight dark bands, 19 medium bands and 14 light bands. The bands can be used as intrachromosomal markers for regional assignment of genes in detailed in situ hybridization mapping or cytogenetic studies. As the pachytene stage progresses, condensation of the chromosome bivalents is accompanied by fusion of adjacent bands.