Small-scale Isolation of Meiocytes from Arabidopsis Thaliana Anthers for Cytological Analyses

In flowering plants, the meiocytes develop within nourishing tissues – the anthers or the ovaries. Although most plants bear one or more anthers in each bisexual or male flower, the size of anthers that undergo meiosis is relatively small, and each anther contains only a small fraction of male meiocytes. As a consequence, the isolation of plant meiocytes is very challenging due to their relative inaccessibility. Efficient manual dissection of meiocytes from young Arabidopsis anthers for cytological and cytogenetic analyses is technically challenging. Indirect detection of cellular or nuclear phenotypes is impaired by poor penetration of the reagents, and cell type-specific imaging is deminished by the lack of clarity in wholemount tissues. Here, we describe a method that allows the efficient isolation of male meiocytes of Arabidopsis. Anthers undergoing meiosis were dissected and collected into appropriate isolation/labeling buffer, depending on the downstream application. Meiocytes were released by crushing with a cover slip on a glass slide. The isolated meiocytes could be analyzed using a microscope. The technical skills required are simple, and the procedure does not require any special equipment. Isolated meiocytes are suitable for a variety of dowstream applications, such as cell morphological investigation, reporter gene assays, chromosome analyses, and callose staining.