INVESTIGATION OF MAIZE MEIOCYTE DEVELOPMENT THROUGH SINGLE-CELL RNA SEQUENCING

Meiosis is a fundamental part of the life cycle for sexually reproducing organisms. In plants, meiosis marks the shift from sporophytic stage of life to the gametophytic stage. During this shift in the life cycle, single differentiated cells undergo major transcriptional and cellular changes as they transition into a multicellular gametophyte. Studying these cells has been a challenge historically as reproductive cells are rare and are embedded within somatic tissue. Here, we describe a method to isolate male meiotic cells and pollen grains for analysis using single-cell RNA sequencing (scRNA-seq) and describe library preparation and independent staging using 4′,6-diamidino-2-phenylindole (DAPI) staining. This method is useful in investigating meiosis through the use of mutants, such as the absence of first division (afd1) in which a protein integral to the cohesin complex is not formed. This research highlights the potential for extensive investigation into the meiotic process using the methods provided.