THE SPECIAL WALLS AROUND GAMETES IN CERATOPTERIS RICHARDII AND AULACOMNIUM PALUSTRE: USING IMMUNOCYTOCHEMISTRY TO EXPOSE STRUCTURE, FUNCTION, AND DEVELOPMENT

AN ABSTRACT OF THE DISSERTATION OF RENEE A LOPEZ SWALLS, for the Doctor of Philosophy degree in PLANT BIOLOGY, presented on June 27, 2016, at Southern Illinois University Carbondale. TITLE: THE SPECIAL WALLS AROUND GAMETES IN CERATOPTERIS RICHARDII AND AULACOMNIUM PALUSTRE: USING IMMUNOCYTOCHEMISTRY TO EXPOSE STRUCTURE, FUNCTION, AND DEVELOPMENT MAJOR PROFESSOR: Dr. Karen S. Renzaglia Gametes are arguably the most important cells formed during the sexual life cycle of plants. The ancestral condition of gametes in land plants is the production of small motile sperm cells and larger non-motile eggs. Unique walls or cell matrices are formed during the development of these highly specialized cells, and are integral to their proper development and maturation. Yet, the polysaccharide composition, structural function, and metabolic processes of these special gamete cell walls remain unexplored beyond examination in the light microscope. Utilizing histochemical techniques coupled with immunocytochemical localizations with monoclonal antibodies (MAbs), I give a detailed survey of AGP and cell wall polymer distribution during male and female gametogenesis in the model fern, Ceratopteris richardi, and for comparison with both Ceratopteris and seed plants, I examined the same cell wall polymers during spermatogenesis in Aulacomnium palustre, a moss species. AGPs are abundant in the extraprotoplasmic matrix that surrounds differentiating sperm and egg cells in the fern, Ceratopteris richardii. During spermatogenesis, AGPs are speculated to regulate growth of flagella and cell morphogenesis through cell signaling via Ca+2 oscillations. Immunogold localizations revealed that AGPs are differentially expressed in the egg envelope in C. richardii. These glycoproteins are extremely abundant prior to fertilization but decrease substantially after fusion of the male gamete with the egg cytoplasm. Contrary to the AGP-filled matrix surrounding developing spermatids, developing eggs are bathed in (1,5)-α-L-arabinan pectins and not AGPs. Lastly, I examined the unique cell walls that are integral to sperm cell differentiation and release in both Ceratopteris and Aulacomnium. The preponderance of callose and hemicelluloses in the walls of the male gametes of Ceratopteris and Aulacomnium, respectively, and the importance of these polysaccharides in development are discussed. Taken together, the studies that comprise this dissertation advance significantly our understanding of cell wall dynamics during gametogenesis in early land plants