Further Studies on Microtubule Organizational Changes During Megagametogenesis in Rice Embryo Sac

Changes in the pattern of microtubule distribution and organization during megagametogenesis in the embryo sac of rice (Oryza sativa L. cv. IR36) were re-examined using a modified polyethylene glycol sectioning technique before immuno-fluorescence staining of microtubules. In the sectioned materials the pattern of distribution and structural organization of the microtubule cytoskeleton were quite well preserved. Fine details of the patterns of structural changes and re-organization of the microtubule cytoskeleton in the major stages of development during embryo sac megagametogenesis (viz. functional megaspore, uni-nucleate, 2-nucleate, 4-nucleate, 8-nucleate and mature stage) could be clearly observed and easily followed. Some new organizational patterns of microtubules associated with the probable movement and positioning of the polar nuclei were observed.

Abnormalities Occurring during Female Gametophyte Development Result in the Diversity of Abnormal Embryo Sacs and Leads to Abnormal Fertilization in indica/japonica Hybrids in Rice

Embryo sac abortion is one of the major reasons for sterility in indicaljaponica hybrids in rice. To clarify the causal mechanism of embryo sac abortion, we studied the female gametophyte development in two indicaljaponica hybrids via an eosin B staining procedure for embryo sac scanning using confocal laser scanning microscope. Different types of abnormalities occurred during megasporogenesis and megagametogenesis were demonstrated. The earliest abnormality was observed in the megasporocyte. A lot of the chalazal-most megaspores were degenerated before the mono-nucleate embryo sac stage. Disordered positioning of nucleus and abnormal nucellus tissue were characteristics of the abnormal female gametes from the mono-nucleate to four-nucleate embryo sac stages. The abnormalities that occurred from the early stage of the eight-nucleate embryo sac development to the mature embryo sac stage were characterized by smaller sizes and wrinkled antipodals. Asynchronous nuclear migration, abnormal positioning of nucleus, and degeneration of egg apparatus were also found at the eight-nucleate embryo sac stage. The abnormalities that occurred during female gametophyte development resulted in five major types of abnormal embryo sacs. These abnormal embryo sacs led to abnormal fertilization. Hand pollination using normal pollens on the spikelets during anthesis showed that normal pollens could not exclude the effect of abnormal embryo sac on seed setting.