Molecular characterization and identification of genes involved in maize female gametophyte development release_zibuqfz2z5g4zk6qhn6laped4q

by Kanok-orn Srilunchang

Published by Universität Regensburg.

2009  

Abstract

Megagametophyte development involves many fundamental phenomena, such as nuclear migration and fusion, polarity, cell death, asymmetric cell division, and cell fate specification. So far, very little is known about genes or gene products that regulate these cellular processes during female gametophyte (FG) development. To identify genes expressed in the FG, several advanced molecular approaches have been used. However, the establishment of powerful forward and genetic methods combined with several molecular markers toolkit now allows to study the processes that control FG development also in more detail in maize. By using several molecular techniques, such as RNA interference (RNAi) I could correlate defective phenotypes of maize FG development with various gene activities. Moreover, I have established a histological method to monitor embryo sac development defects occurring at early and late FG stages of transgenic RNAi plants. Subcellular protein localization studies complemented these experiments to visualize the various pattern during development and during the cell cycle. Following FG development in maize which produces an eight-nucleate embryo sac, the functional megaspore first has to undergo three nuclear divisions. This suggests that the basic cell cycle machinery is required, and any mutation in these cell cycle genes would likely disrupt nuclear division. Chapter 4 and Chapter 5 describe the different stages of maize FG development comparing wild-type with mutant phenotypes. In one study, I could show that knock-down of the MATH-BTB domain containing ZmMAB1 gene (Chapter 4) is required for the first nuclear divisions since embryo sac are arrested at the two-nucleate stage in mab1 mutants. Besides, the two nuclei were not properly segregated to each pole. This suggests that ZmMAB1 is essential for the second nuclear division and proper nuclei positioning. The knock-down of the diSUMO-like ZmDSUL gene (Chapter 5) causes the embryo sac to arrest at the eight-nucleate stage (FG5) and the immature embryo sa [...]
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