Graduate Thesis Or Dissertation
 

Molecular and physiological aspects of maize embryo maturation

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  • Experiments were performed to assess regulatory factors governing maize embryo maturation and vivipary. Both visual and molecular markers of embryo development were used to examine the roles of the hormones abscisic acid (ABA) and gibberellins (GAs), as well as water stress in governing transit from early embryogeny to maturation-phase development. A differential screen identified cDNAs whose expression is impaired in maize viviparous mutants which fail to undergo maturation and instead precociously germinate. The cDNAs isolated in this screen absolutely required both ABA and the Viviparousl (Vpl) gene product for expression both in vivo and in vitro. Two novel clones were isolated: a maize homologue of the wheat metallothionein gene E[subscript]c and a second clone which may encode a novel seed storage protein of maize. In a separate screen, a maize cDNA encoding a Lea group 3 protein was isolated. Like many maturation-associated genes, maize Lea 3 was shown to ABA-inducible but is also expressed in response to water stress in the absence of ABA or the Vp 1 gene. We examined whether gibberellins might also be a factor modulating precocious germination. Gibberellin inhibitors applied to cultured wildtype embryos suppressed precocious germination and enhanced anthocyanin accumulation in a developmentally specific manner. These behaviors mimicked the effect of ABA and they were reversed by the addition of exogenous GA₃. Vivipary in vivo resulting from diminished ABA levels could be suppressed by either chemical or genetic reduction of GA levels in immature kernels and resulted in desiccation-tolerant seed. In contrast, reduction of endogenous gibberellins did not suppress vivipary of the ABA-insensitive mutant vp1. Temporal analysis of gibberellin accumulation in developing kernels revealed the accumulation of two bioactive species (GA₁ and GA₃) during a developmental window just prior to peak ABA levels. It is suggested that these species stimulate a developmental program leading to vivipary in the absence of sufficient levels of ABA and that reduction of GA levels reestablishes a hormone balance appropriate for suppression of germination and induction of maturation in ABA-deficient kernels. The failure to suppress vivipary via reduction of GA levels in the ABA-insensitive mutant vp1 suggests that the wildtype gene product functions downstream of the sites of GA and ABA action in regulation of maturation versus germination.
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