Thesis (Ph. D.) University of Missouri-Columbia, 2006. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Title from title screen of research.pdf file (viewed on July 31, 2007) Includes bibliographical references.

Changes in the concentration of some carbohydrates and in amylolytic activity have been followed during germination of Zea. Mays L. var. Hickory King and var. Early Pearl. Assay techniques have been developed which permitted assay of individual grains. Thus during the investigation both groups and individual grains were used as samples. The use of groups permitted control of assay technique. Length of radicle, coleoptile and lateral roots were recorded in order to permit quantitative estimation of correlation between growth and the concentration of the various carbohydrates. Initially, during the study of changes in the carbohydrate concentration in Hickory King grains, total reducing sugar, sucrose and dextrin concentrations were estimated. However, the results obtained for changes in dextrin concentration, although reproduceable, thereby indicating reliable assay technique, presented a confusing picture and, in view of the apparent importance of sucrose and reducing sugar concentration, assay of dextrin concentration was discontinued in a subsequent study of Early Pearl. Instead changes in total reducing sugar, sucrose and glucose concentrations were followed. The results revealed that there is very considerable variability in physiological activity between grains subjected to the same germination conditions. However, all, irrespective of variety, follow the same basic metabolic pattern during germination. The trends observed were: (i) Reducing sugar accumulates slowly during the first 72-96 hours germination, but thereafter accumulation is very rapid, although concentration may decrease towards the end of the gestation period. (ii) Glucose follows a similar pattern to reducing sugar, accumulating slowly during the early stages of germination, followed by a period of rapid increase in concentration, which may decrease towards the end of the germination period. (iii) Sucrose concentration in dormant grains is fairly high, but it decreases markedly during the first 96 hours germination. This is followed by a phase of sucrose accumulation. (iv) Dextrin concentration shows two peaks. Initial level is low, but it accumulates rapidly during the first 72 hours. The level decreases between 72 and 120 hours but increases when the germination period is increased to 192 hours, after which there is a marked decrease. It was impossible, from the data relating to the study of individual grains, to discern a trend in dextrin concentration. With the exception of dextrin, about which there is little information, the results are in general agreement with the literature. Investigation of correlation between the various carbohydrates and between these and growth revealed that: (i) reducing sugar concentration and growth are positively correlated ; (ii) glucose concentration and growth are positively correlated; (iii) sucrose and reducing sugar concentrctions are negatively correlated during the initial stages of germination; (iv) sucrose and glucose concentrations are negatively correlated during early germination; (v) glucose and reducing sugar are positively correlated; (vi) in general, correlation between growth and concentration of the carbohydrates studied, decreases during the later periods of germination. These observations suggested that growth was, at least during the early stages of germination, dependent on the level of reducing sugar, and more particularly on the level of glucose, and that sucrose is the principal source of reducing sugar during this period. The relationship between amylase activity (total alpha- and beta-amylase activity) and reducing sugar concentration tends to be curvilinear, which suggests that amylolytic activity produces relatively little reducing sugar during early gennination, even though amylase activity and growth may be positively correlated. The results suggest, contrary to the observations of previous workers, that alpha-amylolytic activity may be present in dormant grains and that maize is not characterised by low levels of beta-amylase activity during germination. From the observations it is concluded that the initinl accumulati on of reducing sugar is the result of sucrose hydrolysis, and therefore sucrose is an importnnt metabolite durjng early germination. Amylolytic activity contributes little reducing sugar durlng the initia1 stages of germinatIon but that after approximately 72 hours it represents the major source of reducing sugar.

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Typescript (photocopy). / Digitized by Kansas Correctional Industries

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 GA���. 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 (GA��� and GA���) 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. / Graduation date: 1995

D8 originally designated as the dominant stature mutant of maize, was characterized and shown to be incompletely dominant. The study included morphological measurements, cytology, hormone studies, and enzyme and protein analysis. The effect of the D8 mutation can be detected after 40 hours of germination of the coleoptile. Dwarf (D8/d8) seedling length is 1/2 of the normal sib length for coleoptile, first leaf, and mesocotyl. The cell measurements indicate that cell elongation and cell division are involved in the size discrepancy. Mature dwarf plants have shorter internodes and the shorter, wider leaves are a darker green than the normal plant. The homozygous D8/D8 displays normal meiotic division and pollen formation is normal upto the 2 nucleate stage. Pollen viability of the homozygote is low and no seed was obtained in crosses involving this genotype. Several biological stains were used to test pollen viability with the results indicating greater than 85% viability for the heterozygote and less than 15.6% viability for the homozygote. The examination of the pachytene chromosomes of heterozygotes indicates a loop on a large chromosome. This loop is only found in the D8 heterozygote and implies a duplication or deficiency may be involved with the D8 phenotype. Avena straight growth bioassay for auxin displayed no significant difference in auxin production between dwarf and normal coleoptile tips. The D8 dwarf seedlings responded to the exogenous application of auxin, kinetin, and casamino acids in the same pattern as the normal seedlings, but never attained normal stature. Gibberellic acid (GA) and cyclic adenosine monophosphate (cAMP) exogenous applications displayed a difference in dwarf and normal response patterns and implies that the utilization or destruction of these substances may be involved. The investigation of Laemmli gel patterns for the three genotypes failed to show a difference. The soluble proteins formed 27 bands from the coleoptiles of each phenotype. Adh-1 gel patterns and pollen staining was utilized to examine the possibility of a deletion overlapping this locus. The Adh-1 locus has been mapped proximal to the D8 locus. The results indicate the Adh locus is not included in the putative D8 deletion.

The yield of short-season hybrids is lower than long-season hybrids, mainly as a result of the low final leaf area developed by the short-season plants. This is due to the smaller plant stature and smaller leaf number and size of short-season adapted hybrids. In addition, in short-season areas the thermal-time available may be insufficient to mature grain of current maize hybrids. Maize hybrids which accumulate leaf area quickly, mature earlier, yield well and tolerate higher population densities better than the currently available hybrids would be more suitable for production in short-season areas. The "Leafy reduced-stature" maize hybrids, which have only recently been developed, have traits which address these criteria. There has been no previous effort to evaluate the effects of more than two plant population densities or the effects of planting patterns on the yield, yield components and vegetative growth of these hybrids. in 1995, field experiments were conducted at two sites near Montreal to compare the response of leafy reduced-stature (LRS), non-leafy normal stature (NLNS), and non-leafy reduced-stature (NLRS) maize hybrids to plant population densities and planting patterns. LRS maize hybrids showed the most rapid growth of the first ear, and the highest yield per single plant and per hectare at high plant population densities in paired rows. LRS hybrids also had longer grain filling periods, lower grain moisture contents and higher harvest indices than conventional (NLNS) hybrids. Rapid growth of the first ear and a higher harvest index are indications that LRS hybrids should be more tolerant of higher population densities than currently available hybrids. Therefore, LRS hybrids show promise for production in short-season areas at high plant population densities where maize cultivation is not currently economical due to shortness of the growing-season.

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