Maize is susceptible to biotic and abiotic stresses. The most important abiotic stresses in Africa are drought and low soil fertility. Aflatoxin contamination is a potential problem in areas facing drought and low soil fertility. Three studies were conducted to evaluate maize germplasm for tolerance to stress. In the first study, fifteen maize inbred lines crossed in a diallel were evaluated under drought, low N stress, and well-watered conditions at six locations in three countries to estimate general (GCA) and specific combining ability (SCA), investigate genotype x environment interaction, and estimate genetic diversity and its relationship with grain yield and heterosis. GCA effects were not significant for grain yield across environments. Lines with good GCA effect for grain yield were P501 and CML258 across stresses. Lines CML339, CML341, and SPLC7-F had good GCA effects for anthesis silking interval across stresses. Additive genetic effects were more important for grain yield under drought and well-watered conditions. Heterosis estimates were highest in stress environments. Clustering based on genetic distance calculated using marker data from AFLP, RFLP, and SSRs grouped lines according to origin. Genetic distance was positively correlated with grain yield and specific combining ability. In the second study, synthetic hybrids were evaluated at seven locations in three countries to estimate GCA and SCA effects under low N stress and optimal conditions and investigate genotype x environment interaction. GCA effects were significant for all traits across low N stress and optimal conditions. The highest yielding synthetic hybrids involved synthetics developed from stress tolerant lines. Synthetics 99SADVIA-# and SYNA00F2 had good GCA for grain yield across low N stress conditions. Heterosis was highly correlated with grain yield. Optimal environments explained more variation than stress environments. The third study evaluated the agronomic performance and aflatoxin accumulation of single and three-way cross white maize hybrids at five locations in Texas. Inbreds CML343, Tx601W, and Tx110 showed positive GCA effects for grain yield. Significant GCA effects for reduced aflatoxin concentration were observed in lines CML269, CML270, and CML78 across locations. Differences in performance between single and three-way crosses hybrids were dependent mostly on the inbred lines.

Malaria, an infectious disease caused by Plasmodium parasites, is one of the major world-scale health problems. Despite the efforts aimed at finding an effective way to control the disease, the success has been thwarted by the emergence of parasite drug resistance and mosquito resistance to insecticides. This thesis focuses on the genetic analysis of resistance to murine malaria induced by the lethal Plasmodium berghei ANKA using a wild-derived-inbred strain (WDIS). The aim of this thesis was to exploit the genetic diversity represented among WDIS for identifying loci contributing to resistance/susceptibility to murine malaria. The work included a genome-wide polymorphism survey using microsatellite markers performed on 10 WDIS. Comparisons of these strains to laboratory inbred strains confirmed a higher rate of polymorphism among the WDIS. We conclude that these WDIS represent repositories of unique naturally occurring genetic variability that may prove to be invaluable for the study of complex phenotypes. Next, we used the WDIS to search for novel phenotypes related to malaria pathogenesis. Whereas most laboratory strains were susceptible to experimental cerebral malaria (ECM) after infection with P. berghei ANKA, several WDIS were found to be resistant. To study the genetic inheritance of resistant/susceptibility to P. berghei ANKA infection we analysed backcross and F2 cohorts derived from crossing the WLA wild-derived strain with a laboratory mouse strain (C57BL/6). A novel phenotype represented by the cure of infection, clearance of parasitaemia and establishment of immunological memory was observed in the F2 progeny. The backcross progeny was used to genetically map one locus on chromosome 1 (Berr1) and one locus on chromosome 11 (Berr2) that mediate control of resistance to ECM induced by P. berghei ANKA. Genetic mapping using the F2 progeny showed that a locus on chromosome 1 (Berr1) and a locus on chromosome 9 (Berr3) were contributing to control survival time after infection with lethal Plasmodium. Finally, we identified, a locus on chromosome 4 (Berr4) that appears to control time of death due to hyperparasitaemia. This thesis underlines the value of using WDIS to reveal genetic factors involved in the aetiology of disease phenotypes. The characterisation of the genetic factors represented by the malaria resistance loci identified here are expected to provide a better understanding of the malaria pathology.

Stem rust and Yellow rust diseases are the two major wheat fungal diseases causing considerable yield losses in Turkey and all around the world. There are studies which are carried out to identify and utilize resistance sources in order to obtain resistant lines of wheat. However, virulent pathotypes are continuously being important threats to wheat production and yield. For that reason, new approaches for rapid identification are needed. The aim of this study was to investigate and to understand the structural and functional differences between the resistant and sensitive durum wheat cultivars to the plant fungal diseases of stem and yellow (stripe) rusts. To aim this, forty durum wheat recombinant inbred lines (RILs), which were previously determined to be resistant or sensitive to stem and yellow rust diseases, were investigated by the noninvasive Attenuated Total Reflectance Fourier Transform Infrared (ATR-FTIR) Spectroscopy. Also, classification of the resistant and sensitive lines depending on the structural and functional differences has been attempted. The FTIR spectra for stem rust disease showed that, resistant durum wheat lines had a significant increase in the population of unsaturation in acyl chains of lipid molecules, an increase in lipid and in total protein content and also an increase in carboxylic acids and alcohols. For yellow rust disease, resistant lines had a significant increase in hydrogen bonding and they had also a more ordered membrane structure. In Principal Component Analysis for stem rust disease, according to 3700-650 cm-1 region, amide III band (1213-1273 cm-1 region) and C-H stretching region (3020- 2800 cm-1), the resistant and sensitive groups were separated successfully. For yellow rust disease, according to 3700-650 cm-1 region, Amide A and Amide III bands, the resistant and sensitive lines were grouped distinctly. FTIR spectroscopy provides a useful approach to determine the differences in molecular structure of durum wheat RILs regarding resistance of lines to fungal diseases. However, further research is still needed to ensure if the structural and functional differences in biomolecules of the samples could be used as molecular markers for discrimination of rust resistant materials from rust sensitive ones.

Maize is susceptible to biotic and abiotic stresses. The most important abiotic stresses in Africa are drought and low soil fertility. Aflatoxin contamination is a potential problem in areas facing drought and low soil fertility. Three studies were conducted to evaluate maize germplasm for tolerance to stress. In the first study, fifteen maize inbred lines crossed in a diallel were evaluated under drought, low N stress, and well-watered conditions at six locations in three countries to estimate general (GCA) and specific combining ability (SCA), investigate genotype x environment interaction, and estimate genetic diversity and its relationship with grain yield and heterosis. GCA effects were not significant for grain yield across environments. Lines with good GCA effect for grain yield were P501 and CML258 across stresses. Lines CML339, CML341, and SPLC7-F had good GCA effects for anthesis silking interval across stresses. Additive genetic effects were more important for grain yield under drought and well-watered conditions. Heterosis estimates were highest in stress environments. Clustering based on genetic distance calculated using marker data from AFLP, RFLP, and SSRs grouped lines according to origin. Genetic distance was positively correlated with grain yield and specific combining ability. In the second study, synthetic hybrids were evaluated at seven locations in three countries to estimate GCA and SCA effects under low N stress and optimal conditions and investigate genotype x environment interaction. GCA effects were significant for all traits across low N stress and optimal conditions. The highest yielding synthetic hybrids involved synthetics developed from stress tolerant lines. Synthetics 99SADVIA-# and SYNA00F2 had good GCA for grain yield across low N stress conditions. Heterosis was highly correlated with grain yield. Optimal environments explained more variation than stress environments. The third study evaluated the agronomic performance and aflatoxin accumulation of single and three-way cross white maize hybrids at five locations in Texas. Inbreds CML343, Tx601W, and Tx110 showed positive GCA effects for grain yield. Significant GCA effects for reduced aflatoxin concentration were observed in lines CML269, CML270, and CML78 across locations. Differences in performance between single and three-way crosses hybrids were dependent mostly on the inbred lines.

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 PDF of title page (University of Missouri--Columbia, viewed on January 5, 2010). Vita. Thesis advisor: Habib Zaghouani. Includes bibliographical references.

Retinol (vitamin A) and its active metabolite, all-trans retinoic acid, signal through nuclear retinoic acid and retinoid X receptor (RAR/RXR) heterodimers. These complexes regulate the expression of genes involved in developmental processes such as limb development. In excess, retinoids are potent teratogens and cause marked reductive effects on the developing limb. The goal of this thesis was to elucidate the molecular mechanisms underlying retinoid-induced limb dysmorphogenesis. Specifically, using an in vitro limb culture system, I examined the involvement of one RAR isoform, RARgamma, in mediating retinoid insult. / My first objective was to examine how limbs deficient in RARgamma responded to exogenous retinoid exposure. I showed that RARgamma-null limbs (on an RARalpha1-null background) exhibited less severe limb defects following retinoid insult when compared to their wild-type counterparts. Additionally, the absence of RARgamma abolished the retinoid-induced misregulation of genes important for chondrogenesis (Sox9 and Col2a1 ) and limb outgrowth (Meis-1 and -2). / The next objective set out to determine how pharmacological activation of RARgamma affected limb development. The RARgamma-selective agonist (BMS-189961) caused limb dysmorphology (namely, effects on cartilage) that was comparable to pan-RAR activation with all-trans retinoic acid. A chondrogenesis-focused gene array analysis identified Mgp and Gdf10 as two RARgamma-responsive genes that may mediate retinoid-induced limb insult. / Subsequently, I assessed the functional involvement of Mgp in mediating retinoid teratogenicity. Limbs were treated with all- trans retinoic acid and warfarin (an inhibitor of MGP); warfarin co-treatment rescued limbs from retinoid-induced insult. / My final objective was to determine the importance of Gdf10 in mediating limb development. Recombinant human Gdf10-soaked beads were implanted into distal limb structures; ectopic overexpression of Gdf10 in the web (but not the digital ray) resulted in marked proximal limb malformations. / Collectively, these studies have illustrated the importance of RARgamma in retinoid teratology and have identified several potential mechanisms by which retinoids cause limb defects.

Epigenetic modification of DNA plays an important role in gene regulation. During gametogenesis and early embryogenesis epigenetic states are reset to ensure embryonic-specific gene expression patterns after fertilization. However, certain genomic regions may resist epigenetic reprogramming. This may result in transgenerational epigenetic inheritance. Earlier, a grandparental origin dependent (GPO) transmission ratio distortion (TRD) of alleles in the distal region of mouse chromosome 12 had been found (Croteau et al ., 2002). The distorted region overlaps with the imprinted region of chromosome 12. The mechanism underlying this TRD is unknown, and we hypothesized that it was due to failure to reset imprints in the imprinted region in a proportion of germ cells. Such an imprint resetting failure would represent a particular case of transgenerational epigenetic inheritance. DNA (Cytosine-5) methyltransferase 1 (DNMT1) plays a key role in the maintenance of epigenetic states in mammalian genomes. To test the role of DNA methylation and DNMT1 in the genesis of TRD and its relationship to epigenetic inheritance we investigated the effect of Dnmt1 loss-of-function mutations using two mouse models: GPO (grandparental origin dependent)-TRD (transmission ratio distortion) and epigenetic inheritance at the agouti locus. Here, we report that Dnmt1 mutations have a modifying parental effect on the transmission of grandparental chromosome 12 alleles. However, the same Dnmt1 mutation did not affect the agouti coat color inheritance patterns in mice that inherited the Avy (agouti viable yellow) mutant allele from the father. Our results suggest that Dnmt1 is a trans-acting modifier of allelic transmission and support the role of epigenetic states in the genesis of TRD.

No description available.

Reactive airways dysfunction syndrome (RADS), a form of irritant-induced asthma (IIA) has been observed in humans following acute chlorine (Cl 2) gas exposure in occupational and domestic settings. Following Cl 2 injury, subepithelial fibrosis, mucous hyperplasia, and non-specific airway hyperresponsiveness have been reported. Based on the disease profile, we hypothesized that pulmonary damage may be oxidative in nature. / The aim of this work was to develop a murine model of irritant-induced asthma in order to investigate the pathogenic processes and potential oxidative mechanisms involved in response to Cl2 exposure, with a secondary aim of examining the role of iNOS in response to Cl2 inhalation. / A/J, C57BI/6J (wild type) and iNOS-1- mice exposed to various concentrations of Cl2 were mechanically ventilated for measurement of lung mechanics and responses to i.v. methacholine (MCh). Bronchoalveolar lavage was performed to examine total protein, cell populations and nitrate/nitrates. Tissues were harvested for histology and immunocytochemistry for iNOS, 3NT and carbonyl residues. To examine the role of iNOS, a subset of animals were treated with a selective iNOS inhibitor (1400W) and non-selective NOS inhibitor LNAME. / Chlorine exposure caused airway hyperresponsiveness, which appeared to be mitigated by iNOS blockade with 1400W, however this was not the case in iNOS-1- mice. Cl2 exposure also caused increases in total BAL protein, total cells, NOx, neutrophils, iNOS, 3NT and carbonyl residues. / In conclusion, chlorine exposure causes lung injury, similar to reactive airways dysfunction syndrome, characterized by airway hyperresponsiveness, epithelial sloughing, inflammatory cell influx, oxidative injury and increases in both the activity and expression of iNOS. Chlorine-induced airway hyperresponsiveness is mitigated, in part, by selective blockade of iNOS with the use of pharmacological intervention.

Previously seed coat color in flax has been used as a phenotypic marker for specialty quality traits and currently there is an increasing demand to use seed coat color in flax to market flax for human and animal nutrition uses. Seed coat color was studied to 1) understand the inheritance of seed coat color conditioned by the b1 locus, to 2) understand the relationship of other important flax traits with seed coat color as well as to 3) identify markers that are linked to seed coat color for future marker assisted selection of seed coat color. Spearman’s rank correlation and an allelism test was used to show the inheritance of the alleles at the b1 locus. Bulked segregant analysis (BSA) was used to identify putatively linked markers with the b1 locus, these were then screened on the CDC Bethune x M96006 recombinant inbred line population. Furthermore, the CDC Bethune x M96006 and CDC Bethune x USDA-ARS Crystal recombinant inbred line populations were used to identify any important flax traits that had a significant relationship with seed coat color. It was shown that seed coat color conditioned by the b1 locus was stably inherited and that b1vg and b1 are allelic to one another. The results of the BSA showed that there were 17 candidates for linkage but when these markers were screened on the population only the Lu456 from linkage group (LG) six was identified to have linkage (χ²=3.90; P<0.05) with the b1 locus. Additionally, it was shown that the b1 seed coat color allele of the b1 locus had a pleiotropic effect on flower color and flower shape and that seed coat color was associated with linolenic fatty acid content. None of the traits examined were found to be associated with the b1vg allele of this locus. These results show that the b1 locus is likely present on linkage group six, more marker coverage on linkage group six of markers that are polymorphic between the two seed coat color parents would increase the accuracy of detection. Lastly, this study showed that plant breeders should consider using the b1vg allele that conditions the variegated seed coat color to mark unique lines with important combinations of traits because it sorted independently for seed quality traits. Whereas, the yellow seed coat color conditioned by the b1 allele was found to be associated with higher linolenic fatty acid content and the semi-lethality of this allele would make it not suitable for use in parental lines.