Master of Science / Department of Agronomy / Allan K. Fritz / Fusarium head blight (FHB), caused by Fusarium graminearum, is an important cereal disease in humid and semi-humid wheat growing regions. In recent FHB epidemics in the USA, FHB dramatically reduced wheat yields and grain quality due to mycotoxin contamination. Five types of FHB resistance have been reported, but resistance to disease spread within a spike (Type II) and low deoxynivalenol (DON) accumulation in infected kernels (Type III) have drawn the most attention. A Chinese Spring-Sumai3 chromosome 7A substitution line (CS-SM3-7ADSL) was reported to have a high level of Type II resistance, but quantitative trait locus (QTL) on chromosome 7A has never been mapped. To characterize QTL on chromosome 7A, we developed 191 Chinese Spring-Sumai3-7A chromosome recombinant inbred lines (CRIL) from a cross between Chinese Spring and CS-SM3-7ADSL and evaluated the CRIL in a greenhouse for both types of resistance in three experiments. Two major QTL with Sumai 3 (SM3) origin, conditioning Type II and Type III resistance were mapped in chromosomes 3BS and 7AC. QTL on chromosome 3BS corresponds to Fhb1, previously reported from SM3, whereas 7AC QTL, designated as Fhb5, is a novel QTL identified from SM3 in this study. Fhb5 explains 22% phenotypic variation for Type II resistance and 24% for Type III resistance. Marker Xwmc17 is the closest marker to Fhb5 for both types of resistance. Fhb1 and Fhb5 were additive and together explained 56% variation for Type II and 41% for Type III resistance and resulted in 66% reduction in FHB severity and 84% in DON content. Both QTL showed significant pleiotropy effects on Type II and Type III resistance, suggesting both types of resistance may be controlled by the same gene(s). Haplotype analysis of SM3’s parents revealed that Fhb5 originated from Funo, an Italian cultivar. A survey of worldwide germplasm collection of 400 accessions showed that Fhb5 is present mainly in Chinese cultivars, especially in Funo-related accessions. Further, Fhb5 is the second major QTL from SM3 and have potential to be used in improving wheat cultivars for both types of resistance.

Os índices de seleção foram inicialmente propostos para a seleção simultânea de diversos caracteres e, por requerem a obtenção de estimativas de variâncias e covariâncias, são mais apropriados para programas de seleção recorrente. Posteriormente, desenvolveram-se outros índices, que por não requererem o conhecimento de tais estimativas, podem ser aplicados tanto em programas de seleção recorrente, como para a seleção de genótipos já fixados, como clones, linhagens e híbridos. Recentemente, Garcia (1998) desenvolveu um índice específico para a seleção de genótipos fixados, que se baseia no cálculo da distância de cada genótipo a um ideótipo. As principais vantagens são que este índice permite a aplicação de testes de comparações de médias, bem como a identificação de genótipos com performances aquém daquela exigida para os cultivares comerciais para um dado caráter. O objetivo deste trabalho foi avaliar a eficiência do índice de Garcia (1998) para a seleção de linhagens de soja na fase de pré-cultivares. O material experimental consistiu de 88 linhagens (Geração F7), derivadas pelo método S.S.D. e oriundas do cruzamento entre os genitores BR-80-14853 x PI-123439, e três testemunhas comerciais: IAC-12, IAC-Foscarin-31 e IAS-5. Os experimentos foram conduzidos no delineamento em blocos casualizados em seis anos agrícolas, com diferentes números de repetições por ano: 3 (1995/6), 4 (1996/7), 6 (1998/9), 6 (1999/0), 10 (2000/1) e 10 (2001/2). Os caracteres avaliados foram: produção de grãos (PG: g/parcela) altura das plantas na maturação (AM: cm/planta), e acamamento (AC: notas de 1 a 5). No cálculo das distâncias utilizaram-se a Distância Euclidiana e a Distância de Mahalanobis. Os resultados obtidos mostraram que os índices baseados tanto na Distância Euclidiana quanto na de Mahalanobis foram eficientes para a discriminação dos genótipos, embora os melhores resultados tenham sido obtidos com a Distância de Mahalanobis. Devido a isso, sugere-se o uso da Distância de Mahalanobis para identificar e selecionar pré-cultivares de soja e também, como proposto por Garcia (1998), que esta seleção seja acompanhada da avaliação de caracteres individuais, para evitar que genótipos promissores sejam descartados por apresentarem pequenos defeitos em caracteres de importância secundária. / Selection indices were first proposed for the selection of several traits at the same time, and because of the requirement of variances and covariances estimates, they are useful for recurrent selection programs. Later on, other indices were proposed, which do not require the knowledge of these estimates, and thus are useful for both recurrent selection programs and for the selection of fixed genotypes, such as clones, inbred lines and hybrids. Recently, Garcia (1998) proposed a new index for the selection of fixed genotypes which is based on the distance of each genotype to and ideotype. The main advantages of this index is that multiple comparison tests can be done and also the identification of genotypes that perform below than those required by commercial cultivars for a given trait. The objective of this work was to evaluate the efficiency of Garcia (1998)´s index for the selection of pre-cultivars in soybeans. Experimental material comprised 88 lines (F7 generation) derived by the S.S.D. method from the cross between the BR-80-14853 and PI-123439 parents, and three cultivars as checks: IAC-12, IAC-Foscarin-31 and IAS-5. The evaluation trials were carried out in randomized block designs across six growing seasons, with different number of replications each: 3 (1995/6), 4 (1996/7), 6 (1998/9), 6 (1999/0), 10 (2000/1) and 10 (2001/2). The following traits were evaluated: grain yield (PG: g/plot), plant height at maturity (AM: cm/plant) and lodging (AC: scale: 1 to 5). For the calculation of the distance of each genotype to the ideotype, the Euclidean and Mahalanobis methods were used. General results have shown that the index based on both Euclidean and Mahalanobis distances were efficient to discriminate the genotypes, but the best results were reached by using Mahalanobis distance. Consequently, in order to identify and select fixed genotypes of soybeans we suggest the use of Mahalanobis distance, and as proposed by Garcia (1998), that the selection be based on the index and also on the evaluation of each trait, in order to avoid the discard of superior genotypes showing small problems in traits of minor importance.

Les cellules T invariantes associées aux muqueuse (MAIT) sont des lymphocytes innés caractérisés par l'expression d'un récepteur des cellules T semi-invariant (iTCR) et restreints par la molécule du complexe majeur d'histocompatibilité de classe Ib, MR1. Chez l'homme, les cellules MAIT sont abondantes dans le sang (1 à 10%), l'intestin (3 à 5%) et le foie (20 à 40%) et réagissent contre des métabolites microbiens. En raison de leur rareté dans les souris de laboratoire classiques, les études sur les cellules MAIT murines ont été principalement effectuées sur des souris transgéniques (Tg) pour des TCR MAIT. Cependant, ces cellules MAIT Tg ne récapitulent pas de manière adéquate le phénotype des cellules MAIT humaines. Ici, nous décrivons une souche de souris congénique que nous avons générée qui possède des cellules MAIT qui ressemblent aux cellules MAIT humaines. Nous utilisons cet outil pour étudier les caractéristiques des cellules MAIT murines. L'étude de souches de souris consanguines d'origine sauvage montre que la souche CAST/Ei présente une fréquence des cellules MAIT nettement supérieur à celle retrouvée dans la souche C57BL/6. Un seul locus est impliqué et a été localisé dans la région TCRα. Ceci a permis la génération d'une souche "MAIT" congénique, qui ont été en outre croisé à une souris Tg pour un rapporteur GFP du facteur transcriptionnel RORγt sur la base de données antérieurs montrant que les MAITs humaines expriment ce facteur. Grâce à cet outil, nous montrons que les MAITs murines sont CD4−CD8−/lo, ont un phénotype mémoire effecteurs (CD44+) et coexpriment PLZF et RORγt. Ces MAITs murines sont orientées vers une localisation tissulaire (CCR6+CCR7−) et résident préférentiellement dans les tissus non lymphoïdes périphériques, y compris les poumons, le foie et la peau. Après stimulation du TCR, les MAITs produisent des cytokines TH1/2/17 et sont aussi activées par de antigènes bactériens (par exemple semi-purifié fraction bactérienne ou 5-OP-RU) d'une manière dépendant de MR1. Les MAITs ont une forte expression de récepteurs de cytokines (IL-7R, IL-18Rα, IL-12Rβ) et peuvent ainsi répondre à des cytokines innées. Lors d'une infection expérimentale des voies urinaires, les MAITs migrent vers la vessie et ont une activité protectrice anti-bactérienne. Au total, nos résultats démontrent que les cellules MAIT murines ressemblent étroitement à leurs homologues humains. Ce nouveau modèle murin sera un outil puissant pour faire avancer notre compréhension de la biologie des cellules MAIT en situation normale et pathologique. / Mucosal-associated invariant T cells (MAIT) are innate lymphocytes that express a semi-invariant T cell receptor (iTCR) and are restricted by the major histocompatibility complex (MHC) related molecule, MR1. In human, MAIT cells are abundant in the blood (1-10%), gut (3-5%), and liver (20-40%). They react against microbial-derived riboflavin metabolites that are common in bacteria and yeast. Due to the paucity of MAIT cells in classical inbred laboratory mice, studies on mouse MAIT cells were mostly performed in TCR-transgenic (Tg) mice. However, these Tg MAIT cells do not adequately recapitulate the phenotype of human MAIT cells. Herein, we present a recently generated congenic mouse strain harboring MAIT cells that closely resemble human MAIT cells and use this tool to study the characteristics of natural mouse MAIT cell. An analysis of wild-derived inbred mouse strains revealed that CAST/Ei strain has increased frequency of MAIT cells than C57BL/6 mice. This was linked to a locus on the TCRα region. Introduction of such locus into C57BL/6 mice generated a “MAIT” congenic strain, which were further crossed to Rorc(γt)-GfpTG reporter strain based on previous findings of RORγt expression on human MAIT cells. Using this tool, we show that natural mouse MAIT cells are CD4−CD8−/lo, display an effector memory phenotype (CD44+), and coexpress the transcription factors PLZF and RORγt. They exhibit tissue-homing properties (CCR6+CCR7−) and preferentially reside in peripheral non-lymphoid tissues, including lung, liver, and skin. Upon TCR ligation, MAIT cells produce TH1/2/17 type cytokines and react to bacterial-derived antigens (i.e. semi-purified bacterial fraction or 5-OP-RU) in an MR1-dependent manner. They have high expression of cytokine receptors (IL-7R, IL-18Rα, IL-12Rβ) and may respond to the corresponding innate cytokines. During experimental urinary tract infection, MAIT cells migrate to the bladder and display a protective anti-bacterial activity. Altogether, our results demonstrate that mouse MAIT cells resemble their human counterparts more closely than previously recognized and therefore this new mouse model will be a powerful tool for advancing our understanding of MAIT cell biology in health and disease.

Orientador: Madileine Francely Américo / Resumo: A contratilidade do músculo liso e o trânsito gastrintestinal (GI) são importantes parâmetros da motilidade, cujas alterações podem comprometer a absorção de nutrientes e a biodisponibilidade de fármacos. Diversos fármacos apresentam efeitos adversos associados ao trato GI e, mesmo aqueles utilizados para controlar esses efeitos, podem provocar alterações importantes na motilidade ainda pouco conhecidas. Camundongos das linhagens C57BL/6 e BALB/c são amplamente utilizados nas pesquisas biomédicas, entretanto poucos estudos consideram os parâmetros motores GI. Diante de métodos escassos para avaliar a motilidade in vivo e da importância de conhecer a fisiologia básica dos camundongos, bem como o efeito de fármacos sobre o trato GI, objetivou-se: 1) desenvolver protocolo para avaliação da contratilidade duodenal e trânsito gastrintestinal por meio da Biosusceptometria de Corrente Alternada (BAC), e avaliar comparativamente a morfologia e função gastrintestinal de camundongos das linhagens BALB/c e C57BL/6J e, 2) avaliar o efeito GI do tratamento com cisplatina e a influência da dexametasona como antiemético. A partir deste trabalho foi possível implementar um protocolo experimental inédito, por meio da BAC, para avaliar a contratilidade duodenal e o trânsito gastrintestinal em camundongos. Após prévia implantação de marcador magnético no duodeno de camundongos, observou-se uma alta e uma baixa frequência de contração, expressas em ciclos por minuto (cpm). BALB/c e C57BL/6J apre... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The smooth muscle contractility and gastrointestinal (GI) transit are important parameters of motility, whose alterations may compromise nutrient absorption and bioavailability of drugs. Several drugs have side effects associated with the GI tract and, even those used to control these effects may cause significant changes in motility that are still poorly understood. Mice C57BL/6 and BALB/c strains are widely used in biomedical research, but a few studies consider the GI motor parameters. In front of scarce methods to evaluate the in vivo motility and the importance of knowing the basic physiology of the mice, as well as the effect of drugs on the GI tract. Then, the objectives of this study were 1) to develop protocol for the evaluation of duodenal contractility and gastrointestinal transit through Alternating Current Biosusceptometry (ACB), and comparing the morphology and gastrointestinal function of BALB/c and C57BL/6J mice, and 2) to evaluate the GI effect of cisplatin treatment and the influence of dexamethasone as an antiemetic. From this work, it was possible to implement an original experimental protocol, through the BAC, to evaluate the duodenal contractility and the gastrointestinal transit in mice. After the previous implantation of a magnetic marker in the duodenum of mice, a high and low frequency of contraction, expressed in cycles per minute (cpm), was observed. BALB/c and C57BL/6J presented a high frequency of 40 cpm, while the low frequency in BALB/c was 29 ... (Complete abstract click electronic access below) / Doutor

Thesis (M.Sc. Agric. (Agronomy)) --University of Limpopo, 2018 / Two experiments were conducted at Ukulima Root Biology Centre, Waterberg district in Limpopo Province (24°32'58.1"S, 28°06'21.1"E) during 2013-2014 cropping season to select IBM population inbred lines with improved root whorl and stem diameter that can enable them to tolerate nitrogen and phosphorus stress. The experiments were laid out in a Split plot format based on a randomized complete block design with four replicates. The main plot factors were: nitrogen levels (low and high) and phosphorus (low and high), in the respective trials and maize inbred lines (MO345, MO034, MO001, MO199, MO031 and MO196) were in the sub plots. Traits investigated included shoot morphological traits (plant height, leaf area per plant, chlorophyll content, stem diameter, number of leaves), root architectural traits (Whorl angles, root area, average root density, number of adventitious roots, number basal roots, average lateral root length, lateral branching frequency, root top angle, root bottom angle, distance to the first lateral root) and dry biomass. Results showed morphological traits, root architectural traits and biomass were affected by nitrogen fertilizer. Those traits were greater under high nitrogen level. On the other hand nitrogen had no influence on stem diameter size variation and whorl distribution. Plant height, number of leaves and dry biomass were significantly different among the inbred lines. The interaction of inbred and nitrogen fertilizer level had significant effect on leaf width and leaf area per plant. The lowest leaf width was recorded on inbred MO345 under low nitrogen level, while the highest value was recorded on inbred MO345 under high nitrogen level. Inbred MO031 and MO199 had highest values of leaf area per plant under high nitrogen level and inbred MO345 had the lowest value under low nitrogen level. Inbred lines planted under high nitrogen level had relative advantage in leaf growth over inbred lines planted under low nitrogen level. The study showed that nitrogen fertilizers have positive effect on some root architectural traits and growth parameters of maize. Maximum leaf area was obtained by inbred MO031 and MO199 under high nitrogen level. Thus, in order to enhance leaf growth and physiological traits, the use of either MO031 or MO199 is recommended under high nitrogen level while any of these inbred lines MO001, MO034 or MO199 can be used under low nitrogen production as they are highly tolerant to low soil xiv nitrogen. Morphological and root architectural traits correlated positively with dry biomass in both low and high N level. Results from the phosphorus split plot showed that only projected root area was affected by phosphorus level. Chlorophyll content, plant height, 1st whorl angle, 4th whorl angle, root top angle, root bottom angle, average lateral root length and lateral branching frequency differed significantly among the inbred lines. The interaction effect of phosphorus and inbred on root top angle and average lateral root length was significant. Inbred MO199, inbred MO034 and MO031 recorded the shallowest angles under low and high phosphorus level respectively, while inbred MO345 recorded the steeper root top angle at 54.44° under high phosphorus level. Thus to enhance P uptake, inbred MO199 is a potential candidate on low P soils. To improve water and N acquisition efficiency inbred MO345 with high phosphorus level can be used, therefore MO345 with high phosphorus can be recommended for water scarce areas such as Limpopo province. Inbred MO199 had the longest lateral roots of 251.46 mm under lower P level and significantly longer than inbred MO199 and MO001 both at lower phosphorus level. Inbred MO345 (182.88 mm) and MO001 (179.22 mm) were highly tolerant to the low P conditions as the two had shorter lateral roots, a trait vital for uptake of P. Inbred MO199 (251.46 mm) had the longest lateral roots under low P conditions showing higher tolerance to low P conditions. There were positive and significant correlations between dry biomass and morphological traits and root architectural traits on both low and high phosphorus levels. A strongly negative correlation was however observed between biomass and 2nd whorl angle on high phosphorus level. The high significant correlations indicate that selection of high yielding inbreds may be useful based on phosphorus level and biomass. This study showed that several traits have potential under low N and P levels, hence they can be used as selection criteria for inbred lines with improved nutrient use efficiency. Keywords: Maize inbred lines, fertilizer, growth, correlation, root angles

Thesis (Ph. D.)--University of Missouri--Columbia, 2003. / "December 2003." Typescript. Vita. Includes bibliographical references (leaves 146-172). Also issued on the Internet.

Despite the overwhelming evidence for atrophy of the NGF-dependant Basal Forebrain Cholinergic neurons during aging, there is no persuasive evidence towards a decrease in NGF and/or NGF mRNA content in the brain of aged animals. Previous experiments from our laboratory have shown that NGF is released as a precursor protein and cleaved into the mature form in the extracellular space under the influence of a complex protease cascade. These recent findings have lead us to propose that any alterations in levels and/or activity of this maturation/degradation cascade might affect NGF's biological activity and perhaps lead to cognitive impairments in a subset of aged rats. To investigate this possibility, we measured protein and mRNA levels of the protease cascade players (NGF, pro-NGF, tPA, plasminogen, plasmin, MMP-9, neuroserpin). We found significantly decreased levels of both pro-NGF protein and NGF mRNA, but no difference in the remaining elements of the protease cascade, when comparing aged impaired (Al) to the aged unimpaired (AU) animals. Our second objective was to investigate whether animals trained in the Morris Water Maze would preserve their cognitive status in two additional behavioral paradigms, the Novel Object Location (NOL, spatial memory) and Novel Object Recognition (NOR, nonspatial memory) tasks. We found that both AU and AI animals in the MWM were impaired in the NOL when compared to the young controls, with the AI animals performing significantly worse than the AU in this particular task. In the NOR tasks, AI animals performed significantly worse compared to both young and AU animals. In conclusion, further experiments are required to better understand the implication of the complex protease cascade involved in NGF's maturation and degradation as well as its effect on memory of aged animals. In addition, because the segregation of animals (aged impaired/unimpaired) is a crucial step in aging research, we now have additional behavioral paradigms (NOL/NOR) that confirm the cognitive status of these animals.

BACKGROUND Adult bone marrow derived MSCs had been explored to treat myocardial infarction (MI) and heart failure, for which various beneficial paracrine effects had been suggested. Since MSCs in vitro express anti-inflammatory cytokines, we tested the hypothesis that changes in the pro-/anti-inflammatory cytokine ratio in the infarct microenvironment may provide such a paracrine mechanism to improve early cardiac function following acute coronary occlusion. / Methods Rats (n=88) underwent acute left coronary artery ligations and were randomized into groups M and C and then injected with culture media or MSCs, respectively. These rats underwent blinded echocardiography to evaluate left ventricular ejection fractions (LVEF). Real Time PCR was used to compare cytokine gene expression for IL-1beta, IL-6, IL-8 (pro-inflammatory) and IL-10 (anti-inflammatory) at various times. Extra-cellular matrix (ECM) deposition and inflammatory cell infiltration were also analyzed. / Results As early as 12 hours, the ratio of pro-/anti-inflammatory cytokine gene expression in group C was significantly lower than group M. Similar results were found at 24 hours, 1 and 2 weeks, respectively. LVEF improved significantly in group C (M=62% vs C=68% at 12 hours* , M=66% vs C=75% at 24 hours*, M=57% vs C=75% at 1 week *, and M=52% vs C=70% at 2 weeks*, *p<0.01). The ratio of MMP-2/TIMP1 levels was lower in the Group C at all time frames, reaching significance at 12 and 24 hours and 2 weeks. In group C, histopathological analysis revealed significantly less ECM deposition (M=1.95% vs C=0.75% at 24 hours*, M=19.30% vs C=9.36% at 1 week*, M=24.46% vs C=7.57% at 2 weeks*, *p<0.01). This was associated with significantly decreased inflammatory cell infiltration after 24 hours. / Conclusions The current data suggests that MSCs therapy decreases the pro-/anti-inflammatory cytokine ratio in the infarct microenvironment. This is associated with improved cardiac function, reduced ECM deposition, and decreased inflammatory cell infiltration. This paracrine mechanism of MSCs therapy may explain the early functional improvement after MI before cell transdifferentiation or other mechanisms takes place.

Desired base upper half mean length (UHML) of upland cotton (G. hirsutum) in the U.S. has been set a 27.0 mm and is shorter than the standard set by the international community. Upland cotton genotypes from China, South Africa, West Africa, and the U.S. were test crossed to an extra long staple upland (ELSU) and a short staple upland (SSU) and selected genotypes that included both ELSU and MSU phenotypes were crossed in a half-diallel mating scheme to estimate general combing ability (GCA) effects and specific combining ability (SCA) effects. A recombinant inbred line (RIL) population was established to determine the narrow sense heritability (h^2) of AFIS short fiber content by weight (SFCw) and lower half mean length (LHML) and to estimate SFCw using HVI fiber properties. Obsolete cultivars from China are not likely sources for UHML improvement, cultivars from Africa and the U.S. could harbor alleles not being used in current elite short staple cultivars or modern ELSU cultivars. Two ELSU lines used in this study derived through interspecific hybridization with G. barbadense could contain alleles for UHML improvement in modern ELSU cultivars developed without any apparent G. barbadense introgression. A third line D&PL 45-867, might contain alleles for UHML improvement in long staple upland cotton genotypes. Narrow sense heritability estimates indicated a much higher heritability of LHML than AFIS SFCw. Correlation between AFIS SFCw and LHML did not agree with previous studies when using an ELSU X MSU cross. Further study is needed to understand this complex relationship.

Recent initiatives for biofuel production have increased research and development of sweet sorghum. Currently, the initial major limitation to integrating sweet sorghum into existing production systems is the lack of sweet sorghum hybrids adapted to industrial production systems. Hybrid development is now underway, and the application of genetic markers can be used to define the genetic basis of sugar yield and its components, as well as reduce the time required to deliver new sweet sorghum hybrids to market. The purpose of this research was to further characterize the genetic components that influence sweet sorghum productivity, agronomics, and composition. Specifically, a grain x sweet sorghum recombinant inbred line (RIL) population developed for quantitative trait locus (QTL) analysis related to sugar production was evaluated for 24 phenotypic traits including brix, percent moisture, and biomass yield across four environments. The 185 F4 RILs were derived from the parents 'BTx3197' and 'Rio', which are pithy stalk grain and juicy stalk sweet sorghums respectively. Following screening, two genetic maps were constructed with 372 and 381 single nucleotide polymorphisms (SNPs) evaluated using an Illumina GoldenGate assay. Analysis of the data in QTL Cartographer revealed a major and previously reported QTL for soluble solids on chromosome 3, but in contrast to previous studies, this QTL co-localized with other QTLs that have a negative influence on biomass and seed production. Therefore, selection for this QTL may not be advantageous. Because only a few QTLs for percent moisture were found, the results indicated that the pithy stalk phenotype does not have a major effect on percent moisture as measured in this study. Thus, breeding for high or low moisture content will be more challenging than previously expected. The absence of dominance effects indicated that brix must be high in both parents to produce high brix in the hybrid.