Magister Scientiae (Biodiversity and Conservation Biology) / Wheat is a key global commodity in terms of acreage and tradeable value and as a staple in household diets. Many factors including biotic stress conditions have detrimental effects on global wheat production and yield. The increasing prevalence of biotic stress inflicted by fungal species such as Fusarium has significantly reduced yield and quality of cereal crops thus, threatening sustainable agriculture and food security. Interactions between wheat and Fusarium spp. such as Fusarium proliferatum triggers the accumulation of reactive oxygen species (ROS) to levels toxic to the plant thus leading to oxidative damage and ultimate cellular death. In order to maintain redox homeostasis, plants rely on ROS-scavenging antioxidants (enzymatic and non-enzymatic) to control ROS molecules to levels less toxic to plants. This study investigated the impact of F. proliferatum on the physio-biochemical responses of two wheat cultivars (SST 015 and SST 088). Changes in seed germination, growth, biomass, chlorophyll and mineral contents were monitored. Furthermore, changes in ROS accumulation and antioxidant enzyme activity was measured in the shoots of both wheat cultivars.

>Magister Scientiae - MSc / Biotic stress is one of the main causes for agricultural loss of economically important cereal crops. The increasing prevalence of biotic stress inflicted by fungal species such as Fusarium has significantly reduced yields and quality of cereals, threatening sustainable agriculture and food security worldwide. Interactions between wheat and Fusarium spp. such as Fusarium oxysporum promotes the accumulation of reactive oxygen species (ROS). Overproduction of ROS can become toxic to plants depending on the scavenging ability of antioxidant systems to maintain redox homeostasis. This study investigated the effects of F. oxysporum on the physiological and biochemical response of three wheat cultivars namely, SST 056, SST 088 and SST 015. Physiological responses were monitored by measuring changes observed in plant growth parameters including shoot and root growth and biomass, relative water content as well as photosynthetic metabolism and osmolyte content in all three wheat cultivars. Downstream biochemical analysis involved monitoring the accumulation of ROS biomarkers (superoxide and hydrogen peroxide) as well as the detection of enzymatic activity of superoxide dismutase (SOD), ascorbate peroxidase (APX) and peroxidase (POD). These biochemical responses were only monitored on the two wheat cultivars which presented contrasting responses to F. oxysporum infection. Results showed that F. oxysporum significantly reduced plant growth, biomass, chlorophyll pigments and relative water content of all three cultivars, with the highest reduction observed for SST 088 relative to SST 015 and SST056. On the other hand, proline content was significantly enhanced in all three wheat cultivars, with the highest increase observed for SST 015 relative to SST 056 and SST 088. Based on the contrasting physiological results observed for these three cultivars, downstream biochemical analysis was focused on SST 015 and SST 088. F. oxysporum trigged an increased in superoxide and hydrogen peroxide contents in both cultivars, with the highest increase observed for SST 088. A similar trend was observed for the extent of lipid peroxidation, manifested as enhanced MDA levels. Furthermore, F. oxysporum differentially altered antioxidant enzyme activity relative to the control of both wheat cultivars. A Significant increase in SOD activity was observed for both cultivars in response to F. oxysporum. However, contrasting responses in APX and POD activity (as seen for the band intensities of individual isoforms) was observed in these wheat cultivars in response to F. oxysporum. Based on the results obtained in this study we suggest that F. oxysporum infection has varying degrees of severity in different wheat cultivars. In light of the significant reduction of plant development coupled with enhanced ROS accumulation and differential antioxidant capacity for SST 015 relative SST 088, we suggest that SST 015 is more resilient to F. oxysporum. We thus conclude that a direct relationship exists between ROS accumulation and antioxidant scavenging in regulating plant tolerance against F. oxysporum pathogens.

No description available.

Stagonospora nodorum blotch (SNB), is a major foliar disease of wheat in the mid-Atlantic U.S., is caused by the necrotrophic fungus Parastagonospora nodorum. SNB is managed using cultural practices, resistant varieties, and foliar fungicides. There are increasing trends of severity and incidence of SNB in Virginia and the surrounding mid-Atlantic region, but it is not known if changes in the pathogen population are contributing to this trend. The overall goal of this research was to 1) determine the occurrence of quinone outside inhibitor (QoI) resistance in Virginia populations of P. nodorum infecting wheat, 2) quantify the distribution of G143A mutations conferring fungicide resistance in Virginia populations of P. nodorum, and 3) characterize genetic diversity of P. nodorum populations in Virginia and assess influences of cultivars and environments on population structure and SNB severity. For Objective 1, QoI resistant isolates of P. nodorum were identified from Virginia wheat fields, and this was the first report of QoI resistant P. nodorum in the United States. The G143A substitution in the cytochrome b gene of P. nodorum was associated with reduced QoI sensitivity, and in Objective 2, a state-wide, two-year survey of P. nodorum populations in Virginia determined that the G143A mutation was widespread in the state and among sampled fields the frequency ranged from 5-32% (mean = 19%). For Objective 3, P. nodorum was isolated from five different wheat cultivars across seven locations over two years in Virginia. SNB severity varied by cultivar but greater differences in disease severity were observed among locations and years suggesting environment plays an important role in SNB development. Among the necrotrophic effector (NE) genes examined, SnTox1 was predominant followed by SnTox3, and frequencies of NE genes did not vary by cultivar or location. P. nodorum populations in Virginia had high genetic diversity, but there was no genetic subdivision among locations or wheat cultivars from which individuals were isolated. Results also indicated that the P. nodorum population in Virginia undergoes a mixed mode of reproduction, but sexual reproduction made the greatest contribution to population structure. Overall, this work provides insights into the population biology of P. nodorum in Virginia and information on variability in fungicide sensitivity and cultivar susceptibility to SNB that has implications for the current and future efficacy of fungicides and host resistance for management of SNB. / Doctor of Philosophy / Wheat (Triticum aestivum L.) is one of the major cereal crops grown worldwide for food, feed, and other products. However, yields of this crop are often limited by fungal diseases including Stagonospora nodorum blotch (SNB) caused by Parastagonospora nodorum. Increasing trends of severity and incidence of SNB may be due to reduced sensitivity of P. nodorum to fungicides or increased virulence of P. nodorum populations on commonly grown cultivars. Fungicides such as quinone outside inhibitors (QoIs) are one of the major classes of fungicides used for disease control and G143A substitution is the most common point mutation associated with complete resistance to QoIs. Therefore, the overall goal of this research was to better understand genotypic and phenotypic variation in Virginia populations of P. nodorum in the context of fungicide sensitivity and susceptibility of wheat cultivars to SNB. The specific objectives were to 1) determine the occurrence of quinone outside inhibitor (QoI) fungicide resistance in Virginia populations of P. nodorum infecting wheat, 2) quantify the distribution of G143A mutations conferring QoI fungicide resistance in Virginia populations of P. nodorum, and 3) characterize genetic diversity of P. nodorum populations in Virginia and assess influences of cultivars and environments on population structure and SNB severity. Results from this research indicate that QoI fungicide resistance occurs in Virginia populations of P. nodorum due to a target site mutation (G143A substitution in the cytochrome b gene), and this mutation is widespread and relatively common in Virginia wheat fields. Based on a multi-year multilocation study, P. nodorum populations were genetically diverse, but there was no genetic subdivision among locations or wheat cultivars. SNB severity varied by location and cultivar, but disease severity was greatest at site-years with moderate springtime temperatures and high rainfall. Overall, this work contributes to a better understanding of P. nodorum populations including the current efficacy of fungicides and host resistance for management of SNB in the region.

Disease resistance is critical in soft red winter wheat (Triticum aestivum L.) cultivars. Leaf rust caused by Puccinia triticina Eriks and stripe rust caused by Puccinia striiformis Westend. f.sp. tritici Eriks. are destructive pathogens of wheat. From 2014 to 2015 phenotypic data was collected at diverse locations for resistance to leaf rust (North Carolina, Texas, and Virginia) and stripe rust (Arkansas, North Carolina, Georgia, Texas, and Virginia) in a Pioneer ‘25R47’ /‘Jamestown’ (P47/JT) population composed of 186 F5:9 recombinant inbred lines (RILs). Analysis of the P47/JT population identified two quantitative trait loci (QTL) for leaf rust resistance on chromosome 5B and two QTL for stripe rust resistance on chromosomes 3B and 6A. Phenotypic variation (%) explained by the putative leaf rust resistance QTL of Jamestown on 5B was as high as 22.1%. Variation explained by the putative stripe rust resistance QTL of Jamestown on 3B and 6A was as high as 11.1 and 14.3%, respectively. Jamestown is postulated to contain gene Lr18. Seedlings of 186 F5:9 recombinant inbred lines from the P47/JT population and 200 F2 seedlings from eight other crosses including Jamestown and/or the Lr18 host differential line RL6009 (Thatcher*6/Africa 43) were screened with P. triticina race TNRJJ. Genetic analysis of the populations was conducted to validate the presence of Lr18 in Jamestown. Results of linkage analysis identified SNP maker IWB41960 linked within 5 cM of gene Lr18 in all three populations. From 2016 to 2017 phenotypic data was collected at diverse locations for resistance to leaf rust (Illinois, North Carolina, and Virginia) in a ‘2013412’ (PI 667644) / VA10W-21 (PI 676295) population (412/21) composed of 157 doubled haploid (DH) lines. The 412/21 DH lines were genotyped via genotyping by sequence (GBS). Analysis of the 412/21 population identified one quantitative trait loci (QTL) region associated with adult plant resistance to leaf rust on chromosome 1B. Phenotypic variation (%) explained by the putative leaf rust resistance QTL of 2013412 on 1B was as high as 40.1%. Kompetitive allele-specific (KASP) markers KASP_S1B_8414614 and KASP_S1B_8566239 were developed as markers for use in marker assisted selection. / Ph. D. / Disease resistance to leaf rust and stripe rust is important when growing soft red winter wheat. Genetic resistance can have a benefit to cost ratio of up to 27:1, considerably better than that of fungicide treatments. From 2013 to 2017 disease data was collected across multiple locations spanning the eastern United States (Arkansas, Georgia, Illinois, North Carolina, Texas, and Virginia). DNA molecular markers were used to identify specific chromosome regions containing genes associated with leaf and stripe rust resistance. DNA markers associated with genes conferring resistance to leaf rust resistance were identified in three chromosome regions, and genes in two regions were associated with stripe rust resistance. These genes and molecular markers associated with them can be used by scientists to further enhance resistance in wheat cultivars. Another study was conducted to determine if Lr18, a gene for leaf rust resistance that has a large effect, is present in the Virginia Tech soft red winter wheat breeding material. This gene (Lr18) is known to have been introduced from an ancestral species highly related to wheat. Wheat seedlings derived from crosses between lines postulated to carry Lr18 with susceptible lines were tested for resistance to a specific strain of leaf rust lacking virulence to Lr18. Genetic analysis of the ratio of resistant versus susceptible seedlings and association between DNA molecular markers and resistant seedlings were conducted to validate the presence of gene Lr18. A molecular marker linked tightly to gene Lr18 was identified in the study. This gene was found to be widely distributed in soft red winter wheat breeding materials and the molecular marker associated with gene Lr18 will be useful for scientists to further improve resistance in wheat cultivars.

No description available.

La plupart de variétés élites de blé tendre ne peuvent pas être croisées avec des espèces apparentées ce qui restreint considérablement la base génétique qui peut être utilisée pour l'introgression de nouveaux allèles dans les programmes de sélection. L'inhibition de l'hybridation entre le blé et les espèces apparentées (e.g. seigle, orge) est génétiquement contrôlée. Un certain nombre de QTLs ont été identifiés à ce jour, y compris les gènes Kr1 sur le chromosome 5BL et SKr, un QTL majeur identifié au laboratoire en 1998 sur le bras court du chromosome 5B, tous deux impliqués dans l'inhibition du croisement entre le blé tendre et le seigle. Dans cette étude, nous avons utilisé une population recombinante SSD provenant d'un croisement entre la variété Courtot non croisable et la lignée MP98 croisable pour caractériser l'effet majeur dominant de SKr. Le gène a ensuite été cartographié génétiquement sur la partie distale du chromosome 5BS à proximité du locus GSP (Grain Softness Protein) dont l'homéologue sur le chromosome 5D est impliqué dans la dureté du grain (locus Ha). Les relations de colinéarité avec l'orge et le riz ont été utilisées pour saturer la région de SKr par de nouveaux marqueurs et établir des relations orthologues avec une région de 54 kb sur le chromosome 12L de riz. Au total, 6 marqueurs moléculaires ont été cartographiés dans un intervalle génétique de 0,3 cM, et 400 kb de contigs physiques de BAC ont été établis des deux cotés du gène afin de jeter les bases du clonage positionnel de SKr. De nouvelles populations de grands effectifs ont été développées pour la localisation précise du gène SKr sur les cartes génétiques et physiques. 223 individus d'une population HIF (SSD254.14) ont été testés pour leur aptitude au croisement avec le seigle et génotypés avec les marqueurs proches du gène pour confirmer les données obtenues dans la population de départ. Les résultats montrent que SKr est localisé dans une région hautement recombinante et que les relations entre distances génétiques et distances physiques sont favorables aux dernières étapes de clonage positionnel du gène. Enfin, deux marqueurs SSR complètement liés au gène SKr ont été utilisés pour évaluer une collection de descendances de blé aptes au croisement avec le seigle originaires d'un programme de sélection de triticale primaire. Les résultats confirment l'effet majeur de SKr sur l'aptitude au croisement et l'utilité des deux marqueurs pour introgresser l'aptitude au croisement interspécifique dans des variétés élites de blé tendre.

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No. of bitstreams: 2 dissertacao_felipe_koch.pdf: 799099 bytes, checksum: 5d61cb7ae229b25b533d3f35001c2acc (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-09-28 / Sem bolsa / Este trabalho teve por objetivo avaliar o crescimento de plantas de trigo submetidas a aplicação de regulador de crescimento vegetal e adubação para altos rendimentos, assim como, a composição química e a qualidade fisiológica de sementes produzidas. A cultivar utilizada foi a OR Topázio e o regulador de crescimento foi o trinexapac-ethyl, aplicado via foliar. No capítulo I, o experimento foi conduzido em casa de vegetação e o delineamento experimental foi o inteiramente casualisado, em esquema fatorial 5x8 (cinco doses e oito épocas de coleta) com quatro repetições. Foram avaliados a matéria seca total, a taxa de produção de matéria seca, as taxas de crescimento relativo e assimilatória liquida, a razão de área e massa foliar, o índice de área foliar. Foi determinada a eficiência de conversão da energia solar, a partição de assimilados, o índice de colheita, a altura das plantas, a emergência e o índice de velocidade de emergência de plântulas. No capítulo II, o experimento foi conduzido em delineamento experimental inteiramente casualisado com cinco tratamentos e quatro repetições, sendo avaliados a germinação e primeira contagem de germinação, índice de velocidade de germinação, matéria seca de plântulas, condutividade elétrica e envelhecimento acelerado. No capítulo I, a alocação de matéria seca total, a taxa de crescimento relativo, a altura de plantas e o índice de colheita foram reduzidos, com intensidade variável, conforme a dose do regulador de crescimento. A taxa assimilatória líquida foi reduzida em plantas submetidas a doses de 400, 600 e 800 mL ha-1, a razão de massa e de área foliar foram superiores quando utilizadas as doses de 400 e 800 mL ha-1. A emergência e índice de velocidade de emergência não foram afetados pela ação do regulador de crescimento. No capítulo II, a primeira contagem de germinação aumentou até a maior dose. A condutividade elétrica após três horas de embebição foi reduzida, a partir da dose de 600 mL ha-1. Os teores de amido aumentaram até a dose de 448 mL ha-1; o de aminoácidos até a dose 243 mL ha-1 e os teores de proteína até a dose de 417 mL ha-1. Houve redução no teor de açúcares solúveis totais ao aumentar a dose do regulador. Portanto, plantas submetidas à aplicação do trinexapac-ethyl apresentam respostas diferenciais no crescimento e partição de assimilados, menor índice de colheita e altura de plantas. Já, as sementes possuem maior vigor e teores de amido, proteína e aminoácidos, contudo, menor teor de açúcares solúveis totais. / This work aimed to evaluate the growth of wheat plants subjected to the application of growth regulator and fertilization for high yields, as well as, chemical composition and physiological quality of seeds produced. The variety used was OR Topázio and the growth regulator was trinexapac-ethyl, via leaf application. On chapter I, the experiment was performed on greenhouse and the experimental design was completely randomized, factorial 5x8 (five doses and eight harvest dates) with four repetitions. Total dry matter, dry matter production rate, relative growth and net assimilation rates, leaf mass ratio, leaf area ratio and leaf area index were evaluated. The conversion efficiency of solar energy, assimilate partitioning, harvest index, plant height, seedling emergence and speed of emergence index were determined. On chapter II, the experiment was performed under completely randomized experimental design using five treatments and four repetitions, being evaluated germination and first count of germination, speed of germination index, seedling dry matter, electrical conductivity and accelerated aging. On chapter I, total dry matter allocation, relative growth rate, plant height and harvest index were reduced, with variable intensity, according to the growth regulator dose. The net assimilation rate was reduced in plants subjected to 400, 600 e 800 mL ha-1; leaf mass ratio and leaf area ratio were superior when doses of 400 and 800 mL ha-1 were used. The seedling emergence and the speed of emergence index were not affected by the action of the growth regulator. On chapter II, germination and first count of germination increased until the higher dose. Electrical conductivity after three hours of imbibition was reduced starting from the dose of 600 mL ha-1. The starch content increased until the dose of 448 mL ha-1; the amino acids up to the dose of 243 mL ha-1 and the protein content to the dose of 417 mL ha-1. There was a decrease on total soluble sugars content with the increase of growth regulator dose. Therefore, plants subject to the application of trinexapac-ethyl feature differential responses on growth and assimilate partitioning, lower harvest index and plant height. Seeds produced by these plants possess higher germination, vigor expression and starch, protein and amino acids contents, however, lower content of total soluble sugars.

No description available.

The incorporation of cereal straw/stubble often immobilises nitrogen (N). This can help conserve N in soil in organic forms, thus reducing loss through leaching over dormant winter periods. However, N-depressions that arise during decomposition can reduce crop yield. The inclusion of a legume green manure can supply fixed-N, thus alleviating the low N availability to crops. In this study, the effect of lupin (Lupinus angustifolius L.) green manure incorporation on barley (Hordeum vulgare L.) straw/stubble decomposition, and N availability was investigated. A field experiment was used to determine the effects of the green manure on decomposition. Decomposition of straw/stubble was monitored using the litterbag technique. Following green manure incorporation, soil cores were incubated in a glasshouse to determine mineral-N availability. Though not significant, the inclusion of lupin green manure seemed to increase the decomposition of straw/stubble during the growth period, then slowing it after its incorporation at 110 d. This was described by a logarithmic pattern of loss of - 4.97 g AFDW residue day⁻¹, with 60% remaining after 140 d. Treatments without lupin had a linear decomposition of - 0.12 g AFDW residue day⁻¹, with 49% remaining after 140 d. The loss of cellulose confirmed the differences in decomposition with the inclusion of lupin resulting in 2.79% less cellulose remaining in straw/stubble after 140 d compared to its exclusion. Lupin significantly increased pot oat N uptake and DM yield by 55 % and 46 %, respectively, compared to its exclusion. However, this effect was not observed in field sown wheat yields and the soil mineral-N measurements made. This study showed that the potential of lupin to increase straw/stubble decomposition by improving the retention and availability of N, leading to long-term yield benefits, needed further investigation.