Abiotic stresses like salinity, drought, and extreme temperature are constantly on the rise, posing a very high risk to global agricultural productivity and food security. Hence, understanding stress signaling pathways can help engineer plants that can better withstand stress in unfavorable conditions. The salicylic acid (SA) signaling pathway has been widely studied for its important role in mediating abiotic stress in plants. In tobacco plants, Salicylic Acid Binding Protein 2 (SABP2), a methyl esterase enzyme, catalyzes the conversion of methyl salicylate (MeSA) to SA, which triggers the defense response via the SA-mediated signaling pathway. SIP-428 (SABP2 Interacting Protein-428) is an NAD+ dependent SIR2-like (Silent Information Regulator) deacetylase enzyme that likely interacts with SABP2 during SA biosynthesis. In previous studies, SIP-428 has been shown to be a negative regulator of plant growth under abiotic stress (NaCl and mannitol in vivo). Reactive Oxygen Species (ROS) are oxidizing oxygen products that accumulate under stress conditions, and at high levels can be very harmful to plants. Antioxidant enzymes such as catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX), and superoxide dismutase (SOD) are actively involved in lowering the ROS levels in the cell by combating the oxidative stress. The objective of this study was to analyze the regulatory functions of SIP-428 in ROS signaling of tobacco plants through the biochemical quantification of POD and CAT activities. We investigated the SIP-428 RNAi-silenced tobacco plants for the POD and CAT enzyme activities in Osmotic (Mannitol) and Salinity (NaCl) stressed plants. Our results showed that SIP-428 plays a significant role in modulating antioxidant enzymes in stressed plants. This study has improved our understanding of some regulatory roles of SIP428, and its application can be used to enhance stress tolerance via the use of synthetic biology.

Lipid transfer proteins (LTPs) belong to the pathogenesis-related protein family (PR-14) and are thought to participate in plant defense mechanisms. In this study, we characterize the function of an Arabidopsis thaliana mutant ltp12 (AT3G51590), a homologous lipid transfer protein to SIP470 from Nicotiana tabacum for its role in abiotic and biotic stress. SIP470, a lipid transfer protein, was found to interact with SABP2 in a yeast-two hybrid screen. SABP2 in tobacco is required for inducing a robust SAR response. The objective of this research is to understand the role of LTP12 in mediating abiotic stress as salicylic acid plays an important role in both abiotic and biotic stress in plants. For this research, stressor chemicals, NaCl (salinity), mannitol (osmotic stress), and drought (no water or PEG) will be used. Seedlings were initially germinated and grown on artificial plant growth MS media. The similar-sized young seedlings were transferred to MS media plates supplemented with or without stressor chemicals. Oxidative stress analysis of various antioxidant enzymes, such as catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD) will be performed. The Na+ homeostasis for salinity stress will be studied using CoroNaTM dye and confocal microscopy. Our lab has T-DNA insertion knockout mutants of LTP12 that we will be used in the proposed studies. Here, we hypothesize that mutant ltp12 plants will be hypersensitive to abiotic stressors like NaCl, mannitol, and drought, while wildtype Col-0 will be markedly more tolerant. Reports also suggest that knockout lines of other lipid transfer proteins show a defective growth phenotype and lower expression of systemic acquired resistance (SAR). Moreover, to gain a better understanding of both lines' responses to abiotic stress, we need to carry out further studies on the soil as well. The study will also discuss the subcellular localization of ltp12 in Arabidopsis, which will provide an idea of its functional mechanism. Understanding the role of lipid transfer proteins can lead to the development of transgenic plants that are more tolerant to abiotic stresses and climate change.

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Lipid transfer proteins (LTPs) constitute a pervasive class of small proteins implicated in many biological and physiological processes, including seed development, germination, cuticle formation, and abiotic stress responses. In this study, we explored the role of Arabidopsis LTP12 protein in mitigating environmental stresses. To address this, we analyzed the T-DNA knockout mutant ltp12, focusing on its responses to salinity and osmotic stress. Utilizing antioxidant enzyme assays, phenotypic analyses (including water loss, chlorophyll content, seed germination rates, root length, and relative water content), and lipid profile analysis via Thin Layer Chromatography (TLC), we found that ltp12 mutants showed reduced catalase and peroxidase activities and poorer hydration, chlorophyll content, germination, and growth under stress, compared to wild-type Col-0 plants. Mutants delayed vegetative-to-floral phase transition compared to wild-type. Additionally, lipid analysis indicated that the wild-type plants had increased phospholipids under high osmotic stress, suggesting LTP12's involvement in lipid reorganization during stress responses.

As climate change exacerbates heat stress during the summer season, it becomes increasingly critical to develop effective strategies to safeguard the productivity of tomato plants (Solanum Lycopersicon L.). This research delves into the tools and techniques aimed at enhancing the cultivation of summer tomatoes in the coastal region of Virginia. The study explores the optimization of transplant dates, the implementation of reflective mulches, and the application of kaolin clay particle films. Field trials spanning two seasons were carried out, comparing different planting dates in May, June, and July, as well as the use of reflective, black, and white plastic mulches, both with and without foliar kaolin sprays. The findings of this study underscore the impact of transplanting tomatoes in May, demonstrating a substantial increase in yields when compared to transplanting in June and July. Reflective mulches enhanced plant height and fruit production relative to the conventional black plastic mulch. The combination of kaolin clay sprays with standard black mulch, resulting in yield increases of over 35%, rivaling the outcomes achieved with reflective and white mulch treatments. The application of kaolin did not significantly affect leaf-level physiological processes. These results highlight the significant potential of strategic early planting and the adoption of emerging heat mitigation technologies, such as kaolin clay films, in sustaining and enhancing the productivity of summer tomatoes. This becomes particularly relevant as growing conditions continue to evolve due to rising temperatures and the increasing extremity of weather events resulting from climate change. / Master of Science in Life Sciences / With the challenge of hotter summers due to climate change, finding effective ways to grow tomatoes is more crucial than ever. In our two-season study in Virginia's coastal region, we experimented with various methods to improve tomato growth in these warmer conditions. What we discovered was quite promising. Planting tomatoes in early May resulted in significantly better yields than later planting times. Using reflective mulch was beneficial too; it helped the plants grow taller and produce more fruit compared to traditional black mulch. However, the most impressive result came from combining kaolin clay spray with black mulch. This approach led to a matching of the performance of black plastic plus the combination of kaolin clay against reflective and white mulches. It's interesting to note that the kaolin spray didn't alter the basic functioning of the plants at the leaf level. These findings are encouraging. They suggest that early planting and innovative approaches like kaolin clay sprays can effectively boost tomato production, even as we contend with rising temperatures and evolving climate patterns. Embracing these strategies could be key to successful tomato farming in an era of climate change

Systems biology methods can provide novel insight into the responses of an organism to a suboptimal environment, an infection or exposure to a xenobiotic. In the interaction of salmon and salmon lice, there are several areas requiring further research. These include the impacts of lice infection on wild salmon, response mechanisms of different salmon species or life stages to lice infections, effects of environmental conditions on lice stress, and mechanisms underlying the emergence of resistance to important parasiticidal chemicals. Here, I combine global gene expression analyses with phenotypic and physiological responses of salmon or salmon lice to further our understanding of these topics. In the first chapter, I introduce the work by discussing relevant background material on the current knowledge of salmon and salmon lice interactions, salmon immunity, the state of salmon and louse genomics and the emerging field of ecological genomics. I also discuss how these approaches are applied to the study of non-model organisms and sustainable aquaculture development and fisheries conservation. In the second chapter, I present the first large-scale transcriptome profiling of a Pacific salmon to a salmon lice infection, identifying transcript signatures associated with an infection in a sensitive life stage of pink salmon Oncorhynchus gorbuscha. In the third chapter, I present the results of multiple co-habitation infections of three species of Pacific and Atlantic salmon to compare physiological and transcriptomic responses at the local (skin) and systemic levels (anterior kidney). In the fourth chapter, I explore louse transcriptome functioning during temperature and salinity perturbations to characterize the molecular stress response and coping strategies of lice, as well as provide stressor context to response genes. In the fifth chapter, I evaluate sensitive Pacific and resistant and sensitive Atlantic lice responses to emamectin benzoate, an important compound for louse control which has recently been evaded by the louse through resistance development in multiple regions worldwide. In the sixth and final chapter, I conclude with a synthesis of what was learned about knowledge gaps discussed above and how to best apply this information by providing some approaches for future research to address remaining challenges. / Graduate / 0369 / 0792 / 0718 / bensutherland7@gmail.com

Breeding for abiotic stress tolerance is one of main topics in plant breeding. Oilseed rape breeding programs were for a long time focused on morphological and physiological parameters. In this thesis few experiments focused on identification of genes involved in abiotic stress reaction were performed using RT-qPCR (quantitative reverse transcription PCR). Simultaneously SPR (surface plasmon resonance) method were used as modern optical method facilitating very low native protein concentration even in presence of other substances. This method facilitates quantification of concrete proteins by binding them to specific antigen and in oilseed rape research it was not used by now. ERD10 protein was identified by SPR as protein involved in cold stress reaction (or acclimation). The results show ERD10 accumulation in standard conditions affects dynamics of its accumulation change during cold stress. In case we are searching for genotypes great in acclimation ability even during short and warm autumn SPR method should be suitable method for fast, easy and relatively cheap screening of large number of genotypes in breeding collections. Also genes LTI78, RCI2A, NRP1 and two genes for hypothetical proteins were analysed. Their relative expression during cold stress was markedly increased too. Very little is known about these genes and proteins nowadays therefor it will be interesting topic of our oncoming experiment. Relative expression of genes picked according to MALDI-TOF/TOF analysis results was also tested in microspore embryo regenerants stressed by simulated drought. Genes for lactoylglutathione lyase I, phospholipase D 1 and peroxiredoxin antioxidase were tested. In tolerant cultivar was markedly decreased gene expression of peroxiredoxin antioxidase in standard conditions and early stress. These gene will be subject for next research as potential marker for more tolerant genotypes selection.

Nos últimos 100 anos, a temperatura média global aumentou aproximadamente 0,6ºC e deverá continuar a subir a uma taxa rápida podendo atingir 2ºC. No entanto, não se sabe como as plantas responderão a essa mudança climática, pois existem poucas informações sobre as respostas fisiológicas das plantas a incrementos de temperatura. Este trabalho tem por objetivo avaliar os efeitos de duas condições luminosas e duas condições de temperatura (com interação entre eles), em perspectiva futura, sobre o comportamento morfológico, fisiológico e bioquímico de quatro espécies da Floresta Mesófila Semidecídua: Aspidosperma cylindrocarpon e Aspidosperma polyneuron (Apocynaceae); Cariniana estrellensis e Cariniana legalis (Lecythidaceae). As plantas foram submetidas aos seguintes tratamentos: 100% de radiação e temperatura ambiente (100%RTA), 100% de radiação e temperatura elevada em +2ºC (100%RTE), 20% de radiação (por sombreamento) e temperatura ambiente (20%RTA), e 20% de radiação e temperatura elevada em +2ºC (20%RTE). Aspidosperma polyneuron apresentou-se como a espécie mais sensível a radiação intensa, seguida de C. legalis ambas já ameaçadas de extinção. Os parâmetros morfológicos confirmam a teoria de que plantas cronicamente fotoinibidas continuam crescendo, porém, em menores taxas. Os dados bioquímicos reforçam os fisiológicos mostrando fragilidade das espécies secundárias à radiação intensa e os efeitos principalmente negativos do aquecimento aplicado. / In the last 100 years the average global temperature increased by 0.6ºC and it might increase in a fast rate reaching up to 2ºC. However, little is known about how plants might respond to the climate change because there is not enough information about the physiological alterations plants might suffer due to increased temperature. We evaluated morphological, physiological and biochemical alterations under the effects of two light/temperature conditions and its interactions in four different plant species of the Semideciduous Mesophitic Forest: Aspidosperma cylindrocarpon and Aspidosperma polyneuron (Apocynaceae); Cariniana estrellensis and Cariniana legalis (Lecythidaceae). All plants suffered the following treatments: 100% radiation and environment temperature (100%RTA), 100% radiation and environment temperature +2ºC (100%RTE), 20% radiation (shade) and environment temperature (20%RTA), and 20% radiation (shade) and environment temperature +2ºC (20%RTE). Aspidosperma polyneuron was shown to be the most sensible specie to high radiation, followed by C. legalis, both threatened with extinction. Morphological parameters confirmed the theory that chronically photo-inhibited plants keep growing, but at lower rates. Biochemical findings also confirmed our physiological data showing increased fragility of the secondary species to high radiation and heating.

O milho de segunda safra, também conhecido como milho safrinha, é definido como aquele semeado entre os meses de janeiro e março. Esta modalidade de cultivo atingiu no ano agrícola de 2013/2014 uma área plantada de 9,18 milhões de hectares, superior a área cultivada com milho primeira safra, que no mesmo período foi de 6,61 milhões de hectares. Na segunda safra, há alto risco de instabilidades climáticas, principalmente em decorrência de baixas temperaturas, geadas, má distribuição de chuvas e redução do fotoperíodo. Todos estes fatores prejudicam a atividade fotossintética do milho, reduzindo sua produtividade. No entanto, dada a importância deste cultivo, empresas públicas, privadas e universidades vêm buscando incrementar a produtividade e a estabilidade. Para isso, alguns caracteres são especialmente preconizados. Devido ao alto risco de perda por adversidades ambientais, muitos produtores investem pouco em adubação, principalmente adubação nitrogenada. Neste contexto, o desenvolvimento de plantas mais eficientes no uso e, ou, tolerantes ao estresse por nitrogênio, resultaria em maior segurança para o produtor. Não obstante, a precocidade tem elevada importância, já que materiais precoces reduzem o risco de perdas neste período. No entanto, a mesma deve estar sempre associada a alta produtividade. Assim, para a seleção simultânea destes caracteres, pode-se lançar mão de índices per se de resposta das plantas ao estresse, análises gráficas e, ou, índices de seleção simultânea. Adicionalmente, os valores genotípicos das linhagens para essas características, além de serem preditos via REML/BLUP single-trait (análise univariada), também podem ser preditos via REML/BLUP multi-trait (análise multivariada). Dessa forma, os valores genotípicos são corrigidos pela covariância existente entre os caracteres. Assim, o objetivo deste trabalho foi verificar a possibilidade de seleção simultânea para eficiência no uso e tolerância ao estresse por nitrogênio, além de plantas precoces e produtivas. Para isto, linhagens de milho tropical foram cultivadas e avaliadas para estes caracteres. Foram então simulados diversos cenários de seleção simultânea. A partir destes resultados, observou-se que o índice per se de resposta das plantas ao estresse Média Harmônica da Performance Relativa (MHPR) foi o mais eficiente na seleção de plantas eficientes no uso e tolerantes ao estresse por nitrogênio. Isto ocorreu devido a forte correlação desfavorável entre os índices que estimam a eficiência e a tolerância, além da superioridade e em acurácia, herdabilidade e ganhos com a seleção deste índice per se. Já para a seleção simultânea da produtividade e precocidade, o índice Aditivo de seleção simultânea, utilizando os valores genotípicos preditos via REML/BLUP single-trait se mostrou o mais eficiente, já que obteve ganhos satisfatórios em todos os caracteres e há a possibilidade de modular, de forma mais satisfatória, os ganhos em cada caractere. Conclui-se que a seleção simultânea tanto para eficiência no uso e tolerância ao estresse por nitrogênio, quanto para produtividade e precocidade são possíveis. Além disso, a escolha do melhor método de seleção simultânea depende da magnitude e do sentido da correlação entre os caracteres. / Second growing season maize, also known as winter maize, is the maize sowed in Brazil between January and March. This growing modality reached 9.18 million hectares in 2013/2014, higher than the area cultivated in first growing season that was 6.61 million hectares in the same period. In the second season, there is a high risk of climate instabilities, mainly due to low temperatures, frost, poor rainfall distribution and reduction of photoperiod. All these factors harm photosynthetic activity, reducing the maize yield. However, because of the recent plant area increasing, public, private companies and universities have sought increased yield and stability of the second growing season maize. For this, some traits are mainly in the selection process. With the high risk of yield loses due to environmental adversities, many farmers have done little investment in fertilizers, especially nitrogen fertilization. In this context, the development of plants that are nitrogen use efficient and nitrogen stress tolerant could result in a safer activity for the farmers. In addition, the earliness is highly important, since early materials reduce the risk of losses during this period. However, the earliness must always be associated with a high yield. This way, simultaneous selection of these traits can be made by per se responses indexes of stressed plants, graphical analysis and simultaneous selection indexes. Additionally, the genotypic values of the genotypes for the traits can be predicted not only by REML/BLUP single-trait (univariate analysis), but also by REML/BLUP multi-trait (multivariate analysis). In the second, the genotypic values are adjusted considering the covariance between the traits. This way, the objective of this study was to investigate the possibility of simultaneous selection for nitrogen use efficiency and nitrogen stress tolerance, as well as early and high yielding plants. For this, tropical maize lines were grown and evaluate. By these data, it was simulated several simultaneous selection sets. It was observed that Harmonic Mean of the Relative Performance (HMRP) is the most efficient in the selection for nitrogen use efficient and nitrogen stress tolerance. This probably occurs due to the strong unfavorable correlation between the indexes that estimate the efficiency and the tolerance, as well as the superiority in accuracy, heritability and selections gains of HMRP. In case of simultaneous selection for yield and earliness, the additive simultaneous selection index using the genotypic values predicted by REML/BLUP single-trait proved the most efficient selection, because it got satisfactory gains in all the traits and, this index allows the possibility to modulate the gains in each trait. It was concluded that the simultaneous selection for nitrogen use efficiency and nitrogen stress tolerance, as well as for yield and earliness are possible. Furthermore, the choice of the best simultaneous selection method depends on the magnitude and direction of the correlation between the traits.