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A Genome-Wide Characterization of Differentially Expressed Genes Encoding mRNAs and miRNAs and Methylation Analysis of Phytochrome Genes in a Cotton Phytochrome A1 RNAi lineMiao, Qing 09 December 2016 (has links)
Silencing phytochrome A1 gene (PHYA1) by RNA interference in upland cotton (Gossypium hirsutum L. cv. Coker 312) had generated PHYA1 RNAi lines with increased fiber length, strength and low micronair (finer fiber). In order to identify and characterize mRNAs and miRNAs that are differentially expressed in the RNAi plants, transcriptome and miRNAome analyses via high-throughput RNA sequencing were performed. Total RNA isolated from 10-DPA (days post anthesis) fibers and small RNAs isolated from 5-, 10-, and 15-DPA fibers of RNAi and Coker 312 lines were used to construct 6 RNA libraries and 18 small RNA libraries, respectively, which were sequenced using the Illumina HiSeq system. A total of 142 differentially expressed genes (DEGs) were identified in PHYA1 RNAi compared to Coker 312. GO analysis showed that these DEGs were mainly involved in metabolic pathways, binding and regulating enzymes (hydrolase, transferase, and oxidoreductase activities), and cell structures which were reported to play important roles in fiber development. Twenty-eight KEGG pathways were mapped for 142 DEGs, and the pathways related to glycolysis/gluconeogenesis and pyruvate metabolism were the most abundant, followed by cytochrome P450-involved pathways. Sixty-one conserved miRNA families and thirtyive novel miRNAs were identified in upland cotton. The targets of 6 conserved miRNAs, which expressed differentially in the RNAi line, were reported to participate in primary cell wall synthesis and phytohormone signaling pathways. The 35 novel miRNAs were identified in cotton for the first time, and their target genes were predicted. Nine novel miRNAs were identified to target cytochrome P450 TBP. Together, the results imply that miRNAs involved in fine-tune gene regulation might confer to the phenotype of the RNAi line with improved fiber quality. Besides characterizing mRNAs and miRNAs, the CpG site methylation status within coding regions of phytochrome genes in RNAi line in leaves and 10-DPA fibers was determined using bisulfite genomic sequencing. The PHYA1, PHYC and PHYE in RNAi line had higher methylation levels in leaves than those in Coker 312, but PHYB had lower methylation levels. In fibers, the methylation levels of PHYB also decreased in RNAi plants. However, the methylation of other phytochrome genes showed no significant changes.
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Characterization and Functional Analysis of a Cotton RING-type Ubiquitin Ligase (E3) GeneHo, Meng-Hsuan 11 December 2009 (has links)
A cotton fiber cDNA, GhRING1, and its corresponding gene have been cloned and characterized. The GhRING1 gene encodes a RING-type ubiquitin ligase (E3) containing 337 amino acids (aa). The GhRING1 protein contains a RING finger motif with conserved cysteine and histine residues at the C-terminus and is classified as a C3H2C3-type RING protein. Blast searches show that GhRING1 has the highest homology to At3g19950 from Arabidopsis. Real time RT-PCR analysis indicates that the GhRING1 gene is highly expressed in cotton fiber in a developmental manner. The transcript level of the GhRING1 gene reaches a maximum in elongating fibers at 15 DPA. In vitro auto-ubiquitination assays using wheat germ extract and a reconstitution system demonstrate that GhRING1 has the ubiquitin E3 ligase activity. A fiber specific lipid transfer protein 4 (FSltp4) is identified as the target substrate of GhRING1 by using a bacterial two-hybrid system. The binding of GhRING1 and FSltp4 is confirmed by using an in vitro pull down assay and a yeast two-hybrid system. The histochemical GUS assay was performed to analyze tissue specificity of the GhRING1 and At3g19950 promoters in transgenic Arabidopsis plants. The GUS assay shows that the promoter of At3g19950 is highly activated in leaves, roots, trichomes and also in anthers and stigma of flowers. In contrast, the GUS expression directed by the promoter of GhRING1 is only located at stipules and anthers and stigma of flowers. The GhRING1 is the first ubiquitin E3 gene isolated and studied from cotton. Based on the expression pattern of GhRING1, FSltp4, and GhUBC E2s and the identification of a fiber-specific target protein, FSltp4, we propose that protein ubiquitination occurs in fiber and the ubiquitin-proteasome pathway regulates fiber development.
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Mitigation of herbicide resistance development among weed species in cotton and peanutCalhoun, Justin S 10 December 2021 (has links)
Herbicide resistance development among weed populations in cotton and peanut is becoming increasingly difficult to manage. If resistant populations continue to persist, weed control practices for producers will become less efficient and more costly. The objective of this research was to evaluate alternative weed control techniques designed to mitigate herbicide resistance development for their agronomic and economic impact on weed management systems. Studies were conducted in 2019, 2020, and 2021 at multiple locations in Mississippi and Arkansas investigating multiple techniques including the addition of soil surfactants in herbicide tank mixtures, increasing SOAs utilized in peanut herbicide programs, applying non-labeled herbicides to cotton with post-directed spray placement, and applying complete residual herbicide programs in cotton. Our results suggests that some novel strategies incorporated into existing weed management programs, can provide sufficient control of troublesome weed species and conserve crop yield and profit returns. For example, the use of post-directed application placement allowed for non-labeled herbicides to be applied to cotton without detrimental effects, thus increasing potential options for POST weed control within that crop. Additionally, weed control, seed cotton yield, and net returns were not affected when only residual herbicides were applied in season-long weed control programs as opposed to the standard of mixed, foliar and residual programs. This indicates that high selection pressure associated with foliar chemistries which leads to resistance development, can be alleviated through the adoption of alternative strategies.
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Diapause biology, dispersal capabilities and insecticide use for Lygus lineolaris in Mid-Atlantic cotton systemsSchepis, John Philip 03 June 2024 (has links)
Cotton (Gossypium hirsutum L.), is cultivated in the United States, primarily in regions characterized by long, hot summers to optimize plant growth. Virginia is the northernmost state where cotton is grown, with approximately 84,000 acres annually. The unique challenges of cultivating cotton in Virginia stems from its relatively short season due to its geographical location, lack of large contiguous acreage, and distinctive issues with pests. A significant pest of this region is the tarnished plant bug, Lygus lineolaris (Palisot de Beauvois), which emerged as a major threat to mid-Atlantic cotton during the late 2010s. L. lineolaris utilize a variety of wild and cultivated hosts to survive the winter months. The overwintering success and distribution of diapause survival L. lineolaris was measured on cover crops and weeds common in the Mid-Atlantic. Densities varied between weed and cultivated hosts, with L. lineolaris exhibiting increased survival in legumes compared to grains. Carbohydrate, lipid and protein levels were measured within diapausing and non-diapausing L. lineolaris specimens. Overwintering specimens usually had elevated level of carbohydrates and lipids, while containing decreased concentrations of protein. Nutrient quantification provided an effective tool in selecting for diapause status in L. lineolaris. Through the results from this study, an alternative method to dissection for determining diapause status in L. lineolaris has been identified. In the spring, movement of L. lineolaris throughout the landscape is highly dependent on host senesce. Flight analysis, behavioral assays and nutritional quantification assays on L. lineolaris populations from different weed hosts were performed to assess the flight capacity of specimens fed from different hosts. While weed hosts type provided populations with differing internal nutrient levels, sustained flight was not different between populations. When dispersal of L. lineolaris into cotton occurs, insecticide treatments following scouting are often necessary to prevent economic damage to the plant. Insecticide experiments were conducted aiming to assess the impact of different active ingredients on L. lineolaris, secondary pests, and natural enemy populations. Findings indicated that insecticides used to control L. lineolaris were successful at lowering pest populations and acephate was found to impact natural enemy populations. Plots applied with acephate experienced secondary pest outbreaks, highlighting the crucial role of natural enemies. / Doctor of Philosophy / Cotton is cultivated throughout the southern United States, extending up the east coast and into North Carolina and Virginia. Virginia poses unique challenges for cotton growers due to specific pest issues and need for region-specific research. The tarnished plant bug is a key pest of mid-summer cotton across this region. Overwintering studies were conducted to investigate host preferences on cover crop and weed hosts, as well as diapause survival and termination timing on cover crops. These studies revealed that hairy vetch and deadnettle species may harbor larger populations compared to other weed hosts, and that cover crops such as hairy vetch and crimson clove led to greater overwintering survival compared to grains. During diapause, metabolic changes occur that can lead to increased fat body to aid in overwintering survival. Diapausing and non-diapausing tarnished plant bugs were used in assays to quantify differences in the nutrients commonly found in this fat body. This study successfully found differences in carbohydrates, lipids and proteins levels between the diapausing and non-diapausing populations. This may allow for the identification of reproductive status of tarnished plant bugs based on nutrient levels. Flight capacity, activity levels and nutrient levels in tarnished plant bug populations taken from spring and summer weed hosts were investigated, with the objective to understand the ability of these populations to transition to cotton in the mid-summer. There were some indicators that flight initiation may be influenced by weed host, while all populations had similar flight potentials. Once in cotton, tarnished plant bugs are primarily treated with chemical insecticides which may produce unknown consequences to the system as a whole. Experiments were conducted to assess the impact of insecticides on non-target species within cotton were conducted in the field. Certain insecticides were identified to be harmful to non-target and potentially beneficial insect species, which contributed to outbreaks of other secondary pest species.
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Utilização de sensores de dossel para adubação nitrogenada no algodoeiro / Use canopy sensors for nitrogen fertilization in cottonVilanova Junior, Natanael de Santana 21 June 2016 (has links)
A maioria dos solos tropicais e subtropicais apresenta disponibilidade insuficiente de nitrogênio (N) para atender a demanda da cultura do algodão visando à obtenção de elevados rendimentos. O N nas plantas pode ser avaliado através da utilização de sensores de espectrometria óptica ativa, nos quais se destaca o equipamento comercial utilizado neste trabalho (N-SensorTM ALS), capaz de determinar a dose de N a ser aplicada em cobertura em tempo real. O objetivo geral deste trabalho foi avaliar o desempenho de um sensor óptico ativo, sonares e clorofilômetro para predizer parâmetros de planta e testar estratégias de adubação nitrogenada baseada nas leituras do sensor óptico para aumento da produtividade do algodoeiro em sistemas de cultivo adensado e convencional. Os experimentos foram realizados durante três anos sendo: a) em Chapadão do Céu, GO nas safras 2012/13 e 2013/14; b) e em Campo Verde, MT na safra 2014/15. Os parâmetros de planta avaliados foram: altura, massa seca, massa fresca e nitrogênio acumulado na biomassa. Estes parâmetros foram analisados nas áreas correspondentes à adubação em taxa fixa e correlacionados com o índice de vegetação gerado pelos sensores. Os resultados demonstraram que o sensor ativo de dossel utilizado é altamente eficiente em estimar altura de planta, massa fresca, massa seca e nitrogênio acumulado na cultura do algodoeiro. A utilização de clorofilômetro para monitorar o teor de N foliar no algodoeiro não demonstrou ser uma técnica eficiente. O uso de sensor ultrassônico (sonar) para estimar parâmetros de altura de planta, massa seca, massa fresca e nitrogênio acumulado na planta do algodoeiro é uma estratégia promissora que, embora não tenha a mesma eficiência de sensores de dossel, demonstrou bons resultados na detecção da variabilidade nas lavouras. As diferentes estratégias de adubação nitrogenada no algodoeiro orientadas pelo sensor ativo de dossel não resultaram em efeitos claros na produtividade. A produtividade de cada tratamento mostrou-se mais dependente da localização das repetições dentro da área do que da estratégia de adubação adotada. / Most tropical and subtropical soils has insufficient availability of nitrogen (N) to meet the demand of the cotton crop in order to obtain high yields. The N plants may be evaluated using active optical spectroscopy sensors as the one used in this investigation (N-SensorTM ALS). The equipment is capable of determining in real time the amount of N to be applied spread in the field. The aim of this study was to evaluate the performance of an active optical sensor, sonar and chlorophyll sensor to predict plant parameters and test nitrogen fertilization strategies based on the optical sensor readings in order to increase cotton yield in narrow and conventional farming systems. The experiments were conducted in three years: a) in Chapadão do Céu - GO 2012/13 and 2013/14; b) in Campo Verde, MT - 2014/15. The evaluated parameters were plant height, dry weight, fresh and accumulated nitrogen in the biomass. These parameters were analyzed in the areas corresponding to fertilization in fixed rate and correlated with the vegetation index generated by the sensors. The results showed that the active sensor used is highly efficient in estimating plant height, fresh weight, dry weight and accumulated nitrogen on cotton crop. The use of chlorophyll sensor to monitor the leaf nitrogen content in cotton was not effective. The use of ultrasonic sensor (sonar) to estimate plant height parameters, dry pasta, fresh pasta and accumulated nitrogen on cotton plant is a promising strategy. Although it has not the same efficiency as canopy sensors, it showed good results in the detection of variability in crops. The different nitrogen fertilization strategies in cotton-driven asset canopy sensor resulted in no clear effects on productivity. The productivity of each treatment was more dependent on the location of the repetitions within the area than the adopted fertilization strategy.
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Avaliação da morfologia interna e do vigor de sementes de algodão utilizando técnicas de análise de imagens / Evaluation of the internal morphology and vigor of cotton seeds using techniques of image analysisCorrêa, Plínio Duarte 15 August 2017 (has links)
A avaliação da qualidade de sementes por meio de análise de imagens vem ganhando espaço no mercado sementeiro, devido a sua praticidade, objetividade e rapidez. Assim, o presente trabalho teve como objetivos estudar a utilização do teste de raios X em sementes de algodão do cultivar FM975WS, para avaliação da morfologia interna e a utilização da análise computadorizada de imagens de plântulas (SVIS®) e a sua relação com os testes tradicionais, para avaliação do vigor de sementes. O trabalho foi divido em duas etapas. Na primeira etapa, foram utilizadas imagens radiográficas para avaliar possíveis danos mecânicos, danos por deterioração de tecidos e danos por malformação das sementes e sua relação com a germinação. Na outra etapa, foram utilizados os testes tradicionais para avaliação do vigor de sementes por meio dos testes de primeira contagem de germinação, envelhecimento acelerado, germinação a baixa temperatura, emergência e índice de velocidade de emergência de plântulas em areia, emergência de plântulas em campo e a análise computadorizada de plântulas. Concluiu- se que a análise de imagens, utilizando o teste de raios X, é eficiente na avaliação da morfologia interna de sementes de algodão, com identificação de suas partes internas e na avaliação de danos causados nas sementes, entre eles, danos por deterioração de tecidos, danos por malformação da semente e danos mecânicos, os quais permitem relacionar a ocorrência dos mesmos com a germinação das sementes e que a utilização do SVIS®, como uma técnica de análise computadorizada de imagens de plântulas, é uma alternativa para avaliação do vigor de sementes de algodão. / The evaluation of the physiological quality of seeds by means of image analysis has been gaining space in the seed market due to its practicality, objectivity and speed. The objective of this study was to study the use of the X-ray test in cotton seeds of the cultivar FM975WS, to evaluate the internal morphology and the use of the computerized analysis of seedlings images (SVIS®) and its relation with the traditional tests to evaluate seed vigor. The work was divided into two stages. In the first stage, radiographic images were used to evaluate possible mechanical damages, damages due to deterioration of tissues and damages due to seed malformation and its relation with germination. In the other step, the traditional tests were used to evaluate the vigor of seeds by means of tests of first count of germination, accelerated aging, germination at low temperature, emergence and speed index of emergence of seedlings in sand, emergence of field seedlings and computerized analysis of seedlings. It is concluded that the image analysis, using the X-ray test, is efficient in the evaluation of the internal morphology of cotton seeds, with identification of its internal parts and in the evaluation of damages caused to the seeds, among them damages due to tissues deterioration, damage due to seed malformation and mechanical damages, which allow to relate their occurrence to the germination of the seeds and the use of SVIS®, as a technique of computerized analysis of seedlings images, is an alternative for evaluating the vigor of cotton seeds .
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Sensores ativos de dossel na gestão da adubação nitrogenada e da aplicação de fitorreguladores no algodoeiro / Crop canopy sensor-based management of nitrogen and plant growth regulators in cottonTrevisan, Rodrigo Gonçalves 02 December 2016 (has links)
Devido à existência de variabilidade espacial nos fatores que afetam o desenvolvimento do algodoeiro, faz-se necessário investigar a hipótese de que existem diferenças na dose de máximo retorno econômico da aplicação de nitrogênio (N) em cobertura e de fitorreguladores em áreas de cultivo de algodão. Assim, esse trabalho teve como objetivo investigar o potencial da utilização de sensores ativos de dossel para detecção da variabilidade espacial do desenvolvimento das plantas e da nutrição nitrogenada existente em áreas de produção de algodão, bem como estabelecer estratégias de gestão da variabilidade com o uso de taxas variáveis na adubação nitrogenada de cobertura e na aplicação de fitorreguladores no algodoeiro. Foram conduzidos experimentos em áreas comerciais de cultivo do algodoeiro localizadas em Chapadão do Céu - GO nas safras 2012/2013 e 2013/2014 e em Campo Verde - MT na safra 2014/2015. Avaliou-se o desempenho de dois modelos de sensores ativos de dossel em predizer a altura de plantas e o acúmulo de biomassa e N no algodoeiro, considerando dois sistemas de produção (espaçamento convencional e adensado) e vários estádios de desenvolvimento da cultura. Foram instaladas parcelas para calibração das curvas de dose resposta do algodoeiro ao nitrogênio em cobertura em diferentes condições de desenvolvimento da cultura relacionadas a variações no solo ou no histórico de produtividades no interior do talhão. Com base nos resultados dos experimentos foram desenvolvidos e validados algoritmos para aplicação em taxas variáveis de N e fitorreguladores, adotando-se o conceito de experimentos em nível de fazenda, com o modelo estatístico considerando a condutividade elétrica do solo como uma covariável e uma matriz de variância-covariância espacial, que foi ajustada pelo método da máxima verossimilhança. O desempenho geral dos sensores foi satisfatório, sendo capazes de predizer a altura de plantas e o acúmulo de biomassa e N no algodoeiro com valores de R2 acima de 0,90. Observou-se interação significativa com o espaçamento das fileiras e a fase de desenvolvimento da cultura, com performance inferior em situações de pequena variabilidade na cultura ou de acúmulo de biomassa muito grande. A resposta do algodoeiro ao nitrogênio aplicado em cobertura variou de acordo com as condições de cultivo de cada experimento. Maiores respostas foram observadas em áreas de médio e baixo potencial produtivo, existindo talhões e partes de talhões em que a aplicação de N não proporcionou incremento de produtividade, enquanto que em outras áreas observou-se aumento de produtividade expresso por uma relação quadrática entre a dose aplicada e a produtividade. No talhão com as maiores diferenças, a dose de máximo retorno econômico foi de 0 kg ha-1 na região de alto potencial produtivo, 165 kg ha-1 na região de médio potencial produtivo e 280 kg ha-1 na região de baixo potencial produtivo. Na análise dos experimentos em nível de fazenda, o melhor ajuste foi obtido no modelo que considera a dependência espacial dos resíduos, evidenciando que em experimentos dessa dimensão não se deve assumir que os princípios da estatística clássica são sempre satisfeitos. Em três áreas não foram observadas diferenças significativas entre a aplicação do N em cobertura em taxa fixa ou usando os algoritmos para aplicação em taxa variável, devido à homogeneidade da área e da interferência de outros fatores. Em uma das áreas observou-se vantagem para a aplicação em taxas variáveis usando o algoritmo que recomenda aplicação de maiores doses de N nos locais em que as plantas apresentam menor desenvolvimento vegetativo, com produtividade cerca de 100 kg ha-1 que a aplicação em taxa fixa e cerca de 300 kg ha-1 superior à obtida com a aplicação em taxas variáveis usando o algoritmo que recomenda aplicação de maiores doses de N nos locais em que as plantas apresentam-se mais vigorosas. A aplicação de fitorreguladores em taxas variáveis não apresentou efeito significativo na produtividade de algodão em caroço, entretanto o volume consumido dos produtos foi em média 18% menor na aplicação em taxa variável, principalmente em função das áreas que não foram aplicadas por apresentarem desenvolvimento vegetativo reduzido. / Due to the existence of spatial variability in factors affecting cotton development, it is necessary to investigate the hypothesis that there are differences in the economic optimum fertilizer rates of top dressing nitrogen (N) and plant growth regulators (PGR) applications in cotton fields. Thus, the objective of this study was to investigate the potential of using active crop canopy sensors to detect the spatial variability of plant development and nitrogen nutrition in cotton fields and to establish strategies for managing spatial variability using variable rate technology to apply N and PGR in cotton. We conducted experiments in commercial cotton production fields located in the state of Goiás, in 2012/2013 and 2013/2014 seasons, and in the state of Mato Grosso, in 2014/2015 season. We evaluated the performance of two models of active crop canopy sensors to predict the height of plants and the accumulation of biomass and N in cotton, considering two production systems (conventional and narrow row spacing) and various growth stages. Within-field variations in the response curve to the increasing doses of nitrogen was evaluated under different crop development conditions related to variations in soil and historical yields. Based on the results of the experiments we developed and validated algorithms for variable rate application of N and PGR, using on-farm experiments. The statistical model considered the soil electrical conductivity as a covariate and a spatial variance-covariance matrix, which was adjusted by maximum likelihood. The overall performance of the sensors was satisfactory, predicting the height of plants and the accumulation of biomass and N in cotton with R2 values above 0.90. There was a significant interaction with the row spacing and the development stage of the crop, with lower performance in situations of low variability or very large biomass accumulation. The cotton response to top dressing nitrogen varied according to growing conditions of each experiment. Major responses were observed in areas of medium and low yield potential. There were fields and plots within fields in which the application of N did not increased yields, while in other areas there was a quadratic relationship between the dose applied and yield. In the field with the biggest differences, the economic optimum fertilizer rates of top dressing N was 0, 165 and 280 kg ha-1 in the high, medium and low yield potential zones, respectively. In the analysis of on-farm experiments, the best fit was obtained in the model that considered the spatial dependence of the residues, showing that experiments of this size we should not assume that the principles of classical statistics are always satisfied. In three areas, there was no significant differences between the application of N in fixed rate or using the algorithms for variable rate application, due to the homogeneity of the areas and the interference of external factors. In one of the areas, there was an advantage for application at variable rates using the algorithm that recommends application of higher doses of N in places where the plants have low vegetative development. The yields were about 100 kg ha-1 higher than the application of a fixed rate and 300 kg ha-1 higher than that obtained with the application of variable rates using the algorithm that recommends applying higher doses of N in places where the plants were more vigorous. The variable rate application of PGR had no significant effect on cottonseed yield, however the amount of products used was on average 18% lower in the variable rate application, mainly due to the not applied areas.
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Genetic study of topping-induced cotton/Gossypium hirsutum/ L. plant defense reactions, combining : Bioinformatics, VOC capture and genic expression / Etude génétique des réactions de défense induites par l'écimage chez les cotonniers/Gossypium hirsutum/ L., combinant : bioinformatique, capture de COV et expression géniqueVillamar Torres, Ronald Oswaldo 22 November 2018 (has links)
Gossypium hirsutum, le coton Upland, représente plus de 95% de la fibre de coton produite annuellement dans le monde et est cultivé dans environ 40 pays. La protection des champs de coton contre l’attaque des arthropodes herbivores nécessite des quantités importantes d’insecticides de synthèse, environ 18% de la consommation mondiale en 2000, bien que cela ait beaucoup diminué grâce au coton Bt et aux programmes d’éradication de certains ravageurs. Les composés organiques volatils (COV) naturellement émis par les plantes cultivées peuvent réduire les attaques d'insectes, les COV ayant une influence sur le comportement des arthropodes herbivores et des arthropodes auxiliaires. La recherche scientifique sur les COV des plantes a beaucoup augmenté depuis deux ou trois décennies. La stimulation des émissions de COV dans les champs de coton est désormais recommandée par les entomologistes du Cirad en tant que composante de la stratégie de protection. L’écimage (topping en anglais), c'est-à-dire couper la tête des cotonniers au cours de la floraison, stimule l'émission de COV de défense, une protection écologique qui limite les risques pour la santé dus aux insecticides synthétiques, pour les agriculteurs et les personnes des environs. L'objectif de la thèse était d'améliorer nos connaissances sur les bases génétiques des émissions de COV après l’écimage. La combinaison de plusieurs disciplines telles que la bio-informatique, l'écologie chimique et la génétique moléculaire nous a permis de: 1) analyser les séquences génomiques des gènes des voies de biosynthèse des COVs terpènes et terpénoïdes et des facteurs de transcription (TF) liés à la réponse au stress, à l'aide des bases de données publiées sur les génomes de trois espèces de coton, G. raimondii, G. arboreum (cotons diploïdes) et G. hirsutum (coton tétraploïde), 2) étudier les émissions de COV par les feuilles de coton des plants de G. hirsutum en réponse à l’écimage, en capturant ces molécules en serre et en caractérisant leurs profils cinétiques par chromatographie en phase gazeuse-spectrométrie de masse (GC-MS), incluant des mesures quantitatives, et 3) étudier les modifications de l'expression ARN des plants de coton G. hirsutum après l’écimage, pour 44 gènes impliqués dans la biosynthèse des COV, et également par une comparaison du transcriptome complet au moyen d'une analyse RNA-seq. Les résultats des trois domaines scientifiques, bio-informatique, analyse chimique et expression des gènes, ont pu être liés dans notre thèse de recherche: par ex., deux des gènes initialement identifiés par la bio-informatique, correspondant à deux enzymes, TPS50 (EC: 4.2.3.106 - (E ) bêta-ocimène synthase) et TPS16 (EC: 4.2.3.111 - alpha-terpinéol synthase), ont montré une augmentation de leur expression après l’écimage, et l'analyse GC-MS montre une modification correspondante des profils d'émission de COV. Ces mêmes composés avaient déjà été caractérisés dans d'autres plantes en réponse aux dégâts d’arthropodes. Ce travail de thèse constitue une première exploration des bases génétiques des émissions défensives de COV par les cotonniers cultivés G. hirsutum. La variabilité des comportements d’expression génique observés entre les trois génotypes étudiés de coton Upland africain G. hirsutum, permet de supposer qu’une diversité génétique est présente pour les émissions défensives de COV, ce qui pourrait permettre d’améliorer et d’adapter ces mécanismes de défense naturels et leur réponse à l’écimage, dans la perspective d'une protection naturelle plus efficace des champs de coton. / Gossypium hirsutum, the Upland cotton, represents more than 95% of the cotton fiber annually produced worldwide and is grown in about 40 countries. The protection of cotton fields against the attack of herbivorous arthropods needs important quantities of synthetic insecticides, around 18% of the world consumption for the year 2000, although this decreased very much thanks to Bt cotton and eradication programs for some pests. Volatile organic compounds (VOCs) naturally emitted by crop plants can reduce insect attacks through the influence of VOCs on the behaviors of herbivorous arthropods and auxiliary arthropods. Scientific research about plant VOCs has been increasing much since two or three decades. The stimulation of VOCs emissions by cultivated cotton plants is now recommended by entomologists of CIRAD as a component of the cotton fields protection strategy. “Topping", that is, cutting the head of the cotton plants during the useful floriferous period, stimulates the emission of defense VOCs. It is an environmentally friendly method and it limits health hazards due to the use of synthetic insecticides for the farmers and the surrounding human populations. The objective of the thesis was to improve our knowledge about the genetic bases of VOCs emissions after topping. The combination of several disciplines such as bioinformatics, chemical ecology and molecular genetics allowed us to: 1) analyze the genomic sequences of VOCs genes of the terpene and terpenoid biosynthesis pathways and transcription factors (TF) related to stress response, using the published genome databases of three cotton species, G. raimondii, G. arboreum (both diploid cottons) and G. hirsutum (tetraploid cotton), 2) study the VOCs emissions by cotton leaves of G. hirsutum plants in response to topping, by capturing these molecules in greenhouse and then characterizing their kinetic profiles by means of gas-chromatography mass-spectrometry (GC-MS), with quantitative measurements, and, 3) study the modifications of the RNA expression of G. hirsutum cotton plants after topping, for genes involved in VOCs biosynthesis through quantitative PCR measurements on 44 targeted genes and also by means of a whole-transcriptome comparison through an RNA-seq analysis. The results from the three different fields, bioinformatics, chemical analysis and gene expression, could be interrelated in our research thesis: e.g., two of the genes initially identified by bioinformatics, corresponding to two enzymes, TPS50 (EC: 4.2.3.106 - (E) -beta-ocimene synthase) and TPS16 (EC: 4.2.3.111 - alpha-terpineol synthase), were shown to increase their expression after topping, while the GC-MS analysis showed an modification of the corresponding VOCs in emission profiles. These compounds have been already characterized in other organisms in response to wounds produced by herbivorous insects. This thesis work is a first exploration of the genetic bases of defensive VOCs emission by G. hirsutum cultivated cottons. The variability of genic expression behaviors observed amongst the three genotypes of African Upland cotton G. hirsutum that were studied permits to hypothesize that a genetic diversity is present for defensive VOCs emissions, that could permit to improve and adapt by breeding these natural defense mechanisms and the response to topping, in perspective of a more efficient natural protection of cotton fields.
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Utilização de sensores de dossel para adubação nitrogenada no algodoeiro / Use canopy sensors for nitrogen fertilization in cottonNatanael de Santana Vilanova Junior 21 June 2016 (has links)
A maioria dos solos tropicais e subtropicais apresenta disponibilidade insuficiente de nitrogênio (N) para atender a demanda da cultura do algodão visando à obtenção de elevados rendimentos. O N nas plantas pode ser avaliado através da utilização de sensores de espectrometria óptica ativa, nos quais se destaca o equipamento comercial utilizado neste trabalho (N-SensorTM ALS), capaz de determinar a dose de N a ser aplicada em cobertura em tempo real. O objetivo geral deste trabalho foi avaliar o desempenho de um sensor óptico ativo, sonares e clorofilômetro para predizer parâmetros de planta e testar estratégias de adubação nitrogenada baseada nas leituras do sensor óptico para aumento da produtividade do algodoeiro em sistemas de cultivo adensado e convencional. Os experimentos foram realizados durante três anos sendo: a) em Chapadão do Céu, GO nas safras 2012/13 e 2013/14; b) e em Campo Verde, MT na safra 2014/15. Os parâmetros de planta avaliados foram: altura, massa seca, massa fresca e nitrogênio acumulado na biomassa. Estes parâmetros foram analisados nas áreas correspondentes à adubação em taxa fixa e correlacionados com o índice de vegetação gerado pelos sensores. Os resultados demonstraram que o sensor ativo de dossel utilizado é altamente eficiente em estimar altura de planta, massa fresca, massa seca e nitrogênio acumulado na cultura do algodoeiro. A utilização de clorofilômetro para monitorar o teor de N foliar no algodoeiro não demonstrou ser uma técnica eficiente. O uso de sensor ultrassônico (sonar) para estimar parâmetros de altura de planta, massa seca, massa fresca e nitrogênio acumulado na planta do algodoeiro é uma estratégia promissora que, embora não tenha a mesma eficiência de sensores de dossel, demonstrou bons resultados na detecção da variabilidade nas lavouras. As diferentes estratégias de adubação nitrogenada no algodoeiro orientadas pelo sensor ativo de dossel não resultaram em efeitos claros na produtividade. A produtividade de cada tratamento mostrou-se mais dependente da localização das repetições dentro da área do que da estratégia de adubação adotada. / Most tropical and subtropical soils has insufficient availability of nitrogen (N) to meet the demand of the cotton crop in order to obtain high yields. The N plants may be evaluated using active optical spectroscopy sensors as the one used in this investigation (N-SensorTM ALS). The equipment is capable of determining in real time the amount of N to be applied spread in the field. The aim of this study was to evaluate the performance of an active optical sensor, sonar and chlorophyll sensor to predict plant parameters and test nitrogen fertilization strategies based on the optical sensor readings in order to increase cotton yield in narrow and conventional farming systems. The experiments were conducted in three years: a) in Chapadão do Céu - GO 2012/13 and 2013/14; b) in Campo Verde, MT - 2014/15. The evaluated parameters were plant height, dry weight, fresh and accumulated nitrogen in the biomass. These parameters were analyzed in the areas corresponding to fertilization in fixed rate and correlated with the vegetation index generated by the sensors. The results showed that the active sensor used is highly efficient in estimating plant height, fresh weight, dry weight and accumulated nitrogen on cotton crop. The use of chlorophyll sensor to monitor the leaf nitrogen content in cotton was not effective. The use of ultrasonic sensor (sonar) to estimate plant height parameters, dry pasta, fresh pasta and accumulated nitrogen on cotton plant is a promising strategy. Although it has not the same efficiency as canopy sensors, it showed good results in the detection of variability in crops. The different nitrogen fertilization strategies in cotton-driven asset canopy sensor resulted in no clear effects on productivity. The productivity of each treatment was more dependent on the location of the repetitions within the area than the adopted fertilization strategy.
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Histologia, bioquímica e herança da resistência do genótipo de algodoeiro TX25 a Meloidogyne incognita, raça 3 / Histology, biochemistry and resistance inheritance of cotton genotype TX25 to Meloidogyne incognita, raça 3Alves, Gleina Costa Silva 31 January 2014 (has links)
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Previous issue date: 2014-01-31 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Meloidogyne incognita race 3 is a limiting factor for cotton crop, and the genetic resistance is one of the most desirable control measures for being environmentally and economically suitable. The objective of this research was to study the resistance inheritance of cotton genotype TX 25 to M. incognita race 3 and identify the resistance mechanism. Two experiments for phenotyping were conducted under greenhouse conditions, using two parental genotypes (susceptible FM 966 and resistant TX 25), backcross, generation F1 and generation F2. The plants were maintained for 120 days after inoculation (DAI). Plants were evaluated for gall index, egg mass index and reproduction factor. Histopathological study was conducted and the root systems evaluated at 2, 4, 7, 9, 11, 16, 18, 21 , 23, 29, 34 and 45 DAI. The roots werestained with acid fuscin to assess the nematode penetration. Rootlet cuts were made to observe the changes caused by the presence of the nematode in the root system of susceptible and resistant cotton cultivar. Biochemical tests were also conducted with the two parental genotypes. In this essay, the plants were kept in the greenhouse and inoculated with eggs and J2 of M. incognita. Assessments occurred at eight, 24 and 35 days after inoculation. The genotypes were compared biochemically as to total phenols, flavonoids, liquid chromatography and magnetic resonance imaging. The genotype TX 25 showed resistance to M. incognita race 3 when attacked by the nematode with the plants producing a hypersensitivity reaction. The crosses derived from TX 25 and FM 966 showed olygogenic resistance. TX 25 also shows glycosylated flavonoids and sugars that provide resistance to Meloidogyne incognita race 3. / Meloidogyne incognita, raça 3 é um dos fatores limitantes da cultura do algodoeiro, e a resistência genética é uma medida de controle das mais desejáveis por ser ambientalmente preservadora e não resultar em gastos adicionais ao produtor. O objetivo da presente pesquisa foi estudar a herança da resistência do genótipo de algodão TX 25 a M. incognita, raça 3, e identificar o mecanismo de ação. Foram conduzidos dois experimentos de fenotipagem em casa de vegetação, com dois parentais, um suscetível FM 966 e um resistente TX 25, retrocruzamento, geração F1 e geração F2, onde as plantas foram mantidas por 120 dias após serem inoculadas com o nematóide. Posteriormente avaliou-se índice de galhas, índice de massa de ovos e fator de reprodução. Experimento histopatológico foi conduzido onde as raízes de algodoeiro TX 25 e FM 966 foram avaliadas aos 2, 4, 7, 9, 11, 16, 18, 21, 23 e 29, 34 e 45 dias após a inoculação, com coloração do sistema radicular com fuscina ácida para avaliar a penetração. Foram feitos cortes nas radicelas para observar as alterações causadas pela presença do nematóide no sistema radicular da cultivar suscetível e genótipo resistente. Ensaios bioquímicos com os dois parentais também foram realizados. Nesse ensaio as plantas foram mantidas em casa de vegetação e inoculadas com ovos e J2 de M. incognita. As avaliações ocorreram aos oito, 24 e 35 dias após a inoculação. Nessas avaliações os parentais foram comparados bioquimicante quanto aos fenóis totais, flavonóides, cromatografia líquida e ressonância magnética. O genótipo TX 25 apresenta resistência a M. incognita raça 3, quando incitado por esse nematóide a planta produz reação de hipersensibilidade. A geração F2 oriundas de TX 25 e FiberMax 966 apresentou uma herança de caráter provavelmente oligogênica. O genótipo TX 25 apresenta flavonóides glicolisados e açúcares, que conferem resistência a Meloidogyne incognita, raça 3. O TX 25 é um material indicado como fonte de resistência para os programas de melhoramento genético.
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