Efeito da restrição hídrica sobre o crescimento de clones de eucalipto

O estabelecimento e o crescimento inicial de espécies florestais no campo são fortemente afetados pela disponibilidade de água no solo e pela época de plantio, por isso, o presente trabalho estuda o impacto do déficit hídrico no crescimento de mudas de dois clones do híbrido Eucalyptus grandis x Eucalyptus urophylla, ambos submetidos a 4 níveis de déficit hídrico, em duas épocas de plantio. O estudo foi realizado na área experimental do Núcleo de Estudos e Difusão de Tecnologia em Florestas, Recursos Hídricos e Agricultura Sustentável (NEDTEC), do Centro de Ciências Agrárias da Universidade Federal do Espírito Santo (CCA-UFES), localizado no município de Jerônimo Monteiro. O trabalho foi realizado em duas épocas distintas, sendo a primeira no período de 09 de fevereiro a 09 de junho de 2009 e a segunda no período de 11 de julho a 07 de novembro de 2009, visando à realização das observações em diferentes condições de regime de radiação, déficit de pressão do vapor do ar, temperatura, umidade relativa do ar e velocidade do vento. O delineamento experimental utilizado foi o inteiramente ao acaso em parcelas subdivididas 2 x 4, alocando-se os 4 níveis de déficits hídricos na parcela principal e as 2 épocas nas subparcelas, com três repetições. Os manejos hídricos aplicados foram: Déficit 0 (D0) sem déficit, Déficit 1(D1) corte da irrigação aos 30 dias de experimentação, permanecendo até o final do experimento, Déficit 2 (D2) corte da irrigação aos 30 dias de experimentação, suspensão da irrigação por 60 dias e posterior retomada da irrigação por mais 30 dias; Déficit 3 (D3) corte da irrigação aos 60 dias de experimentação, prolongando até o final do experimento. Os dados experimentais foram submetidos à análise de variância, e quando significativas, as médias foram comparadas pelo teste de média Tukey a 5% de probabilidade, para cada clone estudado. Com este trabalho, foi possível avaliar o impacto de diferentes déficits hídricos, no crescimento inicial das plantas, em duas épocas do ano e avaliar o incremento no desenvolvimento das plantas durante a aplicação dos tratamentos, com retiradas de amostras médias de cada tratamento a cada 30 dias. As variáveis medidas nos dois experimentos foram altura total da planta, diâmetro ao nível do coleto, número de folhas, área foliar, matéria seca de folhas, matéria seca de haste e ramos, matéria seca de raízes e matéria seca total. Foram avaliadas as variáveis climáticas durante todo o período experimental, nas duas épocas, a fim de determinar a condição do clima em cada época. Para os dois clones estudados, em geral, os déficits hídricos promoveram a redução das variáveis morfológicas estudadas e a época experimental foi o fator que mais influenciou a redução do crescimento das plantas. Sendo que a Época 1 foi a que proporcionou resultados superiores, e a Época 2 foi a que prejudicou mais o desenvolvimento das plantas, reduzindo significativamente todas as variáveis morfológicas em todos os déficits hídricos, inclusive o D0.

Efeito da restrição hídrica sobre o crescimento de clones de eucalipto

O estabelecimento e o crescimento inicial de espécies florestais no campo são fortemente afetados pela disponibilidade de água no solo e pela época de plantio, por isso, o presente trabalho estuda o impacto do déficit hídrico no crescimento de mudas de dois clones do híbrido Eucalyptus grandis x Eucalyptus urophylla, ambos submetidos a 4 níveis de déficit hídrico, em duas épocas de plantio. O estudo foi realizado na área experimental do Núcleo de Estudos e Difusão de Tecnologia em Florestas, Recursos Hídricos e Agricultura Sustentável (NEDTEC), do Centro de Ciências Agrárias da Universidade Federal do Espírito Santo (CCA-UFES), localizado no município de Jerônimo Monteiro. O trabalho foi realizado em duas épocas distintas, sendo a primeira no período de 09 de fevereiro a 09 de junho de 2009 e a segunda no período de 11 de julho a 07 de novembro de 2009, visando à realização das observações em diferentes condições de regime de radiação, déficit de pressão do vapor do ar, temperatura, umidade relativa do ar e velocidade do vento. O delineamento experimental utilizado foi o inteiramente ao acaso em parcelas subdivididas 2 x 4, alocando-se os 4 níveis de déficits hídricos na parcela principal e as 2 épocas nas subparcelas, com três repetições. Os manejos hídricos aplicados foram: Déficit 0 (D0) sem déficit, Déficit 1(D1) corte da irrigação aos 30 dias de experimentação, permanecendo até o final do experimento, Déficit 2 (D2) corte da irrigação aos 30 dias de experimentação, suspensão da irrigação por 60 dias e posterior retomada da irrigação por mais 30 dias; Déficit 3 (D3) corte da irrigação aos 60 dias de experimentação, prolongando até o final do experimento. Os dados experimentais foram submetidos à análise de variância, e quando significativas, as médias foram comparadas pelo teste de média Tukey a 5% de probabilidade, para cada clone estudado. Com este trabalho, foi possível avaliar o impacto de diferentes déficits hídricos, no crescimento inicial das plantas, em duas épocas do ano e avaliar o incremento no desenvolvimento das plantas durante a aplicação dos tratamentos, com retiradas de amostras médias de cada tratamento a cada 30 dias. As variáveis medidas nos dois experimentos foram altura total da planta, diâmetro ao nível do coleto, número de folhas, área foliar, matéria seca de folhas, matéria seca de haste e ramos, matéria seca de raízes e matéria seca total. Foram avaliadas as variáveis climáticas durante todo o período experimental, nas duas épocas, a fim de determinar a condição do clima em cada época. Para os dois clones estudados, em geral, os déficits hídricos promoveram a redução das variáveis morfológicas estudadas e a época experimental foi o fator que mais influenciou a redução do crescimento das plantas. Sendo que a Época 1 foi a que proporcionou resultados superiores, e a Época 2 foi a que prejudicou mais o desenvolvimento das plantas, reduzindo significativamente todas as variáveis morfológicas em todos os déficits hídricos, inclusive o D0.

Efeitos dos ácidos húmicos provenientes de aterro sanitário em plantas

O processo de tratamento biológico dos lixiviados de aterros sanitários resulta na geração de grandes quantidades de lodo, caracterizados por conterem altas taxas de matéria orgânica. Por meio do fracionamento químico da matéria orgânica são obtidos os ácidos húmicos (AH), fração de comprovada eficiência sobre o crescimento vegetal, promovendo melhorias no desenvolvimento das plantas. Este trabalho teve como objetivo caracterizar quimicamente os AH extraídos do lodo de lixiviado de aterro sanitário e avaliar os efeitos da aplicação de diferentes doses dos AH por meio de análises biológicas em plantas, visando minimizar os potenciais riscos da utilização do lodo in natura. Por meio de caracterizações químicas, o ácido húmico apresentou elevados teores de carbono e nitrogênio, podendo constituir uma importante fonte de nutrientes para as plantas. Além disso, foram observadas alterações nas taxas de absorção, na bioconcentração e na translocação de alguns nutrientes. Com relação à análise das enzimas antioxidantes, foi possível observar aumento na atividade de algumas enzimas com a aplicação de diferentes doses de AH. Além disto, foram constatadas alterações citogenéticas por meio da análise de células meristemáticas e F1 de Allium cepa. Influências sobre o crescimento da planta também são reportadas, por meio de aumentos expressivos na área radicular e na altura de Zea mays. Em geral, os dados de crescimento revelaram um maior investimento da planta na parte aérea, provavelmente associado com a melhor eficiência do sistema radicular. Além disso, também foram reportadas alterações na espessura da epiderme. Neste contexto, apesar dos benefícios nutricionais e da comprovada atuação dos AH sobre o metabolismo vegetal, os seus efeitos biológicos sobre enzimas do estresse oxidativo e a sua capacidade citotóxica precisam ser melhor investigados. Devido à complexidade do resíduo, a utilização de análises químicas, genéticas, enzimáticas, fisiológicas e anatômicas foi uma importante ferramenta para a avaliação da possível aplicação dos ácidos húmicos em plantas.

Initial performance of maize in response to NPK fertilization combined with Herbaspirillum seropedicae

The inoculation with plant growth-promoting bacteria can be a technological approach useful for increasing the production of maize. The objective of this study was to evaluate the initial performance of maize in response to application of doses of NPK combined with the inoculation of the diazotrophic bacteria Herbaspirillum seropedicae in an greenhouse experiment. The experiment consisted of six fertilizer levels: 0, 25, 50, 75, 100 and 200% of the recommended dose of NPK applied to maize inoculated and non-inoculated with H. seropedicae. At 30 days after the treatment application, the growth characteristics and nutritional status of the plants were evaluated. Plant development was influenced by fertilization, but it was enhanced by combination with the bacteria, which resulted in significant increases in the dry mass of shoots (7%) and leaf area (9%) when compared with non-inoculated plants. The results showed increases in the concentration of N (11%), P (30%) and K (17%) of maize plants in response to bacterial inoculation together with NPK compared with plants that were applied fertilize only. The greater consistency and stability response of the host plant to bacterization in the presence of chemical fertilizer indicate a promissory biotechnological approach for improving the initial growth and adaptation of maize to the cultivation environment.

Gladiolus development in response to bulb treatment with different concentrations of humic acids

Gladiolus is an ornamental species produced for cut flowers and propagated by corms. The early flowering and increase in the number of flower buds, besides the production of commercial corms are constant challenges to be addressed in the crop improvement. Commercial production of ornamentals is technologically accelerated by means of growth regulators. Among them, the auxins stand out for their key role in the adventitious rooting and cell elongation. Alternatively, the humic substances present in the organic matter also have biostimulating effect, which is very similar to the auxinic effect. Therefore, this work aimed to study the growth and development of gladiolus in response to application of different concentrations of humic acids (HA) isolated from vermicompost. Corms were soaked for 24 hours in solutions containing 0, 10, 20, 30 and 40 mmol L-1 of C from HA. The corms were planted in 10-dm³ plastic bags filled with substrate and kept in a greenhouse. Growth of shoots and roots was evaluated. The results showed that the use of HA accelerates growth, and anticipates and increases flowering of Gladiolus.

Heavy metal toxicity in rice and soybean plants cultivated in contaminated soil

Heavy metals can accumulate in soil and cause phytotoxicity in plants with some specific symptoms. The present study evaluated the specific symptoms on rice and soybeans plants caused by excess of heavy metals in soil. Rice and soybean were grown in pots containing soil with different levels of heavy metals. A completely randomized design was used, with four replications, using two crop species and seven sample soils with different contamination levels. Rice and soybean exhibited different responses to the high concentrations of heavy metals in the soil. Rice plants accumulated higher Cu, Mn, Pb and Zn concentrations and were more sensitive to high concentrations of these elements in the soil, absorbing them more easily compared to the soybean plants. However, high available Zn concentrations in the soil caused phytotoxicity symptoms in rice and soybean, mainly chlorosis and inhibited plant growth. Further, high Zn concentrations in the soil reduced the Fe concentration in the shoots of soybean and rice plants to levels considered deficient.

Performance of pineapple slips inoculated with diazotrophic phosphate-solubilizing bacteria and rock phosphate

Besides fixing N2, some diazotrophic bacteria or diazotrophs, also synthesize organic acids and are able to solubilize rock phosphates, increasing the availability of P for plants. The application of these bacteria to pineapple leaf axils in combination with rock phosphate could increase N and P availability for the crop, due to the bacterial activity of biological nitrogen fixation and phosphate solubilization. The objectives of this study were: (i) to select and characterize diazotrophs able to solubilize phosphates in vitro and (ii) evaluate the initial performance of the pineapple cultivars Imperial and Pérola in response to inoculation with selected bacteria in combination with rock phosphate. The experiments were conducted at Universidade Estadual do Norte Fluminense Darcy Ribeiro, in 2009. In the treatments with bacteria the leaf contents of N, P and K were higher than those of the controls, followed by an increase in plant growth. These results indicate that the combined application of diazotrophic phosphate-solubilizing bacteria Burkholderia together with Araxá rock phosphate can be used to improve the initial performance of pineapple slips.

Effect of biostimulant application on production and flavonoid content of marigold (Calendula officinalis L.)

The production of medicinal plants as raw material for industry must associate quality with biomass formation and, with this purpose, the application of plant growth regulators has been studied in these crops. The objective of this study was to evaluate the effect of a biostimulant on growth, inflorescence production and flavonoid content in marigold. The experiment was conducted in a greenhouse and the treatments consisted of increasing doses of the biostimulant (0, 3, 6, 9, 12 and 15 mL L-1) applied by foliar spraying in ten consecutive applications. The experiment was arranged in a completely randomized design, with six treatments and ten repetitions. The number of leaves and flowerheads and dry matter of roots increased linearly with increasing doses of the growth promoter, with 20%, 36.97% and 97.28% increases, respectively, compared with the control. The total dry mass and shoot dry mass showed maximum values at the highest dose tested of 15 mL L-1 (with increases of 40.09% and 46.30%, respectively). Plant height and flavonoid content reached the highest values at a dose of 6 mL L-1. The biostimulant promoted the development of marigold and positively influenced the synthesis of the secondary compound of medicinal interest. Among the tested doses, the application of rates between 6 and 9 mL L-1 of the biostimulant is recommended for more efficient large-scale production of marigold.

Acclimation of croton and hibiscus seedlings in response to the application of indobultiric acid and humic acid for rooting

The vegetative propagation of ornamental plants can be accelerated by applying plant growth regulators. Amongst them, the use of auxins, plant hormones with physiological effects on cell elongation and rooting have stood out. Alternatively, the application of humic acids, bioactive fraction of soil organic matter, also results in increases in rooting cuttings of ornamental plants. The objective of this work was to study the growth characteristics and the nutritional contents of croton and hibiscus plants during acclimation of seedlings in response to different concentrations of indolebutyric acid (IBA) and humic acid (HA) applied to cuttings for rooting. The experiment was conducted in greenhouse, and the apical stem cuttings were treated with solutions with concentrations of 0, 250, 500, 1000 and 2000 mg L-1of IBA and 0, 10, 20, 30 and 40 mg L-1 of C from HA. At 45 days of rooting in carbonized rice husk, they were individually transferred to plastic bags of 2.0 dm3 containing a mixture of soil: sand: manure (2: 1: 1) as substrate. At 90 days of acclimation, the plants were collected for measurement of growth and nutritional variables. The results showed that the application of the IBA stimulates the absorption of nutrients and growth of croton cuttings and transplanted hibiscus, contributing to formation of vigorous seedlings. A similar response occurred with the application of HA in hibiscus cuttings

Co-inoculation of Rhizobium tropici and Azospirillum brasilense in common beans grown under two irrigation depths

ABSTRACT The alternative technique of co-inoculation or mixed inoculation with symbiotic and non-symbiotic bacteria has been studied in leguminous plants. However, there are few field studies with common beans and under the influence of the amount of irrigated water. Thus, the objective of this study was to evaluate the efficiency of inoculation and co-inoculation of common beans with Rhizobium tropici and Azospirillum brasilense under two irrigation depths. The experiment was carried out in the winter of 2012 and 2013, in Selvíria, state of Mato Grosso do Sul. The experimental design was composed of randomized blocks in split-plot scheme with two irrigation depths in the plots (recommended for common beans and 75% of the recommended) and five forms of nitrogen (N) supply in the split-plots (control non-inoculated with 40 kg ha- 1 of N in topdressing, 80 kg ha- 1 of N in topdressing, A. brasilense inoculation with 40 kg ha-1 of N in topdressing, R. tropici inoculation with 40 kg ha-1 of N in topdressing, and co-inoculation of A. brasilense and R. tropici with 40 kg ha- 1 of N in topdressing) with four repetitions. Co-inoculation increased nodulation in the second year of cultivation. None of the evaluated treatments increased the grain yield in relation to non-inoculated control with 40 kg ha-1 of nitrogen in topdressing, which presented average yield of 2,200 kg ha-1. The use of 75% of the recommended irrigation depth provides similar grain yield to the recommended irrigation depth in common beans cropped in winter.

Responses of higher plants to boron deficiency

Dissertation presented to obtain the degree of Doctorate in Biochemistry by Instituto de Tecnologia Química e Biológica of Universidade Nova de Lisboa

Serpentine bacteria influence metal translocation and bioconcentration of Brassica juncea and Ricinus communis grown in multi-metal polluted soils

The aim of this study was to assess the effects of inoculation of rhizosphere or endophytic bacteria (Psychrobacter sp. SRS8 and Pseudomonas sp. A3R3, respectively) isolated from a serpentine environment on the plant growth and the translocation and accumulation of Ni, Zn, and Fe by Brassica juncea and Ricinus communis on a multi-metal polluted serpentine soil (SS). Field collected SS was diluted to 0, 25, 50, and 75% with pristine soil in order to obtain a range of heavy metal concentrations and used in microcosm experiments. Regardless of inoculation with bacteria, the biomass of both plant species decreased with increase of the proportion of SS. Inoculation of plants with bacteria significantly increased the plant biomass and the heavy metal accumulation compared with non-inoculated control in the presence of different proportion of SS, which was attributed to the production of plant growth promoting and/or metal mobilizing metabolites by bacteria. However, SRS8 showed a maximum increase in the biomass of the test plants grown even in the treatment of 75% SS. In turn, A3R3 showed maximum effects on the accumulation of heavy metals in both plants. Regardless of inoculation of bacteria and proportion of SS, both plant species exhibited low values of bioconcentration factor (<1) for Ni and Fe. The inoculation of both bacterial strains significantly increased the translocation factor (TF) of Ni while decreasing the TF of Zn in both plant species. Besides this contrasting effect, the TFs of all metals were <1, indicating that all studied bacteria–plant combinations are suitable for phytostabilization. This study demonstrates that the bacterial isolates A3R3 and SRS8 improved the growth of B. juncea and R. communis in SS soils and have a great potential to be used as inoculants in phytostabilization scenarios of multi-metal contaminated soils.

The hyperaccumulator Sedum plumbizincicola harbors metal-resistant endophytic bacteria that improve its phytoextraction capacity in multi-metal contaminated soil

Endophyte-assisted phytoremediation has recently been suggested as a successful approach for ecological restoration of metal contaminated soils, however little information is available on the influence of endophytic bacteria on the phytoextraction capacity of metal hyperaccumulating plants in multi-metal polluted soils. The aims of our study were to isolate and characterize metal-resistant and 1-aminocyclopropane-1-carboxylate (ACC) utilizing endophytic bacteria from tissues of the newly discovered Zn/Cd hyperaccumulator Sedum plumbizincicola and to examine if these endophytic bacterial strains could improve the efficiency of phytoextraction of multi-metal contaminated soils. Among a collection of 42 metal resistant bacterial strains isolated from the tissues of S. plumbizincicola grown on Pb/Zn mine tailings, five plant growth promoting endophytic bacterial strains (PGPE) were selected due to their ability to promote plant growth and to utilize ACC as the sole nitrogen source. The five isolates were identified as Bacillus pumilus E2S2, Bacillus sp. E1S2, Bacillus sp. E4S1, Achromobacter sp. E4L5 and Stenotrophomonas sp. E1L and subsequent testing revealed that they all exhibited traits associated with plant growth promotion, such as production of indole-3-acetic acid and siderophores and solubilization of phosphorus. These five strains showed high resistance to heavy metals (Cd, Zn and Pb) and various antibiotics. Further, inoculation of these ACC utilizing strains significantly increased the concentrations of water extractable Cd and Zn in soil. Moreover, a pot experiment was conducted to elucidate the effects of inoculating metal-resistant ACC utilizing strains on the growth of S. plumbizincicola and its uptake of Cd, Zn and Pb in multi-metal contaminated soils. Out of the five strains, B. pumilus E2S2 significantly increased root (146%) and shoot (17%) length, fresh (37%) and dry biomass (32%) of S. plumbizincicola as well as plant Cd uptake (43%), whereas Bacillus sp. E1S2 significantly enhanced the accumulation of Zn (18%) in plants compared with non-inoculated controls. The inoculated strains also showed high levels of colonization in rhizosphere and plant tissues. Results demonstrate the potential to improve phytoextraction of soils contaminated with multiple heavy metals by inoculating metal hyperaccumulating plants with their own selected functional endophytic bacterial strains.

Proteomic and Transcriptomic Approaches to Abiotic Stress Tolerance in Rice (Oryza sativaL.)

Dissertação para a obtenção de grau de doutor em Biologia pelo Instituto de Tecnologia Química e Biológica da Universidade Nova de Lisboa

Towards modification of Medicago truncatula epigenome: genome editing with engineered nucleases

Periodic drought is the primary limitation of plant growth and crop yield. The rise of water demand caused by the increase in world population and climate change, leads to one of the biggest challenges of modern agriculture: to increase food and feed production. De novo DNA methylation is a process regulated by small interfering RNA (siRNAs), which play a role in plant response and adaptation to abiotic stress. In the particular case of water deficit, growing evidences suggest a link between the siRNA pathways and drought response in the model legume Medicago truncatula. As a first step to understand the role of DNA methylation under water stress, we have set up several bioinformatics and molecular methodologies allowing the design of Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 systems and the assembly of TALENs (transcription activator-like effector nucleases), to target both dicer-like 3 (MtDCL3) and RNA-Dependent RNA polymerase (MtRDR2), enzymes of the RNA-directed DNA methylation pathway. TALENs efficiency was evaluated prior to plant transformation by a yeast-based assay using two different strategies to test TALENs activity: Polyacrylamide gel electrophoresis (PAGE) and Single strand conformation polymorphisms (SSCP). In this assay, yeast cells triple transformation emerged as good and rapid alternative to laborious yeast mating strategies. PAGE analysis might be a valuable tool to test TALENs efficacy in vivo if we could increase TALENs activity. SSCP-based approach proved to be ineffective due to the generation of several false positives. TALENs and CRISPR/Cas9 system constructed and designed in this work will in the future certainly enable the successful disruption of DCL3 and RDR2 genes and shed the light on the relationship between plant stress resistance and epigenetic regulation mediated by siRNAs in M.truncatula.