Patrícia de Fátima Pereira Goulart

Tolerance of Sesbania virgata plants to flooding

Sesbania virgata (Cav.) Pers. is a bush, pioneer and occurs naturally in Brazil. It belongs to the Fabaceae family and it is indicated for recovery of degraded areas because of its rusticity and capacity to tolerate flooding. The present research was carried out to investigate the ability of S. virgata plants to adapt to flooding conditions. Plants containing six expanded leaves were placed in masonry tanks and were subjected to the following conditions: control (well watered), soil-waterlogging (water to the setup level of 1 cm above the soil surface – roots and parts of the stems flooded) and complete submergence (whole plant flooded). The evaluations were conducted on the day of the stress induction and after 7, 14, 21, 28, 35, 42, 49 and 56 days of the treatment. After 15 days of return to normoxic environment, plant survival was assessed. Growth (height, dry mass of shoots and roots), vigour, carbohydrate content and the activity of enzymes involved in anaerobic metabolism (lactate dehydrogenase, pyruvate decarboxylase and alcohol dehydrogenase) were also evaluated. Our results suggested that sesbania plants are tolerant to flooding, because they can survive being submerged for 56 days. The reasons for this tolerance include the accumulation and use of carbohydrates in the leaves and roots, maintenance of growth and the activation of anaerobic metabolism, particularly in steps catalysed by the enzymes pyruvate decarboxylase and alcohol dehydrogenase.

Purification of polyphenoloxidase from coffee fruits

Polyphenoloxidase (PPO) was purified from coffee beans at the pinhead stage. Purification was carried out by precipitation of contaminating proteins with 30% (NH4)2SO4, dialysis of the supernatant and chromatography on phenyl sepharose and DEAE columns. A purification factor of 364 was obtained. Two bands (29 and 64 kDa) of the purified PPO were detected after SDS-PAGE which presented activity in partially denaturing SDS-PAGE and gave a positive response when probed in Western blots with an apple 27 kDa PPO antibody. It is suggested that the 29 kDa band was a cleavage product of the 64 kDa PPO. The N-terminal sequencing of the 29 kDa protein did not show similarity with others PPOs. Other reports have shown that the 64 kDa is a pre-protein which is converted to a 45 kDa mature form by action of a chloroplastid protease. The 45 kDa was not detected in pinhead fruits but usually detected in the leaves and endosperm of coffee. It is suggested that the appearence of the 45 kDa form might be controlled by the developmental stage of the fruit. Chlorogenic acid was the preferred substrate for the purified PPO.

Aspectos fisiológicos do desenvolvimento de mudas de café cultivadas sob telas de diferentes colorações

Abstract – The objective of this work was to evaluate the physiological aspects of the development of coffee seedlings grown under colored screens with different spectral characteristics. Seedlings of Catucai Amarelo 2SL, in the stage known as “orelha de onca”, were arranged in a randomized block design, with five replicates, under structures individually covered with blue, white, gray, black or red screens with 50% shade. Four months after, evaluations were done for seedling growth, pigment content of the leaves, total soluble sugars and starch contents of the leaves and roots. The red screen was the most effective in promoting growth in four out of the seven studied traits: plant height, leaf area and leaf dry weight and total dry matter. For the other characteristics, there was no difference among the screens. The pigment analysis showed that, except for the gray screen, the other ones did not differ for this trait. In leaves, the red screen promoted higher levels of carbohydrates and starch. At the root, carbohydrate contents were higher under the red and black screens. Among the five screen colors, the red one was the most efficient in the production of coffee seedlings with higher vigor and quality, with outstanding carbohydrate contents and biomass.Index terms:

Stomatal behavior and components of the antioxidative system in coffee plants under water stress

Coffee (Coffea arabica) plants show a positive relationship between stomatal closure and formation and accumulation of H2O2. However, for coffee plants under water restriction such relationship has never been studied. The objective of the present study was evaluate the stomatal movement and the antioxidant capacity of coffee seedlings under different water regimes. Eight months old coffee seedlings of cv. Catuai IAC 99 were submitted to field capacity, gradual and total suspension of irrigation during a period of 21 days. Evaluations of leaf water potential (Ψw) were performed in the beginning of the morning, and stomatal resistance, transpiration rate and vapor pressure deficit were determined at 10 am and 5 pm. All biochemical and enzymatic determinations were performed in leaves collected at 5 pm. Evaluations and samplings were performed at three days intervals. There was no variation in Ψw during the evaluated period for plants in field capacity. However, an expressive decrease of Ψw following day 12, reaching values near -2.5 MPa at the end of the experiment was observed for plants submitted to gradual suspension of irrigation. For plants submitted to total suspension of irrigation, Ψw decreases after the sixth day, reaching -2.5 MPa at day 15. The decay of Ψw in plants submitted to gradual and total suspension of irrigation reflected in increased stomatal resistance and in a decreased transpiration rate leading to an increase in hydrogen peroxide formation and, on final stages, increase in lipid peroxidation. As a conclusion, an increase in the activity of antioxidant enzymes as well as in the levels of ascorbate and dehydroascorbate was observed, which act in the detoxification of free radicals formed as result of the water stress.

Características fisiológicas e anatômicas de plantas de sibipiruna submetidas à hipoxia

Plants in their natural environment are subject to adverse conditions, and the stress periods can be short or long. Some species develop mechanisms that make them bear the adverse conditions, allowing their survival. The aim of this study was to evaluate the anatomic and physiological responses in the initial development of Sibipiruna plants submitted to hypoxia. Plants with three to four leaves fully expanded were subjected to three treatments: plants without flooding, flooded on roots and totally flooded. Evaluations were carried out in the starch, total soluble sugars and reducing sugars levels in the leaves and roots collected every five days, for 30 days. Anatomical characteristics were evaluated 15 days after the beginning of each treatment. The largest reduction in starch level in leaves and in roots was observed in plants completely flooded. For total soluble sugars and reducing sugars were observed behavior similar to that achieved in starch However, the plants which had flooded roots, in some periods of assessment, had levels, mainly in the roots, higher than the plants without flooding. These plants also showed higher stomatal density than the other treatments. For the thickness of the cortex and the cylinder vascular roots, a reduction was observed for the flooded treatment, which completely flooded plants had a higher apoplastic barrier imposed by thickening in phi, which is characterized by the formation of septa in cells near the endodermis.

Antioxidative responses and morpho-anatomical adaptations to waterlogging in Sesbania virgata

Sesbania virgata (Leguminosae) is tolerant of long periods of soil inundation. However, its morphological adaptations to anoxia and its response to possible damage from oxidative stress are still unknown. Here, we provide new information that helps to explain the ability of S. virgata plants to grow in flooded environments. Plants containing six expanded leaves were placed in masonry tanks and were subjected to the following conditions: control (well watered), soil waterlogging (water to the setup level of 1 cm above the soil surface—roots and parts of the stems flooded), and complete submergence (whole plant flooded). Plants exposed to flooding (soil waterlogging and complete submergence) significantly increased their production of hydrogen peroxide (H2O2), indicating the extent of oxidative injury posed by stress conditions. We demonstrate that plants exposed to flooding develop an efficient scavenger of ROS (generated during stress) in the roots through the coordinated action of nonenzymatic ascorbic acid (Asc) and dehydroascorbate (DHA) as well as the enzymatic antioxidants superoxide dismutase (SOD), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), and glutathione reductase (GR) that are present in the tissues. Moreover, we observed the development of morpho-anatomical structures such as adventitious roots, lenticels, and cracks in the stem of plants under soil waterlogging. The secondary root of plants under soil waterlogging showed a thinner cortex and larger number of elements of small diameter vessels. Numerous aerenchymas were observed in the newly formed in the adventitious roots. We conclude that these antioxidative responses and morpho-anatomical adaptations in the roots are part of a suite of adaptations that allow S. virgata plants to survive long periods of flooding, notably under waterlogged conditions.

Respostas fisiológicas em cultivares de soja submetidas ao alagamento em diferentes estádios

A soja e um dos produtos agricolas de maior importância no Brasil, e segundo maior produtor mundial. A tolerância ao excesso de umidade do solo e da rapida recuperacao com o termino do alagamento sao caracteristicas desta cultura, porem, a deficiencia de oxigenio, causada pelo excesso de agua, e um dos fatores determinantes que pode reduzir sua produtividade. O objetivo deste trabalho foi verificar o comportamento de tres cultivares de soja ao alagamento em diferentes estadios de desenvolvimento, por meio de analise da producao de massa seca, metabolismo de carboidratos e atividade das enzimas do metabolismo anaerobio. O experimento foi desenvolvido na Universidade Federal de Lavras (MG), em 2007. As cultivares BRS 267, BRS 257 e BRS 213 foram submetidas a 15 dias de alagamento, nos estadios V6 e V8 e 11 dias no estadio R4. Os resultados revelaram reducao na producao de massa seca nos tratamentos alagados no estadio V8 para a cultivar BRS 267 e no estadio V6 para BRS 257 e BRS 213; esta ultima foi a mais tolerante ao alagamento, pois obteve o menor decrescimo de massa seca em relacao ao controle. O alagamento tambem induziu aumento nos teores de amido e acucares soluveis totais nas folhas, assim como maior atividade das enzimas alcool desidrogenase e piruvato descarboxilase para as tres cultivares, quando comparadas a seus respectivos controles. Ja para a enzima lactato desidrogenase, essa diferenca foi menos expressiva. Esses resultados revelam comportamentos distintos comparando as tres cultivares.

Isoflavone and protein content in soybeans grains submitted to flooding at different stages of development

The stress imposed on plants by soil flooding constitutes a major barrier to growth and productivity. The identification of soybean varieties that produce higher levels of isoflavones, is necessary as soybeans have been used as human food to reduce risks of chronic diseases. Thus, this study was conducted with the objective of quantifying proteins and isoflavones in soybean cultivars subjected to flooding at various stages of development. The cultivars ‘BRS267’, ‘BRS257’ and ‘BRS213’ were subjected to 15 days of flooding, starting at the stages V6 and V8 and 11 days under stress starting at the stage R4. The proteins in the grain were extracted and quantified and analyzed by SDS-PAGE electrophoresis. Isoflavones were extracted, separated and quantified on HPLC. The electrophoretic analysis of the three cultivars under study revealed the same pattern of banding relative to the total protein regardless of the treatment. However, it was noted that flooding led to an increase in the total contents of isoflavones in the BRS 267 plants flooded in stage R4, remaining constant in other cultivars.

MECANISMOS DE TOLERÂNCIA DA VARIEDADE DE MILHO “SARACURA”

Com o objetivo de verificar a multiplicacao, expressao de sintomas foliares e efeito dos molicutes na producao de diferentes genotipos de milho, fitoplasma e Spiroplasma kunkelii foram inoculados, isolada ou simultaneamente, em cinco cultivares, mantidas em vasos ate a producao. Plantas sadias de todas as cultivares foram utilizadas como controle. Em cada vaso, foram cultivadas duas plantas, sendo uma utilizada para a deteccao dos molicutes na ultima folha completamente expandida, aos 30, 60 e 100 dias apos a inoculacao. A outra planta foi utilizada para deteccao dos molicutes nas folhas inferiores, medianas e apicais, aos 100 dias apos a inoculacao. O percentual de reducao causado pelos molicutes na altura e na producao de graos foi determinado nas duas plantas, em relacao as plantas sadias. A expressao de sintomas foliares e os resultados positivos obtidos na deteccao dos molicutes foram mais evidentes aos 100 dias apos a inoculacao. O periodo de enchimento de graos foi considerado a fase mais adequada para a deteccao desses patogenos. Os molicutes foram detectados nas folhas inferiores, medianas e apicais, observando-se maior frequencia nas folhas apicais. Verificou-se predominância de infeccao por espiroplasma e maior efeito detrimental desse patogeno no crescimento e producao das plantas em relacao ao fitoplasma. Nao foi detectado efeito sinergistico significativo dos dois molicutes sobre o crescimento e producao das plantas. Observou-se que o efeito prejudicial desses patogenos sobre o crescimento e producao das cultivares foi determinado pela frequencia de plantas infectadas.Apesar do amplo uso de deltametrina para controlar pragas do milho, no Brasil, existem poucas informacoes sobre o seu impacto nos artropodes associados a cultura. Este trabalho teve como objetivo avaliar os efeitos da pulverizacao de deltametrina em atropodes pragas e predadores mais comuns na parte aerea do milho. O delineamento foi em blocos casualizados, com quatro repeticoes. No estadio de dez folhas expandidas, o inseticida (Decis 25 CE, 7,5 gi.a./ha) foi pulverizado no cartucho das plantas e amostragens realizadas em dez plantas/parcela (1 dia antes, 7, 10, 14, 21 e 28 dias apos a pulverizacao) determinaram a densidade populacional ou a ocorrencia dos artropodes e a percentagem de plantas atacadas por Spodoptera frugiperda . O inseticida controlou a infestacao de S. frugiperda ate sete dias apos a pulverizacao e, nesse periodo, tambem reduziu em 64% a densidade populacional da cigarrinha Dalbulus maidis nas parcelas tratadas. O inseticida nao afetou negativamente o complexo de artropodes predadores associados a parte aerea do milho, mostrando toxicidade seletiva em favor de ninfas e adultos de Doru luteipes , de um grupo nao identificado de formigas e de aranhas. Esses resultados sao favoraveis ao uso de deltametrina em programas de manejo de S. frugiperda na cultura do milho, principalmente nos agroecossistemas com elevada abundância de predadores.Com o objetivo de estudar o efeito da omissao de P no meio de cultura nas caracteristicas fisiologicas de absorcao de P e no comprimento de raizes de oito genotipos de milho, quatro previamente classificados como eficientes e quatro como ineficientes para P, foram desenvolvidos dois experimentos. No primeiro, sementes dos oito hibridos de milho foram germinadas em rolos de papel-toalha e transplantadas para recipientes de plastico contendo solucao nutritiva completa de Steinberg, pH 5,5. Sete dias apos o transplante, o P foi retirado de metade das bandejas. Apos tres dias de omissao de P, realizou-se a cinetica de absorcao de P, iniciando-se a retirada de aliquotas de 10 ml de cada vaso, a cada 30 minutos, durante oito horas. No segundo experimento, aos seis dias apos a omissao do P no meio de cultivo, raizes das plantas dos tratamentos com e sem omissao de fosforo foram coletadas para avaliacao do comprimento total do sistema radicular. A omissao de fosforo por tres dias aumentou significativamente a velocidade maxima de absorcao de fosforo (Vmax) e a taxa de influxo em seis genotipos de milho, quatro ineficientes e dois eficientes para P. O Km (Constante de Michaelis) foi significativamente modificado pela omissao de fosforo em apenas dois genotipos. Conclui-se que aumentos na Vmax e taxa de influxo de P com a omissao de P no meio de cultivo e um mecanismo geral de resposta das plantas a deficiencia de P, mas que nao confere eficiencia diferencial entre genotipos de milho. A omissao de P na solucao nutritiva, por seis dias, aumentou significativamente o comprimento radicular em tres dos genotipos eficientes para P. Nos genotipos ineficientes para P, nao houve modificacao significativa dessas caracteristicas, indicando que o crescimento diferencial do sistema radicular pode ser um dos possiveis mecanismos da eficiencia para fosforo em milho.

Adubação do cafeeiro com nitrato de potássio via solo e folha, no outono-inverno e primavera-verão: efeitos na atividade da redutase do nitrato, no crescimento das plantas e na produção

Objetivou-se verificar o efeito da adubacao de 100 g de N/ano/planta, em tres epocas (outono-inverno, primavera-verao e outono-inverno/primavera-verao) e tres modos de aplicacao (folha, solo e folha/solo), no desenvolvimento, na producao e na atividade da redutase do nitrato (RN) da cultivar Rubi-MG com quatro anos de idade. A analise conjunta entre o modo e epoca de aplicacao mostra que o padrao de RN nao se altera, sendo no frio maior nas raizes e no calor maior nas folhas. O mesmo aconteceu com o padrao de crescimento, rapido na estacao quente/chuvosa e lento no periodo seco/frio. Em funcao da epoca de adubacao, concluiu-se que, no outono-inverno, deve-se dar preferencia pela aplicacao na folha ou folha-solo. Esses modos de adubacao foram os mais eficientes na retomada do crescimento. Quando as adubacoes ocorrerem nos dois periodos, qualquer um dos tres modos pode ser utilizado. Apesar da inviabilidade de varias pulverizacoes, eventualmente, uma ou mais pulverizacoes com KNO3 podem substituir eficientemente a adubacao no solo.