Marcel Giovanni Costa França

A novel patatin-like gene stimulated by drought stress encodes a galactolipid acyl hydrolase.

A cDNA (Vupat1) encoding a predicted 43 kDa protein was isolated from drought‐stressed cowpea (Vigna unguiculata) leaves. It has homology with patatin, a potato tuber storage protein with lipolytic acyl hydrolase activity. The recombinant protein VUPAT1 expressed in the baculovirus system displays preferentially galactolipid acyl hydrolase activity. Phospholipids are very slowly hydrolyzed and apparently triacylglycerols are not deacylated. Vupat1 promoter contains putative drought‐inducible sequences. Northern blots showed that gene expression is stimulated by drought stress and is more pronounced in a drought‐sensitive cultivar than in a drought‐tolerant one. An involvement in drought‐induced galactolipid degradation is proposed for VUPAT1.

Nodulation in Dimorphandra wilsonii Rizz. (Caesalpinioideae), a Threatened Species Native to the Brazilian Cerrado

The threatened caesalpinioid legume Dimorphandra wilsonii, which is native to the Cerrado biome in Brazil, was examined for its nodulation and N2-fixing ability, and was compared with another, less-threatened species, D. jorgei. Nodulation and potential N2 fixation was shown on seedlings that had been inoculated singly with five bradyrhizobial isolates from mature D. wilsonii nodules. The infection of D. wilsonii by two of these strains (Dw10.1, Dw12.5) was followed in detail using light and transmission electron microscopy, and was compared with that of D. jorgei by Bradyrhizobium strain SEMIA6099. The roots of D. wilsonii were infected via small transient root hairs at 42 d after inoculation (dai), and nodules were sufficiently mature at 63 dai to express nitrogenase protein. Similar infection and nodule developmental processes were observed in D. jorgei. The bacteroids in mature Dimorphandra nodules were enclosed in plant cell wall material containing a homogalacturonan (pectic) epitope that was recognized by the monoclonal antibody JIM5. Analysis of sequences of their rrs (16S rRNA) genes and their ITS regions showed that the five D. wilsonii strains, although related to SEMIA6099, may constitute five undescribed species of genus Bradyrhizobium, whilst their nodD and nifH gene sequences showed that they formed clearly separated branches from other rhizobial strains. This is the first study to describe in full the N2-fixing symbiotic interaction between defined rhizobial strains and legumes in the sub-family Caesalpinioideae. This information will hopefully assist in the conservation of the threatened species D. wilsonii.

Electrolyte leakage and chlorophyll a fluorescence among castor bean cultivars under induced water deficit

We evaluated leaf fragments of three castor bean cultivars after being subjected to water stress. Leaf discs were exposed to polyethylene glycol (PEG-6000) solutions for tissue dehydration at various water potentials. After water-stress imposition, electrolyte leakage and chlorophyll a fluorescence were used jointly on the same leaf fragments cut from the same plant leaf. Furthermore, these two experimental procedures were adapted to unequivocally distinguish cultivars’ responses to water stress. Electrolyte leakage, ion efflux, membrane injury index and maximum quantum yield of photosystem II showed genotypic differences between cultivars. Despite these genotypic differences, the photosystem II electron transport rate was not significantly affected by water stress. The membrane injury shown may have been transient, probably due to a disarrangement in the phospholipid bilayer. The use of the two experimental procedures on the same leaf samples was less time-consuming and allowed for more reliable results. Furthermore, the procedures proved efficient for selection of physiological water-stress tolerance traits and could be employed in other plant experimental models.

Morphophysiological differences in leaves of Lavoisiera campos-portoana (Melastomataceae) enhance higher drought tolerance in water shortage events

Lavoisieracampos-portoana Barreto (Melastomataceae) has two kinds of leaves, pubescent and glabrous, and branches may have one or both types of leaves at the same moment. The plant is endemic to high altitude rocky fields in Brazil where rainfall is very seasonal. We predicted that these two leaf types are adaptations to different regimes of water availability. In experimental conditions of drought stress, we measured relative water content (RWC), photosynthetic pigments, chlorophyll a fluorescence and osmotic potential, and we counted stomates and measured stomatal conductance on both sides of leaves and compared these between the two leaf types. Stomatal conductance and electron transport rate at a given photosynthetic photon flux were greater in pubescent leaves than in glabrous leaves, and both declined during drought stress. Excessive photon flux density in glabrous leaves was greater during stress and after rehydration. Photosynthetic pigment content and RWC did not change between leaves, and values reduced during the stress period. Both types of leaves showed osmotic adjustment capacity, which occurred earlier in glabrous ones. These morphophysiological differences highlight the adaptation strategies of this plant to withstand drought, since the glabrous portion of the plant presents a preventive behavior, while the pubescent portion only shows the same responses in more advanced stages of drought stress.

Different Nitrogen Sources Affect Biomass Partitioning and Quality of Potato Production in a Hydroponic System

Crop production systems should reduce nitrogen application costs and assure that the appropriate form of nitrogen is used. Thus, three potato cultivars in a hydroponic system were supplied with two different nitrogen sources to determine the effect on biomass accumulation and partitioning, and total plant nitrogen content. Cultivars Agata, Atlantic and Bintje received, on alternate days, nutritive solutions differing only in either calcium nitrate or urea. Urea stimulated biomass accumulation and total nitrogen in shoots. Moreover, urea reduced the ratios tuber:shoot and tuber:root, and increased the ratio shoot:root, indicating competition for biomass partitioning between shoots and tubers. Urea stimulated greater tuber volume in the Atlantic cultivar, and increased tuber protein content, which is undesirable for industry. These results suggested that salts containing nitrate could be more appropriate for hydroponic potato cultivation, since urea compromised tuber quality and biomass partitioning in all cultivars studied.ResumenLos sistemas de producción deberían reducir los costos de aplicación de nitrógeno y asegurar que este sea usado en su forma apropiada. Por lo tanto, se suministraron, a tres cultivares de papas, en un sistema hidropónico, dos diferentes fuentes de nitrógeno para determinar su efecto sobre la acumulación y partición de biomasa y el contenido total de nitrógeno en la planta. Los cultivares Agata, Atlantic y Bintje recibieron, en días alternados, soluciones nutritivas que se diferenciaron, únicamente, por contener nitrato de calcio o urea. La urea estimuló la acumulación de biomasa y de nitrógeno total en la parte aérea. Además, redujo las relaciones tubérculo:brote y tubérculo:raíz y aumentó la relación brote:raíz, indicando competición en la partición de biomasa entre parte aérea y tubérculos. En el cultivar Atlantic, la urea estimuló el volumen de tubérculo y aumentó su contenido proteico, lo cual es indeseado para propósitos industriales. Estos resultados sugieren que las fuentes nítricas pueden ser mas apropiadas para el cultivo hidropónico de la papa, una vez que la urea comprometió tanto la partición de biomasa cuanto la calidad del tubérculo en todos los cultivares estudiados.

Cloning and characterization of drought-stimulated phosphatidic acid phosphatase genes from Vigna unguiculata

Under environmental stresses, several lipolytic enzymes are known to be activated and to contribute to membrane lipid turnover and generation of second messengers. In animal cells, phosphatidic acid phosphatase (PAP, EC 3.1.3.4), which dephosphorylates phosphatidic acid generating diacylglycerol, is long known as an enzyme involved in lipid synthesis and cell signalling. However, knowledge on PAP in plants remains very limited. The aim of this work was to isolate and characterize PAP genes in the tropical legume Vigna unguiculata (cowpea), and to study their expression under different stress conditions. Two cDNAs designated as VuPAPalpha and VuPAPbeta were cloned from the leaves of cowpea. Both proteins share sequence homology to animal type 2 PAP, namely, the six transmembrane regions and the consensus sequences corresponding to the catalytic domain of the phosphatase family, like the recently described Arabidopsis LPP (Lipid Phosphate Phosphatase) proteins. The recombinant protein VuPAPalpha expressed in Escherichia coli cells was able to convert phosphatidic acid into diacylglycerol. Unlike VuPAPbeta, VuPAPalpha has an N-terminal transit peptide and was addressed to chloroplast in vitro. Both genes are expressed in several cowpea organs and their transcripts accumulate in leaves in response to water deficit, including progressive dehydration of whole plants and rapid desiccation of detached leaves. No changes in expression of both genes were observed after wounding or by treatment with jasmonic acid. Furthermore, the in silico analysis of VuPAPalpha promoter allowed the identification of several putative drought-related regulatory elements. The possible physiological role of the two cloned PAPs is discussed.

Crescimento inicial de Dimorphandra wilsonii (Fabaceae - Caesalpinioideae) em diferentes condições de fertilidade em solo de cerrado

Dimorphandra wilsonii Rizz. is a critically endangered tree species. It has been described for sites of higher soil fertility in the Cerrado Domain. However, it occurs nowadays in isolated areas of low natural fertility. To study species adaptation to oligotrophic conditions, plants were grown in a greenhouse in soils from oligotrophic sites but with different levels of fertility. After soil fertility analysis, the macronutrients (NPK) and lime doses to be used in each treatment were defined, resulting in a combination of two liming levels and three fertilization levels. After 220 days, the treatments that significantly increased seedling growth were the one whose substrate was limed and fertilized with twice the prescribed NPK dosage, and the one with the indicated dosage of NPK but no liming. Liming itself did not promote growth enhancement, but favored increased growth response to increasing levels of nutrient availability. Root nodules were found in all the treatments, and the results led to the conclusion that D. wilsonii nutritional requirements are consistent with its original distribution sites in the Cerrado Domain. It was also possible to highlight nitrogen biological fixation, an unusual feature in Caesalpinioideae, as an adaptive strategy for survival in areas of low nutrient availability.

Nitric oxide increases tolerance responses to moderate water deficit in leaves of Phaseolus vulgaris and Vigna unguiculata bean species.

Drought stress is one of the most intensively studied and widespread constraints, and nitric oxide (NO) is a key signaling molecule involved in the mediation of abiotic stresses in plants. We demonstrated that a sprayed solution of NO from donor sodium nitroprusside increased drought stress tolerance responses in both sensitive (Phaseolus vulgaris) and tolerant (Vigna unguiculata) beans. In intact plants subjected to halting irrigation, NO increased the leaf relative water content and stomatal conductance in both species. After cutting leaf discs and washing them, NO induced increased electrolyte leakage, which was more evident in the tolerant species. These leaf discs were then subjected to different water deficits, simulating moderate and severe drought stress conditions through polyethylene glycol solutions. NO supplied at moderate drought stress revealed a reduced membrane injury index in sensitive species. In hydrated discs and at this level of water deficit, NO increased the electron transport rate in both species, and a reduction of these rates was observed at severe stress levels. Taken together, it can be shown that NO has an effective role in ameliorating drought stress effects, activating tolerance responses at moderate water deficit levels and in both bean species which present differential drought tolerance.

Root growth and proton efflux rates of rice seedlings in a greenhouse environment

Foram avaliados o crescimento e o efluxo de protons (EH+) em raizes de plântulas de arroz (Oryza sativa cv. IAC 47) cultivadas sob as condicoes de luz de uma casa de vegetacao. Quinze dias apos a emergencia, plântulas foram transplantadas para potes contendo 3,7 L de uma solucao nutritiva com 1,43 mmol N-NH4+ L-1 como fonte exclusiva de nitrogenio, visando induzir quatro ciclos sequenciais de acidificacao da solucao, a partir de um valor de pH inicial de 5,3. Os efluxos medios de protons foram calculados com base em periodos de crescimento radicular de 48 h (de 0 a 192 h). As estimativas de EH+ foram obtidas a partir dos valores calculados para as Taxas de Crescimento Relativo (TCR), expressas com base na area radicular (AR), ou massa fresca radicular (MF), e das pendentes da regressao linear da quantidade de protons livres na solucao (UH+) sobre MF ou AR. Os gradientes de acidificacao (DUH+/DMF) foram melhor associados quando relacionados com MF do que com AR. As taxas de crescimento das raizes foram afetadas por flutuacoes de radiacao fotossintetica e temperatura dentro da casa de vegetacao e nao se relacionaram claramente com os valores de EH+. Todavia, foi concluido que quando existe uma relacao linear entre UH+ e MF, as avaliacoes de EH+ expressas por unidades de MF, podem ser mais confiaveis do que aquelas expressas por unidades de AR.

Different salt concentrations induce alterations both in photosynthetic parameters and salt gland activity in leaves of the mangrove Avicennia schaueriana

Mangrove sites are constantly under tidal regimes, where there is variation of dissolved oxygen and saline content in water. This study evaluated the effects of varying salinity in Avicennia schaueriana, an abundant species in the sea-mangrove-river interface. Seedlings of A. schaueriana were harvested at the riverbank and placed in different saline and nutrient solutions in a greenhouse in order to simulate a saline flow. After 7 acclimatization days, plants were exposed to the following saline concentrations: 0, 50, 170 and 250mM NaCl, for a period of 30 days. After this, leaf gas exchange, chlorophyll a fluorescence, Falker chlorophyll index, leaf ultrastructure and micromorphology were evaluated. The highest saline concentration affected mainly the CO2 assimilation, internal CO2 concentration and the intrinsic efficiency of water use. The chlorophyll a fluorescence and Falker index were not significantly affected by any of the saline conditions, however chloroplast was damaged. Concomitantly, leaf salt glands clearly evidenced a higher salt secretion, when plants were submitted to the greatest saline concentration. The results indicated that A. schaueriana tolerates different salt concentration in the mangrove and excretes salt excess by salt gland, and this can attenuate the saline stress. However, when exposed to longer periods of higher salt fluxes the plant growth can be affected.