Aluminum effects on citric and malic acid excretion in roots and calli of rice cultivars

Citric and malic acid excretion in the medium and malic acid accumulation in seedling roots and embryo-derived calli as possible mechanisms of aluminum (Al) resistance and the effects of a 17-h Al stress period on root growth in Oryza sativa have been studied. Four-day-old seedlings and embryo-derived calli of Al-resistant (IRAT 112 and IR6023) and Al-sensitive (Aiwu and IKP) cultivars were treated with 250 and 500 µM {Al2(S04)3.18H20 }of total aluminum or without Al for 36 hours. After 3 to 36 hours of stress, seedlings and calli were removed from the flasks and concentration of citric and malic acids was estimated in the Al and control solutions. Malic acid was also assayed in roots tips and in callus tissues. After 17-h of Al stress, inhibition of root growth was a typical effect of Al in rice and the extent of the inhibition depended on both cultivar and Al concentration. At 500 µM of Al, strong reduction of root elongation occurred in all cultivars while at 250 µM of Al, only IRAT was unaffected, when compared to their control. In the absence of Al, all varieties excreted comparable amounts of citric and malic acid. Al treatments, were without effect upon citrate excretion in both Al-resistant and Al-sensitive cultivars. Al treatment, for periods from 3 to 24h, slightly stimulated the excretion of malic acid from seedlings, in all cultivars. Malic acid concentrations in root apices, in the presence or absence of aluminum, were not correlated with aluminum resistance. No differences in malic excretion and internal concentrations were detected between Al-treated and untreated rice calli of the same four cultivars. It is therefore concluded that, in our experimental conditions, differences in Al resistance in our rice cultivars cannot be attributed to citric and malic acids. Further research needs to be carried out to examine other possible mechanisms of Al-resistance in rice and to determine whether organic acids such as succinic and oxalic acid are implicated.

Absorção e acúmulo de cádmio e seus efeitos sobre o crescimento relativo de plantas de aguapé e de salvínia

A absorção e acúmulo de Cd e seus efeitos sobre o crescimento relativo foram estudados em aguapé (Eichhornia crassipes (Mart.) Solms) e salvínia (Salvinia auriculata Aubl.), em solução nutritiva. O aguapé absorveu quantidade significativamente maior de Cd do que a salvínia, tendo a diferença aumentado com o tempo de exposição ao Cd. O Km da absorção de Cd, estatisticamente igual nas duas espécies aquáticas quando as folhas contatavam a solução nutritiva, tornou-se maior em salvínia quando se impediu fisicamente que as folhas contatassem a solução de absorção. O Vmax de absorção de Cd, por outro lado, foi sempre maior em salvínia, independentemente do contato ou não das folhas com a solução nutritiva. A quantidade de Cd adsorvido e absorvido aumentou com a elevação da concentração de Cd na solução de cultivo, nas duas espécies estudadas, tendo sido maior em salvínia, possivelmente em razão da participação da parte aérea no processo de absorção. O acúmulo de Cd nas plantas, elevado nas primeiras 12 h de exposição, decresceu rapidamente com o tempo de exposição, estabilizando após três dias de exposição. Salvínia apresentou maior acúmulo diário de Cd por unidade de peso mas aguapé maior acúmulo total por unidade experimental. Os teores de Cd aumentaram com o aumento do tempo de exposição e da concentração de Cd na solução nutritiva, nas duas partes da planta das duas espécies, especialmente nas raízes. Cerca de 80% de todo o Cd absorvido acumulou-se nas raízes de aguapé, e a distribuição não foi afetada pelo contato das folhas com a solução nutritiva. Em salvínia, quando se impediu o contato das folhas com a solução nutritiva observou-se uma distribuição do Cd aproximadamente igual à de aguapé. Quando, porém, se permitiu o contato das folhas com a solução nutritiva a parte aérea passou a contribuir com quase 50% do Cd absorvido. As taxas de crescimento relativo decresceram intensamente com o aumento da concentração de Cd na solução nutritiva nas duas espécies aquáticas, especialmente em salvínia. As concentrações de Cd para promover redução de 25% no crescimento relativo foram, em média, doze vezes menores em salvínia do que em aguapé, em razão de sua maior sensibilidade a este metal pesado.

Plant growth and solute accumulation and distribution in two sorghum genotypes, under NaCl stress

Seedlings of two sorghum (Sorghum bicolor (L.) Moench) genotypes with differential tolerance to salinity were exposed to 0 and 100 mM NaCl, gradually added in increments of 25 mM every 12 hours, in nutrient solution. Seven days after starting the salt treatment the growth of the shoot and root system and the inorganic and organic solutes contents were determined. Salinity reduced the dry matter yield and length of the shoot and root system in both sorghum genotypes, specially in the sensitive one. In general, it was observed an increase in Na+ and Cl- transfer to the shoot, in Na+ and Cl- accumulation and in the Na+/Cl- ratio but a decrease in the K+ and Ca2+ transfer to shoot and in the K+ and Ca2+ contents in the shoot, always with higher intensity in sensitive genotype. Apparently, the tolerance to high saline concentrations in sorghum seems to be related to the genotype ability to avoid accumulation of harmful levels of Na+ and Cl- and, or to maintain adequate levels of K+ and Ca2+, specially in the shoot. The soluble carbohydrates and amino acids constituted together over 98% of the total organic solutes and showed the greatest absolute increase in concentration during saline stress. Probably, the soluble carbohydrates were the most important organic solutes to contribute to the osmotic adjustment in the leaves and the amino acids in the roots. Under saline stress there was an expressive increase in proline contents, specially in the oldest leaves of sensitive genotype. The proline contents, however, even under salt stress, did not reach the levels of other organic solutes. Contrary to the general acceptance, proline does not seem to have an important role in the mechanism of salt tolerance, at least for these genotypes and under the experimental conditions applied here.

Modifications in vapor diffusion resistence of leaves and water relations in barbados cherry plants under water stress

Young sexually and assexually propagated Barbados cherry plants were submitted to water deficit (20 days without irrigation). During this period the accumulation of proline, water potential of branches, osmotic potential, the relative water content of leaves, the leaf diffusive resistance, the transpiration rate and leaf temperature in the cuvette were determined. In addition, photosynthetically active radiation (PAR) and vapor pressure deficit (VPD) were measured in the porometer cuvette. The concentration of proline for both types of plants began to increase on the fifth day without watering, and reached 38.1 times the concentration in the control plants grown from seeds and 26.4 times the concentration in grafted plants on the tenth day without watering. The lowest levels of leaf water potential in the plants suffering from severe water stress varied from -4.5 to -5.7 MPa, the lowest values being observed in the sexually propagated plants. These plants also showed the highest values for transpiration (0.9 mmol.m-2.s-1) and proline concentration (20.42 mg.g-1 DM), the lowest for relative water content of the leaves (38.4%) and diffusive resistance (940 s.m-1) at the end of the experiment. The Barbados cherry plants developed strategies for surviving drought, with differences between various characteristics, resulting from prolonged stress, which significantly influenced the parameters evaluated, with the exception of leaf temperature.

Acúmulo e distribuição de metais pesados nas raízes, caule e folhas de mudas de árvores em solo contaminado por rejeitos de indústria de zinco

Avaliaram-se em casa-de-vegetação o acúmulo e a distribuição de metais pesados nas raízes, caule e folhas de mudas de vinte espécies arbóreas tropicais crescendo em solo contaminado com metais pesados. As mudas com tamanho ideal para plantio foram transferidas para vasos contendo 3,3 kg de solo sem contaminação ou contaminado com Zn, Cd, Cu e Pb, onde permaneceram por 90 dias. As espécies que cresceram em solo contaminado apresentaram teores foliares elevados, variando de (mg kg-1): Zn = 154 a 1177; Cd = 0,6 a 54,6; e Cu = 2,8 a 134. Essas concentrações foram, em muitos casos, superiores aos níveis críticos de toxidez, ao passo que os teores foliares de Pb variaram de 0,1 a 4,3 mg kg-1, não atingindo níveis considerados fitotóxicos. Em algumas espécies como Machaerium nictidans, Myroxylon peruiferum, Piptadenia gonoacantha, Senna macranthera e Trema micrantha, que tiveram o crescimento muito inibido pela contaminação, houve elevada translocação de Zn e/ou Cd para parte aérea. Dendropanax cuneatum, que foi pouco afetada pelos metais, também apresentou índice elevado de translocação de Zn e Cd, mas reteve esses elementos no caule. Acacia mangium, Copaifera langsdorffi e Cedrella fissilis também apresentaram baixa sensibilidade à contaminação e apresentaram elevado acúmulo de Zn e Cd nas raízes, indicando que a baixa translocação desses elementos para a parte aérea está envolvida na tolerância dessas espécies ao excesso de metais pesados no solo. O padrão de distribuição proporcional de Zn e Cd nas raízes e parte aérea das plantas se relaciona com o comportamento dessas ao excesso de metais pesados no solo.

Analysis of proteins associated with storage root formation in cassava using two-dimensional gel electrophoresis

Two-dimensional gel electrophoresis analysis was performed on adventitious and storage root in cassava (Manihot esculenta Crantz). Adventitious root lacking swelling formation and swelled storage root were obtained from the accession WU104 grown in the green house of the Department of Biology in Washington University in St. Louis (MO-USA). Saline buffer-soluble proteins were extracted, separated in a high-resolution 2-D electrophoresis system, visualized with silver staining gel procedure, and digital image generate for further analysis. Quantitative and qualitative protein spots analysis was performed with a computer assisted image software system. Results revealed large variation in the complexity of the gel protein profile between the two root systems. About 90% of the protein spots appeared in the pI range value of 4.0 to 6.5 and between 14 to 80 Kda of molecular mass. Detailed computer assisted analysis of this gel allowed us to establish 5 distinct classes of protein based on spot quantification that could be associated with swelling and non-swelling roots. Variation in the complexity of protein pattern was related with different type of root. Whereas the adventitious root showed a more simple profile related to primary growth, the storage root showed to be a more complex profile related to secondary growth and starch accumulation.

Nitrate reductase and glutamine synthetase activities in S1 endogamic families of the maize populations Sol da Manhã NF and Catetão

The possibility of improving nitrogen use efficiency in maize was investigated using S1 endogamic families of the populations Sol da Manhã NF and Catetão. A simple 10 X 10 lattice design was adopted and the trials carried out at the experimental field of MITLA AGRÍCOLA LTDA, in Uberlândia, State of Minas Gerais, during the 1994/95 planting season. Based on grain production figures, the three best and three worst performing S1 endogamic families were selected for this study. These were pooled to form four sub-populations denominated NFB, NFR (the best and worst families, respectively, of the Sol da Manhã NF variety), CATB and CATR (the best and worst families, respectively, of the Catetão variety). Each of these sub-populations was evaluated under greenhouse conditions. The experimental design was factorial with treatments arranged in randomized blocks. Sample replicates consisted of pots with four plants. Feeding with modified Hoagland’s nutrient solution began on the seventh day after sowing. The study involved four nitrogen regimes, where varying proportions of NO3- and NH4+ were formulated, such that the nutrient solution contained the following mixtures: 75% NO3- : 25% NH4+; 25% NO3- : 75% NH4+; 50% NO3- : 50% NH4+ (all high N mixtures) and 5% NO3- : 5% NH4+ (low N mixture). Twenty-five days after planting, the activities of the enzymes nitrate reductase and glutamine synthetase (transferase and synthetase assays) were determined for the leaves using the third topmost expanded leaf of the four plants in each pot. The data show that glutamine synthetase (transferase assay) and nitrate reductase activities were efficient in discriminating the S1 endogamic families and could therefore be useful biochemical parameters in breeding programs seeking nitrogen use efficiency.

Isolation and partial characterization of a lectin from Bauhinia pentandra (bong) vog. Ex. Steua.

Bauhinia pentandra (Bong) Vog. ex. Steua seeds were investigated with respect to phenologic aspects (size, mass, hilum and length) and with respect to their chemical composition. The total nitrogen content of the seed flour was determined, and the flour was extracted in different pH values. A lectin was isolated from the seeds by Sepharose-4B affinity chromatography. The homogeneity of the lectin was demonstrated by SDS-PAGE in the presence of beta-mercaptoethanol. Only one protein band with an apparent molecular mass of 30 kDa was found. The B. pentandra lectin showed a carbohydrate specificity for D-galactose, a requirement for divalent metal cations (Ca2+ and Mn2+) for full activity and amino acid composition with a high content of aspartic acid, glutamic acid and alanine and low levels of methionine, cysteine and tryptophan. The lectin agglutinated rabbit and human A group erythrocytes and was relatively stable to heat treatment, retaining half of its original activity after 60 min at 70 ºC.

Temperature-dependent germination and endo-beta -mannanase activity in sesame seeds

The effects of temperature on germination and endo-beta-mannanase activity in seeds of Sesamum indicum was investigated. The minimum germination temperature (Tmin) lies between 12.8°C and 13.2°C while the maximum temperature (Tmax) is located between 45.5°C and 46°C. Germinabilities are statistically not different from estimated viability (88%) between 18.8°C and 43.2°C. The Mann-Whitney test indicated the interval 31.9°C to 35.1°C as the optimum temperature (Topt) range for germination rate. When seeds incubated at temperatures at or below the Tmin and close to or above the Tmax were transferred to 30°C, those incubated at lower temperatures achieved high germinability. On the other hand, the higher the pre-incubation temperature above Tmax, the lower the germinability achieved near Topt. Seed endosperm cell wall was found to contain mannose as the main monosaccharide. An increase in endo-beta-mannanase activity in the micropylar endosperm prior to seed germination was observed only at supra-optimum temperature.

Metabolismo do nitrogênio em alfafa nodulada sob supressão e ressuprimento de fósforo

Estudaram-se os efeitos da supressão e do ressuprimento de Pi sobre o metabolismo do nitrogênio, mediante a fixação biológica do nitrogênio atmosférico (N2) e atividade da GS, Fd-GOGAT e NADH-GOGAT, em plantas de alfafa, cv. Flórida 77, inoculadas com Sinorhizobium meliloti, em diferentes estádios do desenvolvimento vegetativo (V3 e V4) e reprodutivo (R6 e R8). O ensaio foi conduzido em casa-de-vegetação e as plantas cultivadas em solução nutritiva. O período de supressão de Pi por dez dias reduziu os teores de Pi nas folhas e nos nódulos em todos os estádios do desenvolvimento, ao passo que nas raízes essa redução somente foi observada nos estádios vegetativos. Após o ressuprimento de Pi por igual período, ou seja, dez dias, foi observada a recuperação nos estádios R6 e R8 para folhas, V3 e V4 para raízes e V3 para os nódulos. A supressão de Pi reduziu significativamente os teores de aminoácidos totais nas folhas e nas raízes em todos os estádios do desenvolvimento, indicando que houve alteração no processo de fixação do N2. Após o ressuprimento, os teores de aminoácidos totais nas raízes, em todos os estádios do desenvolvimento, alcançaram valores similares àqueles das plantas do tratamento-controle, ao passo que nas folhas isso só ocorreu nos estádios vegetativos. A composição de aminoácidos na seiva do xilema não foi alterada em função da supressão e do ressuprimento de Pi. As atividades da GS e Fd-GOGAT, nos nódulos, foram reduzidas apenas nos estádios V3 e R8, enquanto com o ressuprimento não houve recuperação da Fd-GOGAT. Contudo, houve a recuperação da atividade da GS em R8. Nas folhas, a atividade da GS foi significativamente alterada em todos os estádios, enquanto a da Fd-GOGAT foi alterada somente em V3 e V4. Após o ressuprimento, não houve a recuperação da atividade da GS e Fd-GOGAT em V3. A NADH-GOGAT nas folhas e nos nódulos foi significativamente reduzida nos tratamentos de supressão de Pi em todos os estádios avaliados, não havendo recuperação após o ressuprimento. A NADH-GOGAT foi muito sensível à baixa disponibilidade de Pi. A supressão do Pi parece exercer controle na atividade dessas enzimas durante os diferentes estádios do desenvolvimento.