ABSTRACT To mitigate the deleterious effects of abiotic stress, the use of plant growth-promoting bacteria along with diazotrophic bacteria has been increasing. The objectives of this study were to investigate the key enzymes related to nitrogen and carbon metabolism in the biological nitrogen fixation process and to elucidate the activities of these enzymes by the synergistic interaction between Bradyrhizobium and plant growth-promoting bacteria in the absence and presence of salt stress. Cowpea plants were cultivated under axenic conditions, inoculated with Bradyrhizobium and co-inoculated with Bradyrhizobium sp. and Actinomadura sp., Bradyrhizobium sp. and Bacillus sp., Bradyrhizobium sp. and Paenibacillus graminis, and Bradyrhizobium sp. and Streptomycessp.; the plants were also maintained in the absence (control) and presence of salt stress (50 mmolL-1 NaCl). Salinity reduced the amino acids, free ammonia, ureides, proteins and total nitrogen content in nodules and increased the levels of sucrose and soluble sugars. The co-inoculations responded differently to the activity of glutamine synthetase enzymes under salt stress, as well as glutamate synthase, glutamate dehydrogenase aminating, and acid invertase in the control and salt stress. Considering the development conditions of this experiment, co-inoculation with Bradyrhizobium sp. and Bacillus sp. in cowpea provided better symbiotic performance, mitigating the deleterious effects of salt stress.

Abstract Soil salinity is an important abiotic stress worldwide, and salt-induced oxidative stress can have detrimental effects on the biological nitrogen fixation. We hypothesized that co-inoculation of cowpea plants with Bradyrhizobium and plant growth-promoting bacteria would minimize the deleterious effects of salt stress via the induction of enzymatic and non-enzymatic antioxidative protection. To test our hypothesis, cowpea seeds were inoculated with Bradyrhizobium or co-inoculated with Bradyrhizobium and plant growth-promoting bacteria and then submitted to salt stress. Afterward, the cowpea nodules were collected, and the levels of hydrogen peroxide; lipid peroxidation; total, reduced and oxidized forms of ascorbate and glutathione; and superoxide dismutase, catalase and phenol peroxidase activities were evaluated. The sodium and potassium ion concentrations were measured in shoot samples. Cowpea plants did not present significant differences in sodium and potassium levels when grown under non-saline conditions, but sodium content was strongly increased under salt stress conditions. Under non-saline and salt stress conditions, plants co-inoculated with Bradyrhizobium and Actinomadura or co-inoculated with Bradyrhizobium and Paenibacillus graminis showed lower hydrogen peroxide content in their nodules, whereas lipid peroxidation was increased by 31% in plants that were subjected to salt stress. Furthermore, cowpea nodules co-inoculated with Bradyrhizobium and plant growth-promoting bacteria and exposed to salt stress displayed significant alterations in the total, reduced and oxidized forms of ascorbate and glutathione. Inoculation with Bradyrhizobium and plant growth-promoting bacteria induced increased superoxide dismutase, catalase and phenol peroxidase activities in the nodules of cowpea plants exposed to salt stress. The catalase activity in plants co-inoculated with Bradyrhizobium and Streptomyces was 55% greater than in plants inoculated with Bradyrhizobium alone, and this value was remarkably greater than that in the other treatments. These results reinforce the beneficial effects of plant growth-promoting bacteria on the antioxidant system that detoxifies reactive oxygen species. We concluded that the combination of Bradyrhizobium and plant growth-promoting bacteria induces positive responses for coping with salt-induced oxidative stress in cowpea nodules, mainly in plants co-inoculated with Bradyrhizobium and P. graminis or co-inoculated with Bradyrhizobium and Bacillus.

Low temperatures negatively impact the metabolism of orange trees, and the extent of damage can be influenced by the rootstock. We evaluated the effects of low nocturnal temperatures on Valencia orange scions grafted on Rangpur lime or Swingle citrumelo rootstocks. We exposed six-month-old plants to night temperatures of 20ºC and 8ºC under controlled conditions. After decreasing the temperature to 8ºC, there were decreases in leaf CO2 assimilation, stomatal conductance, mesophyll conductance and CO2 concentration in the chloroplasts, in plant hydraulic conductivity and in the maximum electron transport rate driven ribulose-1,5-bisphosphate (RuBP) regeneration in plants grafted on both rootstocks. However, the effects of low night temperature were more severe in plants grafted on Rangpur rootstock, which also presented reduction in the maximum rate of RuBP carboxylation and in the maximum quantum efficiency of the PSII. In general, irreversible damage due to night chilling was found in the photosynthetic apparatus of plants grafted on Rangpur lime. Low night temperatures induced similar changes in the antioxidant metabolism, preventing oxidative damage in citrus leaves on both rootstocks. As photosynthesis is linked to plant growth, our findings indicate that the rootstock may improve the performance of citrus trees in environments with low night temperatures, with Swingle rootstock improving the photosynthetic acclimation in leaves of orange plants.

Um dos efeitos da salinidade causado por excesso de Na+ nos solos é a redução dos níveis de K+ nos tecidos vegetais. O trabalho objetivou caracterizar as vias de absorção de K+ de alta e baixa afinidade em raízes de cajueiro na ausência e presença de concentrações de Na+. Os estudos de cinética de absorção de K+ na ausência e presença de NaCl foram realizados com raízes destacadas de plântulas de cajueiro cultivadas sob privação de K+. O excesso de Na+ no meio externo causou redução da absorção de K+ pelos sistemas de alta e de baixa afinidade. A redução da absorção de K+, causada pelo excesso de Na+ no meio, nas raízes de cajueiro ocorre por mecanismo competitivo. Esse tipo de competição foi evidenciado pelo aumento dos valores de Km para a absorção de K+ induzido pelo Na+ externo, sem alterar os valores de Vmáx. Os resultados mostram que a salinidade pode comprometer a aquisição e a utilização de K+ em plantas de cajueiro.

One effect of the salinity caused by an excess of Na+ in soils, is the reduction in levels of K+ in plant tissue. The study aimed to characterize the process of high and low-affinity absorption of K+ in cashew roots, in both the absence and presence of concentrations of Na+. Studies of the kinetics of K+ absorption in the absence and presence of NaCl were carried out using detached roots from cashew seedlings grown under K+ deprivation. The excess of Na+ in the external environment caused a reduction in K+ absorption by both the high and low-affinity systems. The reduction in the absorption of K+ in the roots of the cashew tree, caused by the excess Na+ in the environment, happens through competitive means. This type of competition was evidenced by the increase in Km values for the K+ absorption induced by the external Na+, without changing the values of Vmax. The results show that salinity may affect the acquisition and use of K+ in cashew plants.

O objetivo do trabalho foi avaliar os efeitos de combinações de nitrato e amônio no meio, na acumulação de compostos nitrogenados e carboidratos, na presença e ausência de salinidade em plantas de sorgo. Sementes do genótipo CSF 20 foram cultivadas em casa de vegetação, em solução nutritiva com três concentrações de nitrato e amônio (100% NO3-: 0% NH4+, 75% NO3-: 25% NH4+ e 0% NO3-: 100% NH4+), na presença e ausência de salinidade. A salinidade reduziu a concentração de nitrato nas folhas e colmo, nos tratamentos 100:0 e 75:25. O amônio acumulou-se mais nas plantas com NaCl, nas folhas e colmo. Os teores de aminoácidos livres totais nas folhas não se alteraram, mas, no colmo, aumentaram na presença de sal e aumento do amônio. A concentração de prolina livre nas folhas e colmo elevou-se nas plantas sob salinidade. O conteúdo de nitrogênio total foi maior nas folhas, entretanto não houve grandes alterações com a presença do sal. A concentração de carboidratos solúveis totais foi maior nas plantas tratadas com sal, exceto no colmo 0:100. Nas folhas, o teor de sacarose foi reduzido na presença do NaCl e aumento de amônio, enquanto, no colmo, foi maior no tratamento 75:25 com NaCl. O teor de amido não sofreu alterações com as diferentes combinações nas formas de N e salinidade. Conclui-se que combinações de NO-3 e NH4+ no meio afetam a acumulação de compostos nitrogenados e carboidratos em folhas e colmo, com presença ou ausência de salinidade, em plantas de sorgo forrageiro.

The objective of this study was to evaluate the effects of combinations of nitrate and ammonium in the medium, on the accumulation of nitrogen compounds and carbohydrates, in the presence and absence of salinity in sorghum plants. CSF 20 genotype sorghum seeds, were grown in the greenhouse and supplied with a nutritive solution at three different concentrations of nitrate and ammonium (100% NO3-: 0% NH4+, 75% NO3-: 25% NH4+ e 0% NO3-: 100% NH4+) both in the presence and absence of NaCl. Salinity reduced the nitrate concentration in both leaves and stalks in treatments at 100:0 and 75:25. The accumulation of ammonium was higher in those plants, in both leaves and stalk, in the presence of NaCl. The levels of free amino acids in the leaves showed no change, whereas those in the stalk increased in the presence of salt and with the increase of ammonium. The concentration of free proline in both leaves and stalk was higher in plants under salinity. The total nitrogen content was higher in the leaves without, however, showing much change in the presence of salt. The concentration of total soluble carbohydrates was higher in those plants treated with salt, except in the stalk 0:100. Sucrose levels in the leaves were lower in the presence of NaCl and the increase in ammonium, while they were higher in the stalk at 75:25 NaCl. The starch content did not alter much with the different levels of nitrogen and salinity. It is concluded that NO-3 and NH4+ combinations in the medium affect the accumulation of nitrogen compounds and carbohydrates in the leaves and stalks of sorghum plants, both in the presence and absence of salinity.

O objetivo deste trabalho foi avaliar a resistência à salinidade de genótipos de cajueiro dos grupos anão e gigante, em mudas em fase de enxertia, com o uso de indicadores fisiológicos. A germinação e o crescimento inicial de plântulas de dez genótipos de porta-enxertos foram avaliados sob duas condições de salinidade: irrigação do substrato com água destilada ou com solução de NaCl 50 mmol L-1, até o estádio de oito folhas maduras (28 dias após o plantio). As mudas foram irrigadas com solução nutritiva de Hoagland & Arnon por mais dez dias. Os indicadores fisiológicos de resistência - concentrações de Na+ e K+ e relação K+/Na+, em folhas e raízes - não diferiram entre os genótipos, na ausência ou na presença de salinidade. Os indicadores e as concentrações de prolina não se correlacionaram com a massa acumulada em raízes e folhas. Sob salinidade, os genótipos do grupo anão apresentaram maior massa radicular em comparação aos do grupo gigante, que tiveram maior alocação de massa em folhas. Houve baixa variabilidade genética dentro de cada grupo. Quanto ao crescimento de raízes e folhas, houve variabilidade entre os grupos, nas duas condições de salinidade.

The objective of this work was to evaluate the resistance to salinity of cashew genotypes from dwarf and giant groups, with seedlings in pregrafting phase, using physiological indicators. Germination and seedling initial growth of ten rootstock genotypes were evaluated under two salinity conditions: substrate irrigated with distilled water or with 50-mmol L-1 NaCl solution, until the eight mature leaves stage (28 days after sowing). The seedlings were irrigated with Hoagland & Arnon nutrient solution, for ten more days. Physiological indicators of resistance - Na+ and K+ contents and K+/Na+ relation, in leaves and roots - did not differ between genotypes, in the absence or presence of salinity. The indicators and prolin contents were not correlated to the accumulated mass in roots and leaves. Under salinity conditions, the genotypes from the dwarf group showed higher root mass in comparison to those from the giant group, which had greater mass allocation in leaves. There was low genetic variability within each group. Regarding root and leaf growth, there was variability between groups, in both salinity conditions.

O objetivo deste trabalho foi caracterizar as respostas das trocas gasosas de plantas jovens de cajueiro à densidade de fluxo de fótons fotossintético (DFFF), temperatura, déficit de pressão de vapor (DPV) e concentração interna de CO2 (Ci), sob condições controladas. Cursos diários das trocas gasosas e de variáveis de fluorescência da clorofila a foram obtidos sob condições naturais. A taxa de assimilação de CO2 máxima foi de cerca de 13 mmol m-2 s-1, com saturação pela luz em torno de 1.000 mmol m-2 s-1. As temperaturas das folhas entre 25ºC e 35ºC corresponderam à faixa ótima para a fotossíntese. Os estômatos mostraram-se sensíveis ao CO2 e se fecharam com aumentos de Ci. Aumentos do DPV exerceram pequeno efeito sobre a taxa de assimilação de CO2, com uma pequena diminuição acima de 2,5 kPa. Os estômatos, no entanto, foram fortemente afetados pelo DPV, com fechamento gradativo acima de 1,5 kPa. As condutâncias estomáticas, reduzidas em altos DPVs, foram eficientes em restringir as perdas de água pela transpiração e demonstraram adaptação da espécie a ambientes secos. Sob irradiância natural, a taxa de assimilação de CO2 apresentou saturação no início da manhã, variando posteriormente com as variações da DFFF. Decréscimos transientes da razão Fv/Fm foram registrados em torno de 11h, o que indica a ocorrência de fotoinibição. Decréscimos da eficiência de captura da energia de excitação, decréscimos da eficiência efetiva do fotossistema II e aumentos da extinção não-fotoquímica foram consistentes com a ocorrência de fotoproteção sob irradiância excessiva.

The aim of this study was to characterize gas exchange responses of young cashew plants to varying photosynthetic photon flux density (PPFD), temperature, vapor-pressure deficit (VPD), and intercellular CO2 concentration (Ci), under controlled conditions. Daily courses of gas exchange and chlorophyll a fluorescence parameters were measured under natural conditions. Maximum CO2 assimilation rates, under optimal controlled conditions, were about 13 mmol m-2 s-1 , with light saturation around 1,000 mmol m-2 s-1. Leaf temperatures between 25ºC and 35ºC were optimal for photosynthesis. Stomata showed sensitivity to CO2, and a closing response with increasing Ci. Increasing VPD had a small effect on CO2 assimilation rates, with a small decrease above 2.5 kPa. Stomata, however, were strongly affected by VPD, exhibiting gradual closure above 1.5 kPa. The reduced stomatal conductances at high VPD were efficient in restricting water losses by transpiration, demonstrating the species adaptability to dry environments. Under natural irradiance, CO2 assimilation rates were saturated in early morning, following thereafter the PPFD changes. Transient Fv/Fm decreases were registered around 11h, indicating the occurrence of photoinhibition. Decreases of excitation capture efficiency, decreases of effective quantum yield of photosystem II, and increases in non-photochemical quenching were consistent with the occurrence of photoprotection under excessive irradiance levels.

O uso de mudas enxertadas uniformiza o crescimento de plantas e antecipa o início da produção. Os porta-enxertos regulam aspectos, como taxa fotossintética e relações hídricas das mudas, e distúrbios sobre os mesmos afetam o vigor geral das mudas. Este trabalho objetivou comparar os níveis de resistência dos porta-enxertos CCP06 e CCP09, e das mudas enxertadas CCP76/06 e CCP76/09, submetidas a estresses hídrico e salino, através de algumas características bioquímicas e biofísicas. A comparação entre as mudas CCP76/06 e CCP76/09 mostrou comportamentos diferentes. As mudas CCP76/6 reproduziram o comportamento de abertura estomática do porta-enxerto CCP06, que foi mais resistente aos efeitos dos estresses hídrico e salino do que o CCP09. Portanto, deve ter propiciado uma melhor adaptação ao enxerto CCP76/06 sob aqueles tipos de estresse. Alguns mecanismos de controle do porta-enxerto na absorção de íons e trocas gasosas são também discutidos.

The use of grated plants increases uniformity and anticipate the yield. Rootstocks regulate photosynthetic and water relations of the plant and any disruptions of their metabolism may affect the plant vigor. This work aimed to compare resistance levels of dwarf cashew tree rootstocks CCP06 and CCP09, and grafted plants CCP76/06 and CCP76/09 to water and saline stresses through biochemical and biophysics characteristics. The comparison among the grafted plants CCP76/06 and CCP76/09 showed different behaviors. The grafted CCP76/06 reproduced the behavior of stomatal opening of the rootstock CP06, which was more resistant to the effects of the water and saline stresses than the CCP9 and, therefore, it should have a better adaptation to CCP76/6 under those stresses. The preponderant participation of the rootstock in the regulation of the íons absorption and gas exchanges in dwarf cashew nut tree are also discussed.

Este estudo foi feito com a finalidade de estabelecer efeitos comparativos da seca e da reidratação na assimilação do nitrato e atividade do nódulo relacionado com a fixação de N2 em planta de caupi [Vigna unguiculata L. (Walp.)] previamente inoculada com Bradyrhizobium spp., estirpe BR-3256 (CB-756), na presença de NO-3 (5 mol m-3). Aos 28 dias após a emergência, as plantas noduladas foram submetidas à seca, durante 4 dias sucessivos e, depois, reidratadas com solução nutritiva durante 2 dias. A seca causou um aumento rápido no conteúdo de nitrato da raiz e uma acentuada redução na atividade da redutase do nitrato de folhas (NR). Em contraste, no nódulo esta atividade foi aumentada ligeiramente pelo déficit de água. Concomitantemente, nos nódulos das plantas estressadas, observou-se uma redução progressiva na concentração de leghemoglobina, atividade de glutamina sintetase (GS) e na concentração de ureídeos na seiva do xilema. A atividade da NR nas folhas aumentou rapidamente após a reidratação enquanto que o conteúdo de nitrato da mesma decresceu. Em contraste, a atividade de GS e a concentração de proteínas solúveis nos nódulos continuaram diminuindo nas plantas reidratadas. A concentração de leghemoglobina apresentou uma boa recuperação, enquanto que o conteúdo de ureídeos sofreu um leve aumento após a reidratação. Apesar da assimilação de nitrato em folhas e a atividade do nódulo terem sido severamente afetados pelo déficit de água, a rápida recuperação da atividade de redutase do nitrato nas folhas sugere que o primeiro processo seja menos susceptível ao ciclo de seca/reidratação quando plantas de caupi são noduladas em presença de nível moderado de nitrato.

This study was carried out to establish comparative effects of drought and recovery on the nitrate assimilation and nodule activity related to N2 fixation in cowpea plants [Vigna unguiculata L. (Walp.)] previously inoculated with Bradyrhizobium spp. BR-3256 (CB-756) strain in the presence of 5 mol m-3 NO-3. Twenty-eight-day-old nodulated plants were submitted to water deprivation during 4 consecutive days and afterwards resupplied with nutrient solution during 2 days. The water deprivation caused a rapid increase in the nitrate content in root and a marked reduction in leaf nitrate reductase (NR) activity. In contrast nodule NR activity was slightly increased by water deprivation. Concomitantly, in nodules of water stressed plants, leghemoglobin and glutamine synthetase (GS) activity declined and a progressive reduction in ureide-N concentration in xylem sap was observed. Leaf-NR activity increased rapidly after rehydration while leaf nitrate content declined. In contrast both GS activity and soluble protein content in the nodule continued to decline in rewatered plants. In addition the concentration of leghemoglobin recovered well, while the xylem ureide-N content experienced a slight increase after rehydration. Despite the nitrate assimilation in leaves and the nodule activity had been both severely affected by water stress, the rapid recovery of nitrate reductase activity suggests that the nitrate assimilation process is less sensitive to drought/rehydration cycle when cowpea plants are nodulated in presence of moderate nitrate level.

Foram avaliados, em dois híbridos simples de milho (Zea mays L.), Pioneer 3072 e IAC 701 x B, submetidos à deficiência hídrica, o comportamento da fotossíntese, da remobilização de N e de açúcares durante o período de crescimento dos grãos. As plantas foram cultivadas sob condições naturais, em doze tanques de alvenaria (4 m de comprimento x 0,5 m de largura x 0,6 m de profundidade) contendo terra. A deficiência hídrica foi induzida entre o 5.º e o 15.º dia após a floração, suspendendo-se a irrigação. Sem deficiência hídrica, ambos os híbridos apresentaram comportamento semelhante quanto aos parâmetros avaliados. Sob deficiência hídrica, a remobilização de açúcares e de N, a partir dos órgãos vegetativos para os grãos, foi mais intensa que nas plantas-controle, sem deficiência hídrica, e mais acentuada, tanto em açúcares como em N, no híbrido IAC 701 x B. Sob deficiência hídrica, a taxa de fotossíntese diminuiu em ambos os híbridos, sobretudo no IAC 701 x B.

Photosynthesis, nitrogen and sugar remobilization during the grain filling period were examined in two maize (Zea mays L.) hybrids (Pioneer 3072 and IAC 701 x B) submitted to water stress. The plants were grown under natural conditions in 12 boxes (4 m x 0,5 m x 0.6 m each) containing soil. Water stress was imposed by with holding irrigation during the 5.th and 15.th days after anthesis. Both hybrids in control treatment (without water stress) showed no differences in relation to the avaliated parameters. Under water stress the sugar and N remobilization from vegetative organs to the grains, were more intense than of the control plants. The sugar and N remobilization were larger in IAC 701 x B. The photosynthesis rates were reduced in both hybrids under water stress, but the reduction was larger in IAC 701 x B.