RESUMO

The legume-rhizobia symbiosis is an important process in agriculture because it allows the biological nitrogen fixation (BNF) which contributes to increasing the levels of nitrogen in the soil. Nitric oxide (⋅NO) is a small free radical molecule having diverse signaling roles in plants. Here we present and discuss evidence showing the role of ⋅NO during different stages of the legume-rhizobia interaction such as recognition, infection, nodule development, and nodule senescence. Although the mechanisms by which ⋅NO modulates this interaction are not fully understood, we discuss potential mechanisms including its interaction with cytokinin, auxin, and abscisic acid signaling pathways. In matures nodules, a more active metabolism of ⋅NO has been reported and both the plant and rhizobia participate in ⋅NO production and scavenging. Although ⋅NO has been shown to induce the expression of genes coding for NITROGENASE, controlling the levels of ⋅NO in mature nodules seems to be crucial as ⋅NO was shown to be a potent inhibitor of NITROGENASE activity, to induce nodule senescence, and reduce nitrogen assimilation. In this sense, LEGHEMOGLOBINS (Lbs) were shown to play an important role in the scavenging of ⋅NO and reactive nitrogen species (RNS), potentially more relevant in senescent nodules. Even though ⋅NO can reduce NITROGENASE activity, most reports have linked ⋅NO to positive effects on BNF. This can relate mainly to the regulation of the spatiotemporal distribution of ⋅NO which favors some effects over others. Another plausible explanation for this observation is that the negative effect of ⋅NO requires its direct interaction with NITROGENASE, whereas the positive effect of ⋅NO is related to its signaling function, which results in an amplifier effect. In the near future, it would be interesting to explore the role of environmental stress-induced ⋅NO in BNF.

RESUMO

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.

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.

RESUMO

Potato tubers were transformed with a chimeric gene made by the fusion of the soybean leghemoglobin encoding gene (lba) with the chloroplastic targeting sequence from Rubisco. This construct was placed under the control of the strong constitutive 35S promoter and the 3' nontranslated region of Rubisco from pea. Leghemoglobin expression on kanamycin-resistant plants was monitored by RT-PCR. Furthermore, immunodetection of subcellular fractions of transgenic plants revealed that leghemoglobin was imported and correctively processed inside the organelle. In addition, analysis of transgenic plants revealed reduced growth and decreased tuber production compared with the untransformed plants. It is suggested that leghemoglobin expression in potato chloroplasts interferes with aerobic metabolism, leading to physiological and morphological changes.