Leandro Azevedo Santos

Isoforms of plasma membrane H+-ATPase in rice root and shoot are differentially induced by starvation and resupply of NO3− or NH4+

The goal of this study was to evaluate the effects of nitrogen starvation and resupply in 10 PM H+-ATPase isoforms and the expression of NO₃⁻ and NH₄+ transporters in rice. The net uptake of both forms of NO₃⁻-N or NH₄+-N was increased with its resupply. Resupply of NO₃⁻ resulted in induction of the following PM H+-ATPase isoforms, OsA1, OsA2, OsA5 and OsA7 in the shoots and OsA2, OsA5, OsA7 and OsA8 in the roots. Resupply of NH₄+ resulted in the induction of the following OsA1, OsA3 and OsA7 isoforms in the roots while OsA1 was induced in the shoots. It was observed that increased PM H+-ATPase activity also resulted in increased net uptake of NO₃⁻ and NH₄+. In the roots, OsNRT2.1 and OsNRT2.2 were induced by NO₃⁻ resupply, while OsAMT1.1 and OsAMT1.2 were induced by NH₄+ deficiency. The results showed that the expression of PM H+-ATPase isoforms is related to NO₃⁻ and NH₄+ transporters as well as in which section of the plant it takes place. PM H+-ATPase isoforms OsA2 and OsA7 displayed the strongest induction in response to N resupply, therefore indicating that these genes could be involved in N uptake in rice.

Humic matter elicits proton and calcium fluxes and signaling dependent on Ca 2+ -dependent protein kinase (CDPK) at early stages of lateral plant root development

BackgroundThe humic acid (HA) fraction of soil organic matter (SOM) exerts an effective plant growth promotion through a complex mechanism involving a coordinated activation of several key ion transport and signaling systems. We investigated the effects of HA on H+ and Ca2+ cellular dynamics at the early stages of lateral plant root development.ResultsEmergence of lateral root in rice seedlings were related to specific H+ and Ca2+ fluxes in the root elongation zone underlying an activation of the plasma membrane H+-ATPase and of the Ca2+-dependent protein kinase (CDPK). The latter was coupled with an increased expression of the voltage-dependent OsTPC1 Ca2+channels and two stress responsive CDPK isoforms, such as OsCPK7 and OsCPK17.ConclusionsHA act as molecular elicitors of H+ and Ca2+ fluxes, which seem to be upstream of a complex CDPK cell-signaling cascade. These findings shed light on the first ion signaling events involved in the mechanism of action of HA on plant growth and development.

NITRATE UPTAKE KINETICS AND METABOLIC PARAMETERS IN TWO RICE VARIETIES GROWN IN HIGH AND LOW NITRATE

A study was conducted on the effect of nitrate (NO3) levels on nitrate uptake kinetics and nitrogen (N) metabolism in two rice varieties, Piaui (landrace) and IAC-47 (improved). At 27 days after germination (DAG) N supply was suspended for 72 h, and then restored as 0.2 or 2 mM nitrate. The nitrate uptake kinetics was determined by the depletion method. Plants were harvested at 0, 6, and 24 h. Plants of the Piaui variety under 0.2 mM nitrate showed higher Vmax and lower KM, indicating higher efficiency of nitrate uptake at low supply. In the sheaths of both varieties, there was a greater accumulation of nitrate and lower activities of nitrate reductase and glutamine synthetase. The V-H+-ATPase activity increased between 0 and 6 h accompanying the nitrate influx, suggesting that the activity of this proton pump is necessary for the antiport activity (H+/2NO− 3) involved in the accumulation of nitrate into vacuoles.

Involvement of Hormone- and ROS-Signaling Pathways in the Beneficial Action of Humic Substances on Plants Growing under Normal and Stressing Conditions

The importance of soil humus in soil fertility has been well established many years ago. However, the knowledge about the whole mechanisms by which humic molecules in the rhizosphere improve plant growth remains partial and rather fragmentary. In this review we discuss the relationships between two main signaling pathway families that are affected by humic substances within the plant: one directly related to hormonal action and the other related to reactive oxygen species (ROS). In this sense, our aims are to try the integration of all these events in a more comprehensive model and underline some points in the model that remain unclear and deserve further research.

OsAMT1.3 expression alters rice ammonium uptake kinetics and root morphology

Abstract High-affinity ammonium transporters (AMT1) are responsible for ammonium (NH4+) acquisition and/or perception in the micromolar range, and their expressions can be differentially regulated by nitrogen (N) availability. The present study characterised the functions of the rice (Oryza sativa) OsAMT1.3 transporter to understand its contribution to NH4+ acquisition and plant adaptation to environments with low N availability. Transgenic rice plants were obtained to study the activity of the OsAMT1.3 promoter (POsAMT1.3:GFP:GUS) and the overexpression of the OsAMT1.3 gene (UBIL:OsAMT1.3:3xHA) in plants. The OsAMT1.3 promoter activity was induced strongly in the absence of N and occurred primarily in the zones of lateral root emission and root tips. Anatomical sections of the segment of root tips and the middle third showed a differential pattern of OsAMT1.3 activity. Analysis of the OsAMT1.1–1.3 transporter expression profiles indicated that overexpression of OsAMT1.3 positively affected OsAMT1.2 expression. When subjected to a low N supply, plants overexpressing OsAMT1.3 showed lower KM and Cmin values. Additionally, these lines showed longer roots with a higher area, volume, and number of tips. The data suggested that OsAMT1.3 is involved in the ability of rice plants to adapt to low NH4+ supplies.

Vermicompost humic acids modulate the accumulation and metabolism of ROS in rice plants

This work aims to determine the reactive oxygen species (ROS) accumulation, gene expression, anti-oxidant enzyme activity, and derived effects on membrane lipid peroxidation and certain stress markers (proline and malondialdehyde-MDA) in the roots of unstressed and PEG-stressed rice plants associated with vermicompost humic acid (VCHA) application. The results show that the application of VCHA to the roots of unstressed rice plants caused a slight but significant increase in root ROS accumulation and the gene expression and activity of the major anti-oxidant enzymes (superoxide dismutase and peroxidase). This action did not have negative effects on root development, and an increase in both root growth and root proliferation occurred. However, the root proline and MDA concentrations and the root permeability results indicate the development of a type of mild stress associated with VCHA application. When VCHA was applied to PEG-stressed plants, a clear alleviation of the inhibition in root development linked to PEG-mediated osmotic stress was observed. This was associated with a reduction in root ROS production and anti-oxidant enzymatic activity caused by osmotic stress. This alleviation of stress caused by VCHA was also reflected as a reduction in the PEG-mediated concentration of MDA in the root as well as root permeability. In summary, the beneficial action of VCHA on the root development of unstressed or PEG-stressed rice plants clearly involves the modulation of ROS accumulation in roots.

Effects of nitrogen stress on proton-pumping and nitrogen metabolism in rice.

ABSTRACT Three rice (Oryza sativa L.) varieties: Caiapó (bred for upland crop), Piauí (a landrace), and IAC-47 (an improved variety) were used to study the effects of nitrogen (N) suppression in plant metabolism. Nitrogen suppression increased root growth in Caiapó plants. This growth was simultaneous with increases in hydrogen (H+)-pyrophosphatase (H+-PPase) and vacuolar H+-ATPase (V-H+-ATPase) activities in the roots of Piauí plants. The SDS-PAGE shows 73, 67, and 53 kDa fractions proteins accumulating in the tonoplast what agrees with the observed activities of H+-PPases and V-H+-ATPases. Piauí and IAC-47 varieties exhibited different responses from one another at the beginning of N-Stress: there was an increase in P-H+-ATPase activity for Piauí and a decrease for IAC-47. Piauí plants had a higher content of nitrate (NO3 −) in roots and sheaths, notwithstanding it mobilized less of this NO3 −-N during this growth period than did IAC-47. These results suggest that Piauí plants are better adapted to nutrient-poor environments.

Contribution of dark septate fungi to the nutrient uptake and growth of rice plants

The use of dark septate fungi (DSE) to promote plant growth can be beneficial to agriculture, and these organisms are important allies in the search for sustainable agriculture practices. This study investigates the contribution of dark septate fungi to the absorption of nutrients by rice plants and their ensuing growth. Four dark septate fungi isolates that were identified by Internal transcribed spacer phylogeny were inoculated in rice seeds (Cv. Piauí). The resulting root colonization was estimated and the kinetic parameters Vmax and Km were calculated from the nitrate contents of the nutrient solution. The macronutrient levels in the shoots, and the NO3−-N, NH4+-N, free amino-N and soluble sugars in the roots, sheathes and leaves were measured. The rice roots were significantly colonized by all of the fungi, but in particular, isolate A103 increased the fresh and dry biomass of the shoots and the number of tillers per plant, amino-N, and soluble sugars as well as the N, P, K, Mg and S contents in comparison with the control treatment. When inoculated with isolates A103 and A101, the plants presented lower Km values, indicating affinity increases for NO3−-N absorption. Therefore, the A103 Pleosporales fungus presented the highest potential for the promotion of rice plant growth, increasing the tillering and nutrients uptake, especially N (due to an enhanced affinity for N uptake) and P.

Development and nitrate reductase activity of sugarcane inoculated with five diazotrophic strains

Diazotrophs are able to stimulate plant growth. This study aimed at evaluating the effect of inoculation of five diazotrophic strains on growth promotion and nitrate reductase (NR, EC 1.7.1.1) activity in sugarcane. An experiment was carried out from three stages of cultivation: sprouting, tubes, and in hydroponics. On the first two stages, seven treatments were adopted: uninoculated control; mixed inoculation with five strains; and individual inoculation with Gluconacetobacter diazotrophicus (Gd), Herbaspirillum rubrisubalbicans (Hr), Herbaspirillum seropedicae (Hs), Nitrospirillum amazonense (Na), and Paraburkholderia tropica (Pt). The four treatments showing the best performance were transferred to the hydroponic system for analysis of NR activity. Hs, Pt, and the mixture of all strains led to the highest seedling biomass in tubes, followed by Hr. In hydroponics, the mixture and the strain Hr had the highest growth-promoting effect. NR activity was influenced by inoculation only under low N supply conditions, with positive effect of Hr, Pt, and the mixture.

Dark septate endophytic fungi help tomato to acquire nutrients from ground plant material.

Dark septate endophytic (DSE) fungi are facultative biotrophs that associate with hundreds of plant species, contributing to their growth. These fungi may therefore aid in the search for sustainable agricultural practices. However, several ecological functions of DSE fungi need further clarification. The present study investigated the effects of DSE fungi inoculation on nutrient recovery efficiency, nutrient accumulation, and growth of tomato plants fertilized with organic and inorganic N sources. Two experiments were carried out under greenhouse conditions in a randomized blocks design, with five replicates of tomato seedlings grown in pots filled with non-sterile sandy soil. Tomato seedlings (cv. Santa Clara I-5300) inoculated with DSE fungi (isolates A101, A104, and A105) and without DSE fungi (control) were transplanted to pots filled with 12 kg of soil which had previously received finely ground plant material [Canavalia ensiformis (L.)] that was shoot enriched with 0.7 atom % 15N (organic N source experiment) or ammonium sulfate-15N enriched with 1 atom % 15N (mineral N source experiment). Growth indicators, nutrient content, amount of nitrogen (N) in the plant derived from ammonium sulfate-15N or C. ensiformis-15N, and recovery efficiency of 15N, P, and K by plants were quantified 50 days after transplanting. The treatment inoculated with DSE fungi and supplied with an organic N source showed significantly higher recovery efficiency of 15N, P, and K. In addition, the 15N, N, P, K, Ca, Mg, Fe, Mn, and Zn content, plant height, leaf number, leaf area (only for the A104 inoculation), and shoot dry matter increased. In contrast, the only positive effects observed in the presence of an inorganic N source were fertilizer-K recovery efficiency, content of K, and leaf area when inoculated with the fungus A104. Inoculation with A101, A104, and A105 promoted the growth of tomato using organic N source (finely ground C. ensiformis-15N plant material).