Potassium transport in salt-stressed barley roots

Salinization of the medium inhibits both K⁺ uptake by excised barley (Hordeum vulgare L.) roots and K⁺ release from their stele, as measured by short-term ⁸⁶Rb uptake and xylem exudation, respectively. Although inhibition was not specific to chloride, mannitol caused a different response from that of inorganic sodium salts, indicating that inhibition was at least partly the result of an ion effect. In roots previously exposed to low levels of NaCl, NaCl stress directly affected stelar K⁺ release, whereas in low-sodium roots stelar K⁺ release was much less salt-sensitive than K⁺ uptake.Abbreviation chCl choline chloride     

Photosynthetic response of drought- and salt-stressed tomato and turnip rape plants to foliar-applied glycinebetaine

The effect of foliar application of glycinebetaine (50 and 100 m M ) on photosynthesis, stomatal conductance, photorespiration and transpiration in tomato cv. Bos 3155 ( Lycopersicon esculentum Mill.) and summer turnip rape cv. Kulta ( Brassica rapa L. ssp. oleifera ) plants subjected to drought and salinity are reported. Glycinebetaine application increased net photosynthesis of stressed plants. This was mostly due to increased stomatal conductance following glycinebetaine application as there were no marked changes in light and CO₂ saturated rates of O₂ evolution. Moreover, glycinebetaine application resulted in a significant decrease of photorespiration both in drought- and salt-stressed plants.     

Changes in physiology and protein abundance in salt-stressed wheat chloroplasts

Leaves are the final site of salinity perception through the roots. To better understand how wheat chloroplasts proteins respond to salt stress, the study aimed to the physiochemical and comparative proteomics analysis. Seedlings (12-days-old) were exposed to 150 mM NaCl for 1, 2, or 3 days. Na(+) ions were rapid and excessively increase in roots, stems and leaves. Photosynthesis and transpiration rate, stomatal conductance, and relative water content decreased whereas the level of proline increased. Statistically significant positive correlations were found among the content of hydrogen peroxide, activity of catalase, and superoxide dismutase under salt stress in wheat. Protein abundance within the chloroplasts was examined by two-dimensional electrophoresis. More than 100 protein spots were reproducibly detected on each gel, 21 protein spots were differentially expressed during salt treatment. Using linear quadruple trap-Fourier transform ion cyclotron resonance (LTQ-FTICR) hybrid mass spectrometry, 65 unique proteins assigned in the differentially abundant spots. Most proteins were up-regulated at 2 and 3 days after being down-regulated at 1 day. Others showed only slight responses after 3 days of treatment, including Rubisco, glutamate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, isocitrate dehydrogenase, photosystem I, and pyridoxal biosynthesis protein PDX1.2 and PDX1.3. The ATP synthase (α, β, and γ) and V-type proton ATPase subunits were down-regulated resulting showed negative impact by Na(+) on the photosynthetic machinery. This ephemeral increase and subsequent decrease in protein contents may demonstrate a counterbalancing influence of identified proteins. Several proteins such as cytochrome b6-f (Cyt b6-f), germin-like-protein, the γ-subunit of ATP synthase, glutamine synthetase, fructose-bisphosphate aldolase, S-adenosylmethionine synthase, carbonic anhydrase were gradually up-regulated during the period of treatment, which can be identified as marker proteins.     

Effects of sodium, potassium and calcium on salt-stressed barley

We grew barley ( Hordeum vulgare L. CM 72) for a 28-day period and sequentially harvested plants every 3 or 4 days. Plants were salt-stressed with either NaCl or KCl (125 m M ) with or without supplemental Ca (10 or 0.4 m M final concentration, respectively). We determined tissue concentrations of Na, Ca, Mg, K. S, P, Fe, Mn, Cu and Zn for each harvest date by inductively coupled plasma spectrometry. Uptake (specific absorption rate) was calculated from the element content and growth rates. Salinity had significant effects on the uptake and concentrations of most elements. Mg and Mn concentrations declined with time. The concentrations of all other elements determined increased over time. Element uptake on a root dry weight basis declined with time. Three variables were significantly affected by salinity and correlated with growth; 1) the Ca concentration, 2) the total sum of the cation concentration (TC), and 3) the Mn concentration of the shoot. Salinity reduced Ca uptake and concentrations. Supplemental Ca increased Ca concentrations and was positively correlated with growth during salt stress. Salinity doubled TC, which was negatively correlated with relative growth rate (RGR). Relative growth rate declined at TC values above 150 m M . Salinity reduced the uptake and concentration of Mn. Manganese concentrations in the shoot were highly correlated with RGR. Relative growth rate declined at Mn concentrations below 50 nmol (g fresh weight)⁻¹.     

Analysis of the complete genome sequence of Brevibacterium frigoritolerans ZB201705 isolated from drought- and salt-stressed rhizosphere soil of maize

Purpose

To analyze the complete genome sequence of the Brevibacterium frigoritolerans ZB201705, a Brevibacterium strain was isolated from the maize rhizosphere in drought- and salt-stressed soil, and the activity of the strain under simulated drought and high salt conditions was assessed.

Methods

We used a combination of the PacBio RS and Illumina sequencing platforms to obtain the complete genome sequence of B. frigoritolerans ZB201705.

Results

The genome consists of 5,475,560 bp in a linear chromosome with no gaps, 4,391 protein-coding sequences, 39 ribosomal RNAs, and 81 transfer RNAs. The genome analysis revealed many putative gene clusters involved in defense mechanisms. In addition, an activity analysis of the strain under high-salt and simulated drought conditions helped clarify its potential tolerance to these abiotic stresses.

Conclusions

Our data revealed the complete genome sequence of the new isolated strain, and showed that it produces many proteins involved in drought and salt stress responses, suggesting that B. frigoritolerans ZB201705 may be a potential factor to increase crop yield under abiotic stresses. The information provided here on the genome of B. frigoritolerans ZB201705 provides valuable insight into rhizobacteria-mediated plant salt and drought tolerance and rhizobacteria-based solutions for agriculture under abiotic stress.

     

Systematic identification of mRNAs recruited to argonaute 2 by specific microRNAs and corresponding changes in transcript abundance

microRNAs (miRNAs) are small non-coding RNAs that regulate mRNA stability and translation through the action of the RNAi-induced silencing complex (RISC). Our current understanding of miRNA function is inferred largely from studies of the effects of miRNAs on steady-state mRNA levels and from seed match conservation and context in putative targets. Here we have taken a more direct approach to these issues by comprehensively assessing the miRNAs and mRNAs that are physically associated with Argonaute 2 (Ago2), which is a core RISC component. We transfected HEK293T cells with epitope-tagged Ago2, immunopurified Ago2 together with any associated miRNAs and mRNAs, and quantitatively determined the levels of these RNAs by microarray analyses. We found that Ago2 immunopurified samples contained a representative repertoire of the cell's miRNAs and a select subset of the cell's total mRNAs. Transfection of the miRNAs miR-1 and miR-124 caused significant changes in the association of scores of mRNAs with Ago2. The mRNAs whose association with Ago2 increased upon miRNA expression were much more likely to contain specific miRNA seed matches and to have their overall mRNA levels decrease in response to the miRNA transfection than expected by chance. Hundreds of mRNAs were recruited to Ago2 by each miRNA via seed sequences in 3'-untranslated regions and coding sequences and a few mRNAs appear to be targeted via seed sequences in 5'-untranslated regions. Microarray analysis of Ago2 immunopurified samples provides a simple, direct method for experimentally identifying the targets of miRNAs and for elucidating roles of miRNAs in cellular regulation.     

Root plasma membrane transporters controlling K+/Na+ homeostasis in salt-stressed barley

Plant salinity tolerance is a polygenic trait with contributions from genetic, developmental, and physiological interactions, in addition to interactions between the plant and its environment. In this study, we show that in salt-tolerant genotypes of barley (Hordeum vulgare), multiple mechanisms are well combined to withstand saline conditions. These mechanisms include: (1) better control of membrane voltage so retaining a more negative membrane potential; (2) intrinsically higher H(+) pump activity; (3) better ability of root cells to pump Na(+) from the cytosol to the external medium; and (4) higher sensitivity to supplemental Ca(2+). At the same time, no significant difference was found between contrasting cultivars in their unidirectional (22)Na(+) influx or in the density and voltage dependence of depolarization-activated outward-rectifying K(+) channels. Overall, our results are consistent with the idea of the cytosolic K(+)-to-Na(+) ratio being a key determinant of plant salinity tolerance, and suggest multiple pathways of controlling that important feature in salt-tolerant plants.     

Supplemental manganese improves the relative growth, net assimilation and photosynthetic rates of salt-stressed barley

Previous results in our laboratory indicated that a reduced Mn concentration in the leaves of barley was highly correlated with the reduced relative growth and net assimilation rates of salt-stressed plants. If Mn deficiency limits the growth of salt-stressed barley, then increasing leaf Mn concentrations should increase growth. In the present study, the effect of supplemental Mn on the growth of salt-stressed barley ( Hordeum vulgare L. cv. CM 72) was tested to determine if a salinity-induced Mn deficiency was limiting growth. Plants were salinized with 125 mol m⁻³ NaCl and 9.6 mol m⁻³ CaCl₂. Supplemental Mn was applied in 2 ways: 1) by increasing the Mn concentration in the solution culture and 2) by spraying Mn solutions directly onto the leaves. Growth was markedly inhibited at this salinity level. Dry matter production was increased 100% in salt-stressed plants treated with supplemental Mn to about 32% of the level of nonsalinized controls. The optimum solution culture concentration was 2.0 mmol m⁻³, and the optimum concentration applied to the leaves was 5.0 mol m⁻³. Supplemental Mn did not affect the growth of control plants. Further experiments showed that supplemental Mn increased Mn concentrations and uptake to the shoot. Supplemental Mn increased the relative growth rate of salt-stressed plants and this increase was attributed to an increase in the net assimilation rate; there were no significant effects on the leaf area ratio. Supplemental Mn also increased the net photosynthetic rate of salt-stressed plants. The data support the hypothesis that salinity induced a Mn deficiency in the shoot, which partially reduced photosynthetic rates and growth.     

Effects of sodium, potassium and calcium on salt-stressed barley. I. Growth analysis

Barley ( Hordeum vulgare L. cv. CM 72) was grown for a 28-day period and stressed with treatments of 125 mol m⁻³ NaCl or KC1 with low Ca²⁺ (0.4 mol m⁻³ Ca²⁺) or high Ca²⁺ (10 mol m⁻³ Ca²⁺). Plants were harvested periodically so that relative growth rate (RGR), net assimilation rate (NAR) and leaf area ratio (LAR) could be calculated using the functional approach to plant growth analysis. Relative growth rate declined with time for all treatments, including controls. Salinity inhibited RGR relative to control values by day 10. High Ca²⁺ improved the growth of salt-stressed plants in both NaCl-salinity and KCl-salinity. KC1 proved more toxic than NaCl, especially for KCI-salinity plants with low Ca²⁺, which died by day 28. Net assimilation rate, but not LAR, was highly correlated with RGR for all treatments. This indicates that the photosynthetic-assimilatory machinery was limiting RGR and not the leaf area of the plant.     

The biophysics of leaf growth in salt-stressed barley. A study at the cell level

Biophysical parameters potentially involved in growth regulation were studied at the single-cell level in the third leaf of barley (Hordeum vulgare) after exposure to various degrees of NaCl stress for 3 to 5 d. Gradients of elongation growth were measured, and turgor pressure, osmolality, and water potentials (psi) were determined (pressure probe and picoliter osmometry) in epidermal cells of the elongation zone and the mature blade. Cells in the elongation zone adjusted to decreasing external psi through increases in cell osmolality that were accomplished by increased solute loads and reduced water contents. Cell turgor changed only slightly. In contrast, decreases in turgor also contributed significantly to psi adjustment in the mature blade. Solute deposition rates in the elongation zone increased at moderate stress levels as compared with control conditions, but decreased again at more severe NaCl exposure. Growth-associated psi gradients between expanding epidermal cells and the xylem were significant under control and moderate stress conditions (75 mM NaCl) but seemed negligible at severe stress (120 mM NaCl). We conclude that leaf cell elongation in NaCl-treated barley is probably limited by the rate at which solutes can be taken up to generate turgor, particularly at high NaCl levels.