Angel M. Zamarreño

Arbuscular mycorrhizal symbiosis influences strigolactone production under salinity and alleviates salt stress in lettuce plants.

Arbuscular mycorrhizal (AM) symbiosis can alleviate salt stress in plants. However the intimate mechanisms involved, as well as the effect of salinity on the production of signalling molecules associated to the host plant-AM fungus interaction remains largely unknown. In the present work, we have investigated the effects of salinity on lettuce plant performance and production of strigolactones, and assessed its influence on mycorrhizal root colonization. Three different salt concentrations were applied to mycorrhizal and non-mycorrhizal plants, and their effects, over time, analyzed. Plant biomass, stomatal conductance, efficiency of photosystem II, as well as ABA content and strigolactone production were assessed. The expression of ABA biosynthesis genes was also analyzed. AM plants showed improved growth rates and a better performance of physiological parameters such as stomatal conductance and efficiency of photosystem II than non-mycorrhizal plants under salt stress since very early stages - 3 weeks - of plant colonization. Moreover, ABA levels were lower in those plants, suggesting that they were less stressed than non-colonized plants. On the other hand, we show that both AM symbiosis and salinity influence strigolactone production, although in a different way in AM and non-AM plants. The results suggest that AM symbiosis alleviates salt stress by altering the hormonal profiles and affecting plant physiology in the host plant. Moreover, a correlation between strigolactone production, ABA content, AM root colonization and salinity level is shown. We propose here that under these unfavourable conditions, plants increase strigolactone production in order to promote symbiosis establishment to cope with salt stress.

Arbuscular mycorrhizal symbiosis induces strigolactone biosynthesis under drought and improves drought tolerance in lettuce and tomato

Arbuscular mycorrhizal (AM) symbiosis alleviates drought stress in plants. However, the intimate mechanisms involved, as well as its effect on the production of signalling molecules associated with the host plant-AM fungus interaction remains largely unknown. In the present work, the effects of drought on lettuce and tomato plant performance and hormone levels were investigated in non-AM and AM plants. Three different water regimes were applied, and their effects were analysed over time. AM plants showed an improved growth rate and efficiency of photosystem II than non-AM plants under drought from very early stages of plant colonization. The levels of the phytohormone abscisic acid, as well as the expression of the corresponding marker genes, were influenced by drought stress in non-AM and AM plants. The levels of strigolactones and the expression of corresponding marker genes were affected by both AM symbiosis and drought. The results suggest that AM symbiosis alleviates drought stress by altering the hormonal profiles and affecting plant physiology in the host plant. In addition, a correlation between AM root colonization, strigolactone levels and drought severity is shown, suggesting that under these unfavourable conditions, plants might increase strigolactone production in order to promote symbiosis establishment to cope with the stress.

Involvement of plant endogenous ABA in Bacillus megaterium PGPR activity in tomato plants

BackgroundPlant growth-promoting rhizobacteria (PGPR) are naturally occurring soil bacteria which benefit plants by improving plant productivity and immunity. The mechanisms involved in these processes include the regulation of plant hormone levels such as ethylene and abscisic acid (ABA). The aim of the present study was to determine whether the activity of Bacillus megaterium PGPR is affected by the endogenous ABA content of the host plant. The ABA-deficient tomato mutants flacca and sitiens and their near-isogenic wild-type parental lines were used. Growth, stomatal conductance, shoot hormone concentration, competition assay for colonization of tomato root tips, and root expression of plant genes expected to be modulated by ABA and PGPR were examined.ResultsContrary to the wild-type plants in which PGPR stimulated growth rates, PGPR caused growth inhibition in ABA-deficient mutant plants. PGPR also triggered an over accumulation of ethylene in ABA-deficient plants which correlated with a higher expression of the pathogenesis-related gene Sl-PR1b.ConclusionsPositive correlation between over-accumulation of ethylene and a higher expression of Sl-PR1b in ABA-deficient mutant plants could indicate that maintenance of normal plant endogenous ABA content may be essential for the growth promoting action of B. megaterium by keeping low levels of ethylene production.

The importance of nitrate in ameliorating the effects of ammonium and urea nutrition on plant development: the relationships with free polyamines and plant proline contents

In order to investigate the possible involvement of free polyamines and proline in the mechanism underlying the action of nitrate in correcting the negative effects associated with ammonium and urea nutrition in certain plant species, we studied plant contents of free polyamines and proline associated with nitrogen nutrition involving different nitrogen forms (nitrate, ammonium, urea) in two plant species, wheat and pepper. The results showed that ammonium nutrition and, to a lesser extent, urea nutrition were associated with significant increases in plant putrescine content that were well correlated with reductions in plant growth. These negative effects of ammonium and urea nutrition were corrected by the presence of nitrate in the nutrient solution; the presence of nitrate was also related to a significant decrease in the plant putrescine content. These results are compatible with a specific effect of nitrate reducing ammonium accumulation through the improvement of ammonium assimilation. As for the plant proline content, in pepper a slight increase in this parameter was associated with ammonium and urea nutrition, but it was also decreased by the presence of nitrate in the nutrient solution. These changes, however, were not so clearly related to the variations in plant growth as in the case of putrescine content. These results are compatible with the hypothesis that putrescine biosynthesis might be related to proline degradation by a specific pathway related to ammonium detoxification.

Auxin: A major player in the shoot-to-root regulation of root Fe-stress physiological responses to Fe deficiency in cucumber plants

The aim of this study was to investigate the effects of IAA and ABA in the shoot-to-root regulation of the expression of the main Fe-stress physiological root responses in cucumber plants subjected to shoot Fe functional deficiency. Changes in the expression of the genes CsFRO1, CsIRT1, CsHA1 and CsHA2 (coding for Fe(III)-chelate reductase (FCR), the Fe(II) transporter and H+-ATPase, respectively) and in the enzyme activity of FCR and the acidification capacity were measured. We studied first the ability of exogenous applications of IAA and ABA to induce these Fe-stress root responses in plants grown in Fe-sufficient conditions. The results showed that IAA was able to activate these responses at the transcriptional and functional levels, whereas the results with ABA were less conclusive. Thereafter, we explored the role of IAA in plants with or without shoot Fe functional deficiency in the presence of two types of IAA inhibitors, affecting either IAA polar transport (TIBA) or IAA functionality (PCIB). The results showed that IAA is involved in the regulation at the transcriptional and functional levels of both Fe root acquisition (FCR, Fe(II) transport) and rhizosphere acidification (H+-ATPase), although through different, and probably complementary, mechanisms. These results suggest that IAA is involved in the shoot-to-root regulation of the expression of Fe-stress physiological root responses.

Enhancement of root hydraulic conductivity by methyl jasmonate and the role of calcium and abscisic acid in this process.

The role of jasmonic acid in the induction of stomatal closure is well known. However, its role in regulating root hydraulic conductivity (L) has not yet been explored. The objectives of the present research were to evaluate how JA regulates L and how calcium and abscisic acid (ABA) could be involved in such regulation. We found that exogenous methyl jasmonate (MeJA) increased L of Phaseolus vulgaris, Solanum lycopersicum and Arabidopsis thaliana roots. Tomato plants defective in JA biosynthesis had lower values of L than wild-type plants, and that L was restored by addition of MeJA. The increase of L by MeJA was accompanied by an increase of the phosphorylation state of the aquaporin PIP2. We observed that MeJA addition increased the concentration of cytosolic calcium and that calcium channel blockers inhibited the rise of L caused by MeJA. Treatment with fluoridone, an inhibitor of ABA biosynthesis, partially inhibited the increase of L caused by MeJA, and tomato plants defective in ABA biosynthesis increased their L after application of MeJA. It is concluded that JA enhances L and that this enhancement is linked to calcium and ABA dependent and independent signalling pathways.

Arbuscular mycorrhizal symbiosis and methyl jasmonate avoid the inhibition of root hydraulic conductivity caused by drought

Hormonal regulation and symbiotic relationships provide benefits for plants to overcome stress conditions. The aim of this study was to elucidate the effects of exogenous methyl jasmonate (MeJA) application on root hydraulic conductivity (L) of Phaseolus vulgaris plants which established arbuscular mycorrhizal (AM) symbiosis under two water regimes (well-watered and drought conditions). The variation in endogenous contents of several hormones (MeJA, JA, abscisic acid (ABA), indol-3-acetic acid (IAA), salicylic acid (SA)) and the changes in aquaporin gene expression, protein abundance and phosphorylation state were analyzed. AM symbiosis decreased L under well-watered conditions, which was partially reverted by the MeJA treatment, apparently by a drop in root IAA contents. Also, AM symbiosis and MeJA prevented inhibition of L under drought conditions, most probably by a reduction in root SA contents. Additionally, the gene expression of two fungal aquaporins was upregulated under drought conditions, independently of the MeJA treatment. Plant aquaporin gene expression could not explain the behaviour of L. Conversely, evidence was found for the control of L by phosphorylation of aquaporins. Hence, MeJA addition modified the response of L to both AM symbiosis and drought, presumably by regulating the root contents of IAA and SA and the phosphorylation state of aquaporins.

Changes in the C/N balance caused by increasing external ammonium concentrations are driven by carbon and energy availabilities during ammonium nutrition in pea plants: the key roles of asparagine synthetase and anaplerotic enzymes

An understanding of the mechanisms underlying ammonium (NH(4)(+)) toxicity in plants requires prior knowledge of the metabolic uses for nitrogen (N) and carbon (C). We have recently shown that pea plants grown at high NH(4)(+) concentrations suffer an energy deficiency associated with a disruption of ionic homeostasis. Furthermore, these plants are unable to adequately regulate internal NH4(+) levels and the cell-charge balance associated with cation uptake. Herein we show a role for an extra-C application in the regulation of C-N metabolism in NH(4)(+) -fed plants. Thus, pea plants (Pisum sativum) were grown at a range of NH(4)(+) concentrations as sole N source, and two light intensities were applied to vary the C supply to the plants. Control plants grown at high NH(4)(+) concentration triggered a toxicity response with the characteristic pattern of C-starvation conditions. This toxicity response resulted in the redistribution of N from amino acids, mostly asparagine, and lower C/N ratios. The C/N imbalance at high NH(4)(+) concentration under control conditions induced a strong activation of root C metabolism and the upregulation of anaplerotic enzymes to provide C intermediates for the tricarboxylic acid cycle. A high light intensity partially reverted these C-starvation symptoms by providing higher C availability to the plants. The extra-C contributed to a lower C4/C5 amino acid ratio while maintaining the relative contents of some minor amino acids involved in key pathways regulating the C/N status of the plants unchanged. C availability can therefore be considered to be a determinant factor in the tolerance/sensitivity mechanisms to NH(4)(+) nutrition in plants.

The Symbiosis with the Arbuscular Mycorrhizal Fungus Rhizophagus irregularis Drives Root Water Transport in Flooded Tomato Plants

It is known that the presence of arbuscular mycorrhizal fungi within the plant roots enhances the tolerance of the host plant to different environmental stresses, although the positive effect of the fungi in plants under waterlogged conditions has not been well studied. Tolerance of plants to flooding can be achieved through different molecular, physiological and anatomical adaptations, which will affect their water uptake capacity and therefore their root hydraulic properties. Here, we investigated the root hydraulic properties under non-flooded and flooded conditions in non-mycorrhizal tomato plants and plants inoculated with the arbuscular mycorrhizal fungus Rhizophagus irregularis. Only flooded mycorrhizal plants increased their root hydraulic conductivity, and this effect was correlated with a higher expression of the plant aquaporin SlPIP1;7 and the fungal aquaporin GintAQP1. There was also a higher abundance of the PIP2 protein phoshorylated at Ser280 in mycorrhizal flooded plants. The role of plant hormones (ethylene, ABA and IAA) in root hydraulic properties was also taken into consideration, and it was concluded that, in mycorrhizal flooded plants, ethylene has a secondary role regulating root hydraulic conductivity whereas IAA may be the key hormone that allows the enhancement of root hydraulic conductivity in mycorrhizal plants under low oxygen conditions.

The signal effect of nitrate supply enhances active forms of cytokinins and indole acetic content and reduces abscisic acid in wheat plants grown with ammonium

Ammonium can result in toxicity symptoms in many plants when supplied as a sole nitrogen source. Nitrate reduces the negative effects caused by ammonium and promotes plant growth. In order to explore the mechanism responsible of this beneficial effect, we investigated whether nitrate application causes significant changes in the indoleacetic acid (IAA)- and cytokinin-plant distribution and abscisic acid (ABA) accumulation in wheat (Triticum aestivum L.) plants grown with ammonium. Two different doses of nitrate were supplied to ammonium-fed plants (100 microM and 5mM), to determine whether the effects of nitrate require significant doses (nutritional character), or can be promoted by very low doses (signal effect). The results showed that the presence of NO(3)(-) was associated with clear increases in the active forms of cytokinins (zeatine (Z), trans-zeatine riboside (tZR), isopentenyl adenosine (IPR)) and reduction of the levels of the lower active forms (cis-zeatine riboside (cZR)), independently of the dose applied. Likewise, the presence of nitrate also enhanced IAA shoot content, which correlated with higher cytokinin levels and a tendency toward lower ABA concentration. This study presents further evidence that the possible signal effect of NO(3)(-) involved in its beneficial effect on the growth of wheat plants fed with NH(4)(+) could be mediated by a coordinated action of the levels of cytokinins, IAA and ABA in the shoot.