Ricardo Aroca

Regulation of root water uptake under abiotic stress conditions

A common effect of several abiotic stresses is to cause tissue dehydration. Such dehydration is caused by the imbalance between root water uptake and leaf transpiration. Under some specific stress conditions, regulation of root water uptake is more crucial to overcome stress injury than regulation of leaf transpiration. This review first describes present knowledge about how water is taken up by roots and then discusses how specific stress situations such as drought, salinity, low temperature, and flooding modify root water uptake. The rate of root water uptake of a given plant is the result of its root hydraulic characteristics, which are ultimately regulated by aquaporin activity and, to some extent, by suberin deposition. Present knowledge about the effects of different stresses on these features is also summarized. Finally, current findings regarding how molecular signals such as the plant hormones abscisic acid, ethylene, and salicylic acid, and how reactive oxygen species may modulate the final response of root water uptake under stress conditions are discussed.

Regulation by arbuscular mycorrhizae of the integrated physiological response to salinity in plants: new challenges in physiological and molecular studies

Excessive salt accumulation in soils is a major ecological and agronomical problem, in particular in arid and semi-arid areas. Excessive soil salinity affects the establishment, development, and growth of plants, resulting in important losses in productivity. Plants have evolved biochemical and molecular mechanisms that may act in a concerted manner and constitute the integrated physiological response to soil salinity. These include the synthesis and accumulation of compatible solutes to avoid cell dehydration and maintain root water uptake, the regulation of ion homeostasis to control ion uptake by roots, compartmentation and transport into shoots, the fine regulation of water uptake and distribution to plant tissues by the action of aquaporins, the reduction of oxidative damage through improved antioxidant capacity and the maintenance of photosynthesis at values adequate for plant growth. Arbuscular mycorrhizal (AM) symbiosis can help the host plants to cope with the detrimental effects of high soil salinity. There is evidence that AM symbiosis affects and regulates several of the above mentioned mechanisms, but the molecular bases of such effects are almost completely unknown. This review summarizes current knowledge about the effects of AM symbiosis on these physiological mechanisms, emphasizing new perspectives and challenges in physiological and molecular studies on salt-stress alleviation by AM symbiosis.

Salinity stress alleviation using arbuscular mycorrhizal fungi. A review

Salinity is one of the most severe environmental stress as it decreases crop production of more than 20% of irrigated land worldwide. Hence, it is important to develop salt-tolerant crops. Understanding the mechanisms that enable plant growth under saline conditions is therefore required. Acclimation of plants to salinized conditions depends upon activation of cascades of molecular networks involved in stress sensing, signal transduction, and the expression of specific stress-related genes and metabolites. The stress signal is first perceived at the membrane level by the receptors and then transduced in the cell to switch on the stress-responsive genes which mediate stress tolerance. In addition to stress-adaptative mechanisms developed by plants, arbuscular mycorrhizal fungi have been shown to improve plant tolerance to abiotic environmental factors such as salinity. In this review, we emphasize the significance of arbuscular mycorrhizal fungi alleviation of salt stress and their beneficial effects on plant growth and productivity. Although salinity can affect negatively arbuscular mycorrhizal fungi, many reports show improved growth and performance of mycorrhizal plants under salt stress conditions. These positive effects are explained by improved host plant nutrition, higher K+/Na+ ratios in plant tissues and a better osmotic adjustment by accumulation of compatible solutes such as proline, glycine betaine, or soluble sugars. Arbuscular mycorrhizal plants also improve photosynthetic- and water use efficiency under salt stress. Arbuscular mycorrhizal plants enhance the activity of antioxidant enzymes in order to cope with the reactive oxygen species generated by salinity. At the molecular level, arbuscular mycorrhizal symbiosis regulates the expression of plant genes involved in the biosynthesis of proline, of genes encoding aquaporins, and of genes encoding late embryogenesis abundant proteins, with chaperone activity. The regulation of these genes allows mycorrhizal plants to maintain a better water status in their tissues. Gene expression patterns suggest that mycorrhizal plants are less strained by salt stress than non-mycorrhizal plants. In contrast, scarce information is available on the possible regulation by the arbuscular mycorrhizal symbiosis of plant genes encoding Na+/H+ antiporters or cyclic nucleotide-gated channels. These genes encode proteins with a key role in the regulation of uptake, distribution and compartimentation of sodium and other ions within the plant, and are major determinants for the salt sensitiveness of a plant. Thus, we propose that investigating the participation of cation proton antiporters and cyclic nucleotide-gated channels on arbuscular mycorrhizal symbiosis under salinity is a promising field that should shed further light on new mechanisms involved in the enhanced tolerance of mycorrhizal plants to salt stress.

The Role of Aquaporins and Membrane Damage in Chilling and Hydrogen Peroxide Induced Changes in the Hydraulic Conductance of Maize Roots

When chilling-sensitive plants are chilled, root hydraulic conductance (Lo) declines precipitously; Lo also declines in chilling-tolerant plants, but it subsequently recovers, whereas in chilling-sensitive plants it does not. As a result, the chilling-sensitive plants dry out and may die. Using a chilling-sensitive and a chilling-tolerant maize genotype we investigated the effect of chilling on Lo, and its relationship to osmotic water permeability of isolated root cortex protoplasts, aquaporin gene expression, aquaporin abundance, and aquaporin phosphorylation, hydrogen peroxide (H2O2) accumulation in the roots and electrolyte leakage from the roots. Because chilling can cause H2O2 accumulation we also determined the effects of a short H2O2 treatment of the roots and examined the same parameters. We conclude from these studies that the recovery of Lo during chilling in the chilling-tolerant genotype is made possible by avoiding or repairing membrane damage and by a greater abundance and/or activity of aquaporins. The same changes in aquaporins take place in the chilling-sensitive genotype, but we postulate that membrane damage prevents the Lo recovery. It appears that the aquaporin response is necessary but not sufficient to respond to chilling injury. The plant must also be able to avoid the oxidative damage that accompanies chilling.

PIP aquaporin gene expression in arbuscular mycorrhizal Glycine max and Lactuca sativa plants in relation to drought stress tolerance

Although the discovery of aquaporins in plants has resulted in a paradigm shift in the understanding of plant water relations, the relationship between aquaporins and plant responses to drought still remains elusive. Moreover, the contribution of aquaporin genes to the enhanced tolerance to drought in arbuscular mycorrhisal (AM) plants has never been investigated. Therefore, we studied, at a molecular level, whether the expression of aquaporin-encoding genes in roots is altered by the AM symbiosis as a mechanism to enhance host plant tolerance to water deficit. In this study, genes encoding plasma membrane aquaporins (PIPs) from soybean and lettuce were cloned and their expression pattern studied in AM and nonAM plants cultivated under well-watered or drought stressed conditions. Results showed that AM plants responded to drought stress by down-regulating the expression of the PIP genes studied and anticipating its down-regulation as compared to nonAM plants. The possible physiological implications of this down-regulation of PIP genes as a mechanism to decrease membrane water permeability and to allow cellular water conservation is further discussed.

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.

Mycorrhizal and non-mycorrhizal Lactuca sativa plants exhibit contrasting responses to exogenous ABA during drought stress and recovery

The arbuscular mycorrhizal (AM) symbiosis enhances plant tolerance to water deficit through the alteration of plant physiology and the expression of plant genes. These changes have been postulated to be caused (among others) by different contents of abscisic acid (ABA) between AM and non-AM plants. However, there are no studies dealing with the effects of exogenous ABA on the expression of stress-related genes and on the physiology of AM plants. The aim of the present study was to evaluate the influence of AM symbiosis and exogenous ABA application on plant development, physiology, and expression of several stress-related genes after both drought and a recovery period. Results show that the application of exogenous ABA had contrasting effects on AM and non-AM plants. Only AM plants fed with exogenous ABA maintained shoot biomass production unaltered by drought stress. The addition of exogenous ABA enhanced considerably the ABA content in shoots of non-AM plants, concomitantly with the expression of the stress marker genes Lsp5cs and Lslea and the gene Lsnced. By contrast, the addition of exogenous ABA decreased the content of ABA in shoots of AM plants and did not produce any further enhancement of the expression of these three genes. AM plants always exhibited higher values of root hydraulic conductivity and reduced transpiration rate under drought stress. From plants subjected to drought, only the AM plants recovered their root hydraulic conductivity completely after the 3 d recovery period. As a whole, the results indicate that AM plants regulate their ABA levels better and faster than non-AM plants, allowing a more adequate balance between leaf transpiration and root water movement during drought and recovery.

The arbuscular mycorrhizal symbiosis enhances the photosynthetic efficiency and the antioxidative response of rice plants subjected to drought stress.

Rice (Oryza sativa) is the most important crop for human consumption, providing staple food for more than half of the worlds population. Rice is conventionally grown under flooded conditions for most of its growing cycle. However, about half of the rice area in the world does not have sufficient water to maintain optimal growing conditions and yield is reduced by drought. One possible way to increase rice production in order to meet the rice demand is to improve its drought tolerance by means of the arbuscular mycorrhizal (AM) symbiosis. Thus, AM and non-AM rice plants were maintained under well-watered conditions or were subjected to moderate and severe drought stress for 15d. After that, half of the plants from each treatment were harvested, while the other half were allowed to recover from drought for additional 25d. The results showed that rice can benefit from the AM symbiosis and improve their long-term development after a drought stress period. In fact, at each watering level, AM plants showed about 50% enhanced shoot fresh weight as compared to non-AM plants. The AM symbiosis enhanced the plant photosynthetic efficiency under stress over 40%, induced the accumulation of the antioxidant molecule glutathione and reduced the accumulation of hydrogen peroxide and the oxidative damage to lipids in these plants. Thus, these combined effects enhanced the plant performance after a drought stress period.

Arbuscular mycorrhizal symbiosis increases relative apoplastic water flow in roots of the host plant under both well-watered and drought stress conditions.

BACKGROUND AND AIMS The movement of water through mycorrhizal fungal tissues and between the fungus and roots is little understood. It has been demonstrated that arbuscular mycorrhizal (AM) symbiosis regulates root hydraulic properties, including root hydraulic conductivity. However, it is not clear whether this effect is due to a regulation of root aquaporins (cell-to-cell pathway) or to enhanced apoplastic water flow. Here we measured the relative contributions of the apoplastic versus the cell-to-cell pathway for water movement in roots of AM and non-AM plants. METHODS We used a combination of two experiments using the apoplastic tracer dye light green SF yellowish and sodium azide as an inhibitor of aquaporin activity. Plant water and physiological status, root hydraulic conductivity and apoplastic water flow were measured. KEY RESULTS Roots of AM plants enhanced significantly relative apoplastic water flow as compared with non-AM plants and this increase was evident under both well-watered and drought stress conditions. The presence of the AM fungus in the roots of the host plants was able to modulate the switching between apoplastic and cell-to-cell water transport pathways. CONCLUSIONS The ability of AM plants to switch between water transport pathways could allow a higher flexibility in the response of these plants to water shortage according to the demand from the shoot.

Involvement of abscisic acid in leaf and root of maize (Zea mays L.) in avoiding chilling-induced water stress

In the present study, we investigated the role of abscisic acid (ABA) on chilling tolerance of maize. Two maize genotypes differing in chilling sensitivity (Z7 tolerant and Penjalinan sensitive) were subjected to chilling (5 8C, 12 h photoperiod, 150 mmol m � 2 s � 1 PPFD) for 3 days under two relative humidity (RH) regimes (60 or 100% RH). Some plants were exogenously treated 24 h before chilling with ABA (100 mM). As expected, high humidity (100% RH) or ABA pre-treatment prevented the leaf water deficit induced by chilling at 60% RH in chilling sensitive Penjalinan plants. ABA pre-treatment improved chilling tolerance of Penjalinan plants, mainly by decreasing leaf conductance and by increasing root water flow. At the leaf level, we found a relationship between ABA content and chilling tolerance in both maize genotypes. No relationship between ABA content and leaf conductance was found. Moreover, during chilling, no differences on leaf conductance between the two genotypes were observed, probably indicating that the different water stress suffered by the two genotypes could be linked to differences in the root water uptake. The rise in leaf ABA content during chilling was independent of the leaf water status, so it must be induced by the low temperature per se, and after a longer cold exposure also by the vapour pressure deficit (VPD) (a higher VPD allows more ABA accumulation). At the root level, we did not observe a relationship between the root hydraulic acclimation to chilling and the root ABA content. Z7 plants chilled at 60% RH had the same root ABA content as those which were chilled at 100% RH and as Penjalinan plants; however, the former showed a higher root hydraulic conductance. The rise in the root ABA content in Z7 plants followed the same pattern as observed in the leaves. In Penjalinan plants, the rise in root ABA content was linked only to low temperatures per se, since it increased in the same way in plants chilled under 60 or 100% RH. # 2003 Elsevier Ireland Ltd. All rights reserved.