Hydraulic conductance and water potential gradients in squash leaves showing mycorrhiza-induced increases in stomatal conductance

Stomatal conductance (g _s) and transpiration rates vary widely across plant species. Leaf hydraulic conductance (k _leaf) tends to change with g _s, to maintain hydraulic homeostasis and prevent wide and potentially harmful fluctuations in transpiration-induced water potential gradients across the leaf (ΔΨ _leaf). Because arbuscular mycorrhizal (AM) symbiosis often increases g _s in the plant host, we tested whether the symbiosis affects leaf hydraulic homeostasis. Specifically, we tested whether k _leaf changes with g _s to maintain ΔΨ _leaf or whether ΔΨ _leaf differs when g _s differs in AM and non-AM plants. Colonization of squash plants with Glomus intraradices resulted in increased g _s relative to non-AM controls, by an average of 27% under amply watered, unstressed conditions. Stomatal conductance was similar in AM and non-AM plants with exposure to NaCl stress. Across all AM and NaCl treatments, k _leaf did change in synchrony with g _s (positive correlation of g _s and k _leaf), corroborating leaf tendency toward hydraulic homeostasis under varying rates of transpirational water loss. However, k _leaf did not increase in AM plants to compensate for the higher g _s of unstressed AM plants relative to non-AM plants. Consequently, ΔΨ _leaf did tend to be higher in AM leaves. A trend toward slightly higher ΔΨ _leaf has been observed recently in more highly evolved plant taxa having higher productivity. Higher ΔΨ _leaf in leaves of mycorrhizal plants would therefore be consistent with the higher rates of gas exchange that often accompany mycorrhizal symbiosis and that are presumed to be necessary to supply the carbon needs of the fungal symbiont.     

Contribution of the arbuscular mycorrhizal symbiosis to heavy metal phytoremediation

High concentrations of heavy metals (HM) in the soil have detrimental effects on ecosystems and are a risk to human health as they can enter the food chain via agricultural products or contaminated drinking water. Phytoremediation, a sustainable and inexpensive technology based on the removal of pollutants from the environment by plants, is becoming an increasingly important objective in plant research. However, as phytoremediation is a slow process, improvement of efficiency and thus increased stabilization or removal of HMs from soils is an important goal. Arbuscular mycorrhizal (AM) fungi provide an attractive system to advance plant-based environmental clean-up. During symbiotic interaction the hyphal network functionally extends the root system of their hosts. Thus, plants in symbiosis with AM fungi have the potential to take up HM from an enlarged soil volume. In this review, we summarize current knowledge about the contribution of the AM symbiosis to phytoremediation of heavy metals.     

Mycorrhizal fungi enhancement of growth and gas exchange of micropropagated guava plantlets (Psidium guajava L.) during ex vitro acclimatization and plant establishment

Psidium guajava L.) plantlets was determined during acclimatization and plant establishment. Guava plantlets were asexually propagated through tissue culture and grown in a glasshouse for 18 weeks. Half of the plantlets were inoculated with a mixed endomycorrhiza isolate from Mexico, ZAC-19, containing Glomus diaphanum, G. albidum and G. claroides. Plantlets were fertilized with modified Long Ashton nutrient solution that supplied 11 μg P ml⁻¹. Gas exchange measurements were taken at 2, 4, 8, and 18 weeks after inoculation using a portable photosynthesis system. All micropropagated guava plantlets survived transplant shock. After 6 weeks, mycorrhizal plantlets had greater shoot growth rates and leaf production than non-mycorrhizal plantlets. This also corresponded with increased photosynthetic rates and stomatal conductance of mycorrhizal plants. By 18 weeks, mycorrhizal plantlets had greater shoot length, leaf area, leaf, stem, and root dry mass. However, gas exchange was comparable among treatments, in part because the container size was restricting growth of the larger mycorrhizal plantlets. Non-mycorrhizal plantlets had greater leaf area ratios and specific leaf areas than mycorrhizal plantlets. Increased leaf tissue mineral levels of P, Mg, Cu, and Mo also occurred with mycorrhizal plantlets. Roots of inoculated guava plantlets were heavily colonized with arbuscules, vesicles and endospores. Guava plantlets were highly mycotrophic with a mycorrhizal dependency index of 103%. Accepted: 27 December 1999     

The response of sorghum and sunflower to short-term waterlogging

Summary The effect of waterlogging on water use and nutrient uptake in sunflower and sorghum was investigated in relation to stage of development of the crops and the timing and duration of waterlogging. Waterlogging at the vegetative and floral initiation stages of plant growth induced a reduction in water use of sunflower, with corresponding declines in leaf expansion and leaf water potential; in sorghum, the transpiration rates were much lower than for sunflower and relatively unaffected by waterlogging. Waterlogging at anthesis, however, caused an immediate reduction in water use in sunflower with a similar but delayed effect in sorghum. The differences in response of these two species are discussed in relation to the relative importance of water stress and nutrient uptake. Plant analysis at maturity indicated that waterlogging at any growth stage reduced both total and seed phosphorus in sunflower; similar effects were recorded with sorghum, with the exception of anthesis waterlogging which did not reduce nutrient uptake. Waterlogging effects on plant potassium levels were variable.     

Leaf age and salinity influence water relations of pepper leaves

Plant growth is reduced under saline conditions even when turgor in mature leaves is maintained by osmotic adjustment. The objective of this study was to determine if young leaves from salt-affected plants were also osmotically adjusted. Pepper plants (Capsicum annuum L. cv. California Wonder) were grown in several levels of solution osmotic potential and various components of the plants' water relations were measured to determine if young, rapidly growing leaves could accumulate solutes rapidly enough to maintain turgor for normal cell enlargement. Psychrometric measurements indicated that osmotic adjustment is similar for both young and mature leaves although osmotic potential is slightly lower for young leaves. Total water potential is also lower for young leaves, particularly at dawn for the saline treatments. The result is reduced turgor under saline conditions at dawn for young but not mature leaves. This reduced turgor at dawn, and presumably low night value, is possibly a cause of reduced growth under saline conditions. No differences in leaf turgor occur at midday. Porometer measurements indicated that young leaves at a given salinity level have a higher stomatal conductance than mature leaves, regardless of the time of day. The result of stomatal closure is a linear reduction of transpiration.     

Azospirillum and arbuscular mycorrhizal colonization enhance rice growth and physiological traits under well-watered and drought conditions

The response of rice plants to inoculation with an arbuscular mycorrhizal (AM) fungus, Azospirillum brasilense, or combination of both microorganisms, was assayed under well-watered or drought stress conditions. Water deficit treatment was imposed by reducing the amount of water added, but AM plants, with a significantly higher biomass, received the same amount of water as non-AM plants, with a poor biomass. Thus, the water stress treatment was more severe for AM plants than for non-AM plants. The results showed that AM colonization significantly enhanced rice growth under both water conditions, although the greatest rice development was reached in plants dually inoculated under well-watered conditions. Water level did not affect the efficiency of photosystem II, but both AM and A. brasilense inoculations increased this value. AM colonization increased stomatal conductance, particularly when associated with A. brasilense, which enhanced this parameter by 80% under drought conditions and by 35% under well-watered conditions as compared to single AM plants. Exposure of AM rice to drought stress decreased the high levels of glutathione that AM plants exhibited under well-watered conditions, while drought had no effect on the ascorbate content. The decrease of glutathione content in AM plants under drought stress conditions led to enhance lipid peroxidation. On the other hand, inoculation with the AM fungus itself increased ascorbate and proline as protective compounds to cope with the harmful effects of water limitation. Inoculation with A. brasilense also enhanced ascorbate accumulation, reaching a similar level as in AM plants. These results showed that, in spite of the fact that drought stress imposed by AM treatments was considerably more severe than non-AM treatments, rice plants benefited not only from the AM symbiosis but also from A. brasilense root colonization, regardless of the watering level. However, the beneficial effects of A. brasilense on most of the physiological and biochemical traits of rice plants were only clearly visible when the plants were mycorrhized. This microbial consortium was effective for rice plants as an acceptable and ecofriendly technology to improve plant performance and development.     

Water relations and stomatal characteristics of Mediterranean plants with different growth forms and leaf habits: responses to water stress and recovery

The aim of this study was to extent the range of knowledge about water relations and stomatal responses to water stress to ten Mediterranean plants with different growth forms and leaf habits. Plants were subjected to different levels of water stress and a treatment of recovery. Stomatal attributes (stomatal density, StoD), stomatal conductance (g _s), stomatal responsiveness to water stress (SR), leaf water relations (pre-dawn and midday leaf water potential and relative water content), soil to leaf apparent hydraulic conductance (K _L) and bulk modulus of elasticity (ε) were determined. The observed wide range of water relations and stomatal characteristics was found to be partially depended on the growth form. Maximum g _s was related to StoD and the stomatal area index (SAI), while g _s evolution after water stress and recovery was highly correlated with K _L. Relationships between SR to water deficit and other morphological leaf traits, such as StoD, LMA or ε, provided no general correlations when including all species. It is concluded that a high variability is present among Mediterranean plants reflecting a continuum of leaf water relations and stomatal behaviour in response to water stress.     

Comparing contributions of soil versus root colonization to variations in stomatal behavior and soil drying in mycorrhizal Sorghum bicolor and Cucurbita pepo

In prior studies we learned that colonization of soil can be as important as colonization of roots in determining mycorrhizal influence on the water relations of host plants. Here we use a path analysis modeling approach to test (a) whether quantity of hyphae in soil contributes to variations in stomatal behavior and soil drying, and (b) whether soil colonization or root colonization has a stronger influence on these stomatal and soil drying responses. Experiments were performed on Sorghum bicolor and Cucurbita pepo, with soils and roots colonized by a mixture of Glomus intraradices and Gigaspora margarita. Soil colonization generally made more significant contributions to stomatal conductance than did root colonization. Soil colonization did not make significant direct contributions to soil water potential measures (soil water potential at stomatal closure or soil drying rate), whereas root colonization did contribute a potentially important path to each. The findings further support a role for mycorrhization of the soil itself in contributing to the regulation of stomatal behavior of host plants.     

Effects of drought on non-mycorrhizal and mycorrhizal maize: changes in the pools of non-structural carbohydrates, in the activities of invertase and trehalase, and in the pools of amino acids and imino acids

To study the response of non-mycorrhizal and mycorrhizal maize plants to drought, the changes in the pools of non-structural carbohydrates and amino acids were analysed in leaves and roots of two maize cvs. Plants well colonized by the arbuscular mycorrhizal fungus Glomus mosseae (Nicol. & Gerd.) (60% of root length infected) and comparable non-mycorrhizal plants were subjected to moderate drought stress by reducing the water supply. This stress induced a conspicuous increase in the trehalose pool in the mycorrhizal roots, probably because it was accumulated by the fungal symbiont. Furthermore, glucose and fructose were accumulated in leaves and roots of non-mycorrhizal plants but not in the mycorrhizal ones. Starch disappeared completely from the leaves of both mycorrhizal and non-mycorrhizal plants in response to drought. Activities of soluble acid invertase and trehalase were also measured. Acid invertase activity increased during drought in the leaves of both non-mycorrhizal and mycorrhizal plants whilst in the roots it was unaffected in non-mycorrhizal plants and decreased in the mycorrhizal ones. Without drought stress, trehalase activity was considerably higher in the leaves and roots of mycorrhizal plants than in those of non-mycorrhizal plants. It increased conspicuously during drought, primarily in the leaves of non-mycorrhizal plants. A drought-induced accumulation of amino acids as well as imino acids was found in roots and leaves of both mycorrhizal and non-mycorrhizal plants; leaves of mycorrhizal plants accumulated more imino acids than those of non-mycorrhizal ones. Our results show that drought stress and the presence of a mycorrhizal fungus have a considerable effect on carbon partitioning, imino acid and amino acid accumulation in maize plants.     

Yield and yield components of hybrid corn (Zea mays L.) as affected by mycorrhizal symbiosis and zinc sulfate under drought stress

With respect to the significance of improving hybrid corn performance under stress, this experiment was conducted at the Islamic Azad University, Arak Branch, Iran. A complete randomized block design with three levels of irrigations (at 100%, 75% and 50% crop water requirement), two levels of arbuscular mycorrhizal (AM) fungi (Glumus intraradisis) (including control), and three levels of zinc (Zn) sulfate (0, 25 and 45 kg ha⁻¹), was performed. Results of the 2-year experiments indicated that irrigation treatment significantly affected corn yield and its components at P = 1%. AM fungi and increasing Zn levels also resulted in similar effects on corn growth and production. Although AM fungi did not significantly affect corn growth at the non-stressed irrigation treatment, at moderate drought stress AM fungi significantly enhanced corn quality and yield relative to the control treatment. The combined effects of AM fungi and Zn sulfate at 45 kg ha⁻¹ application significantly affected corn growth and production. In addition, the tripartite treatments significantly enhanced corn yield at P = 1%. Effects of Zn and AM fungi on plant growth under drought stress is affected by the stress level.     

Osmotic solute responses of mycorrhizal citrus (<Emphasis Type="Italic">Poncirus trifoliata</Emphasis>) seedlings to drought stress

This study investigated the accumulation of osmotic solutes in citrus (Poncirus trifoliata) seedlings colonized by Glomus versiforme subjected to drought stress or kept well watered. Development of mycorrhizae was higher under well watered than under drought-stressed conditions. Arbuscular mycorrhizal (AM) seedlings accumulated more soluble sugars, soluble starch and total non-structural carbohydrates in leaves and roots than corresponding non-AM seedlings regardless of soil-water status. Glucose and sucrose contents of well-watered and drought-stressed roots, fructose contents of well-watered roots and sucrose contents of drought-stressed leaves were notably higher in AM than in non-AM seedlings. K⁺ and Ca²⁺ levels in AM leaves and roots were greater than those in non-AM leaves and roots, while AM symbiosis did not affect the Mg²⁺ level. AM seedlings accumulated less proline than non-AM seedlings. AM symbiosis altered both the allocation of carbohydrate to roots and the net osmotic solute accumulations in response to drought stress. It is concluded that AM colonization enhances osmotic solute accumulation of trifoliate orange seedlings, thus providing better osmotic adjustment in AM seedlings, which did not correlate with proline but with K⁺, Ca²⁺, Mg²⁺, glucose, fructose and sucrose accumulation.     

Growth response of major USA cowpea cultivars: II. Effect of salinity on leaf gas exchange

In an effort to elucidate the physiological processes involved in cowpea differential growth response of four major USA cowpea cultivars (CB5, CB27, 8517 and 7964) to increasing salinity, we investigated the effect of salinity on leaf gas exchange of net photosynthetic rate per unit leaf mass (Pnm) and per unit leaf area (Pna), and stomatal conductance (gs) of the four cowpea cultivars. The experiment was set up as a standard split-plot design in which cowpea plants were grown in greenhouse sand tanks irrigated with nutrient solutions. Seven salinities ranging from 2.6 to 20.5 dS m⁻¹ were constructed based on Colorado River water salt composition with NaCl, CaCl₂ and MgSO₄ as the salinization salts. Light-saturated Pnm, Pna and gs of fully expanded trifoliage were examined at the vegetative growth and flowering stages, and the data were analyzed using a split-plot analysis of variance (ANOVA) model. We found a highly significant (P ≤ 0.0001) reduction of Pnm, Pna and gs due to salinity. The responses of Pnm, Pna and gs to salinity could be further described by a general model of log(y) = a₁ + a₂x + a₃x², where y represents either Pnm, Pna, or gs; a₁, a₂ and a₃, empirical constants; x, salinity. We found that Pnm was more sensitive to salinity than Pna. Additionally, we found that increasing stomatal closure with increasing salinity might limit Pnm or Pna. While we did not find any significant difference (P > 0.05) of Pnm and Pna among the four cultivars, we did find a significant difference (P ≤ 0.05) in gs. No significant salt × cultivar interaction effect (P > 0.05) was found with Pnm, Pna and gs indicating that the four cowpea cultivars have the same response pattern of their leaf gas exchange to salinity.     

Field response of wheat to arbuscular mycorrhizal fungi and drought stress

Mycorrhizal plants often have greater tolerance to drought than nonmycorrhizal plants. This study was conducted to determine the effects of arbuscular mycorrhizal (AM) fungi inoculation on growth, grain yield and mineral acquisition of two winter wheat (Triticum aestivum L.) cultivars grown in the field under well-watered and water-stressed conditions. Wheat seeds were planted in furrows after treatment with or without the AM fungi Glomus mosseae or G. etunicatum. Roots were sampled at four growth stages (leaf, tillering, heading and grain-filling) to quantify AM fungi. There was negligible AM fungi colonization during winter months following seeding (leaf sampling in February), when soil temperature was low. During the spring, AM fungi colonization increased gradually. Mycorrhizal colonization was higher in well-watered plants colonized with AM fungi isolates than water-stressed plants. Plants inoculated with G. etunicatum generally had higher colonization than plants colonized with G. mosseae under both soil moisture conditions. Biomass and grain yields were higher in mycorrhizal than nonmycorrhizal plots irrespective of soil moisture, and G. etunicatum inoculated plants generally had higher biomass and grain yields than those colonized by G. mosseae under either soil moisture condition. The mycorrhizal plants had higher shoot P and Fe concentrations than nonmycorrhizal plants at all samplings regardless of soil moisture conditions. The improved growth, yield and nutrient uptake in wheat plants reported here demonstrate the potential of mycorrhizal inoculation to reduce the effects of drought stress on wheat grown under field conditions in semiarid areas of the world.     

Response of photosynthesis to high light and drought for Arabidopsis thaliana grown under a UV-B enhanced light regime

Arabidopsis thaliana grown in a light regime that included ultraviolet-B (UV-B) radiation (6 kJ m⁻² d⁻¹) had similar light-saturated photosynthetic rates but up to 50% lower stomatal conductance rates, as compared to plants grown without UV-B radiation. Growth responses of Arabidopsis to UV-B radiation included lower leaf area (25%) and biomass (10%) and higher UV-B absorbing compounds (30%) and chlorophyll content (52%). Lower stomatal conductance rates for plants grown with UV-B radiation were, in part, due to lower stomatal density on the adaxial surface. Plants grown with UV-B radiation had more capacity to down regulate photochemical efficiency of photosystem II (PSII) as shown by up to 25% lower φ_PSII and 30% higher non-photochemical quenching of chlorophyll fluorescence under saturating light. These contributed to a smaller reduction in the maximum photochemical efficiency of PSII (F _v/F _m), greater dark-recovery of F _v/F _m, and higher light-saturated carbon assimilation and stomatal conductance and transpiration rates after a four-hour high light treatment for plants grown with UV-B radiation. Plants grown with UV-B were more tolerant to a 12 day drought treatment than plants grown without UV-B as indicated by two times higher photosynthetic rates and 12% higher relative water content. UV-B-grown plants also had three times higher proline content. Higher tolerance to drought stress for Arabidopsis plants grown under UV-B radiation may be attributed to both increased proline content and decreased stomatal conductance. Growth of Arabidopsis in a UV-B-enhanced light regime increased tolerance to high light exposure and drought stress.     

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.     

Proline accumulation and nitrate reductase activity in contrasting sorghum lines during mid-season drought stress

Six lines of sorghum ( Sorghum bicolor L. Moench) with differing drought resistance (IS 22380, ICSV 213, IS 13441 and SPH 263, resistant and IS 12739 and IS 12744, susceptible) were grown under field conditions in the semi-arid tropics and analysed for proline and nitrate reductase activity (NRA; EC 1.6.6.1) during a mid-season drought. The resistant lines accumulated high levels of proline, while the susceptible lines showed no significant proline accumulation. Most of the proline was accumulated after growth of the plants had ceased. In a separate greenhouse experiment, most of the proline was found in the green rather than the fired portions of leaves. The levels returned to that of irrigated controls within 5 days of rewatering. Proline levels increased as leaf water potential and relative water content fell, and there was no apparent difference among the different sorghum lines with change in plant water status. Susceptible lines accumulated less proline than resistant lines as leaf death occurred at higher water potentials. Proline accumulation may, however, contribute to the immediate recovery of plants from drought. Leaf NRA reached high levels at about 35 days after sowing in both the stressed and irrigated plants, after which it declined. The decline in NRA was more pronounced in the stressed than in the irrigated plants and closely followed changes in the growth rate. Upon rewatering, NRA increased several-fold in all the lines and, in contrast to proline accumulation, genotypic differences in NRA were small, both during stress and upon rewatering. The high sensitivity of NRA to mild drought stress was reflected in the rapid decline of activity with small changes in leaf water potential and relative water content. The results are discussed in the light of a possible role for proline during recovery from drought, and the maintenance of NRA during stress and its recovery upon rewatering.     

Altered physiology in trehalose-producing transgenic tobacco plants: Enhanced tolerance to drought and salinity stresses

Transgenic tobaccoNicotiana tabacum L. var. SR1) plants that over-express theEscherichia coli trehalose-6-phosphate synthase (TPS) gene(otsA) synthesized small amounts of trehalose (<400 μg g^-1 leaf) while non-transformants produced no detectable trehalose. Some transgenic plants expressing a high level ofotsA exhibited stunted growth and morphologically altered leaves. We tested F₂2 homozygous plants devoid of phenotypic changes to determine their physiological responses to dehydration and salinity stresses. All transgenic plants maintained better leaf turgidity under a limited water supply or after treatment with polyethylene glycol (PEG). Furthermore, fresh weight was maintained at higher levels after either treatment. The initial leaf water potential was higher in transgenic plants than non-transformants, but, in both plant types, was decreased to a comparable degree following dehydration. When grown with 250 mM NaCl, transgenic plants exhibited a significant delay in leaf withering and chlorosis, as well as more efficient seed germination. Our results suggest that either trehalose or trehalose-6-phosphate can act as an osmoprotective molecule without maintaining water potential, in contrast to other osmolytes. Furthermore, both appear to protect young embryos under unfavorable water status to ensure subsequent germination.     

Differential response of two Central-European oak species to single and combined stress factors

We studied the effects of repeated defoliation and drought, which are considered to be most effective in triggering oak decline, on the Central-European oak species Quercus robur L. and Quercus petraea [Matt.] Liebl. Three-year-old saplings were subjected to artificial defoliation in two consecutive years and to drought stress, applied singly or in combination. Differences in the morphological, anatomical and physiological responses to these stress factors among treatments and between species were tested in a 3-year experiment. Drought stress lowered the pre-dawn and the afternoon leaf-water potentials in both species, irrespective of additional defoliation. The relative water content of the leaves was unaffected by drought stress; but, in Q. robur, it decreased in trees subjected to defoliation and to the combination of drought stress and defoliation. When repeated defoliation and drought stress were applied within the same growing season, the leaf-to-air difference in the partial pressure of water vapour (w) explained most of the variation in the daily integral of the stomatal conductance to water vapour (g _{s day sum}) in Q. petraea; however, in Q. robur, the hydraulic conductance of the twigs was the most powerful predictor variable. In the same year, the actual hydraulic conductance was correlated with g _{s day sum} in Q. robur, but not in Q. petraea. The studied parameters of gas exchange and biomass production revealed that Q. robur recovered more rapidly from stress than did Q. petraea. We hypothesize that this is due to Q. roburs lower responsiveness to the environmental variable w and its higher responsiveness to internal hydraulic conductance compared to Q. petraea. This revised version was published online in May 2005 with corrections to Table 4.