Nitrogen fixation control under drought stress. Localized or systemic?

Legume-Rhizobium nitrogen fixation is dramatically affected under drought and other environmental constraints. However, it has yet to be established as to whether such regulation of nitrogen fixation is only exerted at the whole-plant level (e.g. by a systemic nitrogen feedback mechanism) or can also occur at a local nodule level. To address this question, nodulated pea (Pisum sativum) plants were grown in a split-root system, which allowed for half of the root system to be irrigated at field capacity, while the other half was water deprived, thus provoking changes in the nodule water potential. Nitrogen fixation only declined in the water-deprived, half-root system and this result was correlated with modifications in the activities of key nodule's enzymes such as sucrose synthase and isocitrate dehydrogenase and in nodular malate content. Furthermore, the decline in nodule water potential resulted in a cell redox imbalance. The results also indicate that systemic nitrogen feedback signaling was not operating in these water-stressed plants, since nitrogen fixation activity was maintained at control values in the watered half of the split-root plants. Thus, the use of a partially droughted split-root system provides evidence that nitrogen fixation activity under drought stress is mainly controlled at the local level rather than by a systemic nitrogen signal.     

Nitrate uptake ability by maize roots during and after drought stress

The effects of different intensities and durations of soil drought and re-watering on the nitrate uptake ability of maize roots were studied. Plants were grown in split-root containers with one part of the root system subjected to different intensities and durations of soil drought and re-watering while the other part of the root system was continuously watered to 23% (w/w) soil water content (70% water capacity). Experiments were performed in split-root containers to maintain a high growth rate, thus ensuring high nutrient demand of the shoot irrespective of the soil water regime. To avoid limitation of nitrate uptake by transport processes in the dry soil, and to ensure a uniform ¹⁴N/¹⁵N ratio at the root surface, ¹⁵N was applied to the roots by placing them into an aerated nutrient solution with 0.5 mM Ca(¹⁵NO₃)₂. Shoot elongation and biomass were only slightly affected by drought in one root compartment when the soil in the other root compartment was kept wet. Therefore, the growth-related nutrient demand of the shoot remained at a high level. At moderate levels of soil drought (10% w/w water content) the ability of the roots for N-uptake was not affected even after 10 d of drought. N-uptake ability was reduced to about 20% of the well-watered control only when the soil water content was decreased to 5%. Total soluble sugar content of the roots increased with increasing soil drought, indicating that low N-uptake ability of roots subjected to severe soil drought was not caused by low assimilate supply from the shoot. Nitrate uptake ability of roots maintained in very dry soil (5% soil water content w/w) even for a prolonged period of 8 d, recovered within 3 d following re-watering. Root growth increased one day after re-watering. A short-term experiment with excised roots formerly subjected to severe soil drought showed that nitrate uptake ability recovered in old and young root segments after 2 d of re-watering. Obviously, the increase in N-uptake ability after re-watering was caused not only by new root growth but also by recovery of the uptake ability of formerly stressed roots.     

Mycorrhizal colonization and drought stress affect Δ13C in CO2‐labeled lettuce plants

We studied the role of different arbuscular‐mycorrhizal (AM) fungi on lettuce (Lactuca sativa L.) plant carbon metabolism under drought stress. Plants were grown in pots maintained at two levels of soil moisture and labeled during photosynthesis with CO₂. P‐fertilized plants were used as a non‐mycorrhizal control. Well‐watered mycorrhizal plants showed similar growth to that of P‐fertilized plants. The level of mycorrhizal root infection was not significantly affected by fungal species or by water treatment. In contrast, important differences in Δ¹³C between P‐fertilized and AM plants were found in shoot and root tissues as a consequence of both water limitation and fungal presence. Δ¹³C in shoots and roots increased in non‐mycorrhizal treatment as compared with the well‐watered plants, whereas this parameter decreased significantly in mycorrhizal plants. Photosynthetic activity was increased in AM plants in well‐watered and droughted plants. G. deserticola was the most beneficial endophyte for water use efficiency in both water treatments. Transpiration rate was not affected by any of the treatments. On the basis of total C in plant tissues, in AM plants the newly fixed C seemed to be preferentially utilized for fungal activity rather than being stored in roots.     

Charcoal and shrubs modify soil processes in ponderosa pine forests of western Montana

In split-root systems of alfalfa (Medicago sativa L.), already existing nodules or arbuscular mycorrhizal roots suppress further establishment of symbiosis in other root parts, a phenomenon named autoregulation. Roots treated with rhizobial nodulation signals (Nod factors) induce a similar systemic suppression of symbiosis.In order to test the hypothesis that flavonoids play a role in this systemic suppression, split-root systems of alfalfa plants were inoculated on one side of the split-root system with Sinorhizobium meliloti or Glomus mosseae or were treated with Nod factor. HPLC-analysis of alfalfa root extracts from both sides of the split-root system revealed a persistent local and systemic accumulation pattern of some flavonoids associated with the different treatments. The two flavonoids, formononetin and ononin, could be identified to be similarily altered after rhizobial or mycorrhizal inoculation or when treated with Nod factor.Exogenous application of formononetin and ononin partially restored nodulation and mycorrhization pointing towards the involvement of these two secondary compounds in the autoregulation of both symbioses.     

Water Transfer in an Alfalfa/Maize Association : Survival of Maize during Drought

We investigated the possibility of interspecific water transfer in an alfalfa (Medicago sativa L.) and maize (Zea mays L.) association. An alfalfa plant was grown through two vertically stacked plastic tubes. A 5 centimeter air gap between tubes was bridged by alfalfa roots. Five-week old maize plants with roots confined to the top tube were not watered, while associated alfalfa roots had free access to water in the bottom tube (the −/+ treatment). Additional treatments included: top and bottom tubes watered (+/+), top and bottom tubes droughted (−/−), and top tube droughted after removal of alfalfa root bridges and routine removal of alfalfa tillers (−^*). Predawn leaf water potential of maize in the −/+ treatment fell to −1.5 megapascals 13 days after the start of drought; thereafter, predawn and midday potentials were maintained near −1.9 megapascals. Leaf water potentials of maize in the −/− and −^* treatments declined steadily; all plants in these treatments were completely desiccated before day 50. High levels of tritium activity were detected in water extracted from both alfalfa and maize leaves after ³H₂O was injected into the bottom −/+ tube at day 70 or later. Maize in the −/+ treatment was able to survive an otherwise lethal period of drought by utilizing water lost by alfalfa roots.     

Stomatal sensitivity of six temperate, deciduous tree species to non-hydraulic root-to-shoot signalling of partial soil drying

The objective of this study were to (1) characterize stomatal response of six deciduous tree species to non-hydraulic, root-sourced signals of soil drying, and (2) test whether species sensitivity to non-hydraulic signalling is allied with their drought avoidance and tolerance profiles. Saplings were grown with roots divided between two pots. Three treatments were compared: one half of the root system watered and half droughted (WD), one half of the root system watered and half severed (WS), both halves watered (WW). Drying about half of the root system caused non-hydraulic declines in stomatal conductance (gs) in all species, with gs of WD plants reduced to from 40% to 60% of WS controls. Declines in stomatal conductance were closely related to declining soil matric potential (m) between -0.01 and -0.10 MPa. Soil m required to cause declines in gs of WD plants to 80% of WS controls varied from a high of -0.013 to a low of -0.044 MPa. Stomatal inhibition varied somewhat with leaf age in half of the species. Leaf osmotic potentials during soil drying were mostly similar among treatments. Although stomatal sensitivity to the non-hydraulic, root-sourced signal (characterized as decline in gs per unit decline in soil ) was not closely correlated with previously identified lethal leaf water potentials or capacity for osmotic adjustment, species having the highest stomatal sensitivity also had the least hydration tolerance. This suggests that stomatal sensitivity to non-hydraulic root signals may be mechanistically linked to a limited extent with other characteristics defining relative species drought tolerance.     

The influence of predawn leaf water potential on stomatal responses to atmospheric water content at constant Ci and on stem hydraulic conductance and foliar ABA concentrations

Stomata and photosynthetic responses to increasing leaf-to-air vapour pressure difference (V) were investigated in watered and droughted Eucalyptus tetrodonta when either ambient CO2 (Ca) or internal CO2 concentration (Ci) were constant. Stem hydraulic conductance and xylem and foliar abscisic acid levels were measured periodically during the drought period.As V increased, stomatal conductance (gs) declined. Maintaining Ci did not affect the response of Gs to V or predawn leaf water potential (pd). In fully watered plants the decline in gs was insufficient to prevent increased transpiration rates (Et) with increasing V. In contrast, in droughted plants, stomatal closure was sufficient to prevent increasing Et. Stomatal sensitivity to increasing V was increased by drought. As drought developed, the three phases of stomatal responses to V progressively collapsed to one where feedforward mechanisms result in decreasing Et with increasing V. Thus as drought developed the feedforward response of stomata to V entirely dominated.Net photosynthesis (Pn) and gs responded in unison to changes in V and pd. The decline in Pn as V increased, despite Ci being maintained constant, was observed in fully watered plants and in severely droughted plants. Pn was most dependent on gs at large values of V and in droughted plants.As drought progressed and pd declined, stem hydraulic conductance decreased and foliar ABA concentrations increased. The decline in maximum gs was correlated with foliar ABA levels. It is concluded here that hydraulic signals, be they atmospheric water or soil water in origin, and possibly also chemical signals regulate gs, which in turn can limit assimilation rates in seasonally dry savannas.Key words: Eucalyptus tetrodonta, stomatal sensitivity, transpiration, hydraulic conductance, ABA.      

Arbuscular mycorrhizal influence on leaf water potential, solute accumulation, and oxidative stress in soybean plants subjected to drought stress

This study investigated several aspects related to drought tolerance in arbuscular mycorrhizal (AM) soybean plants. The investigation included both shoot and root tissues in order to reveal the preferred target tissue for AM effects against drought stress. Non-AM and AM soybean plants were grown under well-watered or drought-stressed conditions, and leaf water status, solute accumulation, oxidative damage to lipids, and other parameters were determined. Results showed that AM plants were protected against drought, as shown by their significantly higher shoot-biomass production. The leaf water potential was also higher in stressed AM plants (-1.9 MPa) than in non-AM plants (-2.5 MPa). The AM roots had accumulated more proline than non-AM roots, while the opposite was observed in shoots. Lipid peroxides were 55% lower in shoots of droughted AM plants than in droughted non-AM plants. Since there was no correlation between the lower oxidative damage to lipids in AM plants and the activity of antioxidant enzymes, it seems that first the AM symbiosis enhanced osmotic adjustment in roots, which could contribute to maintaining a water potential gradient favourable to the water entrance from soil into the roots. This enabled higher leaf water potential in AM plants during drought and kept the plants protected against oxidative stress, and these cumulative effects increased the plant tolerance to drought.     

The effect of root damage caused by simulated white grub attack on the growth,yield and water-use of groundnut plants

White grubs (larval scarabaeids) are now recognized as being important pests of groundnut (peanut) in many parts of the world because of their ability to damage roots. A method of simulating white grub damage to groundnut plants was developed to obtain an indication of how the feeding activity of these insects influences plant growth. The effect of root cutting and drought stress on water uptake and biomass production was evaluated, with roots being cut at three depths in the late vegetative and early podfilling stages. As groundnut plants are often grown under conditions of drought stress, the effects of which would be accentuated by root damage, this factor was introduced into the experiment. Plant water-use measurements indicated that the root systems of plants cut 30 days after emergence (DAE) and watered twice a week became fully functional again after 40 days. Pod and total biomass production were however significantly less than those of the uncut control plants, with drought stress reducing yields below the well-watered controls, particularly when cut at 10 cm below the soil surface. The root systems of plants cut 51 DAE did not regrow to any appreciable extent, and rates of plant water-use remained less than half of the uncut control plants. Over all treatments, there was a strong positive correlation between total (and pod) biomass and plant water-use. It was concluded that the phenological stage of the plant at which root damage occurred had a profound influence on the subsequent recovery in root growth and function, and ultimately on pod yield.     

Long-term drought stress alters nitrogenase activity and carbon translocation in split-root cultured Alnus incana

Split-root cultured grey alder, Alnus incana (L.) Moench., was grown in sand in cuvettes with a continuous supply of nutrient solution. During the drought treatment for up to 9 days the supply of solution was withheld from one of the split-root halves. After 2–3 days of treatment, soil water became depleted and the unwatered root halves were at a constant drought stress, water potential (Ψ_nodules) = -1.1 to -1.6 MPa. Nitrogenase activity in the dry half decreased to about 70% of the initial value during the first 2–3 days and then stayed at this level. The water supply to the shoot from the wet root half was high and only a temporary slight decrease in photosynthesis and stomatal conductance was found in drought-stressed split-root plants. Labelling studies showed a reduced translocation of photoassimilates to the dry nodules. The fixation of CO₂ in the nodules seemed to be more tolerant to drought than nitrogenase activity. During the drought treatment there was an osmotic adjustment from -0.9 to -1.7 MPa, but no change in the storage of starch in the nodules. In alders where parts of the root system is kept dry these roots acclimate and continue a persistent nitrogenase activity.     

Arbuscular mycorrhizal symbiosis modifies the effects of a nitric oxide donor (sodium nitroprusside;SNP) and a nitric oxide synthesis inhibitor (Nω-nitro-L-arginine methyl ester;L-NAME) on lettuce plants under well watered and drought conditions

Arbuscular mycorrhizal (AM) symbiosis is known to help the host plant to overcome environmental stresses as drought by a combination of multiple mechanisms including enhancing of root water uptake capacity. On the other hand, Nitric oxide (NO) is involved in regulating the response of plants to environmental stresses and colonization process of AM fungi. The objective of this research was to study how AM and non-AM lettuce plants responded to a NO donor (sodium nitroprusside; SNP) or to a NO synthesis inhibitor (Nω-nitro-L-arginine methyl ester hydrochloride; L-NAME) under well watered and drought conditions. Most remarkable results were that L-NAME increased the percentage of AM colonized roots under both water regimes and AM plants modified the shoot:root ratio by both chemicals under well watered conditions. Also, the deleterious effects of SNP treatment were partially prevented by AM symbiosis. Moreover, NO could be involved in the diminution of leaf water content under drought conditions, and SNP treatment seems to favor apoplastic water path inside roots. Therefore, different outcomes of relative water content, stomatal conductance and root hydraulic conductivity observed between AM and non-AM plants could be mediated by NO.     

Identification of proteins regulated by ABA in response to combined drought and heat stress in maize roots

We adopted a proteomics approach to identify and analyze the differential expression of maize root proteins associated with abscisic acid (ABA) regulation under combined drought and heat stress. Using mass spectrometry, we identified 22 major proteins that were significantly up-regulated under combined drought and heat stress. These 22 proteins were classified into 6 functional categories: disease/defense (8), metabolism (3), cell growth/division (3), signal transduction (2), transporters (2) and unclassified (4). Our previous reports showed that ABA regulates the expression of several small heat-shock proteins (sHSPs) in maize leaves subjected to the combination of drought and heat stress; however, no sHSPs were identified among the root proteins up-regulated in this study. RT-PCR and western blot analyses were used to identify six known sHSPs. The maize roots were pretreated with 100 μM of ABA, and subsequently, the expression of the 22 up-regulated proteins and 6 sHSPs was examined. 11 proteins were up-regulated in an ABA-dependent manner, 13 proteins were up-regulated in an ABA-independent manner, and 4 proteins were up-regulated but inhibited by ABA. The up-regulated proteins are interesting candidates for further physiological and molecular investigations of combination stress tolerance in maize.     

Rhizosphere carboxylate concentrations of chickpea are affected by soil bulk density

We investigated whether carboxylate exudation by chickpea (Cicer arietinum L.) was affected by soil bulk density and if this effect was local or systemic. We hypothesised that concentrations of carboxylates would increase with distance from the root apex due to continuous and constitutive accumulation of carboxylates, and that exudate accumulation would be greater in a compacted soil than in a loose soil. Plants were grown in split-root or single cylinders containing loose (1400 kg m (-3)) or compacted (1800 kg m (-3)) soil. Rhizosphere carboxylate concentrations were measured of whole root systems as well as of sections along the root. The root diameter was greatest of plants grown in the compacted soil; however, root diameters were the same for both root halves in the split-root design, whether they grew in loose soil or in compacted soil. Similarly, carboxylate concentrations tended to be lower for the whole root system in the compacted soil, but were the same for both root halves in the split-root design, irrespective of whether the roots were in loose soil or in compacted soil. These results indicate that both root diameter and carboxylate exudation by roots in chickpea is regulated systemically via a signal from the shoot rather than by local signals in the roots. There was no accumulation of carboxylates with increasing distance from the apex, probably because microbial degradation occurred at similar rates as carboxylate exudation. Malonate, previously suggested as deterrent to microorganisms, is likely only a selective deterrent.     

Root growth,aerenchyma development,and oxygen transport in rice genotypes subjected to drought and waterlogging

Soils under field conditions may experience fluctuating soil water regimes ranging from drought to waterlogging. The inability of roots to acclimate to such changes in soil water regimes may result in reduced growth and function thereby, dry matter production. This study compared the root and shoot growth, root aerenchyma development, and associated root oxygen transport of aerobic and irrigated lowland rice genotypes grown under well-watered (control), waterlogged, and droughted soil conditions for 30 days. The aerobic genotypes were as tolerant as the irrigated lowland genotypes under waterlogging because of their comparable abilities to enhance aerenchyma that effectively facilitated O₂ diffusion to the roots for maintaining root growth and dry matter production. Under drought, aerobic genotypes were more tolerant than the irrigated lowland genotypes due to their higher ability to maintain nodal root production, elongation, and branching, thus, less reduction in dry matter production. Aerenchyma was also formed in droughted roots regardless of genotypes, but was resistant to internal O₂ transport under O₂ deficiency. The ability of roots to resist temporal variations in drought and waterlogging stresses might have strong implications for the adaptation of rice growing in environments with fluctuating soil water regimes.     

Effects of Drought on CAM and Water Relations in Plants of Peperomia carnevalii

The mechanisms underlying the drought tolerance of Peperomiacarnevalii Steyermark (Piperaceae), a succulent herb growingin the understorey of seasonally dry forests, were examined.Crassulacean acid metabolism (CAM) was studied in the fieldand laboratory, and measurements of water status were made inplants subjected to drought in the greenhouse. Nocturnal acidaccumulation and day and night-time CO₂assimilation rates weregreatest in watered plants and decreased in drought. The proportionof CO₂recycled through CAM in droughted plants, with nocturnalCO₂uptake close to zero, was higher than in watered plants.Maximum quantum yield of chlorophyll fluorescence remained unchangedduring drought, but the PSII quantum yield at the photosyntheticphoton flux density at which the plants were grown was significantlydecreased. Leaf anatomy consists of a chlorophyll-less hydrenchymalocated beneath the upper epidermis, and a two-layered mesophyll.Leaves nearer to the apex are thinner than those nearer to thebase of the shoot. Drought caused a reduction in leaf thicknessdue to shrinkage of the hydrenchyma, but not of the mesophyll.This was associated with the occurrence of a gradient of osmoticpotential between these tissues. Comparison of water loss fromthin leaves of watered and droughted plants, either partly defoliatedat the lower nodes or intact, suggested that water moved fromthe thick to the thin leaves. This process was related to theoccurrence of a gradient of water potential between the thickand the thin leaves. Drought tolerance in P. carnevalii is achievedby the operation of CAM and the occurrence of water movementwithin and between leaves. Copyright 2000 Annals of Botany Company Crassulacean acid metabolism, fluorescence, hydrenchyma, mesophyll, Peperomia carnevalii, water relations     

水分亏缺下玉米根系ZmPIP1亚族基因的表达

在PEG-6000胁迫条件下,以微管蛋白基因为内参基因、水通道蛋白基因ZmPIP1-1和ZmPIP1-2为检测基因,采用半定量逆转录聚合酶链式反应（RT-PCR）体系检测它们在玉米根系中的表达情况。实验结果是：胁迫条件下,ZmPIP1-1的表达量在杂交F,代‘户单4号’（抗旱）和母本‘天四’（抗旱）根系中增多,它的表达量与品种的抗旱性呈正相关,并且胁迫不同时间段它的表达量有差异;而ZmPIP1-2在3个玉米品种的不同水分处理条件下,表达量均没有明显变化。这提示,水分胁迫条件下根系中某些种类的水通道蛋白基因的表达量增多,并且与品种的抗旱性有关;而另一些水通道蛋白基因的表达不受水分亏缺的影响。     

Heterogeneous distribution of phosphorus and potassium in soil influences wheat growth and nutrient uptake

Heterogeneous distribution of mineral nutrients in soil profiles is a norm in agricultural lands, but its influence on nutrient uptake and crop growth is poorly documented. In this study, we examined the effects of varying phosphorus (P) and potassium (K) distribution on plant growth and nutrient uptake by wheat (Triticum aestivum L.) grown in a layered or split soil culture in glasshouse conditions. In the layered pot system the upper soil was supplied with P and either kept watered or allowed to dry or left P-deficient but watered, whereas the lower soil was watered and fertilised with K. Greater reductions in shoot growth, root length and dry weight in the upper soil layer occurred in −P/wet than in +P/dry upper soil treatment. Shoot P concentration and total P content were reduced by P deficiency but not by upper soil drying. Genotypic responses showed that K-efficient cv. Nyabing grew better and took up more P and K than K-inefficient cv. Gutha in well-watered condition, but the differences decreased when the upper soil layer was dry. In the split-root system, shoot dry weight and shoot P and K contents were similar when P and K were applied together in one compartment or separated into two compartments. In comparison, root growth was stimulated and plants took up more P and K in the treatment with the two nutrients supplied together compared with the treatment in which the two nutrients were separated. Roots proliferated in the compartment applied with either P or K at the expense of root growth in the adjoining compartment with neither P nor K. Heterogeneous nutrient distribution has a direct decreasing effect on root growth in deficient patches, and nutrient redistribution within the plant is unlikely to meet the demand of roots grown in such patches.     

Elevated CO2 enhances plant growth in droughted N2-fixing alfalfa without improving water status

The long-term interaction between elevated CO₂ and soil water deficit was analysed in N₂-fixing alfalfa plants in order to assess the possible drought tolerance effect of CO₂. Elevated CO₂ could delay the onset of drought stress by decreasing transpiration rates, but this effect was avoided by subjecting plants to the same soil water content. Nodulated alfalfa plants subjected to ambient (400 μmol mol^?1) or elevated (700 μmol mol^?1) CO₂ were either well watered or partially watered by restricting water to obtain 30% of the water content at field capacity (ampproximately 0.55 g water cm^?3). The negative effects of soil water deficit on plant growth were counterbalanced by elevated CO₂. In droughted plants, elevated CO₂ stimulated carbon fixation and, as a result, biomass production was even greater than in well-watered plants grown in ambient CO₂. Below-ground production was preferentially stimulated by elevated CO₂ in droughted plants, increasing nodule biomass production and the availability of photosynthates to the nodules. As a result, total nitrogen content in droughted plants was higher than in well-watered plants grown in ambient CO₂. The beneficial effect of elevated CO₂ was not correlated with a better plant water status. It is concluded that elevated CO₂ enhances growth of droughted plants by stimulating carbon fixation, preferentially increasing the availability of photosynthates to below-ground production (roots and nodules) without improving water status. This means that elevated CO₂ enhances the ability to produce more biomass in N₂-fixing alfalfa under given soil water stress, improving drought tolerance.     

Stomatal control in tomato with ABA-deficient roots: response of grafted plants to soil drying

The hypothesis that ABA produced by roots in drying soil is responsible for stomatal closure was tested with grafted plants constructed from the ABA-deficient tomato mutants, sitiens and flacca and their near-isogenic wild-type parent. Three types of experiments were conducted. In the first type, reciprocal grafts were made between the wild type and sitiens or flacca. Stomatal conductance accorded with the genotype of the shoot, not the root. Stomates closed in all of the grafted plants in response to soil drying, regardless of the root genotype, i.e. regardless of the ability of the roots to produce ABA. In the second type of experiment, wild-type shoots were grafted onto a split-root system consisting of one wild-type root grafted to one mutant (flacca or sitiens) root. Water was withheld from one root system, while the other was watered well so that the shoots did not experience any decline in water potential or loss of turgor. Stomates closed to a similar extent when water was withheld from the mutant roots or the wild-type roots. In the third type of experiment, grafted plants with wild-type shoots and either wild-type or sitiens roots were established in pots that could be placed inside a pressure chamber, and the pressure increased as the soil dried so that the shoots remained fully turgid throughout. Stomates closed as the soil dried, regardless of whether the roots were wild type or sitiens. These experiments demonstrate that stomatal closure in response to soil drying can occur in the absence of leaf water deficit, and does not require ABA production by roots. A chemical signal from roots leading to a change in apoplastic ABA levels in leaves may be responsible for the stomatal closure.