Petiole Pithiness in Celery Leaves: Induction by Environmental Stresses and the Involvement of Abscisic Acid

Petiole pithiness in celery, Apium graveolens L., was found to be stimulated by several types of root stress. Flooding of the root zone as well as nutrition deficiency required a prolonged period (25–30 days), but water deprivation had a rapid (2–3 days) effect on pithiness development. Pithiness development induced by water deprivation is not reversible upon rehydration of whole plants but is reversible upon rehydration of affected petiole slices. The stimulation of water stress-related pithiness was found to be associated with an increased level of endogenous free abscisic acid (ABA). Exogenously applied ABA stimulated petiole pithiness of detached leaves in a concentration-dependent fashion. It is suggested that in celery, ABA is an agent which mediates stimulation of petiole pithiness by water stress.     

Identification of different ABA biosynthesis sites at seedling and fruiting stages in Arachis hypogaea L. following water stress

Abscisic acid (ABA) is one of the most important phytohormones involved in abiotic stress responses. ABA transport in plants is important in determining endogenous ABA levels and their resulting physiological responses. However, the regulation of ABA transport remains unclear. In this study, we compared the ABA concentrations and AhNCED1 levels at seedling and fruiting stages in peanut (Arachis hypogaea L.), in response to water stress. At the seedling stage, ABA initially accumulated in roots (1 h), followed by the lower stem (2 h) and finally in the upper stem (4 h). The expression/activity of an ABA biosynthesis rate-limiting enzyme, AhNCED1, showed the same accumulation patterns. In contrast, during the fruiting stage, ABA and AhNCED1 increases were initially detected in the first apical leaf of main stem, followed by the stem, and finally in the root. These results imply that biosynthesis of ABA in peanut plants subject to water deficiency could be dependent on developmental stage with the roots being the initial site of ABA biosynthesis during the seedling stage, whereas during the fruiting stage ABA biosynthesis occurs initially in the leaf. The distribution patterns of ABA in seedling stage peanuts in response to water stress were: root-stem-leaf, while in fruiting stage peanuts the distribution patterns of ABA were: leaf-stem-root. These findings will help to understand plant regulatory water deficit resistance mechanisms at seedling and fruiting stages and to advance our total understanding of the regulation of ABA transport.     

Abscisic acid and cytokinins in the root exudates and leaves and their relationship to senescence and remobilization of carbon reserves in rice subjected to water stress during grain filling

The possible regulation of senescence-initiated remobilization of carbon reserves in rice (Oryza sativa L.) by abscisic acid (ABA) and cytokinins was studied using two rice cultivars with high lodging resistance and slow remobilization. The plants were grown in pots and either well-watered (WW, soil water potential = 0 MPa) or water-stressed (WS, soil water potential = -0.05 MPa) from 9 days after anthesis until they reached maturity. Leaf water potentials of both cultivars markedly decreased at midday as a result of water stress but completely recovered by early morning. Chlorophyll (Chl) and photosynthetic rate (Pr) of the flag leaves declined faster in WS plants than in WW plants, indicating that the water deficit enhanced senescence. Water stress accelerated starch remobilization in the stems, promoted the re-allocation of pre-fixed (14)C from the stems to grains, shortened the grain-filling period and increased the grain-filling rate. Sucrose phosphate synthase (SPS, EC 2.4.1.14) activity was enhanced by water stress and positively correlated with sucrose accumulation in both the stem and leaves. Water stress substantially increased ABA but reduced zeatin (Z) + zeatin riboside (ZR) concentrations in the root exudates and leaves. ABA significantly and negatively, while Z+ZR positively, correlated with Pr and Chl of the flag leaves. ABA, not Z+ZR, was positively and significantly correlated with SPS activity and remobilization of pre-stored carbon. Spraying ABA reduced Chl in the flag leaves, and enhanced SPS activity and remobilization of carbon reserves. Spraying kinetin had the opposite effect. The results suggest that both ABA and cytokinins are involved in controlling plant senescence, and an enhanced carbon remobilization is attributed to an elevated ABA level in rice plants subjected to water stress.     

Scion and Rootstock Effects on ABA-mediated Plant Growth Regulation and Salt Tolerance of Acclimated and Unacclimated Potato Genotypes

Tolerance of salt stress in potato (Solanum tuberosum L.) increased when the plants were pre-exposed to low concentrations of salt (salt acclimation). This acclimation was accompanied by increased levels of abscisic acid (ABA) in the shoot. To further study the role of roots and shoots in this acclimation process, reciprocal grafts were made between a salt-tolerant (9506) and salt-sensitive ABA(−) mutant and its ABA(+) normal sibling potato genotype. The grafted plants were acclimated with 75 or 100 mM NaCl for 3 weeks and then exposed to 150–180 mM NaCl, depending on the salt tolerance of the rootstock. After 2 weeks of exposure to the salt stress, the acclimated and unacclimated plants were compared for physiologic and morphologic parameters. The response to the salt stress was strongly influenced by the rootstock. The salt-tolerant 9506 rootstock increased the salt tolerance of scions of both the ABA-deficient mutant and its ABA(+) sibling. This salt tolerance induced by the rootstock was primarily modulated by salt acclimation and manifested in the scion via increased plant water content, stem diameter, dry matter accumulation, stomatal conductivity, and osmotic potential, and is associated with a reduction in leaf necrosis. There was also a pronounced scion effect on the rootstock. Using 9506 as a scion significantly increased root fresh and dry weights, stem diameter, and root water content of ABA(−) mutant rootstocks. Specific evidence was found of the role of exogenous ABA in the enhancement of water status in grafted plants under salt stress beyond that of grafting alone. This was verified by more positive stomatal conductivity and upward water flow in ABA-treated grafted and nongrafted plants and the absence of upward water flow in nontreated grafted plants through NMR imaging. Grafting using either salt-tolerant scions or rootstocks with inherently high ABA levels may positively modify subsequent responses of the plant under salt stress.     

Metabolism of R,S-[2-14C]abscisic acid by non-stressed and water-stressed detached leaves of wheat (Triticum aestivum L.)

Metabolism of R,S-[2-¹⁴C]abscisic acid (ABA) was studied in detached leaves of six wheat (Triticum aestivum) cultivars, using non-stressed leaves or leaves water stressed by desiccation to 90% of their original fresh weight. The rate constant of ABA metabolism was similar in nonstressed leaves of all cultivars. Water stress resulted in significantly lower rate constants in two cultivars which accumulated high levels of ABA when stressed, the constants decreasing by a factor of about 1.5. Rate constants for the remainder of the cultivars were not significantly different from those for the non-stressed controls. It was calculated that if decreased metabolism was the mechanism for the accumulation of ABA following water stress the rate constants of metabolism would have to be reduced by a factor of between 25 and 70. The results therefore support the hypothesis that enhanced synthesis rather than reduced degradation is the main process by which ABA levels are elevated following experimentally induced water stress. There were differences between the six cultivars in the products of ABA metabolism. Over the time period studied, oxidation to phaseic acid and dihydrophaseic acid as well as to other unidentified metabolites appeared to be the predominant pathway of ABA metabolism, rather than conjugation to ABA glucose ester and other more polar compounds.Abbreviations ABA abscisic acid - ABAGE abscisic-acid glucose ester - DPA dihydrophaseic acid - PA phaseic acid     

Effects of Abscisic Acid and Water Stress on Development and Morphology of Wheat

Experiments were conducted to compare the effects of abscisicacid (ABA) and water stress treatments on leaf morphology andfloral development in a spring wheat. In one experiment injectionsof ABA or a control solution were given twice a week into thebase of the main stem for a period of 3 weeks. In a similarexperiment control plants were watered daily and treated plantswere subjected to water stress by watering only once a week.In both experiments the treated plants produced smaller leavesand fewer spikelets per ear. Analysis of epidermal morphologyusing polystyrene imprints of selected leaf blades from themain stem and a tiller of each plant showed that, compared withcontrol plants, both ABA and water stress decreased the meancell size, reduced the number of stomata per leaf, and increasedthe production of trichomes in all the leaves sampled. Datafor stomatal lengths and stomatal indices showed differencesbetween a main stem leaf and a tiller leaf which were consistentfor both experiments. It is concluded that ABA could mediatemany of the responses of wheat plants to prolonged water stress.The possible adaptive value of these responses is discussed.     

How the roots contribute to the ability of Phaseolus vulgaris L. to cope with chilling-induced water stress

Intact plants and stem-girdled plants of Phaseolus vulgaris grown hydroponically were exposed to 5 degrees C for up to 4 d; stem girdling was used to inhibit the phloem transport from the leaves to the roots. After initial water stress, stomatal closure and an amelioration of root water transport properties allowed the plants to rehydrate and regain turgor. Chilling augmented the concentration of abscisic acid (ABA) content in leaves, roots and xylem sap. In intact plants stomatal closure and leaf ABA accumulation were preceded by a slight alkalinization of xylem sap, but they occurred earlier than any increase in xylem ABA concentration could be detected. Stem girdling did not affect the influence of chilling on plant water relations and leaf ABA content, but it reduced slightly the alkalinization of xylem sap and, principally, prevented the massive ABA accumulation in root tissues and the associated transport in the xylem that was observed in non-girdled plants. When the plants were defoliated just prior to chilling or after 10 h at 5 degrees C, root and xylem sap ABA concentration remained unchanged throughout the whole stress period. When the plants were chilled under conditions preventing the occurrence of leaf water deficit (i.e. at 100% relative humidity), there were no significant variations in endogenous ABA levels. The increase in root hydraulic conductance in chilled plants was a response neither to root ABA accretion, nor to some leaf-borne chemical signal transported downwards in the phloem, nor to low temperature per se, as indicated by the results of the experiments with defoliated or girdled plants and with plants chilled at 100% relative humidity. It was concluded that the root system contributed substantially to the bean's ability to cope with chilling-induced water stress, but not in an ABA-dependent manner.     

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.     

Differential sensitivity of canola (Brassica napus) seedlings to ultraviolet-B radiation,water stress and abscisic acid

Responses of canola (Brassica napus L.) seedlings to three ultraviolet (UV)-B levels [0 (zero), 5 (ambient) and 10 (enhanced) kJ m^?2 d^?1], two watering regimes (well-watered and water-stressed), and two abscisic acid (ABA) levels (with and without application) were investigated. Overall, enhanced UVB and water stress negatively affected plant growth and physiology, but ABA had very little effect. Enhanced UVB decreased stem height, leaf area, plant dry matter, water use efficiency and wax content, but increased concentrations of chlorophyll a, carotenoids and flavonoids, and ethylene evolution. Water stress reduced stem height and diameter, leaf area, plant dry matter, leaf weight ratio and shoot:root weight ratio under zero and ambient UVB. Water stress also reduced chlorophyll a and carotenoids in plants exposed to enhanced UVB. ABA with watering regime had significant interactive effects only on leaf dry matter and wax content. We found that enhanced UVB and water stress adversely affected B. napus seedlings. Interaction between these two factors affected plant performance. In this interaction, ABA had little significant role. Also, optimum vegetative growth and biomass were achieved under ambient UVB.     

Endogenous abscisic acid levels in stems and axillary buds of intact or decapitated broad-bean plants (Vicia faba L.)

Endogenous levels of abscisic acid (ABA) were measured by gas-liquid chromatography (electron capture) in stems and axillary buds of intact or decapitated broad-bean plants ( Vicia faba L. cv. Aguadulce). Endogenous ABA was distributed in the main axis according to a concentration gradient from the apical part of the stem towards the base. Axillary buds contained ABA levels which were from 4 to 9 times higher than those in the corresponding nodes and internodes. Decapitation of the plant was followed within 6 h by a large decrease of ABA levels in all the parts of the main axis. The diminution of ABA content was the most important in axillary buds released from apical dominance. Twenty-four hours after the decapitation, the ABA concentration further decreased in the upper parts of the stem, while no modification was observed in the basal parts of the stem containing the smallest levels of ABA.     

Changes in abscisic acid and proline levels in maize varieties of different drought resistance

Four varieties of maize differing in drought resistance and geographical origin (Swabi White and Shaheen from Pakistan, Garbo and Goldprinz from Germany) were analyzed for their proline and abscisic acid (ABA) accumulation during a prolonged water stress period. Proline levels increased continuously during the stress period in all the four varieties, but to different amounts. The drought-susceptible varieties Shaheen and Goldprinz produced higher levels of proline than the drought-resistant varieties Swabi White and Garbo. A negative correlation was also found between maximal ABA contents and degree of drought resistance during prolonged stress of younger plants. ABA levels did not increase steadily, but reached a maximum long before the end of the stress phase, and then declined. The results are discussed in relation to the possibility of using proline and ABA levels as biochemical indicators of resistance against drought.     

Effect of water stress on proline content and transcript levels in Lathyrus sativus

Response of Lathyrus sativus plants to water stress showed that ABA responsive genes such as PLE 25, TAS 14 and RAB 17 are synthesized constitutively, the levels of which decline gradually with increase in water stress or ABA levels. Proline accumulation was highest in leaves (65-fold) followed by stem (56-fold), root (38-fold) and marginal increase in etiolated seedlings. Proline increase was also observed in plant parts not exposed to light.     

Do brassinosteroids mediate the water stress response?

Brassinosteroids (BRs) have been suggested to increase the resistance of plants to a variety of stresses, including water stress. This is based on application studies, where exogenously applied bioactive BRs have been shown to improve various aspects of plant growth under water stress conditions. However, it is not known whether changes in endogenous BR levels are normally involved in mediating the plant's response to stress. We have utilized BR mutants in pea ( Pisum sativum L.) to determine whether changes in endogenous BR levels are part of the plant's response to water stress and whether low endogenous BR levels alter the plant's ability to cope with water stress. In wild-type (WT) plants, we show that while water stress causes a significant increase in ABA levels, it does not result in altered BR levels in either apical, internode or leaf tissue. Furthermore, the plant's ability to increase ABA levels in response to water stress is not affected by BR deficiency, as there was no significant difference in ABA levels between WT, lkb (a BR-deficient mutant) and lka (a BR-perception mutant) plants before or 14 days after the cessation of watering. In addition, the effect of water stress on traits such as height, leaf size and water potential in lkb and lka was similar to that observed in WT plants. Therefore, it appears that, at least in pea, changes in endogenous BR levels are not normally part of the plant's response to water stress.     

The role of cytokinins in responses to water deficit in tobacco plants over-expressing trans-zeatin O-glucosyltransferase gene under 35S or SAG12 promoters

The impact of water deficit progression on cytokinin (CK), auxin and abscisic acid (ABA) levels was followed in upper, middle and lower leaves and roots of Nicotiana tabacum L. cv. Wisconsin 38 plants [wild type (WT)]. ABA content was strongly increased during drought stress, especially in upper leaves. In plants with a uniformly elevated total CK content, expressing constitutively the trans -zeatin O-glucosyltransferase gene ( 35S::ZOG1 ), a delay in the increase of ABA was observed; later on, ABA levels were comparable with those of WT.
As drought progressed, the bioactive CK content in leaves gradually decreased, being maintained longer in the upper leaves of all tested genotypes. Under severe stress (11 d dehydration), a large stimulation of cytokinin oxidase/dehydrogenase (CKX) activity was monitored in lower leaves, which correlated well with the decrease in bioactive CK levels. This suggests that a gradient of bioactive CKs in favour of upper leaves is established during drought stress, which might be beneficial for the preferential protection of these leaves.
During drought, significant accumulation of CKs occurred in roots, partially because of decreased CKX activity. Simultaneously, auxin increased in roots and lower leaves. This indicates that both CKs and auxin play a role in root response to severe drought, which involves the stimulation of primary root growth and branching inhibition.     

Pithiness in plants: I. The effect of mechanical perturbation and the involvement of ethylene in petiole pithiness in celery

Mechanical perturbation (MP) applied to celery (Appium graveolens L. cv. Florida 683) leaf petioles or ethephon application to the plant did not induce thigmomorphogenesis (inhibition of elongation and increase in thickness of the petiole). However, the two treatments did cause the parenchyma breakdown which leads to pithiness or increased natural pithiness, mainly at the base of the petiole. Nevertheless, MP (but not ethephon) decreased the severity of drought-stress or GA3-induced pithiness. Although MP stimulates ethylene production, mainly at the middle part of the petiole, it seems that the protection by MP of the petiole may not be directly mediated by ethylene production. The exposure of the plant to drought stress brought about an increase in ethylene evolution. Upon reirrigating the plants, the first steps of pithiness were accompanied by a sharp decline in ethylene production. This decrease might be due to membrane disruption. The increase in ethylene production during drought stress may be one of the events which stimulate pithiness of the celery leaf petiole.     

Thigmomorphogenesis: the Effect of Mechanical Perturbation and Ethrel on Stem Pithiness in Tomato [Lycopersicon esculentum (Mill.) Plants]

The stems of ‘Y’-shaped (double stemmed) tomato(Lycopersicon esculentum Mill.) plants were mechanically perturbed(MP) by stroking for 6 successive days. The treatment reducedelongation of the two stems by 40 per cent. When only one branchof the pair was treated, its length was reduced to the sameextent as the two branches in the previous treatment, whilethe elongation of the untreated branch was increased by 60 percent over that of the control. Withholding irrigation induced stem pithiness due to droughtstress in non-MP-plants. However, in MP-pretreated plants, thenumber of pithy internodes was markedly less and the degreeof severity of the disorder was reduced. Ethrel applicationmimicked the effects of MP on pithiness. In some unknown way,the plants are hardened by MP or Ethrel. Lycopersicon esculentum Mill, tomato, drought stress, thigmomorphogenesis, ethylene, pithiness     

Arabidopsis mutants of AtABCG22, an ABC transporter gene, increase water transpiration and drought susceptibility

In plants, water vapour is released into the atmosphere through stomata in a process called transpiration. Abscisic acid (ABA) is a key phytohormone that facilitates stomatal closure through its action on guard cells. Recently, ATP-binding cassette (ABC) transporter genes, AtABCG25 and AtABCG40, were shown to be involved in ABA transport and responses. However, the functions of many other AtABCG family genes are still unknown. Here, we identified another ABCG gene (AtABCG22) that is required for stomatal regulation in Arabidopsis. The atabcg22 mutant plants had lower leaf temperatures and increased water loss, implying elevated transpiration through an influence on stomatal regulation. We also found that atabcg22 plants were more suspectible to drought stress than wild-type plants. AtABCG22 was expressed in aerial organs, mainly guard cells, in which the gene expression pattern was consistent with the mutant phenotypes. Using double mutants, we investigated the genetic relationships between the mutations. The atabcg22 mutation further increased the water loss of srk2e/ost1 mutants, which were defective in ABA signalling in guard cells. Also, the atabcg22 mutation enhanced the phenotype of nced3 mutants, which were defective in ABA biosynthesis. Accordingly, the additive roles of AtABCG22 functions in ABA signalling and ABA biosynthesis are discussed.     

Effect of drought and high solar radiation on 1-aminocyclopropane-1-carboxylic acid and abscisic acid concentrations in Rosmarinus officinalis plants

The endogenous concentrations of ACC and ABA were measured, at predawn and at maximum solar radiation, during a summer drought, and recovery after autumn rainfalls, in rosemary (Rosmarinus officinalis L.), a drought-tolerant species, growing under Mediterranean field conditions. During the summer, plants were subjected to both water deficit and high solar radiation. Plants showed severe reductions in shoot water potential to -3 MPa, which were associated with drastic stomatal closure (73%), a decrease in net photosynthesis, reaching almost zero, and a severe chlorophyll loss (74%). Despite the severity of the stress, plants recovered after the autumn rainfalls. The concentration of ACC was not enhanced by drought, and at predawn these concentrations remained constant at approximately 600 pmol ACC-1 DW throughout the experiment. Thus, ethylene did not regulate the response of rosemary to drought. However, a sharp increase in ACC levels between predawn and midday was observed. This increase was positively correlated to the intensity of the incident solar radiation. ACC levels recorded in June at midday reached 16 000 pmol g DW and in October values of 1000 pmol g-1 DW were observed. In contrast, in drought-stressed plants predawn concentrations of ABA were up to 130-fold those of recovered plants, and the levels of ABA scored at midday were double of those scored at predawn. In conclusion, although drought-stressed rosemary plants showed a relatively moderate ABA accumulation (approximately 500 pmol g-1 DW#, at predawn), it seems to be an essential factor for the regulation of the plant response to stress, thereby enabling a rapid recovery after stress release, although other mechanisms can not be excluded. As drought stress did not induce ACC accumulation, it was concluded that ethylene production was not a major factor in the drought stress resistance of rosemary plants. The increased ACC and ABA concentrations at midday were correlated with day length and light intensity and not with the water status of the plant.