Aquaporin expression in response to different water stress intensities and recovery in Richter-110 (<Emphasis Type="Italic">Vitis</Emphasis> sp.): relationship with ecophysiological status

Aquaporins seem essential for the regulation of plant water status and expenses. Richter-110 is a Vitis hybrid (Vitis berlandieri × rupestris) reputed to be strongly drought-tolerant. Three irrigation treatments were established in Richter-110 plants growing outdoors defined by the resulting maximum stomatal conductance (g _s), and ensuring water stress situations not severe enough as to stop photosynthesis and growth: well-watered plants (g _s about 250 mmol H₂O m⁻² s⁻¹), moderate water stress (g _s about 150 mmol H₂O m⁻² s⁻¹) and severe water stress (g _s about 50 mmol H₂O m⁻² s⁻¹). Plants under water stress were kept at constant water availability for 7 days to check for possible acclimation. Finally, plants were re-watered, and allowed to recover, for 3 days. Stomatal conductance, leaf water potential, xylem abscisic acid (ABA) content and root and stem hydraulic conductivity were determined. The relative amounts of expression of mRNA encoding seven putative aquaporins were determined in roots and leaves by RT-PCR. The decrease in stomatal conductance with moderate and severe water stress was associated with increasing ABA contents, but not with the leaf water potential and hydraulic conductivities, which remained unchanged during the entire experiment. Aquaporin gene expression varied depending on which aquaporin, water stress level and the plant organ. We suggest that aquaporin expression was responsive to water stress as part of the homeostasis, which resulted in constant leaf water potential and hydraulic conductivity.     

Diurnal change in the relationship between stomatal conductance and abscisic acid in the xylem sap of field-grown peach trees

To evaluate whether abscisic acid (ABA) in the xylem sap playsan important role in controlling stomatal aperture of field-grownPrunus persica trees under drought conditions, stomatal conductance(g) and xylem ABA concentrations were monitored both in irrigatedand non-irrigated trees, on two consecutive summer days (threetimes a day). Stomata1 conductance of non-irrigated trees hada morning maximum and declined afterwards. The changes in gduring the day, rather than resulting from variations in theconcentrations of ABA in the xylem sap or the delivery rateof this compound to the leaves, were associated with changesin the relationship between g and xylem ABA. The stomata ofwater-stressed trees opened during the first hours of the day,despite the occurrence of a high concentration of ABA in thexylem sap. However, stomatal responsiveness to ABA in the xylemwas enhanced throughout the day. As a result, a tight inverserelationship between g and the logarithm of xylem ABA concentrationwas found both at midday and in the afternoon. A similar relationshipbetween g and ABA was found when exogenous ABA was fed to leavesdetached from well-watered trees. These results indicate thatABA derived from the xylem may account for the differences ing observed between field-grown peach trees growing with differentsoil water availabilities. Several possible explanations forthe apparent low stomatal sensitivity to xylem ABA in the morning,are discussed, such as high leaf water potential, low temperatureand high cytokinin activity. Key words: Prunus persica L., stomata, xylem ABA, water deficits, root-to-shoot communication     

Hydraulic Lift in Cork Oak Trees in a Savannah-Type Mediterranean Ecosystem and its Contribution to the Local Water Balance

The aim of this study was to identify the sources and depth of water uptake by 15-years old Quercus suber L. trees in southern Portugal under a Mediterranean climate, measuring δ¹⁸O and δD in the soil–plant-atmosphere continuum. Evidence for hydraulic lift was substantiated by the daily fluctuations observed in Ψ_s at 0.4 and 1 m depth and supported by similar δ¹⁸O values found in tree xylem sap, soil water in the rhizosphere and groundwater. From 0.25 m down to a depth of 1 m, δD trends differed according to vegetation type, showing a more depleted value in soil water collected under the evergreen trees (−47‰) than under dead grasses (−35‰). The hypothesis of a fractionation process occurring in the soil due to diffusion of water vapour in the dry soil is proposed to explain the more depleted soil δD signature observed under trees. Hydraulically lifted water was estimated to account for 17–81% of the water used during the following day by tree transpiration at the peak of the drought season, i.e., 0.1–14 L tree⁻¹ day⁻¹. Significant relationships found between xylem sap isotopic composition and leaf water potential in early September emphasized the positive impact of the redistribution of groundwater in the rhizosphere on tree water status.     

Whole-plant ABA flux and the regulation of water loss in Cedrella odorata

Three-month-old Cedrella odorata seedlings were exposed to a soil-drying treatment. During this period, xylem sap was periodically collected from the plant by applying pneumatic pressure to the roots. This also allowed whole-plant water status to be measured by recording the balancing pressure applied. The concentration of ABA in xylem sap (C) was related to the whole-plant transpiration rate (V) which was measured with a sap flow gauge. The analysis of these paired measurements centred on how the reciprocal of C (R) varied with respect to V. This revealed that (1) the observed increases in C could not be explained by the reductions in V alone, (2) initially, decreases in V were associated with proportional increases in the whole-plant ABA flux (M), and (3) this relationship broke down at low values of V since zero flow was associated with a finite value for C estimated to be 41 pmol ABA mmol^?1 H₂O. A simple static model is developed from the observations that is able to explain the data well, and the results are discussed in terms of the effects of ABA on stomatal conductance (g_sw).     

Improvement of <Emphasis Type="Italic">ex vitro</Emphasis> transfer of tobacco plantlets by addition of abscisic acid to the last subculture

Tobacco (Nicotiana tabacum L.) plantlets were grown on Murashige and Skoog medium in ventilated Magenta boxes and for the last subculture 10 μM ABA was added to the medium. After three weeks plantlets were transferred into pots with Perlite moistened with water and grown in controlled conditions (16-h photoperiod, day/night temperature 25/20 °C, air humidity about 45 %) either under low or high irradiance of 150 (LI) and 700 (HI) μmol m⁻² s⁻¹, respectively. Content of endogenous ABA was 271.7 pmol g⁻¹(f.m.) in ABA treated plantlets, while in control plantlets it was only 53.3 pmol g⁻¹(f.m.). After ex vitro transfer, stomatal conductance and transpiration rate decreased considerably in comparison with in vitro grown plantlets and remained lower also 7 d after ex vitro transfer, especially in ABA-treated plants and so wilting of plants was practically eliminated. Net photosynthetic rate also decreased 1 d after ex vitro transfer but after 7 d it was mostly higher than that of in vitro grown plantlets. Water use efficiency significantly increased in ABA-treated plants. Chlorophyll a+b content did not change immediately after ex vitro transfer, nevertheless, after 7 d chlorophyll content was higher in ABA-treated plants. Pool of xanthophyll cycle pigments (XCP) and the degree of their deepoxidation (DEPS), which are connected with harmless dissipation of light energy, increased under high irradiance. Contents of XCP and ABA precursors (neoxanthin and violaxanthin) were lower in ABA-treated plants than in control plants indicating less stress in these plants. Most chlorophyll a fluorescence parameters did not change considerably after ex vitro transfer and so the photoinhibition was not observed even under HI. Slight increase in non-photochemical quenching under HI in ABA-treated plants suggested their better photoprotection. Thus application of ABA to the last subculture can improve acclimatization of in vitro grown plants to ex vitro conditions     

Variations in abscisic acid, indole-3-acetic acid and zeatin riboside concentrations in two Mediterranean shrubs subjected to water stress

Water stress induced an increase in endogenous concentrations of ABA in Lavandula stoechas L. plants to 13100 pmol ABA g^–1 FW, which may contribute to the maintenance of water relations between the second and the third day of water stress treatment. After the third day, a sharp decrease in ABA levels was observed to 2630 pmol ABA g^–1 FW, together with a decrease in water content and water potential and a loss of plant response to water stress. Water deficit did not induce an increase in endogenous ABA concentration, which remained at 514 pmol ABA g^–1 FW in Rosmarinus officinalis L., which is more sclerophyllous than L. stoechas. Nevertheless, the relative water content of Rosmarinus officinalis L. after seven days of water stress decreased more than 40% and reached values of –3.2 MPa. R. officinalis showed lower levels of ABA, but significantly higher levels of IAA and ZR than L. stoechas (4 times and 6 times respectively in well watered-plants). The increase in ABA levels is not a common mechanism in these two Mediterranean shrubs which survive under water stress conditions.Abbreviations ABA abscisic acid - d days of water stress treatment - DW dry weight - FW fresh weight - IAA indole-3-acetic acid - RP Reversed Phase - RWC relative water content - TW turgid weight - WC water content - ZR zeatin riboside - water potential     

Regulation of sulfate uptake and xylem loading of poplar roots (Populus tremula x P. alba)

Sulfate transport processes and its regulation were studied in roots of poplar trees (Populus tremula x P. alba). From the exponential increase in sulfate uptake with temperature an activation energy (E_a) of 9.0±0.8 kJ mol^–1 was calculated. In the concentration range 0.005–10 mM sulfate uptake showed biphasic Michaelis-Menten kinetics with a K_m of 3.2±3.4 M and a V_max of 49±11 nmol SO₄^2– g^–1 FW h^–1 for the high-affinity uptake system (phase 1) and a K_m of 1.33±0.41 mM and a V_max of 255±25 nmol SO₄^2– g^–1 FW h^–1 for the low-affinity system (phase 2). Xylem loading decreased linearly with temperature and remained unchanged within the sulfate concentration range studied. Regulation of sulfate uptake and xylem loading by O-acetyl serine (OAS), Cys, reduced glutathione (GSH), Met and S-methylmethionine (SMM) were tested by perfusion into the xylem sap with the pressure probe and by addition to the incubation medium. When added directly to the transport medium, Cys and GSH repressed, and OAS stimulated sulfate uptake; xylem loading was stimulated by Cys, repressed by GSH and only slightly affected by OAS. When perfused into the xylem, none of the compounds tested affected sulfate uptake of excised roots, but xylem loading was stimulated by SMM and OAS and repressed by Met. Apparently, the site of application strongly determined the effect of regulatory compounds of sulfate transport processes.     

Stomatal control by both [ABA] in the xylem sap and leaf water status: a test of a model for draughted or ABA-fed field-grown maize

A model of maize stomatal behaviour has been developed, in which stomatal conductance is linked to the concentration of abscisic acid ([ABA]) in the xylem sap, with a sensitivity dependent upon the leaf water potential (Ψ₁). It was tested against two alternative hypotheses, namely that stomatal sensitivity to xylem [ABA] would be linked to the leaf-to-air vapour pressure difference (VPD), or to the flux of ABA into the leaf. Stomatal conductance (g_s) was studied: (1) in field-grown plants whose xylem [ABA] and Ψ₁ depended on soil water status and evaporative demand; (2) in field-grown plants fed with ABA solutions such that xylem [ABA] was artificially raised, thereby decreasing g_s and increasing Ψ₁ and leaf-to-air VPD; and (3) in ABA-fed detached leaves exposed to varying evaporative demands, but with a constant and high Ψ₁. The same relationships between g_s, xylem [ABA] and Ψ₁, showing lower stomatal sensitivity to [ABA] at high Ψ₁, applied whether variations in xylem [ABA] were due to natural increase or to feeding, and whether variations in Ψ₁, were due to changes in evaporative demand or to the increased Ψ₁ observed in ABA-fed plants. Conversely, neither the leaf-to-air VPD nor the ABA flux into the leaf accounted for the observed changes in stomatal sensitivity to xylem [ABA]. The model, using parameters calculated from previous field data and the detached-leaf data, was tested against the observations of both ABA-fed and droughted plants in the field. It accounted with reasonable accuracy for changes in g_s (r² ranging from 0.77 to 0.81). These results support the view that modelling of stomatal behaviour requires consideration of both chemical and hydraulic aspects of root-to-shoot communication.     

Uptake of EDTA-complexed Pb, Cd and Fe by solution- and sand-cultured Brassica juncea

Direct plant uptake of metals bound to chelating agents has important implications for metal uptake and the free-ion activity model. Uptake of hydrophilic solutes such as metal–EDTA complexes is believed to occur via bypass apoplastic flow, but many questions remain about the relative importance and selectivity of this pathway. In this study, Brassica juncea (Indian mustard) plants grown in solution- and sand-culture conditions were exposed to metal–EDTA complexes and to PTS, a hydrophilic fluorescent dye previously used as a tracer of apoplastic flow. The results suggest that there are two general phases of solute uptake. Under normal conditions, xylem sap solute concentrations are relatively low (i.e., <0.5% of concentration in solution) and there is a high degree of selectivity among different solutes, while under conditions of stress, xylem sap concentrations are significantly higher (i.e., >3% of concentration in solution) and the selectivity among solutes is less. In healthy plants, xylem sap metal–EDTA concentrations were generally an order of magnitude higher than those of PTS and differences among complexes were observed, with CdEDTA²⁻ exhibiting slightly higher xylem sap concentrations than PbEDTA²⁻ or FeEDTA⁻. Metal–EDTA complexes were found to dominate xylem sap metal speciation and the fraction of metal in xylem sap present as metal–EDTA was greater for non-nutrient metals (Pb, Cd) than for the nutrient metal Fe. Despite differences in root morphology between plants grown under solution- and sand-culture conditions, uptake of solutes was similar under both sets of growth conditions.     

δ13C analysis of phloem sap carbon: novel means of evaluating seasonal water stress and interpreting carbon isotope signatures of foliage and trunk wood of Eucalyptus globulus

A recently described phloem-bleeding technique was used to study seasonal changes in δ¹³C, sugar levels and the amino acid:sugar balance of phloem translocate of 2- to 3-year old trees of Eucalyptus globulus at a rain-fed site (Eulup) and a waste-effluent-irrigated site (Albany) in south-west Australia. δ¹³C of phloem sap from the Eulup site fluctuated widely between winter (−27.6‰) and peak summer stress (−20.2‰), compared with a much smaller range of −28.4 to −26.3 at Albany. Seasonal changes in sugar concentrations in sap fluctuated closely with those of phloem δ¹³C, with highest concentrations and least negative δ¹³C values at times of greatest soil water deficit. Molar ratios of amino acids to sugars in phloem sap were similar between plantations in winter through to early summer. They then remained high at the nitrogen-rich effluent-treated site, but fell dramatically once soils dried out at Eulup. Mature leaf dry matter sampled at peak yearly stress (early autumn) showed more negative δ¹³C values than concurrently harvested phloem sap or recently initiated shoot apex dry matter, presumably because the sampled foliage had laid down its structural carbon earlier under relatively unstressed winter/spring conditions. Differences between Albany and Eulup were much greater for δ¹³C of phloem and new apical dry matter than for dry matter of mature foliage. Comparisons of δ¹³C signatures of phloem sap carbon with those of dry matter of nascent xylem tissues showed seasonal fluctuations in δ¹³C of phloem translocate which were mirrored a month or so later by those for xylem carbon. δ¹³C analyses of trunk growth rings from Eulup and Albany showed well-defined seasonal oscillations over the first 2 or 3 years of growth until irrigation commenced at Albany. Fluctuations in δ¹³C at the latter site then became noticeably less pronounced than at Eulup. Future use of phloem sap δ¹³C and solute analyses for studying seasonal water and nutrient status of E. globulus is discussed. Received: 9 April 1998 / Accepted: 20 August 1998     

Abscisic acid and gibberellin-like substances in roots and root nodules ofGlycine max

Summary The content of endogenous gibberellin (GA)-like substances of roots and root nodules of SOya, and GA production byRhizobium japonicum cultures, were investigated by a combined thin layer chromatographic (TLC)-dwarf pea epicotyl bioassay technique. GAs were more concentrated in root nodules than in the roots, totalling 1.34 and 0.16 nM GA₃ equivalents g⁻¹ dry wt. respectively. GA production byR. japonicum cultures was demonstrated (1.00 nM GA₃ equivalentsl ⁻¹) and comparison of the GA components of plant and bacterial culture medium extracts, suggested that rhizobial GA production may contribute to the nodule GA content. Cis-trans abscisic acid (ABA) was identified in root and nodule extracts by TLC-gas liquid chromatography (GLC), and amounted to 0.18 and 2.21 nM g⁻¹ dry wt. respectively, whereas 0.30 and 4.63 nM ABA equivalents g⁻¹ dry wt. were detected by a TLC-wheat embryo bioassay technique. ABA was not detected in extracts of bacterial cultures.     

Plant regeneration from shoot apical meristems of Melia azedarach L. (Meliaceae)

A protocol was developed for plant regeneration of Melia azedarach L. by in vitro culture of apical meristem (0.5 mm in length). The influence of six clones was investigated. The culture procedure comprised two sequential steps: 1) Induction of shoots by in vitro culture of axillary buds from adult trees (10–15 years old) by culture on Murashige and Skoog (1962) medium (MS) supplemented with 0.5 mg·dm⁻³ BAP (6-benzylaminopurine), 0.1 mg·dm⁻³ IBA (indolebutyric acid), and 0.1 mg·dm⁻³ GA₃ (gibberellic acid). The Multiplication of the regenerated shoots was achieved in MS + 0.5 mg·dm⁻³ BAP + 0.1 mg·dm⁻³ GA₃. 2) In vitro culture of the apical meristems from the regenerated shoots in MS medium (0.7 %) supplemented with various combinations of BAP and IBA. Maximum shoot proliferation was obtained on MS medium supplemented with 0.5 mg·dm⁻³ BAP and 0.1 mg·dm⁻³ IBA. Regenerated shoots were rooted on MS + 3.5 mg·dm⁻³ IBA (4 days) followed by subculture on MS lacking growth regulators (30 days). Complete plants were transferred to soil.     

The effect of phytohormones on growth and artemisinin production in <Emphasis Type="Italic">Artemisia annua</Emphasis> hairy roots

Summary Few studies have focused on the effect of a broad range of phytohormones on growth and secondary metabolism of a single hairy root species. We measured growth, development, and production of the antimalarial drug, artemisinin, in Artemisia annua hairy roots in response to the five main hormones: auxins, cytokinins, ethylene, gibberellins (GA), and abscisic acid (ABA). Single roots grown in six-well plates in medium B5 with 0.01 mgl⁻¹ (0.029 μM) GA₃ produced the highest values overall in terms of the number of lateral roots, length of the primary root, lateral root tip density, total lateral root length, and total root length. When the total root lengths are compared, the best conditions for stimulating elongation appear to be: GA 0.01 mgl⁻¹ (0.029μM)> ABA 1.0 mgl⁻¹ (3.78μM)=GA 0.02 mgl⁻¹ (0.058μM). Bulk yields of biomass were inversely proportional to the concentration of each hormone tested. All cultures provided with ABA yielded the highest amount of biomass. Both 6-benzylaminopurine and 2-isopentenyladenine inhibited root growth, however, only 2-isopentenyladenine stimulated artemisinin production, more than twice that of the B5 controls, and more than any other hormone studied. These results will prove useful in increasing hairy root growth and artemisinin production.     

Plant-growth-promoting compounds produced by two agronomically important strains of <Emphasis Type="Italic">Azospirillum brasilense</Emphasis>, and implications for inoculant formulation

We evaluated phytohormone and polyamine biosynthesis, siderophore production, and phosphate solubilization in two strains (Cd and Az39) of Azospirillum brasilense used for inoculant formulation in Argentina during the last 20 years. Siderophore production and phosphate solubilization were evaluated in a chemically defined medium, with negative results. Indole 3-acetic acid (IAA), gibberellic acid (GA₃), and abscisic acid (ABA) production were analyzed by gas chromatography-mass spectrometry. Ethylene, polyamine, and zeatin (Z) biosynthesis were determined by gas chromatography-flame ionization detector and high performance liquid chromatography (HPLC-fluorescence and -UV), respectively. Phytohormones IAA, Z, GA₃, ABA, ethylene, and growth regulators putrescine, spermine, spermidine, and cadaverine (CAD) were found in culture supernatant of both strains. IAA, Z, and GA₃ were found in all two strains; however, their levels were significantly higher (p < 0.01) in Cd (10.8, 2.32, 0.66 μg ml⁻¹). ABA biosynthesis was significantly higher (p < 0.01) in Az39 (0.077 μg ml⁻¹). Ethylene and polyamine CAD were found in all two strains, with highest production in Cd cultured in NFb plus l-methionine (3.94 ng ml⁻¹ h⁻¹) and Az39 cultured in NFb plus l-lysine (36.55 ng ml⁻¹ h⁻¹). This is the first report on the evaluation of important bioactive molecules in strains of A. brasilense as potentially capable of direct plant growth promotion or agronomic yield increase. Az39 and Cd showed differential capability to produce the five major phytohormones and CAD in chemically defined medium. This fact has important technological implications for inoculant formulation as different concentrations of growth regulators are produced by different strains or culture conditions.     

Phytohormone production by three strains of <Emphasis Type="Italic">Bradyrhizobium japonicum</Emphasis> and possible physiological and technological implications

The aim of this work was to evaluate phytohormone biosynthesis, siderophores production, and phosphate solubilization in three strains (E109, USDA110, and SEMIA5080) of Bradyrhizobium japonicum, most commonly used for inoculation of soybean and nonlegumes in USA, Canada, and South America. Siderophore production and phosphate solubilization were evaluated in selective culture conditions, which had negative results. Indole-3-acetic acid (IAA), gibberellic acid (GA₃), and abscisic acid (ABA) production were analyzed by gas chromatography–mass spectrometry (GC-MS). Ethylene and zeatin biosynthesis were determined by GS–flame ionization detection and high-performance liquid chromatography (HPLC-UV), respectively. IAA, zeatin, and GA₃ were found in all three strains; however, their levels were significantly higher (p < 0.01) in SEMIA5080 (3.8 μg ml⁻¹), USDA110 (2.5 μg ml⁻¹), and E109 (0.87 μg ml⁻¹), respectively. ABA biosynthesis was detected only in USDA110 (0.019 μg ml⁻¹). Ethylene was found in all three strains, with highest production rate (18.1 ng ml⁻¹ h⁻¹) in E109 cultured in yeast extract mannitol medium plus l-methionine. This is the first report of IAA, GA₃, zeatin, ethylene, and ABA production by B. japonicum in pure cultures, using quantitative physicochemical methodology. The three strains have differential capability to produce the five major phytohormones and this fact may have an important technological implication for inoculant formulation.     

Effects of macroelements and nitrogen source on biomass accumulation and withanolide-A production from cell suspension cultures of <Emphasis Type="Italic">Withania somnifera</Emphasis> (L.) Dunal

Withania somnifera is an important medicinal plant that contains withanolides and withaferins, both bioactive compounds. We have tested the effects of macroelements and nitrogen source in W. somnifera cell suspension cultures with the aim of optimizing the production of biomass and withanolide A. The effects of the macroelements NH₄NO_3, KNO_3, CaCl_2, MgSO₄ and KH₂PO₄ at concentrations of 0.0, 0.5, 1.0, 1.5 and 2.0× strength and of the nitrogen source [NH₄ ⁺/NO₃ ⁻ (mM/mM) ratio of: 0.00/18.80, 7.19/18.80, 14.38/18.80, 21.57/18.80, 28.75/18.80, 14.38/0.00, 14.38/9.40, 14.38/18.80, 14.38/28.20, and 14.38/37.60 (mM)] in Murashige and Skoog medium were tested for biomass and withanolide A production. The highest accumulation of biomass [147.81 g l⁻¹ fresh weight (FW) and 14.02 g l⁻¹ (dry weight (DW)] was recorded in the medium containing a 0.5× concentration of NH₄NO₃, and the highest production of withanolide A content was recorded in the medium with 2.0× KNO₃ (4.36 mg g⁻¹ DW). The NH₄ ⁺/NO₃ ⁻ ratio also influenced cell growth and withanolide A production, with both parameters being larger when the NO₃ ⁻ concentration was higher than that of NH₄ ⁺. Maximum biomass growth (110.45 g l⁻¹ FW and 9.29 g l⁻¹ DW) was achieved at an NH₄ ⁺/NO₃ ⁻ ratio of 7.19/18.80, while withanolide A production was greatest (3.96 mg g⁻¹ DW) when the NH₄ ⁺/NO₃ ⁻ ratio was 14.38/37.60 mM.     

Abscisic acid concentrations and fluxes in droughted conifer saplings

We present the first study of abscisic acid (ABA) concentrations and fluxes in the xylem sap of conifers during a drought cycle. In both Pinus sylvestris and Picea sitchensis the concentration of ABA in the sap rose 11-fold as the drought progressed. There were clear diurnal trends in this concentration, which reached its maximum (6–8.ininol ABA m^?3) near the middle of the day. The fluxes of ABA were calculated by multiplying the xylem ABA concentration by the sap flow rate. The ABA fluxes in the droughted plants in the middle of the day were usually no higher than those of the controls, as a result of the very low sap flow in the droughted plants at that time. However, the ABA flux in the droughted plants was higher than in the controls in the morning, and we postulate that the stomata are responding to these ‘morning doses’ Stomatal conductance, g_s, could not be related statistically to leaf turgor or to the ABA flux. However, £s did display a negative exponential relationship with ABA concentration in the xylem. Pinus displayed more acclimation to drought than Picea, Its ABA concentration rose and its stomatal conductance fell at day 6 of the drought, as opposed to day 17 for Picea, and its osmotic potential fell during the drought treatment.     

Silicate-Mediated Alleviation of Pb Toxicity in Banana Grown in Pb-Contaminated Soil

Silicate (Si) can enhance plant resistance or tolerance to the toxicity of heavy metals. However, it remains unclear whether Si can ameliorate lead (Pb) toxicity in banana (Musa xparadisiaca) roots. In this study, treatment with 800 mg kg⁻¹ Pb decreased both the shoot and root weight of banana seedlings. The amendment of 800 mg kg⁻¹ Si (sodium metasilicate, Na₂SiO₃·9H₂O) to the Pb-contaminated soil enhanced banana biomass at two growth stages significantly. The amendment of 800 mg kg⁻¹ Si significantly increased soil pH and decreased exchangeable Pb, thus reducing soil Pb availability, while Si addition of 100 mg kg⁻¹ did not influence soil pH. Results from Pb fractionation analysis indicated that more Pb were in the form of carbonate and residual-bound fractions in the Si-amended Pb-contaminated soils. The ratio of Pb-bound carbonate to the total Pb tended to increase with increasing growth stages. Treatment with 100 mg kg⁻¹ Si had smaller effects on Pb forms in the Si-amended soils than that of 800 mg kg⁻¹ Si. Pb treatment decreased the xylem sap greatly, but the addition of Si at both levels increased xylem sap and reduced Pb concentration in xylem sap significantly in the Si-amended Pb treatments. The addition of Si increased the activities of POD, SOD, and CAT in banana roots by 14.2% to 72.1% in the Si-amended Pb treatments. The results suggested that Si-enhanced tolerance to Pb toxicity in banana seedlings was associated with Pb immobilization in the soils, the decrease of Pb transport from roots to shoots, and Si-mediated detoxification of Pb in the plants.     

Linking xylem diameter variations with sap flow measurements

Measurements of variation in the diameter of tree stems provide a rapid response, high resolution tool for detecting changes in water tension inside the xylem. Water movement inside the xylem is caused by changes in the water tension and theoretically, the sap flow rate should be directly proportional to the water tension gradient and, therefore, also linearly linked to the xylem diameter variations. The coefficient of proportionality describes the water conductivity and elasticity of the conducting tissue. Xylem diameter variation measurements could thus provide an alternative approach for estimating sap flow rates, but currently we lack means for calibration. On the other hand, xylem diameter variation measurements could also be used as a tool for studying xylem structure and function. If we knew both the water tension in the xylem and the sap flow rate, xylem conductivity and/or elasticity could be calculated from the slope of their relationship. In this study we measured diurnal xylem diameter variation simultaneously with sap flow rates (Granier-type thermal method) in six deciduous species (Acer rubrum L., Alnus glutinosa Miller, Betula lenta L., Fagus Sylvatica L. Quercus rubra L., and Tilia vulgaris L.) for 7–91 day periods during summers 2003, 2005 and 2006 and analyzed the relationship between these two measurements. We found that in all species xylem diameter variations and sap flow rate were linearly related in daily scale (daily average R ² = 0.61–0.87) but there was a significant variation in the daily slopes of the linear regressions. The largest variance in the slopes, however, was found between species, which is encouraging for finding a species specific calibration method for measuring sap flow rates using xylem diameter variations. At a daily timescale, xylem diameter variation and sap flow rate were related to each other via a hysteresis loop. The slopes during the morning and afternoon did not differ statistically significantly from each other, indicating no overall change in the conductivity. Because of the variance in the daily slopes, we tested three different data averaging methods to obtain calibration coefficients. The performance of the averaging methods depended on the source of variance in the data set and none of them performed best for all species. The best estimates of instantaneous sap flow rates were also given by different averaging methods than the best estimates of total daily water use. Using the linear relationship of sap flow rate and xylem diameter variations we calculated the conductance and specific conductivity of the soil–xylem–atmosphere water pathway. The conductance were of the order of magnitude 10⁻⁵ kg s⁻¹ MPa⁻¹ for all species, which compares well with measured water fluxes from broadleaved forests. Interestingly, because of the large sap wood area the conductance of Betula was approximately 10 times larger than in other species.     

Thermotolerance and Related Antioxidant Enzyme Activities Induced by Heat Acclimation and Salicylic Acid in Grape (Vitis vinifera L.) Leaves

Thermotolerance and related antioxidant enzyme activities induced by both heat acclimation and exogenous salicylic acid (SA) application were studied in grapevine (Vitis vinifera L. cv. Jingxiu). Heat acclimation and exogenous SA application induced comparable changes in thermotolerance, ascorbic acid (AsA), glutathione (GSH), and hydrogen peroxide (H₂O₂) concentrations, and in activities of the antioxidant enzymes superoxide dismutase (SOD), peroxidase (POD), glutathione reductase (GR), ascorbic peroxidase (APX) and catalase (CAT) in grape leaves. Within 1 h at 38 °C, free SA concentration in leaves rose from 3.1 μg g⁻¹ FW to 19.1 μg g⁻¹ FW, then sharply declined. SA application and heat acclimation induced thermotolerance were related to changes of antioxidant enzyme activities and antioxidant concentration, indicating a role for endogenous SA in heat acclimation in grape leaves.