Water Relations of Pachysandra Leaves during Freezing and Thawing : Evidence for a Negative Pressure Potential Alleviating Freeze-Dehydration Stress

The evergreen herb Pachysandra terminalis becomes moderately frost-hardy in winter. The water relations of its frost-hardy leaves were studied during a freeze-thaw cycle. Leaf water potentials, measured by psychrometry at subfreezing temperatures, were identical with those of ice, indicating equilibrium freezing. Microscopic observations showed extracellular freezing of tissue water. As evidenced by thermal analysis, the freezing process starts with the crystallization of a minor volume which was identified as apoplasmic water. The following long-lasting exotherm indicated slow export of water from the protoplasts driven by extracellular crystallization. In partially frozen leaves, the fractions of liquid water were measured at several subfreezing temperatures by nuclear magnetic resonance spectroscopy. They were consistently greater than those calculated from the osmotic potentials of cellular fluid, and the differences increased with decreasing temperature. About 50% of the differences could be abolished by freeze-killing of the leaf and was thus ascribed to the effect of a (negative) pressure reinforcing the osmotic potential. The persistent part of the differences may have reflected a matric component. At −7°C, the absolute values of both potentials were −1.7 megapascals each. The water relations of Pachysandra leaves clearly indicate nonideal equilibrium freezing where negative pressures and matric potentials contribute to the leaf water potential and thus alleviate freeze-dehydration of the tissue.     

Equilibrium freezing of leaf water and extracellular ice formation in Afroalpine ‘giant rosette’ plants

The water potentials of frozen leaves of Afroalpine plants were measured psychrometrically in the field. Comparison of these potentials with the osmotic potentials of an expressed cellular sap and the water potentials of ice indicated almost ideal freezing behaviour and suggested equilibrium freezing. On the basis of the osmotic potentials of expressed cellular sap, the fractions of frozen cellular water which correspond to the measured water potentials of the frozen leaves could be determined (e.g. 74% at -3.0° C). The freezing points of leaves were found to be in the range between 0° C and -0.5° C, rendering evidence for freezing of almost pure water and thus confirming the conclusions drawn from the water-potential measurements. The leaves proved to be frost resistant down to temperatures between -5° C and -15° C, as depending on the species. They tolerated short supercooling periods which were necessary in order to start ice nucleation. Extracellular ice caps and ice crystals in the intercellular space were observed when cross sections of frozen leaves were investigated microscopically at subfreezing temperatures.Symbols T temperature - water potential Dedicated to Professor Dr. Hubert Ziegler on the occasion of his 60th birthday     

Leaf physiology does not predict leaf habit; examples from tropical dry forest

Leaf structure and physiology are thought to be closely linked to leaf longevity and leaf habit. Here we compare the seasonal variation in leaf hydraulic conductance (k_leaf) and water potential of two evergreen tree species with contrasting leaf life spans, and two species with similar leaf longevity but contrasting leaf habit, one being deciduous and the other evergreen. One of the evergreen species, Simarouba glauca, produced relatively short-lived leaves that maintained high hydraulic conductance year round by periodic flushing. The other evergreen species, Quercus oleoides, produced longer-lived leaves with lower k_leaf and as a result minimum leaf water potential was much lower than in S. glauca (–2.8 MPa vs –1.6 MPa). Associated with exposure to lower water potentials, Q. oleoides leaves were harder, had a higher modulus of elasticity, and were less vulnerable to cavitation than S. glauca leaves. Both species operate at water potentials capable of inducing 20 (S. glauca) to 50% (Q. oleoides) loss of k_leaf during the dry season although no evidence of cumulative losses in k_leaf were observed in either species suggesting regular repair of embolisms. Leaf longevity in the deciduous species Rhedera trinervis is similar to that of S. glauca, although maximum k_leaf was lower. Furthermore, a decline in leaf water potential at the onset of the dry season led to cumulative losses in k_leaf in R. trinervis that culminated in leaf shedding.     

Acclimation of leaf hydraulic conductance and stomatal conductance of Pinus taeda (loblolly pine) to long-term growth in elevated CO2 (free-air CO2 enrichment) and N-fertilization

We investigated how leaf hydraulic conductance (K_leaf) of loblolly pine trees is influenced by soil nitrogen amendment (N) in stands subjected to ambient or elevated CO₂ concentrations (CO₂^a and CO₂^e, respectively). We also examined how K_leaf varies with changes in reference leaf water potential (Ψ_leaf‐ref) and stomatal conductance (g_s‐ref) calculated at vapour pressure deficit, D of 1 kPa. We detected significant reductions in K_leaf caused by N and CO₂^e, but neither treatment affected pre‐dawn or midday Ψ_leaf. We also detected a significant CO₂^e‐induced reduction in g_s‐ref and Ψ_leaf‐ref. Among treatments, the sensitivity of K_leaf to Ψ_leaf was directly related to a reference K_leaf (K_leaf‐ref computed at Ψ_leaf‐ref). This liquid‐phase response was reflected in a similar gas‐phase response, with g_s sensitivity to D proportional to g_s‐ref. Because leaves represented a substantial component of the whole‐tree conductance, reduction in K_leaf under CO₂^e affected whole‐tree water use by inducing a decline in g_s‐ref. The consequences of the acclimation of leaves to the treatments were: (1) trees growing under CO₂^e controlled morning leaf water status less than CO₂^a trees resulting in a higher diurnal loss of K_leaf; (2) the effect of CO₂^e on g_s‐ref was manifested only during times of high soil moisture.     

Freezing avoidance by supercooling in Olea europaea cultivars: the role of apoplastic water,solute content and cell wall rigidity

Plants can avoid freezing damage by preventing extracellular ice formation below the equilibrium freezing temperature (supercooling). We used Olea europaea cultivars to assess which traits contribute to avoid ice nucleation at sub‐zero temperatures. Seasonal leaf water relations, non‐structural carbohydrates, nitrogen and tissue damage and ice nucleation temperatures in different plant parts were determined in five cultivars growing in the Patagonian cold desert. Ice seeding in roots occurred at higher temperatures than in stems and leaves. Leaves of cold acclimated cultivars supercooled down to ?13 °C, substantially lower than the minimum air temperatures observed in the study site. During winter, leaf ice nucleation and leaf freezing damage (LT₅₀) occurred at similar temperatures, typical of plant tissues that supercool. Higher leaf density and cell wall rigidity were observed during winter, consistent with a substantial acclimation to sub‐zero temperatures. Larger supercooling capacity and lower LT₅₀ were observed in cold‐acclimated cultivars with higher osmotically active solute content, higher tissue elastic adjustments and lower apoplastic water. Irreversible leaf damage was only observed in laboratory experiments at very low temperatures, but not in the field. A comparative analysis of closely related plants avoids phylogenetic independence bias in a comparative study of adaptations to survive low temperatures.     

Measurement of Leaf Hydraulic Conductance and Stomatal Conductance and Their Responses to Irradiance and Dehydration Using the Evaporative Flux Method (EFM)

Water is a key resource, and the plant water transport system sets limits on maximum growth and drought tolerance. When plants open their stomata to achieve a high stomatal conductance (g_s) to capture CO₂ for photosynthesis, water is lost by transpiration^1,2. Water evaporating from the airspaces is replaced from cell walls, in turn drawing water from the xylem of leaf veins, in turn drawing from xylem in the stems and roots. As water is pulled through the system, it experiences hydraulic resistance, creating tension throughout the system and a low leaf water potential (Ψ_leaf). The leaf itself is a critical bottleneck in the whole plant system, accounting for on average 30% of the plant hydraulic resistance³. Leaf hydraulic conductance (K_leaf = 1/ leaf hydraulic resistance) is the ratio of the water flow rate to the water potential gradient across the leaf, and summarizes the behavior of a complex system: water moves through the petiole and through several orders of veins, exits into the bundle sheath and passes through or around mesophyll cells before evaporating into the airspace and being transpired from the stomata. K_leaf is of strong interest as an important physiological trait to compare species, quantifying the effectiveness of the leaf structure and physiology for water transport, and a key variable to investigate for its relationship to variation in structure (e.g., in leaf venation architecture) and its impacts on photosynthetic gas exchange. Further, K_leaf responds strongly to the internal and external leaf environment³. K_leaf can increase dramatically with irradiance apparently due to changes in the expression and activation of aquaporins, the proteins involved in water transport through membranes⁴, and K_leaf declines strongly during drought, due to cavitation and/or collapse of xylem conduits, and/or loss of permeability in the extra-xylem tissues due to mesophyll and bundle sheath cell shrinkage or aquaporin deactivation^5-10. Because K_leaf can constrain g_s and photosynthetic rate across species in well watered conditions and during drought, and thus limit whole-plant performance they may possibly determine species distributions especially as droughts increase in frequency and severity^11-14.We present a simple method for simultaneous determination of K_leaf and g_s on excised leaves. A transpiring leaf is connected by its petiole to tubing running to a water source on a balance. The loss of water from the balance is recorded to calculate the flow rate through the leaf. When steady state transpiration (E, mmol • m^-2 • s^-1) is reached, g_s is determined by dividing by vapor pressure deficit, and K_leaf by dividing by the water potential driving force determined using a pressure chamber (K_leaf= E /- Δψ_leaf, MPa)¹⁵.This method can be used to assess K_leaf responses to different irradiances and the vulnerability of K_leaf to dehydration^14,16,17.     

Interpreting Leaf Water Potential Measurements with a Model of the Soil-Plant-Atmosphere Continuum

A water flux model, which assumes that the dynamic functioning of the soil-plant-atmosphere continuum may be described by a series of steady states, was examined as a means for interpreting leaf water potential measurements in ‘Valencia’ orange trees (Citrus sinensis (L.) Osbeck). According to the model, leaf water potential should be related to transpirational flux, which in this experiment was estimated by the ratio of vapor pressure deficit of the atmosphere to leaf diffusion resistance (VPD/r_leaf). Leaf water potentials decreased in a specific relationship with increasing values of VPD/r_leaf provided that soil water was adequate and soil temperature was not too low, but regardless of season of the year or climatic or edaphic differences among 3 field locations. When soil water tensions exceeded 0.3 bar or when soil temperatures were lower than 15°C, deviations from the model occurred in the form of more negative leaf water potentials than predicted by VPD/r_leaf. The model predicts from simple measurements made on intact plants that these differences were due to the modification of flow resistances by cool temperatures and the modification of both resistances and the potential of water at the source in the case of soil water depletion. The model may be a useful tool for interpreting plant water potential data under contrasting environmental conditions.     

Leaf hydraulic safety margin and safety–efficiency trade-off across angiosperm woody species

Leaf hydraulic conductance and the vulnerability to water deficits have profound effects on plant distribution and mortality. In this study, we compiled a leaf hydraulic trait dataset with 311 species-at-site combinations from biomes worldwide. These traits included maximum leaf hydraulic conductance (K_leaf), water potential at 50% loss of K_leaf (P50_leaf), and minimum leaf water potential (Ψ_min). Leaf hydraulic safety margin (HSM_leaf) was calculated as the difference between Ψ_min and P50_leaf. Our results indicated that 70% of the studied species had a narrow HSM_leaf (less than 1 MPa), which was consistent with the global pattern of stem hydraulic safety margin. There was a positive relationship between HSM_leaf and aridity index (the ratio of mean annual precipitation to potential evapotranspiration), as species from humid sites tended to have larger HSM_leaf. We found a significant relationship between K_leaf and P50_leaf across global angiosperm woody species and within each of the different plant groups. This global analysis of leaf hydraulic traits improves our understanding of plant hydraulic response to environmental change.     

Plasticity of vulnerability to leaf hydraulic dysfunction during acclimation to drought in grapevines: an osmotic‐mediated process

Previous studies have reported correlation of leaf hydraulic vulnerability with pressure–volume parameters related to cell turgor. This link has been explained on the basis of the effects of turgor on connectivity among cells and tissue structural integrity, which affect leaf water transport. In this study, we tested the hypothesis that osmotic adjustment to water stress would shift the leaf vulnerability curve toward more negative water potential (Ψ_leaf) by increasing turgor at low Ψ_leaf. We measured leaf hydraulic conductance (K_leaf), K_leaf vulnerability [50 and 80% loss of K_leaf (P₅₀ and P₈₀); |Ψ_leaf| at 50 and 80% loss of K_leaf, respectively), bulk leaf water relations, leaf gas exchange and sap flow in two Vitis vinifera cultivars (Tempranillo and Grenache), under two water treatments. We found that P₅₀, P₈₀ and maximum K_leaf decreased seasonally by more than 20% in both cultivars and watering treatments. However, K_leaf at ?2 MPa increased threefold, while osmotic potential at full turgor and turgor loss point decreased. Our results indicate that leaf resistance to hydraulic dysfunction is seasonally plastic, and this plasticity may be mediated by osmotic adjustment.     

Stem and leaf hydraulics of congeneric tree species from adjacent tropical savanna and forest ecosystems

Leaf and stem functional traits related to plant water relations were studied for six congeneric species pairs, each composed of one tree species typical of savanna habitats and another typical of adjacent forest habitats, to determine whether there were intrinsic differences in plant hydraulics between these two functional types. Only individuals growing in savanna habitats were studied. Most stem traits, including wood density, the xylem water potential at 50% loss of hydraulic conductivity, sapwood area specific conductivity, and leaf area specific conductivity did not differ significantly between savanna and forest species. However, maximum leaf hydraulic conductance (K _leaf) and leaf capacitance tended to be higher in savanna species. Predawn leaf water potential and leaf mass per area were also higher in savanna species in all congeneric pairs. Hydraulic vulnerability curves of stems and leaves indicated that leaves were more vulnerable to drought-induced cavitation than terminal branches regardless of genus. The midday K _leaf values estimated from leaf vulnerability curves were very low implying that daily embolism repair may occur in leaves. An electric circuit analog model predicted that, compared to forest species, savanna species took longer for their leaf water potentials to drop from predawn values to values corresponding to 50% loss of K _leaf or to the turgor loss points, suggesting that savanna species were more buffered from changes in leaf water potential. The results of this study suggest that the relative success of savanna over forest species in savanna is related in part to their ability to cope with drought, which is determined more by leaf than by stem hydraulic traits. Variation among genera accounted for a large proportion of the total variance in most traits, which indicates that, despite different selective pressures in savanna and forest habitats, phylogeny has a stronger effect than habitat in determining most hydraulic traits.     

Diurnal depression of leaf hydraulic conductance in a tropical tree species

Diurnal patterns of hydraulic conductance of the leaf lamina (K_leaf) were monitored in a field‐grown tropical tree species in an attempt to ascertain whether the dynamics of stomatal conductance (g_s) and CO₂ uptake (A_leaf) were associated with short‐term changes in K_leaf. On days of high evaporative demand mid‐day depression of K_leaf to between 40 and 50% of pre‐dawn values was followed by a rapid recovery after 1500 h. Leaf water potential during the recovery stage was less than ?1 MPa implying a refilling mechanism, or that loss of K_leaf was not linked to cavitation. Laboratory measurement of the response of K_leaf to Ψ_leaf confirmed that leaves in the field were operating at water potentials within the depressed region of the leaf ‘vulnerability curve’. Diurnal courses of K_leaf and Ψ_leaf predicted from measured transpiration, xylem water potential and the K_leaf vulnerability function, yielded good agreement with observed trends in both leaf parameters. Close correlation between depression of K_leaf, g_s and A_leaf suggests that xylem dysfunction in the leaf may lead to mid‐day depression of gas exchange in this species.     

Cold tolerance,water relations and accumulation of osmolytes in <Emphasis Type="Italic">Bituminaria bituminosa</Emphasis>

We determined the cold (freezing) tolerance of five Spanish populations of the perennial shrub Bituminaria bituminosa (L.) C.H. Stirton (Fabaceae), as the temperature at which 50 % of leaf electrolytes are released (LT₅₀) using leaves of field-grown plants, obtained in two winters and one spring. The freezing tolerance was greater in winter and reflected the minimum temperatures at the original sites from which the populations were obtained. Tolerance in vitro was related to osmotic adjustment in the leaves; more negative osmotic potential values and more positive pressure potential values (MPa) were associated with greater tolerance. Tolerance and osmotic potential were not related to leaf cation contents but to leaf amino acids, soluble sugar and proline contents.     

美国海滨桤木和薄叶桤木水分生理特性的比较

采取盆栽、人工控水的方式, 研究并比较了美国本土海滨桤木(Alnus maritima)和薄叶桤木(A. incana)的气孔导度(G_s)、叶片水势(ψ_leaf)以及渗透调节能力对土壤水分条件的响应, 以探讨引起两种桤木生态分布差异巨大的生理生态原因。结果表明: 1)正常水分条件下, 海滨桤木的G_s低于薄叶桤木, 其与大气温度、相对湿度和水蒸气亏缺等气象因子的相关性低于薄叶桤木; 干旱胁迫下, 海滨桤木的G_s对其自身ψ_leaf下降信号的敏感度低于薄叶桤木; 复水后, 其G_s恢复更为缓慢。2)正常水分条件下, 海滨桤木的ψ_leaf高于薄叶桤木, 且引起气孔关闭的ψ_leaf临界值较高; 干旱胁迫下, 海滨桤木的ψ_leaf下降幅度高于薄叶桤木。3)正常水分条件下, 海滨桤木和薄叶桤木的渗透调节能力无显著差异; 干旱胁迫下, 尽管两种桤木均表现出饱和状态渗透势(ψ_s^sat)下降、膨压与水势关系的最大变化率降低、初始失膨点渗透势(ψ_s^tlp)增加、细胞渗透调节能力范围(ψ_s^sat-ψ_s^tlp, Dψ_s)减小的趋势, 但与薄叶桤木相比, 海滨桤木的ψ_s^tlp较高, Dψ_s较小。从以上生理生态指标可以看出, 较高的叶片水势、较低的气孔调节能力、干旱下较低的渗透调节能力是造成海滨桤木分布范围狭小的重要原因。     

Daily and seasonal variation in water relations of macchia shrubs and trees in France (Montpellier) and Turkey (Antalya)

Based upon two different research studies in the mediterranean regions of France and Turkey, drought resistance strategies were investigated in a broad group of species. The diurnal and seasonal patterns of the water relations of different lifeforms from the thermo-mediterranean to submediterranean lifezones were compared. Three sites near Montpellier, in Southern France, and five sites near Antalya, Turkey were used for this comparison. Xylem pressure potential and relative stomatal aperture were the key water relations parameters collected in France while these parameters as well as osmotic potential and leaf conductance were studied in Turkey.From the 26 different study species investigated in France, 7 distinct types of stomatal control were observed, with the deciduous lifeforms showing the least control, the sclerophyllous and coniferous evergreens the greatest control and the malacophyllous shrublets intermediate levels of control. Predawn water potential values provided a means of classifying species according to their temporal and spatial utilization of site water reserves. The comparison of turgor potentials (difference between water and osmotic potentials) gave an insight into leaf adaptations to site moisture. Species with high predawn water potentials generally maintain positive turgor even at midday during the summer, whereas species with low predawn values were frequently at zero turgor even at predawn. Phlomis grandiflora was the most extreme species with mid-summer predawns and midday water potentials of –6 MPa and osmotic potentials never more negative than –2.4 MPa.     

Which water potential? Differences between isopiestic thermocouple psychrometer measurements of intact and excised plant materials

Water potentials of leaves from well-watered plants were measured. There were species-specific differences in both the total and the osmotic potentials of pea (Pisum sativum), tradescantia (Tradescantia versicolor), rose (Rosa hybrida), bitter lemon (Citrus aurantium) and olive (Olea europaea). With tradescantia the potential measured after the destruction of turgor by freezing was less negative than before, a result which suggests that the value obtained is not identical with the real osmotic potential of the leaf. detached leaves of all species showed less negative water potential readings, and those of pea even a less negative osmotic potential, when cut into five pieces than when measured intact. Application of vaseline to the cut surface of the leaves reduced this effect with rose and olive, though not with tradescantia and pea. Measurements were also made of the water potentials of comparable leaves of tradescantia and bitter lemon, attached to and detached from their plants; when bitter lemon leaves were detached and watered through their petioles which protruded outside the thermocouple chamber, their potential became considerably less negative than when the same leaves had been attached to well watered plants. However, similar leaves whose cut petioles were introduced into the thermocouple chamber registered an even less negative potential. The results are consistent with the hypothesis that when a leaf is cut off a plant, and even more so when it is cut into sections, the water previously held by matrix forces becomes available to dilute the “spilled” cell sap and to be absorbed by adjacent cells and thereby to increase their turgor and render the net water potential of the leaf less negative. Similarly, the apparent negative turgor of the succulent, tradescantia leaves is likely to be due to dilution of the osmotic component by cell wall water. The discrepancies between the readings of attached and detached leaves indicate a considerable whole-plant matrix component, and the results as a whole suglest that thermocouple psychrometer readings carried out on detached and even more on cut-up leaves may be artifacts and that it is desirable to determine water potentials on leaves attached to their plants. The work was supported by a Government of Israel Fellowship and was conducted at the Department of Pomology and Viticulture, Faculty of Agriculture of the Hebrew University of Jerusalem, Rehovot, Israel.     

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.     

The relationship between leaf water potential ψ leaf and the levels of abscisic acid and ethylene in excised wheat leaves

The amount of diffusible ethylene from excised wheat leaves (Triticum aestivum L. cv. Eclipse) increased when they were subjected to water stress. The quantity of ethylene produced was related to the severity of the stress, reaching a maximum at a leaf water potential _leaf of approximately-12 bars. Irrespective of the severity of the stress, the maximum rate of ethylene production usually occurred between 135–270 min after applying the stress and then the rate declined. Part of the decline may have been due to an oxygen deficiency in the leaf chambers. In excised water-stressed leaves there was a sigmoid relationship between increasing ethylene and abscisic acid (ABA) levels and decreasing leaf water potential values. The two curves were displaced from each other by approximately 1 bar, with ethylene evolution leading that of ABA accumulation. The maximum rate of increase in ethylene occurred between-8 and-9 bars and for ABA between-9 and-10 bars. A significant increase in the levels of these two plant growth regulators was found when the _leaf decreased outside the normal diurnal _leaf range by 1 bar for ethylene and 2 bars for ABA. Because of the sigmoid nature of the curves there was no distinct threshold _leaf value triggering-off an increase in ethylene or ABA, but with ABA the curve became very steep at a _leaf value of-9.3 bars and this could be looked upon as a kind of threshold value.It seems unlikely that the stress-induced ethylene evolution in excised wheat leaves stimulated the accumulation of ABA, because when the leaves were subjected to a substantial water stress (e.g. _leaf bars) ABA increased immediately and at a faster rate than ethylene.Abbreviations ABA abscisic acid - GLC gas-liquid chromatography - RWC relative water content - TLC thin-layer chromatography - _leaf leaf water potential     

Direct measurement of turgor and osmotic potential in individual epidermal cells : independent confirmation of leaf water potential as determined by in situ psychrometry

The pressure probe, which is routinely used to measure the turgor potential (Ψ_p) of individual epidermal cells in Tradescantia virginiana (L.), has also been used to sample small volumes of vacuolar fluid from these same cells (as low as 0.02 nl) for measurement of cellular solute (osmotic) potential (Ψ_s) in a micro freezing point osmometer. The water potential components Ψ_p and Ψ_o have been used to calculate the total water potential of individual epidermal cells (Ψ_cell) which has then been directly compared to the total leaf water potential (Ψ_leaf) measured psychrometrically. The relation of Ψ_leaf and Ψ_cell to leaf transpiration indicates that in T. virginiana, a relatively straightforward relation exists between the level of water flow through the leaf tissue, and the ΔΨ within the leaf, between two points along the water flow pathway. Substantial agreement was found between the two independent, in situ methods of measuring Ψ when extrapolated to zero transpiration conditions. These results are discussed with respect to the thermodynamics of water transport in plant tissues.     

Factors of between- and within-plant distribution of Metopolophium dirhodum (Hom., Aphididae) on small grain cereals

Abstract: The relationship between abundance of rose‐grain aphid Metopolophium dirhodum (Walker) and leaf chlorophyll content of spring wheat, spring oats and winter barley was investigated. Within production stands of each crop 18–25 plots were established, located at places with different plant quality where aphids were counted on particular leaves of 50–100 tillers, and the leaf chlorophyll content and area were determined. In all stands aphid numbers × tiller^?1 increased exponentially with chlorophyll content (r² = 0.783–0.933). This parallel variation may be explained by increased nitrogen content and assimilate production of vigorous chlorophyll‐rich plants. The aphid numbers on leaves of particular order (within‐plant distribution) were also correlated with leaf chlorophyll content (r² = 0.373–0.827). However, in oats and barley the analysis of variance of residuals of log aphids × leaf^?1 versus leaf chlorophyll regression revealed a significant effect of leaf position (the order of the leaf from the top of the plant). The magnitude of residuals was positively related to leaf size and may be related to the intensity of phloem transport from the source leaves to sink organs.     

Freezing tolerance and avoidance in high-elevation Hawaiian plants

Freezing resistance mechanisms were studied in five endemic Hawaiian species growing at high elevations on Haleakala volcano, Hawaii, where nocturnal subzero (°C) air temperatures frequently occur. Extracellular freezing occurred at around -5°C in leaves of Argyroxiphium sandwicense and Sophora chrysophylla, but these leaves can tolerate extracellular ice accumulation to -15°C and -12°C, respectively. Mucilage, which apparently acted as an ice nucleator, comprised 9 to 11% of the dry weight of leaf tissue in these two species. Leaves of Vaccinium reticulatum and Styphelia tameiameiae were also found to tolerate substantial extracellular freezing. Dubautia menziesii, on the other hand, exhibited the characteristics of permanent supercooling; a very rapid decline in liquid water content associated with simultaneous intracellular and extracellular freezing. However, in those species that tolerate extracellular freezing, the decline in liquid water content during freezing is relatively slow. Osmotic potential was lower at pre-dawn than at midday in four of the species studied. Nocturnal production of osmotically active solutes may have helped to prevent intracellular freeze dehydration as well as to provide non-colligative protection of cell membranes. Styphelia tameiameiae supercooled to -9·3°C and tolerated tissue freezing to below -15°C, a unique combination of physiological characteristics related to freezing. Tolerance of extracellular ice formation after considerable supercooling may have resulted from low tissue water content and a high degree of intracellular water binding in this species, as determined by nuclear magnetic resonance studies. The climate at high elevations in Hawaii is relatively unpredictable in terms of the duration of subzero temperatures and the lowest subzero temperature reached during the night. It appears that plants growing in this tropical alpine habitat have been under selective pressures for the evolution of freezing tolerance mechanisms.