Historical biogeography and phylogenetic relationships of the genus Chamaecyparis (Cupressaceae) inferred from chloroplast DNA polymorphism

The genus Chamaecyparis comprises five species and one variety native to Taiwan, Japan, Canada, and USA, which demonstrates a classical eastern Asian, western North American, and eastern North American disjunct distributional pattern. The phylogenetic relationships of the species of Chamaecyparis were inferred by comparing 1130 bp of the combined data set of chloroplast trnV intron and petG-trnP intergenic spacer. The phylogenetic tree shows that Chamaecyparis nootkatensis (Cupressus nootkatensis or Xanthocyparis nootkatensis) is clearly diverged from other Chamaecyparis species. For Chamaecyparis species, C. thyoides is sister to C. formosensis and C. pisifera and these together form a monophyletic group. C. lawsoniana is sister to C. obtusa and C. taiwanensis; and these form another monophyletic group. Homogeneity in evolutionary rates was found among species in these two monophyletic groups. Results indicate the divergent evolution of C. taiwanensis and C. formosensis and molecular evidence in this investigation supports C. taiwanensis as a variety of C. obtusa. Utility of cpDNA intergenic spacer petG-trnP in Chamaecyparis is also discussed. Several biogeographical implications were inferred: (1) at least two divergence events have produced the eastern Asian, and both western and eastern North American disjunct distribution in Chamaecyparis; (2) intercontinental sister species pairs are found in Chamaecyparis; (3) cpDNA divergence between two intercontinental sister pairs of C. thyoides and C. pisifera, and C. lawsoniana and C. obtusa is 2.8% and 1.1%, which suggest an estimated divergence time of 14 and 5.5 million years ago during middle and late Miocene, respectively; (4) cpDNA divergence of two Asian Chamaecyparis groups between C. obtusa and C. taiwanensis, and between C. pisifera and C. formosensis is 0.25% and 0.57%, which suggest an estimated divergence time of 1.3 and 2.9 million years ago during Pleistocene and late Pliocene, respectively; these estimated divergence times suggest a relatively recent migration of Chamaecyparis to Taiwan from the Japanese Archipelago; (5) that climatic deterioration caused the disappearance of Chamaecyparis in continental Asia is probable.     

Foliar nutrient concentrations of small Chamaecyparis in Taiwan

Summary Seedlings ofChamaecyparis lawsoniana, C. formosensis andC. taiwanensis were grown in a forest nursery in Taiwan. Native saplings of the latter two species were sampled in a clearcut and in the forest understory. Foliar nutrient concentrations were mostly in the mid-range of values for other conifers. However, Ca (.59–1.5%) and Cu (7–20 ppm) were almost always high. Concentrations of N, P, and K in field saplings in Taiwan were low enough (.7–1.5%, .07–.15%, .47–.60%, respectively) to indicate that these elements are probably limiting growth.Among the three species,C. lawsoniana usually had the lowest foliar nutrient values. When a difference among the species from Taiwan occurred,C. taiwanensis usually was lower. Foliar nutrient concentration varied among provenances, especially withinC. formosensis. Seasonal variability did not occur consistently; nutrients which did change increased their concentrations from the dormant to the growing season, except for K which decreased.     

RNA Synthesis in Spring and Winter Wheat during Cold Acclimation

The factors responsible for the low transpiration rates of citrus were investigated. Leaf resistance to water vapor exchange by orange seedlings (Citrus sinensis L. cv. Koethen) including a substantial boundary layer resistance, was as low as 1 s cm⁻¹ in humid air. Leaf resistance of well watered plants increased to values as large as 5 s cm⁻¹ when the difference in absolute humidity between leaf and air was increased. Leaf resistance was only slightly influenced by temperature between 20 and 30°C providing the humidity difference between leaf and air was kept constant. Leaf resistance increased when leaf temperature was increased between 20 and 30°C when the absolute humidity external to the leaf was kept constant. Increased humidity differences resulted in greater increases in leaf resistance during initial experiments than when the experiments were repeated with the same leaves indicating acclimation by the plant. It was concluded that the effects of humidity differences on leaf resistance are partially responsible for the low transpiration rates of citrus.     

Effects of water stress on gas exchange and water relations of a succulent epiphyte,Kalanchoë uniflora

Summary Measurements of gas exchange, xylem tension and nocturnal malate synthesis were conducted with well-watered and droughted plants of Kalanchoë uniflora. Corresponding results were obtained with plants grown in 9 h and 12 h photoperiods. In well-watered plants, 50 to 90% of total CO₂-uptake occurred during the light period. Nocturnal CO₂-uptake and malate synthesis were higher and respiration rate was lower in old leaves (leaf pairs 6 to 10) compared to young leaves (leaf pairs 1 to 5). Within four days of drought distinct physiological changes occurred. Gas exchange during the light period decreased and CO₂-uptake during the dark period increased. Nocturnal malate synthesis significantly increased in young leaves.Respiration rate decreased during periods of drought, this decrease being more pronounced in young leaves compared to old leaves. Restriction of gas exchange during the light period resulted in a decrease of transpiration ratio from more than 100 to about 20. The difference between osmotic pressure and xylem tension decreased in young leaves, indicating a reduction in bulk leaf turgor-pressure.We conclude that both the CAM-enhancement in young leaves and the decrease of respiration rate are responsible for the increase of nocturnal CO₂-uptake during water stress. During short drought periods, which frequently occur in humid habitats, the observed physiological changes result in a marked reduction of water loss while net CO₂-uptake is maintained. This might be relevant for plant growth in the natural habitat.Abbreviations LP light period - DP dark period - CAM crassulacean acid metabolism     

Drought and changes in leaf orientation for two California chaparral shrubs: Ceanothus megacarpus and Ceanothus crassifolius

Summary Predawn xylem pressure potentials were measured on two California chaparral shrubs, Ceanothus megacarpus and Ceanothus crassifolius, throughout the winter and spring growing season and into the summer drought. On the days xylem pressure potentials were measured, leaf orientation measurements were made on a population of marked leaves from the same shrubs. Predawn xylem pressure potentials decreased from -0.1 MPa in both species to -7.8 and -6.6 MPa in C. megacarpus and C. crassifolius, respectively, between May and August, 1981. Leaf inclinations became more vertical during this period with the greatest change observed in C. crassifolius. This change in leaf inclination was reversible, and, in the late winter and early spring, one year old leaves became more horizontal. Leaf azimuths were random and did not change seasonally. Simulations of solar radiation interception indicated that the increase in leaf inclination associated with summer drought reduced the absorption of solar radiation in August by 6% for C. megacarpus and 20% for C. crassifolius. Standard leaf energy budget calculations suggest that steep leaf inclinations would result in slightly lower leaf temperatures and transpiration rates under summer conditions.     

Transpiration inhibition by stored xylem sap from well-watered maize plants

There is increasing evidence that a chemical signal exists in xylem sap of plants subjected to water deficits which influences physiological responses in plant shoots. An important method of studying this signal is the transpiration response of excised leaves exposed to xylem sap collected from plants. However, Munns et al [Plant, Cell & Environment 16, 867–877] cautioned that transpiration inhibition is observed when xylem sap collected from wheat and barley is stored before determining physiological activity. The objective of the study reported here was to determine if transpiration inhibition develops in maize sap collected from well-watered plants when the sap is stored under various conditions. It was found that storage of maize sap collected from well-watered plants for only 1 d at -20°C resulted in the development of substantial transpiration inhibition in bioassay leaves. Storage of sap at 4°C resulted in the development of the effect after 2 weeks, while storage at ?86°C showed only small transpiration inhibition after 3 weeks. The major source of the transpiration inhibition was the development of a substance in the stored sap that resulted in physical blockage of the transpiration stream in bioassay leaves. However, a small signal component may also have developed in the stored sap. Because of the possibility of ionic activity under freezing conditions at ?20°C, calcium was studied for its potential involvement in the transpiration inhibition. However, the calcium concentrations found to inhibit transpiration were nearly an order of magnitude larger than the calcium concentrations observed in xylem sap.     

Abscisic acid in apoplastic sap can account for the restriction in leaf conductance of white lupins during moderate soil drying and after rewatering

We studied the effects of drought on leaf conductance (g) and on the concentration of abscisic acid (ABA) in the apoplastic sap of Lupinus albus L. leaves. Withholding watering for 5d resulted in complete stomatal closure and in severe leaf water deficit. Leaf water potential fully recovered immediately after rewatering, but the aftereffect of drought on stomata persisted for 2d. ABA and sucrose were quantified in pressurized leaf xylem extrudates. We assumed that the xylem sucrose concentration is negligible and hence that the presence of sucrose in leaf extrudates indicated that they were contaminated by phloem. To eliminate this interference, the concentration of ABA in leaf apoplast was estimated by extrapolation to zero sucrose concentration, using the regression between ABA and sucrose concentrations. The estimated apoplastic ABA concentration increased by 100-fold with soil drying and did not return to pre-stress values immediately following rewatering. g was closely related to the concentration of ABA in leaf apoplast. Furthermore, the feeding of exogenous ABA to leaves detached from well-watered plants brought about the same degree of depression in g as resulted from the drought-induced increase in ABA concentration. We therefore conclude that the observed changes in the concentration of ABA in leaf apoplast were quantitatively adequate to explain drought-induced stomatal closure and the delay in stomatal reopening following rewatering.     

Environmental Effects on Transpiration and Leaf Water Potential in Avocado

A field study was conducted to determine how atmospheric and edaphic conditions influenced the water relations of avocado trees (Persea americana Mill. cv. Bacon). With high and low levels of incident photosynthetically active radiation (PAR, 400–700 nm wave length), and either wet or dry soil, leaf conductance decreased as the absolute humidity difference from leaf to air increased. For any water stress treatment, conductance was higher at high PAR than at low PAR. Both conductance and transpiration were higher in well-watered trees than in stressed trees, and in prestressed trees levels were intermediate to unstressed and stressed trees. A model for water flux through the soil-plant-atmosphere continuum was used to examine the relationship of leaf xylem pressure potential to transpiration in well-watered trees and in trees stressed by dry soil. There was a close linkage between leaf xylem pressure potential and transpiration in unstressed and previously stressed trees with high or low PAR, i.e. similar potentials occurred with equivalent transpiration regardless of previous treatment or time of day. In stressed trees, xylem pressure potential was lower than in unstressed trees both during the day and night, and at a given transpiration rate the potential was lower after 1400 h than before that time. The model indicated that in stressed trees xylem pressure potential was uncoupled from transpiration, presumably because of altered resistance in the soil-root portion of the transport system.     

Development of a dot immunobinding assay to detect Phytophthora spp. in naturally dark-rooted woody plants

An indirect dot immunobinding assay (DIBA) with a PAb raised against an isolate of P. cinnumomi was evaluated to detect Phytophthora spp. in naturally dark-rooted woody plants. Screening of different modifications of the procedure were done with root samples of Chamaecyparis lawsoniana non-infected and infected with P. cinnamomi. With the optimised DIBA procedure, root infection could be diagnosed by a clearly defined halo around the spot and by a colour change in the spot using Fast Red or Fast Blue substrate. Detection of different Phytophthora species in Chamaecyparis plants from commercial nurseries was successful with the optimised procedure.     

Effect of Evaporative Demand on Water Relations of Citrus limon

Conductance, transpirational flux and xylem pressure potentialwere measured in leaves of well-watered 5-year-old lemon trees(Citrus limon (L.) Burm. f.) subjected to different levels ofevaporative demand. Increased leaf-to-air absolute humiditydifference generally decreased stomatal conductance and increasedxylem pressure potential, with a good correlation between thelast two parameters; but this trend was reversed on days withvery high evaporative demand, when stomata opened in spite ofthe low humidity. Citrus limon (L.) Burm. f., lemon, water stress, stomatal conductance, leaf water potential, transpiration, air humidity     

Hydraulic properties of Pinus halepensis,Pinus pinea and Tetraclinis articulata in a dune ecosystem of Eastern Spain

The hydraulic properties of Pinus pinea, Pinus halepensis and Tetraclinis articulata were studied in a coastal dune area from Eastern Spain. The measured variables include vulnerability to xylem embolism (vulnerability curves), hydraulic conductivity and carbon isotopic discrimination in leaves. Leaf water potentials were also monitored in the three studied populations during an extremely dry period. Our results showed that roots had always wider vessels and higher hydraulic conductivity than branches. Roots were also more vulnerable to xylem embolism and operated closer to their hydraulic limit (i.e., with narrower safety margins). Although it was not quantified, extensive root mortality was observed in the two pines during the study period, in agreement with the high values of xylem embolism (> 75%) predicted from vulnerability curves and the water potentials measured in the field. T. articulata was much more resistant to embolism than P. pinea and P. halepensis. Since T. articulata experienced also lower water potentials, safety margins from hydraulic failure were only slightly wider in this species than in the pines. Combining species and tissues, high resistance to xylem embolism was associated with low hydraulic conductivity and with high wood density. Both relationships imply a cost of having a resistant xylem. The study outlined very different water-use strategies for T. articulata and the pines. Whereas T. articulata had a conservative strategy that relied on the low vulnerability of its conducting system to drought-induced xylem embolism, the two pines showed regulatory mechanisms at different levels (i.e., embolism, root demography) that constrained the absorption of water when it became scarce. This revised version was published online in August 2006 with corrections to the Cover Date.     

Gas exchange and water balance of a mistletoe species and its mangrove hosts

Summary The gas exchange and water relations of the hemiparasite Pthirusa maritima and two its mangrove host species, Conocarpus erectus and Coccoloba uvifera, were studied in an intertidal zone of the Venezuelan coast. Carbon uptake and transpiration, leaf osmotic and total water potential, as well as nutrient content in the xylem sap and leaves of mistletoes and hosts were followed through the dry and wet season. In addition, carbon isotope ratios of leaf tissue were measured to further evaluate water use efficiency. Under similar light and humidity conditions, mistletoes had higher transpiration rates, lower leaf water potentials, and lower water use efficiencies than their hosts. Potassium content was much higher in mistletoes than in host leaves, but mineral nutrient content in the xylem sap of mistletoes was relatively low. The resistance of the liquid pathway from the soil to the leaf surface of mistletoes was larger than the total liquid flow resistance of host plants. Differences in the daily cycles of osmotic potential of the xylem sap also indicate the existence of a high resistance pathway along the vascular connection between the parasite pathway along the vascular connection between the parasite and its host. P. maritima mistletoes adjust to the different physiological characteristics of the host species which it parasitizes, thus ensuring an adequate water and carbon balance.     

The Relative Magnitude of the Partial Resistances to Transpirational Water Movement in Sunflower (Helianthus annuus L.)

The effects on leaf water status of cooling entire root systemsor stem and petiolar segments were examined using hydroponicsunflower plants. Leaf water potential (^I) decreased by up to7 x 10⁵ Pa when root temperature was reduced to 10 °C orbelow; complete recovery occurred subsequently provided freezingwas avoided. Leaf water status was unaffected by cooling stemor petiolar segments unless freezing occurred, when severe irreversiblewater stress developed above the cooled zone. The leaves belowthe cooled zone were unaffected, demonstrating that the stressdid not originate from transmitted effects on the roots. Subsequent measurements using small heads of water applied toexcised petioles demonstrated that petiolar resistance was low(c. 0.04 Pa s cm_–4) except in immature and senescent petioleswhere resistance was up to 10 times larger. This trend reflectedthe developmental stage and condition of the xylem. Abrupt increases in evaporative demand, obtained by rapid reductionof relative humidity from 100% to 60 or 40%, induced transientdecreases in of approximately 4.5 and 2.5 x 10⁵ Pa, respectively,which were accompanied by simultaneous large increases in stomatalresistance. No simultaneous transient effects were observedin the stem xylem, demonstrating that the factor responsiblefor the formation of the transient foliar stress resides withinthe lamina. The results are discussed in relation to the relative magnitudeof the various partial resistances to transpirational watermovement.     

Hydraulic conductance, stomatal conductance, and maximal photosynthetic rate in bean leaves

A positive correlation was found between steady state values of hydraulic (L _pA) and stomatal conductance (g _s) of French bean leaves: both were lower in the dark than in the light and lower in water-deficient plants than in the well-watered ones. The relative rate of stomatal opening after a pressure rise in the xylem was also positively related to L _pA. The L _pA and g _s were both related to the maximal photosynthetic rate at saturating CO2 concentrations. This revised version was published online in June 2006 with corrections to the Cover Date.     

Leaf Water Diffusion Resistance in Clonal Tea (Camellia sinensis L.): Effects of Water Stress, Leaf Age and Clones

Leaf diffusion resistance was influenced by leaf age in well-wateredand water-stressed clonal tea plants. In well-watered plantsand in two of the three clones studied, young leaves showeda significantly lower diffusion resistance than old leaves.In water-stressed plants young leaves always exhibited a higherdiffusion resistance than old leaves. The highest diffusionresistance, irrespective of leaf age and water stress, occurredin clone DN which is known to be the most drought tolerant ofthe three clones studied, suggesting that drought tolerancein clone DN is caused, at least in part, by a stomatal regulationmechanism. Water release characteristic curves for the threeclones indicated differences in the water content-water potentialrelationship between young and old leaves as well as betweenclones. The drought tolerant clones had a higher relative watercontent for a given water potential compared with the drought-susceptibleclone. Camellia sinensis L., tea, diffussion resistance, water stress     

Leaf growth dynamics of two congener gymnosperm tree species reflect the heterogeneity of light intensities given in their natural ecological niche

Chamaecyparis obtusa var. formosana and Chamaecyparis formosensis are congener gymnosperm tree species native to Taiwan cloud forests; occupying different niches there. While the seedlings of C. formosensis occur predominantly under bright conditions in large forest gaps, seedlings of C. obtusa var. formosana are mainly found below the canopy of mature forests or in small gaps. It is well known that congener species occupying different niches typically differ in several ecophysiological and morphological traits, but the differences in growth dynamics of such species are still totally unclear, as the diurnal growth dynamics of gymnosperm leaves have not been investigated before. Modern methods of digital image sequence processing were used in this study to analyse the leaf growth dynamics of the two species. We found that both species show similar base–tip gradients and pronounced diurnal growth rhythms with maxima in the evening. Differences between the two species concerning their growth dynamics correlated closely with their ecological amplitudes and abundances. Chamaecyparis obtusa var. formosana grew faster than C. formosensis in low light intensity, typical for closed-canopy situations, and reacted quickly by increasing or decreasing growth rate when light intensity changed within a range typically found below small canopy gaps. In contrast to this, C. formosensis grew better in light intensities typical for open vegetation situations, but reacted slower towards changes of light intensity. Based on those results, the hypothesis can be developed that fluctuations of leaf growth dynamics reflect heterogeneities of the light environment within the niche occupied by a given species.     

Water availability affects tolerance and resistance to aphids but not the trade-off between the two

This study evaluates the effect of water availability on tolerance and resistance to the aphid Chaitophorus leucomelas by clones of the double hybrid [(Populus trichocarpa × P. maximowiczii Henry) × (P. trichocarpa × P. maximowiczii)] (TM × TM), a hybrid that was previously described as resistant to this aphid. In a 2 × 2 experimental design implemented in a nursery, we were able to assess growth (branch length, number of leaves and branch base diameter) in saplings reared in the natural presence of aphids (natural aphid damage) and aphid-controlled conditions (undamaged) under both well-watered and drought stress conditions. We found that resistance was reduced under drought stress conditions, while tolerance in branch length was increased. Cost of resistance was detected as clones displaying higher tolerance grew less in the absence of aphids, whereas no evidence of costs associated with tolerance was found in any of the growth traits measured, A genetic trade-off between tolerance and resistance was detected, but this trade-off was not affected by water availability. Considering the average response of both defence strategies, well-watered trees seem to allocate more resources to resistance than to tolerance, whereas drought-stressed trees allocate more to tolerance than to resistance. This suggests that tolerance would imply a lower cost than resistance, and the shift to either strategy could be modulated by resource availability.     

Effect of controlled inoculation with specific mycorrhizal fungi from the urban environment on growth and physiology of containerized shade tree species growing under different water regimes

The aim of this work was to evaluate the effects of selected mycorrhiza obtained in the urban environment on growth, leaf gas exchange, and drought tolerance of containerized plants growing in the nursery. Two-year-old uniform Acer campestre L., Tilia cordata Mill., and Quercus robur L. were inoculated with a mixture of infected roots and mycelium of selected arbuscular (maple, linden) and/or ectomycorrhiza (linden, oak) fungi and grown in well-watered or water shortage conditions. Plant biomass and leaf area were measured 1 and 2 years after inoculation. Leaf gas exchange, chlorophyll fluorescence, and water relations were measured during the first and second growing seasons after inoculation. Our data suggest that the mycelium-based inoculum used in this experiment was able to colonize the roots of the tree species growing in the nursery. Plant biomass was affected by water shortage, but not by inoculation. Leaf area was affected by water regime and, in oak and linden, by inoculation. Leaf gas exchange was affected by inoculation and water stress. V _cmax and J _max were increased by inoculation and decreased by water shortage in all species. F _v/F _m was also generally higher in inoculated plants than in control. Changes in PSII photochemistry and photosynthesis may be related to the capacity of inoculated plants to maintain less negative leaf water potential under drought conditions. The overall data suggest that inoculated plants were better able to maintain physiological activity during water stress in comparison to non-inoculated plants.     

Structural and Functional Evidence for Xylem-Mediated Water Transport and High Transpiration in Agrobacterium tumefaciens-Induced Tumors of Ricinus communis*

Tumors induced by the wild-type strain C58 of Agrobacterium tumefaciens in hypocotyls of Ricinus communis L. were investigated structurally and functionally with respect to xylem differentiation, cuticle and stomata development, water pathway and transpiration. Clearing of tissue with lactic acid and staining with lacmoid revealed a continuation of stem xylem into differentiated bundles in the tumor. Under the influence of tumors the host xylem below the tumors increased considerably in size. Transport of negatively-charged dyes, amido black, acid fuchsin and the fluorescent pyrenetrisulfonate demonstrated a continuous water flow through the vessels from the stem into the tumor, and up to its surface. Infrared thermography and quantitative measurements of transpiration revealed that transpiration was about 15 times and 7.5 times higher at the tumor surface in comparison to host leaves and to leaves of non-infected plants, respectively. Leaf CO₂ assimilation rate remained unaffected by tumorisation. Tumor growth caused disruption of the epidermis, which did not regenerate and hence no cuticle developed to protect against water loss. Stomata located at the tumor rim hypertrophied and lost their function. Tumors are thus characterised as being structurally and functionally strong pathological water sinks on their host plant.     

Aluminum,Fe, Ca,Mg, K,Mn, Cu,Zn and P in above- and belowground biomass. I.Abies amabilis andTsuga mertensiana

In a mature mixed subalpine stand ofTsuga mertensiana andAbies amabilis, significantly higher Al levels were found in foliage, branch and root tissues ofT. mertensiana.Tsuga mertensiana had significant increases in Al, Ca and Mn levels with increasing foliage age. In current foliage,T. mertensiana had lower levels of Ca, similar levels of Mg and P, and higher levels of Mn thanA. amabilis. Both tree species had Cu and Fe present at higher levels in branch than foliage tissues. Fine roots had the highest concentrations of Al, Fe and Cu but the lowest Ca and Mn concentrations of all tissues analyzed. In the roots of both species, phloem tissues always had significantly higher Al levels than xylem. Fine roots (< 1 and 1–2 mm) ofT. mertensiana had higher Al levels than were found inA. amabilis. Roots greater than 2 mm in diameter exhibited no significant differences in Al levels in phloem or xylem tissue betweenA. amabilis andT. mertensiana. The two species show a clear difference in their ability to accumulate specific elements from the soil.