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Mechanisms of drought response in Thuja occidentalis L. I. Water stress conditioning and osmotic adjustment.

We investigated the extent of osmotic adjustment and changes in transpiration rate that occur in response to repeated cycles of water deficit stress in 6-year-old Thuja occidentalis L. (eastern white cedar) trees. Groups of trees were water-stress conditioned by repeated exposure to predetermined thresholds of nonlethal water stress by withholding water until the predawn water potential fell to -0.9 (mild conditioning) or -1.4 MPa (moderate conditioning). Both the mild and moderate conditioning treatments resulted in a decrease in osmotic potential of 0.08 to 0.20 MPa and 0.11 to 0.28 MPa, respectively, relative to the well-watered controls. Mildly and moderately conditioned trees exhibited an approximately 35 and 50% reduction in cumulative transpiration, respectively, following at least two stress cycles. Transpiration rates of conditioned trees remained below those of the well-watered controls even when water potential integrals were similar. We conclude that the decrease in transpiration rate was more important than osmotic adjustment as a mechanism of response to repeated water stress.

Journal Article↗

Mechanisms of drought response in Thuja occidentalis L. II. Post-conditioning water stress and stress relief.

We examined the extent of osmotic adjustment and the changes in relative water content (RWC) and transpiration rate (i.e., relative stomatal function) that occur in water-deficit-conditioned 6-year-old Thuja occidentalis L. (eastern white cedar) trees in response to a severe drought. Trees conditioned by successive cycles of mild or moderate nonlethal water stress (conditioning) and nonconditioned trees were exposed to drought (i.e., -2.0 MPa predawn water potential) to determine if water deficit conditioning enhanced tolerance to further drought stress. Following drought, all trees were well watered for 11 days to evaluate how quickly osmotic potential, RWC and transpiration rate returned to preconditioning values. Both nonconditioned trees and mildly conditioned trees exhibited similar responses to drought, whereas moderately conditioned trees maintained higher water potentials and transpiration rates were 38% lower. Both conditioned and nonconditioned trees exhibited a similar degree of osmotic adjustment (-0.39 MPa) in response to drought relative to the well-watered control trees. The well-watered control trees, nonconditioned trees and mildly conditioned trees had similar leaf RWCs that were about 3% lower than those of the moderately conditioned trees. Following the 11-day stress relief, there were no significant differences in osmotic potential between the well-watered control trees and any of the drought-treated trees. Daily transpiration rates and water potential integrals (WPI) of all drought-treated trees approached those of the well-watered control trees during the stress relief period. However, the relationship between cumulative transpiration and WPI showed that previous exposure to drought stress reduced transpiration rates. Leaf RWC of the moderately conditioned trees remained slightly higher than that of the nonconditioned and mildly conditioned trees.

Journal Article↗

In vitro plantlet formation from embryonic explants of eastern white cedar (Thuja occidentalis L.).

A protocol is described for the in vitro production of plantlets from embryonic explants of eastern white cedar (Thuja occidentalis L.). Bud induction was optimal when embryonic explants were cultured for 20-25 days on half-strength Quoirin and Le Poivre mineral salts containing equimolar concentrations (10(-6) M) of N(6)-benzyladenine and 2-isopentyl adenine. Bud development was achieved in phytohormone-free medium in the presence of activated charcoal. Maximal shoot elongation occurred on half-strength Quoirin and Le Poivre salts, whereas shoot multiplication was optimal on half-strength Bornman's MCM salts in the presence of cytokinin. Hardened shoots, dipped in commercial rooting powder containing indole-3-butyric acid, rooted optimally in mist under non-sterile greenhouse conditions. Both rooting and subsequent plantlet growth was best when Redi-Earth((R)) was used as a substrate. Over 250 plantlets per embryo can be produced annually by this technique.

Journal Article↗

Collapse of Water-Stress Emboli in the Tracheids of Thuja occidentalis L.

We report the kinetics of embolus formation and collapse in the tracheids of Thuja occidentalis L. stem segments. Radial wood sections were trimmed to 4 mm long paralleling the tracheids by 1 mm wide and 0.1 mm thick. They were observed under a dissecting microscope at 128x while sections were dehydrated and rehydrated. During dehydration, cavitations resulted in the formation of emboli in tracheids, but we concluded that the cavitated tracheids did not immediately fill with air at atmospheric pressure. This conclusion was based on the time required for the emboli to collapse after the rewetting of the dehydrated segment. By hypothesis, the time for the emboli to collapse should be proportional to the amount of air in the emboli. The time for all the emboli to collapse was a linear function of the dehydration time for times up to 15 min. For dehydrations greater than 80 min, the time for collapse after rewetting was constant, and we concluded that the tracheids have saturated with air by 80 min of dehydration. The kinetics of embolus formation is discussed in terms of the air-seeding hypothesis for cavitation, and collapse is discussed in terms of the physics of gas dissolution and diffusion. Embolus formation and dissolution in intact herbaceous and woody plants should follow the same physical laws.

Journal Article↗

Cavitation Events in Thuja occidentalis L.? : Utrasonic Acoustic Emissions from the Sapwood Can Be Measured.

Ultrasonic acoustic emissions (AE) in the frequency range of 0.1 to 1 megahertz appear to originate in the sapwood of Thuja occidentalis L. The AE are vibrations of an impulsive nature. The vibrations can be transduced to a voltage waveform and amplified. The vibrations of each AE event begin at a large amplitude which decays over 20 to 100 microseconds. Strong circumstantial evidence indicates that the ultrasonic AE result from cavitation events because: (a) they occur only when the xylem pressure potential Psi(xp) is more negative than a threshold level of about -1 megapascal; (b) the rate of AE events increases as Psi(xp) decreases and when the net rate of water loss increases; (c) the AE can be stopped by raising Psi(xp) above -1 megapascal. Ultrasonic AE have been measured in whole terminal shoots allowed to dry in the laboratory, in isolated pieces of sapwood as they dried in the laboratory, and in whole terminal shoots in a pressure bomb when Psi(xp) was decreased by lowering the gas pressure in the pressure bomb.

Journal Article↗

Ultrasonic Acoustic Emissions from the Sapwood of Thuja occidentalis Measured inside a Pressure Bomb.

An improved method of counting acoustic emission (AE) events from water-stressed stems of cedar (Thuja occidentalis L.) is presented. Amplified AEs are analyzed on a real time basis by a microcomputer. The instrumentation counts AE events in a fashion nearly analogous to scintillation counting of radioactive materials.The technique was applied to measuring ultrasonic AEs from the stems of cedar inside a pressure bomb. The shoots were originally fully hydrated. When the shoots are dehydrated in the bomb by application of an overpressure very few AEs were detected. When the bomb pressure is reduced after dehydration of the shoot, AE events could be detected. We conclude that ultrasonic AEs are caused by cavitation events (= structural breakdown of water columns in the tracheids of cedar) and not by the breaking of cellulose fibers in the wood.

Journal Article↗

Thujaplicins from Thuja plicata as iron transport agents for Salmonella typhimurium.

Strains of Salmonella typhimurium which are unable to synthesize their normal iron transport agent, enterobactin, and which must be supported with an exogenous chelator (siderophore) on certain media, were used to examine various types of wood for the presence of chelators. Western red cedar wood, Thuja plicata, was observed to contain large amounts of three substances that in low concentration would serve as chelators for S. typhimurium. The chelators from T. plicata were characterized and found to be alpha-, beta-, and gamma-thujaplicin. Other planar cyclic alpha-hydroxyketones were examined, and several were found to function as chelators for S. typhimurium.

Biological Assay↗

Seedbed and moisture availability determine safe sites for early Thuja occidentalis (Cupressaceae) regeneration.

Regeneration of many late-successional tree species depends on specialized safe sites. The primary objective was to investigate the roles of seedbed and moisture retention as dimensions of safe sites for the early regeneration of drought-sensitive northern white cedar (Thuja occidentalis). We hypothesized that rates of germination, survival, and growth of T. occidentalis are unlikely to differ among seedbed types under conditions of abundant water, but that differences are likely to emerge as water becomes more limited. In a 67-d greenhouse experiment, cedar seeds were sown on logs, leaf litter, and soil of cedar and paper birch (Betula papyrifera) canopy origin. Seedbeds were subjected to three water treatments. Among the water treatments, highest germination rates occurred within the high water treatment, although germination on cedar litter was comparable to that of the low water treatment. Higher germination and survival rates occurred on decayed logs than other natural seedbeds for medium (P = 0.001) and low (P < 0.0001) water treatments. Germination on birch logs occurred at higher rates than on cedar logs within the low water treatment (P = 0.04). Seedling growth for the medium water treatment was lower on leaf litter than any other type of seedbed (P < 0.01). Results generally demonstrated that the interplay between seedbed and moisture retention is a component of safe sites for T. occidentalis regeneration.

Journal Article↗

[Effects of polysaccharide from Thuja occidentale L. on stromal precursor cells of hematopoietic microenvironment in mice].

The effect of high molecular polysaccharide subfraction from Thuja occidentale L. (TPS) on stromal precursor cells of hematopoietic microenvironment under the "steady-state" conditions and after sublethal irradiation was investigated. The stromal precursor cells of different stages of differentiation were detected by the implantation of mouse bone marrow under the renal capsule of syngeneic intact recipients and chimeras. It was shown that TPS did not occur the toxic influence on the stromal precursor cells and provided the defense effect on them under the strong (6 Gy) radiation damages.

Animals↗

Mitogenic activity of high molecular polysaccharide fractions isolated from the Cupressaceae Thuja occidentale L. I. Macrophage-dependent induction of CD-4-positive T-helper (Th+) lymphocytes.

The arborvitae or Thuja occidentale L., one of the Cupressaceae, has rarely been investigated until now. Several authors have demonstrated that allopathic extracts of this plant could be used as strong antiviral agents directed against plant and animal viruses. Polysaccharide fractions with molecular weights ranging between 20,000 and 1,000,000 and higher have been isolated from the aqueous alkaline extract of the herbal parts of T. occidentale L. by ethanol precipitation and fractionation by ultrafiltration. High molecular subfractions of Thujapolysaccharides (TPS) proved to be highly mitogenic on peripheral blood leukocytes. It was demonstrated by the alcalic immune phosphatase-antiphosphatase and Pappenheim staining methods that the mitogenic and cluster-forming activity of TPS cause T cell induction, in particular, of CD 4-positive T-helper/inducer cells as opposed to B cells. The CD-4+ T-helper/inducer cell induction is connected to an increased production of IL-2. The cluster-forming ability and mitogenity of TPS correlates well with the 3H-thymidine uptake and seems to be IL-1 and IFN-gamma dependent as could be shown by blocking the mitogenic effect using anti-IL-1- and anti-IFN antibodies. Whether it is possible to use these polysaccharide fractions as an adjuvant in the therapy of immune deficiency syndromes and cancer must now be further investigated.

Antigens, Differentiation, T-Lymphocyte↗

A reproving of Thuja.

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Plants, Medicinal↗