Temperature-controlled open-top chambers for global change research

To enable experiments on the interactive effects of elevated atmospheric CO₂ and increased air temperature on physiological processes in trees to be carried out, we altered the standard design of open-top chambers by replacing blowers with evaporative coolers and in-line heaters, with a feedback control system to maintain ambient or elevated air temperatures within the chambers. Ambient and elevated (+ 4 °C) temperature regimes were attained consistently and reliably throughout the growing season, with high reproducibility between chambers. From May through December the average of nearly 300,000 temperature measurements was 18.5 °C in ambient air, 18.9 ± 0.6 °C in six ambient chambers, and 22.4 ± 0.9 °C in six elevated temperature chambers. The difference in soil temperature between ambient and elevated chambers was 1.2 °C. Absolute humidity (vapour pressure) in the chambers was higher than that of ambient air, but it was generally similar between temperature treatments. Vapour pressure deficit therefore was higher in elevated temperature chambers than in ambient chambers, and this difference is considered an inseparable part of the temperature treatment. The addition of a temperature control system to open-top chambers removes what has been an important flaw in this important tool for global change research.     

The fine structure of a multiterminal innervation of an insect muscle

The detailed structure of nerve branches, neuromuscular junctions, and muscle fibers of a multiterminal innervation of cockroach abdominal muscle has been studied with the electron microscope. The muscle fiber is of the banded myofibril type; with paired mitochondria and abundant endoplasmic reticulum. The peripheral nerve branches are multiaxonal with large central axon and several small peripheral tunicated axons. Tracheoblasts closely accompany the nerve branches. The multiple neuromuscular junctions show typical axonal vesicles, muscle aposynaptic granules, and close plasma membrane apposition with no interposition of basement membrane material.     

Changes in membrane characteristics of heart muscle during inhibition

Membrane characteristics were studied in isolated muscle strands from auricles of frogs using the "square pulse" technique. Changes in the time course and spatial spread of subthreshold electrotonic potentials were measured. If acetylcholine is applied in concentrations which cause slowing or stoppage of the heart beat, the following changes are produced: (a) the length constant (λ) of the membrane is reduced, (b) the time constant is shortened. The effects are reversible and increase with acetylcholine concentration. The membrane changes caused by acetylcholine dimmish with time. It is concluded that during acetylcholine inhibition, as well as during vagal inhibition, the conductance of the muscle membrane is increased. Appreciable changes in the resting membrane potential need not accompany inhibition.     

Dormancy in Seeds of Charlock: IV. INTERRELATIONSHIPS OF GROWTH, OXYGEN SUPPLY AND CONCENTRATION OF INHIBITOR

Respiration measurements were made over a period of 24 h at25 °C on seeds and excised embryos maintained in Warburgflasks with partial pressures of oxygen ranging from 0 to 1atm. In the initial phase (0 to 4 h), the rate of oxygen uptake(Q_O2 of excised embryos increased linearly with external oxygenconcentration (C_O from 0 to 0.1 atm O₂ from 0.1 to 0.2 atm O₂the relation was curvilinear, and from 0.2 to 1.0 atm O₂ uptakewas independent of concentration. In later stages the relationbetween Q_O2 and C_o changed, and from 20 to 24 h the rate ofoxygen uptake increased with concentration to 1.0 atm O₂. Thechanges with time were associated with increase in rate of respiration,increase in cell size and cell number, and the oxidation offats. The decline in concentration of oxygen from the surfaceto the centre of embryos was calculated to be relatively smallat each external oxygen concentration. Althugh the rate of diffusionfailed to keep pace with consumption, the main parameters whichdetermined the internal oxygen status of the embryos were thesurface concentrations and the permeability of the seed coat.The resistance of the seed coat to diffusion of oxygen was foundto be very high, the coefficient of diffusion being about 10^–7mm² s^–1. The concentration of oxygen and in air were estimatedto be approximately 0.04 and 0.02 atm O₂, respectively. Sincea smaller concentration of oxygen (0.012 atm O₂) in the tissueswas found to be sufficient for growth, the dormancy of the seedswas not due to lack of oxygen. Dormancy appeared to be due tothe activity of growth-inhibiting substances, the concentrationof which increased with decrease in oxygen supply; below 0.1atm O₂ their rate of production increased with decrease in theoxygen concentration of the tissues. They accumulated withinthe testas of dormant seeds and prevented cell elongation. Extractsof the inhibitory substances were partially purifield by partitioningthe aqueous fraction with ether and separating chromatographically.The active principle(s) was not abscisic acid ((+)—AbscisinII, ‘Dormin’) or the mustard oil, allylisothiocyanate.     

Dispersed cuticles of putative non-vascular plants from the Lower Devonian of Britain

Dispersed cuticles with surface ornamentation recovered from the Lower Old Red Sandstone (Gedinnian) of Hereford and Worcester, England, are described, named and placed in an artificial, morphologically based classification system. Four species of the new genus Cosmochlaina are distinguished on the nature of the ornament. The affinities of the plants covered by such cuticles remain unknown: the cuticles are named to facilitate their use in biogeographic, stratigraphic and evolutionary studies.     

A method for measuring radial differences in water content of intact tree stems by attenuation of gamma radiation

Abstract. A technique is described for the nondestructive measurement of water content variations in the stems of trees.
The density of intact tree stems in the forest was estimated using attenuation of gamma radiation. Water content was calculated after subsequent derivation of the volume fraction of solid matrix.
A radial sequence of such measurements was obtained by measuring a sequence of chords, and assuming homogeneity within concentric tori. Analysis of the source of errors showed that the system was a close approximation of an ideal, rigid-geometry, mono-energetic source/detector system, with basic precision dependent on the quantity of gamma radiation measured. Correlation coefficients between the gamma-attenuation technique and subsequent gravimetric estimates of water content for two field experiments reported here were 0.882 and 0.938.
The technique was shown to be capable of describing the radial and diurnal variation in water content at two heights in the stem of a 20-year-old tree of Pinus contorta .     

Dormancy in Seeds of Charlock: II. THE INFLUENCE OF THE SEED COAT

An attempt was made to elucidate those factors associated withthe seed coat which are responsible for maintaining the dormancyof the charlock seed. The rates of water uptake by the seedand by the embryo were measured; the permeability of the coveringlayers to water was much the same as that of parenchymatouscells. The action of certain dormancy-breaking treatments, viz.embryo excision, exposure to concentrated sulphuric acid, alternatingtemperatures, and gibberellic acid, were therefore examined.Fromexperiments with excised embryos and seeds exposed to concentratedsulphuric acid, it appeared that the loss of dormancy followingthe removal of the seed coat resulted from an increase in oxygensupply to the embryo. The germination of a small proportionof seeds at low temperature may also be due to an increase inthe oxygen concentration within the seed. The dormancy-breakingeffect of gibberellic acid, however, is unlikely to be due tochanges in the permeability of the seed coat to oxygen. Estimatesof the rate of oxygen uptake and growth of seeds treated withgibberellic acid at various external oxygen concentrations showthat the resistance of the covering layers to diffusion of oxygenretards, but does not prevent, the initiation of growth.It wouldappear that dormancy is maintained in charlock seeds by theaction of a specific growth-inhibiting substance which is producedat low oxygen concentration in the interior of the seed andwhich diffuses to the meristems.     

Cherry leaf roll virus in the female gametophyte and seed of birch and its relevance to vertical virus transmission

ELISA detected cherry leaf roll virus (CLRV) in infected birch leaf sap diluted 1/320 in buffer, in extracts of one infected leaf with nine healthy leaves and in leaf sap frozen for 2 wk. Similarly, ELISA detected CLRV in mixtures of one infected bud with four virus-free buds. Intensive bioassays and ELISA showed that in some trees CLRV was restricted to only a few branches whereas in others it occurred throughout the tree. The prevalence of CLRV in unmanaged birch populations in Britain was less (3%) than in Midlands street trees (17%). In CLRV-free birch trees that received pollen from infected ones, ELISA indicated that antigen was introduced into, and multiplied within, the embryos but not the seed coat or the pericarp/wings. In one instance, antigen was detected in a branch of an experimentally pollinated tree but not in those parts of the crown that had been exposed to open pollination. The proportion of seed germinating after crosses in which both parents were CLRV-free was greater than when either or both parents were infected but the largest difference occurred with infection in the female parent. Few embryos seemed to escape invasion with CLRV when the maternal tissue was naturally infected. Overall, seed transmission ranged between 0 and 38% (mean 17%) when only females in a cross were infected, and between 11 and 75% (mean 30%) when only the males were infected. Assuming no selective advantage that would help infected plants to achieve reproductive age, we found that CLRV would be lost from a birch population within two generations if transmitted only through seed. Embryos in seeds from CLRV-infected birch that received CLRV-free pollen differed from their healthy counterparts in being shrunken and suspended in a loosely fibrillar matrix that contained numerous virus-like particles in tubular inclusions. In two trees, CLRV-free pollen tended to fertilise a greater percentage of ovules than did CLRV-infected pollen. Seedlings derived from infected seed and cuttings from naturally-infected trees grew less rapidly than their healthy counterparts. In still air, most birch pollen liberated from a height of 3·5 m fell within 3 m of the drop zone and none was detected 10 m from the source. Field observations on the patterns of virus spread as measured by seedling infection were consistent; about 3% of seedlings from a tree 6·9 m away from the nearest source of inoculum were infected but no infected seedlings were detected in more distant trees, even though each was experimentally infectible with CLRV and pollen from the infected tree germinated on their stigmas.