Differentiation and structural decline in the leaves and bark of birch (Betula pendula) under low ozone concentrations

Summary Leaf and bark structure of a birch clone (Betula pendula Roth) continuously exposed to charcoal-filtered air or charcoal-filtered air plus ozone (0.05, 0.075, 0.1 l 1^-1) was investigated throughout one growing season. Increasing ozone dose influenced leaf differentiation by reducing leaf area and increasing inner leaf air space, density of cells developing into stomata, scales and hairs. When approximately the same ozone dose had been reached, macroscopical and microscopical symptoms appeared irrespective of the ozone concentration used during treatment. Structural decline began in mesophyll cells around stomatal cavities (droplet-like exudates on the cell walls), continued with disintegration of the cytoplasma and ended in cell collapse. Epidermal cells showed shrinkage of the mucilaginous layer (related to water loss). Their collapse marked the final stage of leaf decline. When subsidiary cells collapsed, guard cells passively opened for a transitory period before collapsing and closing. With increasing ozone dose starch remained accumulated along the small leaf veins and in guard cells. IIK-positive grains were formed in the epidermal cells. This contrasted with the senescent leaves, where starch was entirely retranslocated. Injury symptoms in stem and petiole proceeded from the epidermis to the cambium. Reduced tissue area indicated reduced cambial activity. In plants grown in filtered air and transferred into ozone on 20 August, injury symptoms developed faster than in leaves formed in the presence of ozone. Results are discussed with regard to O₃-caused acclimation and injury mechanisms.     

Molecular analysis of the Lactococcus lactis subspecies lactis CNRZ270 bidirectional theta replicating lactose plasmid pUCL22

pUCL22 is the lactose protease plasmid of Lactococcus lactis ssp. lactis CNRZ270. The nucleotide sequence of its replication region Rep22 contains a non-transcribed region, the replication origin, followed by a gene encoding a putative 388-amino-acid protein named Rep22A. The promoter regions of the rep22A and pC194 cat genes share strong similarities and the pUCL22 replicon exerted trans or cis negative control on the pC194 cat gene expression in L. lactis. We suggest that Rep22A binds to its own promoter as well as to the pC194 cat promoter and thus is autoregulated. We show that pUCL22 replicates mainly by a bidirectional theta mechanism in L. lactis, and is representative of a widely distributed replicon family, members of which could be co-resident. We propose that compatibility between these closely related replicons results from minor replication protein modifications coupled with base changes in their respective binding sites, supporting the co-existence of numerous related replicons in lactococcal strains.     

CD44 standard and variant isoform expression in normal human skin appendages and epidermis

 CD44 isoforms have been implicated in tumor progression and metastasis formation. This study presents a thorough immunohistochemical analysis of CD44 standard and isoform expression in normal human skin appendages and epidermis applying monoclonal antibodies against CD44s, CD44v3, -v4, -v5, -v6, and -v9. An improved immunohistochemical protocol with microwave-based antigen retrieval in paraffin sections and heavy metal amplification of the diaminobenzidine reaction product provided enhanced resolution and sensitivity as compared to studies on frozen sections. The hair follicle, the seborrheic and eccrine sweat glands were strongly positive for all CD44 isoforms studied. In the latter, the clear cells but not the dark (intercalated) cells were positive. The sudoriferous ducts adjacent to the glands were weakly positive for all CD44 isoforms and strongly positive near the skin surface. In the apocrine glands, the basal cells showed only a moderate positivity. The myoepithelial cells expressed only CD44s. In the epidermis, all CD44 isoforms were detectable, with strongest CD44 immunostaining in the lower third of the stratum spinosum and weaker staining in the stratum basale and the upper two-thirds of the stratum granulosum. The stratum granulosum and corneum were unreactive. Thus, a regional and cell type-specific CD44 expression was revealed. Accepted: 10 May 1996     

Distribution of zooplankton associated with the Endeavour Ridge Hydrothermal Plume

Enhanced zooplankton biomass was found in shallow (<900 m)and deep (>900 m) net samples collected over the EndeavourRidge hydrothermal plume in 1991 and 1992. This enhanced biomasswas manifest in considerably higher abundance values for mostspecies in proximity to the vent field compared with abundancevalues from net samples collected 10–50 km off-axis. Thespecies most enriched in abundance by the hydrothermal plumewere those normally found in the mid-depth scattering layerat 400–900 m depth. These mid-depth animals were alsothe dominant fauna in the deep scattering layers that overliethe hydrothermal plume at depths of 1200–1900 m near themain vent field. The abundance and biomass dominants were specieswith pronounced ontogenetic migration patterns, and their associatedpredators. The abundance of many typically deep species wasalso enhanced over the main vent field. Faimal compositionsof net samples were compared using a similarity measure andaverage linkage rule. Deep fauna in proximity to the vent fieldbut not associated with scattering layers (Group 1), were similarto deep fauna 10–50 km off axis. The fauna of the deepscattering layer over the vent field (Group 2) was most similarto surface and mid- depth scattering layer fauna found withina 50km radius of the vent field. Statistical tests of linkagesobtained using a bootstrap method indicate that the abundanceand taxonomic composition of the two faunal groups were significantlydistinct in 1992, but not in 1991. We conclude that there wasconsiderable infiltration of shallow fauna into the deep scatteringlayers within 2–3 km of the main vent field, less extensiveinfiltration 10–15 km to the noilh and south of the ventfield in 1991, and insignificant infiltration at stations 50kmto the west of the vent field in 1992. A bootstrap analysiscomparing the faunal composition of nets towed above 900 m depthshowed that shallow fauna were not significantly distinct betweenthe two sampling years or up to 50 km away from the vent field.     

Seasonal variation in hydrochloric acid, malic acid, and calciumions secreted by the trichomes of chickpea (Cicer arietinum)

A novel experimental set-up and method of recording of electrical potential differences in plants have been developed which enable continuous, 8-channel monitoring of electrical activity over extended periods of time using inserted, extracellular electrodes. The investigations were carried out on 21- to 23-day-old Helianthus annuus plants, and spontaneously-generated action potentials were recorded during monitoring sessions lasting for 3 days and nights. Characteristics of these spontaneous action potentials were elaborated, adopting as parameters their typical form, amplitude, duration, velocity, direction, and distance of propagation and frequency of occurrence in morphologically different parts of the plant, Variability, similarities, and interdependence of the above parameters in individual plants and in a group of 15 plants were determined. A hypothesis concerning propagation of action potentials in plants along specific impulse-propagating 'columns' is discussed. The frequency of generated impulses is highest at night and lowest in the day and also displays an apparent 24-h rhythm. Presumably this mechanism is under both endogenous and exogenous control and may be partly dependent on a biological clock.     

The Impact of Chlorophyll-Retention Mutations,d1d2 and cyt-G1, during Embryogeny in Soybean

The ultrastructural, physiological, and molecular changes in developing and mature seeds were monitored in a control line (Glycine max [L.] Merr., cv Clark) that exhibited seed degreening and two mutant lines (d1d2 and cyt-G1) that retained chlorophyll upon seed maturation. Ultrastructural studies showed that the control line had no internal membranes, whereas stacked thylakoid membranes were detected in the green seed from the mutant lines. Pigment analyses indicated that total chlorophyll was lowest in the mature seeds of the control line. Mature d1d2 and cyt-G1 seed had elevated Chl a and Chl b levels, respectively. In both control and mutant lines, Lhcb1, Lhcb2, and RbcS mRNAs were abundant in embryos prior to cotyledon filling, declined after the onset of storage protein accumulation, and were barely detectable or undetectable in all later stages of seed development. Therefore, the chlorophyll-retention phenotype must be a result of the alteration of a process that occurs after translation of photosynthesis-related mRNAs to stabilize apoprotein and pigment levels. Furthermore, different elements controlling either the synthesis or turnover of Chl a and Chl b must be impaired in the d1d2 and cyt-G1 lines. No reproducible differences in total leaf, embryonic, and chloroplast protein profiles and plastid DNAs could be correlated with the mutations that induced chlorophyll retention.     

Feeding preference of Zetzellia mali: does absolute or relative abundance of prey matter more?

The importance of the acarine predator, Zetzellia mali, in the control of phytophagous mites in apple orchards is not well understood. Zetzellia mali tends to prefer the eriophyid, Aculus schlechtendali, over the economically more significant tetranychid, Panonychus ulmi, but quite a wide range of preference values have been reported in the literature. In sets of laboratory choice trials, we determined that prey preference of this predator varies with the relative but not absolute density of its prey. We attempt to explain these results in terms of behavioural mechanisms and discuss the potential implications of our results for the effectiveness of Z. mali in the biological control of phytophagous mites in apple orchards.     

Characterization of a novel plant poly(A) polymerase

We have purified and characterized poly(A) polymerases (PAPs) from Pisum sativum, Brassica juncea, and Zea mays. Through chromatography on DEAE-Sepharose and heparin-Sepharose, these PAPs copurified as a single enzyme along with RNPs that could provide RNA substrates for the enzyme. More extensive purification by chromatography on MonoQ resulted in the resolution of the PAPs into as many as three fractions. One of these (PAP-I) contained a 43-kDa polypeptide immunologically related to the yeast PAP, and two others (PAP-II and PAP-III) contained RNAs that could serve as substrates for polyadenylation. These fractions by themselves possessed little PAP activity, but mixtures containing combinations of these displayed substantial activity. Similar PAP factors (PAP-I and PAP-III) were identified after fractionation of extracts prepared from Brassica juncea and Zea mays. The factors from one plant were completely interchangeable with those from different plants. We conclude that the poly(A) polymerases present in vegetative plant tissues consist of more than one component. In this respect, they are substantially different from other reported plant, mammalian, and yeast PAPs.     

Condition factor and hepatosomatic index as estimates of energy status in male three-spined stickleback

Seasonal variation in direct and indirect measures of energy status was examined using estimates of glycogen, lipid and protein levels in a single cohort of male three-spined sticklebacks from an annual population collected each month over one complete year. Condition factor, somatic condition factor and hepatosomatic index (HSI) were calculated as indirect indices of energy status and the accuracy of these indirect measures as predictors of energy status was investigated. Results indicate that both condition factors were significant predictors of energy reserves (lipid, protein, glycogen and total energy), but that the proportion of variance accounted for was small. Both condition factors perform better as predictors of energy content per unit body weight. The HSI was a significant, but a weak predictor of total glycogen levels over the whole year. On a seasonal basis the relationship between HSI and energy reserves was highly variable. These indices are therefore poor predictors of energy reserves in male three-spined sticklebacks.     

PHYLOGENYAND HISTORICAL ECOLOGY OF THE DESMARESTIACEAE (PHAEOPHYCEAE) SUPPORT A SOUTHERN HEMISPHERE ORIGIN1

Phylogenetic relationships in the Desmarestiales (Phaeophyceae) were inferred among the monotypic Arthrocladia (Arthrocladiaceae) and 27 isolates from Desmarestiaceae, representing 17 taxa of Desmarestia and the monotypic Antarctic genera Himantothallus and Phaeurus. Phaeurus and Arthrocladia were used as outgroups. Parsimony analyses of nuclear ribosomal DNA internal transcribed spacer (ITS1 and ITS2) sequences, in which gaps were both included and excluded, yielded well-resolved trees with a consistent general branching pattern. A parallel analysis of nine morphological and life-history characters and three ecological characters yielded a similar tree but provided little resolution in the terminal clades. The position of the monotypic Arthrocladia villosa within the Desmarestiales is consistent with monophyly for the order, but its position as the most primitive desmarestialean is not resolvable from the molecular data set. The basal position of Phaeurus, the Antarctic Desmarestia species, and Himantothallus is consistent with the hypothesis of a Southern Hemisphere origin for the family Desmarestiaceae. The more recent Northern Hemisphere “aculeata” clade evolved from an Antarctic ancestor. A “D. aculeata-like” species was ancestral to a lineage characterized by annual sporophytes with high sulfuric acid content, which radiated into many species, widely distributed in both hemispheres. Mapping of morphological and ecological characters onto the molecular tree confirm the informativeness of sulfuric acid-containing vacuoles and unilocular sporangial types. There is good congruence between phylogenetic tree topology and temperature impints in relation to biogeographic distribution, supporting the theory that temperature tolerance is a conservative trait.