Expression of Tobacco Carbonic Anhydrase in the C4 Dicot Flaveria bidentis Leads to Increased Leakiness of the Bundle Sheath and a Defective CO2-Concentrating Mechanism

Flaveria bidentis (L.) Kuntze, a C₄ dicot, was genetically transformed with a construct encoding the mature form of tobacco (Nicotiana tabacum L.) carbonic anhydrase (CA) under the control of a strong constitutive promoter. Expression of the tobacco CA was detected in transformant whole-leaf and bundle-sheath cell (bsc) extracts by immunoblot analysis. Whole-leaf extracts from two CA-transformed lines demonstrated 10% to 50% more CA activity on a ribulose-1,5-bisphosphate carboxylase/oxygenase-site basis than the extracts from transformed, nonexpressing control plants, whereas 3 to 5 times more activity was measured in CA transformant bsc extracts. This increased CA activity resulted in plants with moderately reduced rates of CO₂ assimilation (A) and an appreciable increase in C isotope discrimination compared with the controls. With increasing O₂ concentrations up to 40% (v/v), a greater inhibition of A was found for transformants than for wild-type plants; however, the quantum yield of photosystem II did not differ appreciably between these two groups over the O₂ levels tested. The quantum yield of photosystem II-to-A ratio suggested that at higher O₂ concentrations, the transformants had increased rates of photorespiration. Thus, the expression of active tobacco CA in the cytosol of F. bidentis bsc and mesophyll cells perturbed the C₄ CO₂-concentrating mechanism by increasing the permeability of the bsc to inorganic C and, thereby, decreasing the availability of CO₂ for photosynthetic assimilation by ribulose-1,5-bisphosphate carboxylase/oxygenase.     

Actin-like protein associated with plasma membranes fromEuglena gracilis

Summary Microtubules are characteristic components of the membrane skeleton ofEuglena gracilis, but whether microfilaments are present has been controversial. We here present evidence that an actin-like protein may indeed be associated with the plasma membrane (PM) ofE. gracilis. Firstly, a 47 kDa, PM-associated, polypeptide was recognized by an anti-amoeba actin antibody. Secondly, this 47 kDa protein seemed to be peripherally attached to PM in much the same way as -tubulin, since both could be released from PM by treatment with 150 mM NaOH but not with ethylene glycol, NaCl, or formamide. Thirdly, the 47 kDa polypeptide and -tubulin were found mainly in the Triton X-1 14-insoluble fraction, indicating that they were part of a protein complex resistant to detergents, such as the cytoskeleton. Finally, DNase I activity was inhibited by a fraction enriched in the 47 kDa polypeptide, a property typical of actin.Abbreviations CP-medium cytoskeleton preparation medium - BNSP-skatole 2-(2-nitrophenylsulfenyl)-3-methyl-3-bromoindolenine - ECL enhanced chemiluminescence - HEPES N-[2-hydroxyethyl]-piperazine-N[2-ethane sulfonic acid] - ICM intracellular membranes - MF mitochondrial and microsomal fraction - PM plasma membrane - PPB potassium phosphate buffer - PVDF polyvinylidene difluoride - SDS sodium dodecyl sulphate - TBS Tris-buffered saline - TBST Tris-buffered saline with Tween 20     

Body size, sex ratio and diet composition of Arctogadus glacialis (Peters, 1874) (Pisces: Gadidae) in the Northeast Water Polynya (Greenland)

Little is known about the biology and feeding ecology of Arctogadus glacialis (Peters, 1874), an endemic gadid fish species occurring circumpolarly in both ice-free and ice-covered Arctic seas. In this study we analysed specimens sampled from bottom trawl and Agassiz trawl catches conducted at eight stations in water depths from 115 to 490 m in the Northeast Water Polynya off Greenland in August 1990. Size composition and sex ratios were assessed for 585 fish with standard lengths ranging from 7.8 to 34.5 cm. Median fish sizes at the stations ranged between 8.6 and 18.8 cm. Quantitative stomach content analyses of a subsample of 175 specimens revealed that in the Northeast Water A. glacialis fed almost exclusively on pelagic prey. Truly benthic organisms were not found in the stomachs, and sympagic species were rarely recorded. Overall, calanoid copepods were the major prey in terms of numbers. However, stomach contents varied considerably between fish size groups. In terms of prey biomass, copepods dominated the diet of small fish (<12 cm), primarily occurring at shallow stations (115–250 m), whereas amphipods and mysids were relatively more important for larger fish (>16 cm), which dominated the catches at deep stations (360–480 m). The broad variety of diet composition indicated an opportunistic pelagic feeding pattern. Received: 6 November 1997 / Accepted: 11 May 1998     

Nitrogen uptake in the infiltration community, an ice algal community in Antarctic pack-ice

An infiltration community was the dominating ice algal community in pack-ice off Queen Maud Land, Southern Ocean, in January 1993. The community was dominated by autotrophic processes, and the most common species were the prymnesiophyte Phaeocystis antarctica and the diatoms Chaetoceros neglectus and Fragilariopsis cylindrus. The concentration of chlorophyll a was 1.3–47.9 μg l⁻¹, and the inner part of the community was nitrate depleted. Uptake rates of nitrate, nitrite, ammonium, urea and amino acids were measured using ¹⁵N. Nitrate was the major nitrogen source for ice algal growth (67 ± 6% nitrate uptake). It is suggested that % nitrate uptake in the infiltration community decreases during the growth season, from 92% during spring (literature data) to 67% during summer. Scalar irradiance in the infiltration community was high and variable. It reached ca. 2000 μmol m⁻² s⁻¹ at some locations, and nitrate uptake rate was potentially photoinhibited at irradiances >500 μmol m⁻² s⁻¹. Nitrate uptake rate in an average infiltration community (0.6 m of snow cover) was lowered by 13% over a 2-week period due to photoinhibition. Received: 16 December 1996 / Accepted: 5 January 1998     

Dorymenia troncosoi sp. nov. (Mollusca Solenogastres: Proneomeniidae), a new species from the South Shetland Islands (Antarctica)

During the Spanish oceanographic expedition BENTART '95, carried out in Antarctic waters off Livingston Island (South Shetland Islands), five small specimens of a new species of Solenogastres-Cavibelonia, Dorymenia troncosoi sp. nov., were collected at a depth of 65–240 m on a silt bottom. The species is characterized by the presence of a pallial cavity with four pouches (a dorsal, two lateral and a ventral pouch), seminal receptacles that open into the spawning ducts through a short duct, radula having 9–11 teeth per row (1 central and 4–5 lateral, all the same size), the end of the copulatory spicules having a cross-section in the shape of a four-pointed star, and abdominal spicules present. These characteristics separate this species from other species of the genus, particularly from Dorymenia profunda, which is the most similar. Received: 9 April 1998 / Accepted: 12 June 1998     

Distribution, standing stock, growth, mortality and production of Strongylocentrotus pallidus (Echinodermata: Echinoidea) in the northern Barents Sea

The regular sea urchin, Strongylocentrotus pallidus (G.O. Sars, 1871), is a widespread epibenthic species in high-Arctic waters. However, little is known about its distribution, standing stock, population dynamics and production. In the northern Barents Sea, S. pallidus was recorded on seabed still photographs at 10 out of 11 stations in water depths of 80–360?m. Mean abundances along photographic transects of 150–300 m length ranged between <0.1 and 14.7?ind. m^?2 yielding a grand average of 3.6?ind.?m^?2. The small-scale distribution along the transects was patchy, with densities varying from nil to an overall maximum of 25.5 ind. m^?2, and exhibited a significant relation to the number of stones present. Sea urchin test diameters, measured on scaled photographs, extended from 7 to 90?mm. Median values at single stations varied from 14 to 46?mm, showing a significant inverse relationship to water depth. Biomass, estimated by combining photographic abundances, size frequencies and a size-mass function established with trawled specimens, ranged between <0.1 and 3.0?g ash-free dry mass m^?2, averaging about 1.0?g ash free dry mass m^?2. An analysis of skeletal growth bands in genital plates was carried out with 143 trawled individuals ranging in test diameter (D) from 4 to 48?mm. Assuming these bands to represent annual growth marks, the ages of the specimens analysed ranged between 3 and 42 years. A von Bertalanffy function was fitted to size-at-age data to model individual growth pattern (D_∞?=?102.3?mm, k?=?0.011 year^?1, t₀?=?0.633?year). The annual mortality rate Z of the population in the northern Barents Sea was estimated from a size-converted catch curve to be 0.08 year^?1. Applying the weight-specific growth rate method, the average P/B ratio and the mean annual production of this population were estimated as 0.07 year^?1 and 0.076?g AFDM m^?2 year^?1, respectively. In conclusion, S. pallidus is characterized by slow growth, low mortality, high longevity and low productivity. Because of its relatively high biomass, it is considered to contribute significantly to total benthic standing stock and carbon flux in the study area.     

Activity of the CaMV 35S promoter in various parts of transgenic early flowering birch clones

Early flowering together with small size would be useful for various biotechnical or genetic studies on trees. We report here the selection and micropropagation of early flowering birch (Betula pendula) clones (BPM1–12) obtained from seeds of birches bred elsewhere for early flowering. Under conditions that accelerate flowering (a high CO₂ level, strong and continuous illumination), the first male inflorescences emerged in 3–5 months, the trees then being 20–80 cm high. Transgenic lines (CaMV 35S-GUS INT) were produced through Agrobacterium-mediated gene transfer from BPM2, BPM5 and JR1/4 (a normally flowering birch). β-Glucuronidase (GUS) activities in the different lines, assayed 1–1.5 years after transformation, varied greatly. During further in vitro culture for 10 months, the activities decreased to 0.3–7% of the original values. GUS activities were detected in all organs studied, including the developing male inflorescences; the highest activity was in the roots. Received: 28 April 1997 / Revision received: 5 September 1997 / Accepted: 30 November 1997     

Phylogenetic characterization and molecular evolution of bacterial endosymbionts in psyllids (Hemiptera: Sternorrhyncha)

Most sternorrhynchan insects harbor endosymbiotic bacteria in specialized cells (bacteriocytes) near the gut which provide essential nutrients for hosts. In lineages investigated so far with molecular methods (aphids, mealybugs, whiteflies), endosymbionts apparently have arisen from independent infections of common host ancestors and co-speciated with their hosts. Some endosymbionts also exhibit putatively negative genetic effects from their symbiotic association. In this study, the identity of endosymbionts in one major sternorrhynchan lineage, psyllids (Psylloidea), was investigated to determine their position in eubacterial phylogeny and their relationship to other sternorrhynchan endosymbionts. Small-subunit ribosomal RNA genes (16S rDNA) from bacteria in three psyllid species (families Psyllidae and Triozidae) were sequenced and incorporated into an alignment including other insect endosymbionts and free-living bacteria. In phylogenetic analysis, all sequences were placed within the gamma subdivision of the Proteobacteria. Three sequences, one from each psyllid species, formed a highly supported monophyletic group whose branching order matched the host phylogeny, and also exhibited accelerated rates of evolution and mutational bias toward A and T nucleotides. These attributes, characteristic of primary (P) bacteriocyte-dwelling endosymbionts, suggested that these sequences were from the putative psyllid P endosymbiont. Two other sequences were placed within the gamma-3 subgroup of Proteobacteria and were hypothesized to be secondary endosymbionts. The analysis also suggested a sister relationship between P endosymbionts of psyllids and whiteflies. Thus, a continuous mutualistic association between bacteria and insects may have existed since the common ancestor of psyllids and whiteflies. Calculations using a universal substitution rate in bacteria corrected for endosymbiont rate acceleration support the idea that this common ancestor was also the ancestor of all Sternorrhyncha. Compared with other P endosymbiont lineages, the genetic consequences of intracellular life for some psyllid endosymbionts have been exaggerated, indicating possible differences in population structures of bacteria and/or hosts.