Tissue culture ofKarwinskia humboldtiana—a plant producing toxins with antitumoural effects

Isolated embryos ofKarwinskia humboldtiana were cultured in vitro. The growth of embryos and development to plantlets on woody plant medium supplemented with indole-3-acetic acid 6.10^-2 mol l^–1, gibberellic acid (GA₃) 3.10^-2 mol l^–1, and 6-benzylaminopurine (BA) 2 mol l^–1 was obtained. Multiplication of shoots and rooting of excised shoots has been achieved. Callus formation on modified Murashige-Skoog medium supplemented with 1-naphthaleneacetic acid 10 mol l^–1, GA₃ 14 mol l^–1, and kinetin 5 mol l^–1 on hypocotyls, or on root cultures on medium supplemented with 2.4-dichlorophenoxyacetic acid 10 mol l^–1 and BA 10 mol l^–1 was induced.Abbreviations BA 6-benzylaminopurine - 2,4-d 2,4-dichlorophenoxyacetic acid - GA₃ gibberellic acid - IAA indole-3-acetic acid - NAA 1-naphthaleneacetic acid - TEM transmission electron microscopy     

The contribution of size-fractionated plankton to biomass and primary production of phytoplankton in saline–alkaline ponds

Primary productivity, biomass and chlorophyll-a of size fractionated phytoplankton (<0.22 m, <3 m, <8 m, <10 m, <40 m, <64 m, <112 m and <200 m) were estimated in 6 ponds and 5 experimental enclosures. The results showed that the planktonic algae less than 10 m are important in the biomass and production of phytoplankton in saline–alkaline ponds. The production of size fractionated phytoplankton corresponding to <112 m, <10 m and <3 m in saline–alkaline ponds were 10.5 ± 6.6 , 8.6 ± 5.4 and 0.33 ± 0.1 mgC l^–1 d^–1, respectively. Mean community respiration rate was 1.80 ± 0.73, 1.69 ± 0.90 and 1.38 ± 1.12 mgC l^–1 d^–1, respectively. The average production of phytoplankton corresponding to micro- (10–112 m), nano- (3–10 m) and pico- (<3 m) were 1.61, 8.30 and 0.33 mgC l^–1 d^–1, respectively. The ratio of those to the total phytoplankton production was 15%, 79% and 3%, respectively. The mean respiration rate of the different size groups was 0.11, 0.31 and 1.38 mgC l^–1 d^–1; the ratio of those to total respiration of phytoplankton was 6%, 17% and 77%, respectively. The production of size-fractionated phytoplankton corresponding to <200 m, <10 m and <3 m in enclosures was 2.19 ± 1.63, 2.08 ± 1.75 and 0.22 ± 0.08 mgC l^–1 d^-1, respectively. Mean community respiration rates were 1.25 ± 1.55, 1.17 ± 1.42 and 0.47 ± 0.32 mgC l^–1 d^–1, respectively. The average production of phytoplankton corresponding to micro- (10–200 m), nano- (3–10 m) and pico- (<3 m) plankton was 0.11, 1.86 and 0.22 mgC l^–1 d^–1, respectively. The ratio of those to the total production of phytoplankton was 5%, 85% and 10%, respectively. The mean respiration rate of different size groups were 0.08, 0.72 and 0.46 mgC l^–1 d^–1, the ratio of those to total respiration of phytoplankton was 6%, 57% and 37%, respectively. The concentrations of chlorophyll-a of the phytoplankton in the corresponding size of micro- (10–112 m), nano- (3–10 m) and pico- (<3 m) plankton in the experimental ponds were 19.3, 98.2 and 11. 9 g l^–1, respectively. The ratio of those to the total chlorophyll-a was 15%, 76% and 9%, respectively. The concentrations of chlorophyll-a of phytoplankton micro- (10–200 m), nano- (3–10 m) and pico- (<3 m) plankton in enclosures were 1.7, 34.3 and 3.0 g l^–1, respectively. The ratio of those to the total chlorophyll-a was 4%, 88% and 8%, respectively.     

Accumulation of silver by growing and non-growing populations of Citrobacter intermedius B6

Summary Accumulation of silver is reported for growing and non-growing populations of Citrobacter intermedius B6. In non-growing cultures a maximum uptake of 4.35% (w/w) was observed at an initial silver concentration of 2111.2 mol Ag⁺ l^-1. In contrast the maximum uptake of silver by growing bacteria was 2.81% (w/w) at an effective concentration of silver of 217.8 mol·l^-1. Silver accumulation rates in both resting (460 mol Ag⁺ g^-1 per hour) and continuously grown (41 mol Ag⁺ g^-1 per hour) bacteria are higher than previously reported. Cell fractionation and electron microscopy of continuously grown bacteria indicated that accumulation of silver was associated with the cell envelope, in the form of dense deposits of macromolecular proportions. This observation discounts simple surface adsorption as the process of accumulation in growing cultures.     

Build-up and decline of summer phytoplankton biomass in the eastern Weddell Sea,Antarctica

The seasonal development and decline of phytoplankton was investigated in the eastern Weddell Sea during summer and fall 1991. During the first half of the study (15 Jan–13 Feb) in an area off Vestkapp, favourable irradiance/mixing regimes initiated net phytoplankton growth in ice-free waters on the shelf and in stretches of open water over the partially ice-covered deep ocean. Chi a concentrations in the upper water column were moderate (0.2–0.8 g l^–1), but significantly above winter values. Later in the season (16 Feb–11 March), a phytoplankton bloom with surface Chl a concentrations ranging from 1.6–2.3 g l^–1 was encountered in an area further to the east. We suggest that the upper water column must have been stratified in this region for time scales of weeks to faciliate bloom development. Bacterial biomass and productivity generally paralleled the seasonal development of the phytoplankton. Nitrate concentrations in the upper mixed layer were substantially lower than would be expected from the existing phytoplankton standing stock, suggesting that heterotrophic consumption of organic matter by bacteria and zooplankton removed a large fraction of the primary production. The shallow seasonal pycnocline was eventually eroded by the passage of a storm, resulting in a homogeneous distribution of phytoplankton biomass over the entire water column, followed by sedimentation and deposition of phytodetritus on the sea floor. After the storm induced destratification, bacterial productivity was particularly high, amounting to more than half of the primary production (range: 10%–120%) in the upper water column. Subsequently, phytoplankton biomass in the upper water column decreased to values <1 g Chl a l^–1. The combination of low incident irradiances and incessant deep mixing prevented the phytoplankton biomass to increase again. During the last week of the investigation, extensive new-ice formation was observed. A major fraction of the residual surface plankton was incorporated into new sea ice, thus terminating the pelagic growth season of the phytoplankton in the eastern Weddell Sea.     

A liquid cytokinin pulse induces adventitious shoot formation from Douglas-fir cotyledons

Summary The effects of high-concentration, 2-h liquid pulses of N⁶-benzylaminopurine (BA) and thidiazuron (TD) on adventitious bud and shoot formation were tested in cotyledons of Douglas-fir (Pseudotsuga menziesii). Seedling age proved important; on average, cotyledons from the youngest seedlings formed 10-fold more buds than cotyledons from the oldest seedlings. Optimal cytokinin concentrations for the youngest cotyledons were 400 and 800 M BA, and 100 and 200 M TD. Shoots developed best from buds induced with 300, 400, and 800 M BA. Four gelling agents were tested; BRL agarose yielded more than three times the number of buds, and Gelrite nearly twice the number of buds, as either Sigma agar or Difco Bacto-Agar. One of the best treatments (400 M BA, agarose) yielded more cotyledons with buds, and more buds per cotyledon, than when cytokinins were incorporated into the growth medium.Abbreviations BA N⁶-benzylaminopurine - TD thidiazuron - SH Schenk and Hildebrandt (1972) - NAA 1-naphthaleneacetic acid Paper 2691 of the Forest Research Laboratory, Oregon State University, Corvallis. Mention of trade names or commercial products does not constitute endorsement or recommendation for use by the authors or Oregon State University.     

High frequency shoot regeneration from leaf explants of muskmelon

Efficient in vitro plant regeneration systems are critical for many purposes including plant transformation. Current regeneration systems for melon (Cucumis melo L.) plants generally utilize cotyledon explants; regeneration from melon leaves has received limited attention. We investigated several factors that influence regeneration from melon leaves including: genotype growth conditions and age of the source plant, leaf age, explant orientation, gelling agent, and the addition of silver nitrate and sulfonylurea herbicide to the culture media. Critical factors that influenced regeneration were preculture conditions of the donor plants, leaf size, and the use of silver nitrate and Phytagel in the medium. The best results were obtained with 3–4 cm diam leaves excised from pot grown greenhouse or growth chamber plants cultured on MS medium with 5 M IAA, 5 M BA, 1 M ABA, 30 M silver nitrate and 2.6 g l^-1 Phytagel. Low concentratons of sulfonylurea herbicide (0.25 mg l^-1 DPX-M 6316) also enhanced regeneration. Under optimized conditions 80–100% of the explants regenerated, with 10–100 shoots per explantAbbreviations ABA abscisic acid - BA benzyladenine - IAA indole-3-acetic acid - MS Murashige and Skoog medium - NAA naphthalene acetic acid     

Somatic embryo induction in Eucalyptus dunnii

Somatic embryogenesis has been reported in three Eucalyptus species so far. Here we report the initial success on the somatic embryo induction of a fourth species, E. dunnii, which is one of two species of choice to be grown in southern Brazil. Induction was performed on three day-old seedlings by means of naphthaleneacetic acid (5.5 or 16.5 M) alone or in combination with 2,4-dichlorophenoxyacetic acid (4.5 M). Either 10% coconut milk (v/v) or 1 g l^-1 hydrolysed casein enriched auxin-free medium with was able to induce the development of the somatic embryos.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - B5 Gamborg et al. (1968) medium - MS Murashige & Skoog (1962) medium - NAA -naphthaleneacetic acid - BA benzyladenine     

Contribution of flow cytometry to estimate picoplankton biomass in estuarine systems

Picoplankton (plankton 3 m) biomass was determined by flow cytometry in three European estuarine systems (Krka Estuary in Croatia, Rhône Delta in France, and Lena Delta and Laptev Sea in Russia). The size of natural phytoplankton groups was obtained by a calibration curve, with different picoplankton's strains (from 1.6 to 3.4 m), measured by a Coulter counter (size) and a flow cytometer (light-scattering). Two natural groups of picoplankton were identified by flow cytometry in the three systems: Synechococcus sp and picoeukaryotes. Picoplankton cells abundance ranged between: 2800 and 42000, 5000 and 37000, 1000 and 50000 cells ml^–1 in the Krka estuary, in the Rhône delta and in the Lena-Laptev system, respectively. In the Krka estuary, picoplankton biomass ranges between 11 and 68 gC l^–1. It can make up as much as 88% of the total photosynthetic plankton population and 50% of total organic particulate carbon. Picoplankton biomass was greater in the summer than in the autumn. At the halocline layer this biomass can attempt ca. 390 gC l^–1during the summer cruise. In the Rhône delta, a lower picoplankton biomass (6–39 gC l^–1) was observed at the end of the winter. These biomass represented between 0.4 and 22% of the particulate organic carbon, which could reach 71% of the total photosynthetic plankton biomass at the marine station. In the Lena-Laptev system, picoplankton biomass varied between 6 and 56 gC l^–1 in surface waters. Picoplankton biomass decreased with depth, but picoeukaryotes were still observed in deep samples (20, 30 m) in the Laptev Sea, showing a considerable autotrophic activity in spite of low temperatures (0–1 °C). Although the widely dispersed estuary geographic distribution and their different estuarine characteristics, the data point out that these small organisms can also play an important role in the transfer of organic carbon from rivers to oceans and that flow cytometry can be able to detect these small cells in turbid systems.     

Arsenic concentrations in water and fish from Chautauqua Lake,New York

Synopsis Arsenic persists in Chautauqua Lake, New York waters 13 years after cessation of herbicide (sodium arsenite) application and continues to cycle within the lake. Arsenic concentrations in lake water ranged from 22.4–114.81 g l^–1, = 49.0 ag l^–1. Well water samples generally contained less than 10 g l^–1 arsenic. Arsenic concentrations in lake water exceeded U.S. Public Health Service recommended maximum concentrations (10 g l^–1) and many samples exceeded the maximum permissible limit (50 g l^–1). Fish accumulated arsenic from water but did not magnify it. Fish to water arsenic ratios ranged from 0.4–41.6. Black crappie (Pomoxis nigromaculatus) contained the highest arsenic concentrations (0.14–2.04 g g^–1 ), X = 0.7 g g^–1) while perch (Perca flavescens), muskellunge (Esox masquinongy) and largemouth bass (Micropterus salmoides) contained the lowest concentrations (0.02–0.13 g g^–1). Arsenic concentrations in fish do not appear to pose a health hazard for human consumers.     

Effects of source of macronutrients and plant growth regulator concentrations on shoot proliferation of Cornus florida

Nodal stem segments of flowering dogwood (Cornus florida L.) were cultured on media containing seven different sources of macronutrients including full- and half-strength Murashige & Skoog (MS) macrosalts, N6, Anderson's (AND), Quoirin & Lepoivre's (LP), Nitsch & Nitsch (NIT), and Woody Plant Medium (WPM). All media contained MS micronutrients, Staba vitamins, 20 g l^-1 sucrose, and 6.5 g l^-1 Difco Bacto-agar, and were supplemented with 2.2 M 6-benzyladenine (BA) and 0.49 M indole-3-butyric acid (IBA). Following three subcultures, the best shoot proliferation was supported on media containing WPM macronutrients. To optimize the proliferation rate, shoots were cultured on WPM macronutrients supplemented with eight combinations of BA and IBA, and 3.3 M BA without IBA was determined to be the best.Abbreviations BA 6-benzyladenine - IBA indole-3-butyric acid