Structure and In Vitro Growth of Zygotic Embryos of Taro (Colocasia esculenta var. antiquorum)

Zygotic embryos of taro, Colocasia esculenta var. antiquorumwere examined using both light and scanning electron microscopyand cultured on Linsmaier-Skoog (LS) medium without the additionof growth regulators. Embryos present within mature seed consistof a hypocotyl-root axis and an undeveloped cotyledon and aresurrounded by two major types of endosperm cells, aleurone andstarchy endosperm. Embryos cultured on LS medium developed intomature plants only in the presence of endosperm tissue. Excisedembryos turned green after 2–4 d in culture and reacheda rapid growth period between days 4 and 6. Culture of taroembryos leading to viable plantlet development depends upon(1) removal of the outer and inner integument, and (2) the presenceof endosperm tissue (including an intact aleurone layer). Colocasia esculenta var. antiquorum, Araceae, taro, embryo culture, integument, endosperm     

The Effects of Bacterial Endotoxin on the Infection of Mice with Trypanosoma cruzi*

Mice (Rockland strain) infected with Trypanosoma cruzi strain Tulahuén were treated with Escherichia coli endotoxin before, simultaneously with, and after inoculation of the parasites. The peak parasitemias of endotoxin-treated mice were higher than those of nontreated infected animals, regardless of the time of endotoxin administration. Peak parasitemias occurred at the same time in infected nontreated mice as in animals given endotoxin before or simultaneously with the trypanosomes. If endotoxin was administered 24 hr after the infection, a delay in the peak parasitemia was noted. Changes in the survival time were not observed unless endotoxin was given 24 hr postinfection. Infected mice had an increasing susceptibility to the lethal effect of endotoxin. The LD₅₀ of endotoxin decreased from 675 μg for normal mice to 230, 92, and 18 μg for infected animals 1, 3, and 8 days after the infection, respectively. In the infected mice, the endotoxin-detoxifying ability of the spleen was found to be impaired.     

Ethanol production from two varieties of henequen (Agave fourcroydes Lem)

This work demonstrates the significant differences in ethanol production of two henequen (Agave fourcroydes Lem) varieties. Yaax ki, or green henequen, surpasses Sac ki, or white henequen, in weight, sugar accumulation capability and ethanol production. The study was carried out with the ‘piña’ (stem and basal part of the leaves that remain attached to it after harvesting the leaves) of 5‐ and 9‐year‐old plants, cultivated in two localities of Yucatán, México. At 5 years of age, Yaax ki piñas are 33% larger than those of Sac ki, and at 9 years this difference can increase to 59%. Juice from 5‐year‐old piñas of Yaax ki contains 15.6% more reducing sugars, whereas in the must it can exceed 67%. Values obtained in the 9‐year‐old plants indicate that the Yaax ki variety accumulates 30.6% more reducing sugars than the Sac ki, whereas in the must the difference is in favor of Yaax ki is 21.7%. To produce 1 L of ethanol at 40% concentration by volume from 5‐year‐old plants, 48 kg of Sac ki piñas and 29 kg of Yaax ki are required, whereas with the 9‐year‐old plants, only 23 and 19 kg are needed, respectively.     

Ecosystem recovery: heathland response to a reduction in nitrogen deposition

Atmospheric deposition of nitrogen is responsible for widespread changes in the structure and function of sensitive seminatural ecosystems. The proposed reduction in emissions of nitrogenous pollutants in Europe under the Gothenburg Protocol raises the question of whether affected ecosystems have the potential to recover to their previous condition and, if so, over what timescale. Since 1998, we have monitored the response of a lowland heathland in southern England following the cessation of a long‐term nitrogen addition experiment, and subsequent management, assessing changes in vegetation growth and chemistry, soil chemistry and the soil microbial community. Persistent effects of earlier nutrient loading on Calluna growth and phenology, and on the abundance of lichens, were apparent up to 8 years after nitrogen additions ceased, indicating the potential for long‐term effects of modest nutrient loading (up to 15.4 kg N ha^?1 yr^?1, over 7 years) on heathland ecosystems. The size and activity of the soil microbial community was elevated in former N‐treated plots, 6–8 years after additions ceased, suggesting a prolonged effect on the rate of nutrient cycling. Although habitat management in 1998 reduced nitrogen stores in plant biomass, effects on belowground nitrogen stores were small. Although some parameters (e.g. soil pH) recover pretreatment levels relatively rapidly, others (e.g. vegetation cover and microbial activity) respond much more slowly, indicating that the ecological effects of even small increases in nitrogen deposition will persist for many years after deposition inputs are reduced. Indeed, calculations suggest that the additional soil nitrogen storage associated with 7 years of experimental nitrogen inputs could sustain the observed effects on plant growth and phenology for several decades. Carry over effects on plant phenology and sensitivity to drought suggest that the persistence of vegetation responses to nitrogen deposition should be integrated into long‐term assessments of the impact of global climate change on sensitive ecosystems.     

Gene Cloning and Biochemical Characterization of an Alcohol Dehydrogenase from Euglena gracilis1

ABSTRACT. Euglena gracilis is a freshwater free‐living organism able to grow with ethanol as carbon source; to facilitate this metabolism several alcohol dehydrogenase (ADH) activities have been detected. We report the gene cloning, over‐expression, and biochemical characterization of a medium‐chain NAD⁺‐dependent ADH from E. gracilis (EgADH). The enzyme's amino acid sequence displayed the highest percentages of similarity and identity with ADHs of bacteria and fungi. In the predicted three‐dimensional model, all the residues involved in Zn²⁺, cofactor, and substrate binding were conserved. A conventional signal peptide for import into mitochondria could not be clearly identified. The protein of 37 kDa was over‐expressed, purified to homogeneity, and kinetically characterized. The enzyme's optimal pH was 7.0 for ethanol oxidation displaying a V_m of 11.7±3.6 U/mg protein and a K_m of 3.2±0.7 mM for this substrate. Isopropanol and isopentanol were also utilized, although with less efficiency. It showed specificity for NAD⁺ with a K_m value of 0.39±0.1 mM and Mg²⁺ or Zn²⁺ were essential for activity. The recombinant EgADH reported here may help to elucidate the roles that different ADHs have on the metabolism of short‐ and long‐chain alcohols in this microorganism.     

Chromosome counts in the genus Festuca section Eskia (Poaceae) in the Iberian Peninsula

Chromosome numbers of taxa of Festuca L. section Eskia Willk. in the Iberian Peninsula are given. The levels of ploidy for five taxa are confirmed. Idiograms and karyotypic formulae of the five taxa are presented for the first time. Two levels of ploidy occur in this section: diploid and tetraploid. One taxon, Festuca elegans ssp. merinoi is tetraploid and two other taxa have diploid and tetraploid populations. The remaining two taxa are solely diploid.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 331–337.     

Associations between water chemistry and fish community composition: a comparison between isolated and connected lakes in northern Sweden

1. The correlation between water chemistry, physical variables and fish community composition was examined in 40 small (≤30 ha) coastal lakes in northern Sweden. Twenty of the 40 lakes were isolated from other water bodies and 20 were connected to the Baltic Sea. Lakes were fished in summer, using three different methods. Water chemistry was sampled in late winter prior to ice‐out and pH was measured additionally in summer. 2. Our central question was whether water chemistry plays a greater role in the composition of fish communities in isolated lakes than in connected lakes, as isolated lakes cannot be recolonised once a species has become extinct. 3. Results indicate that winter anoxia affects community composition only in isolated lakes, whereas acidity is of importance in both connected and isolated lakes. Methane (indicating anoxia), was significantly correlated with variation in fish community composition in isolated lakes, and a group of variables that indicate anoxia (CH₄, pCO₂, inorganic carbon and dissolved oxygen) explained 24–34% of the variation. pH alone explained 12% of the variation in community composition for connected lakes and a group of variables indicating acidity (summer and winter pH and ANC) explained 10–20% of the variation in isolated lakes. Lake area was the most important physical variable, being significantly correlated with the variation in fish community composition in connected lakes. 4. In isolated lakes, the presence of pike (Esox lucius), perch (Perca fluviatilis) and roach (Rutilus rutilus) was associated with low CH₄. The occurrence of crucian carp (Carassius carassius) and roach was positively correlated with pH, and the crucian carp was apparently also affected by predation by pike and perch. In connected lakes the effect of anoxia was low, probably due to the possibility of recolonisation and pockets of oxygenated water, allowing pike and perch to persist and thereby limiting the distribution of crucian carp.     

Responses in organic matter accumulation and processing to an experimental wood addition in three headwater streams

1. Additions of large wood are being used to restore streams that have been subjected to channelization, wood removal or riparian timber harvest. This added wood potentially increases channel stability, habitat complexity and organic matter retention and improves habitat and productivity of higher trophic levels. However, few stream restorations monitor restoration effectiveness after project completion. 2. We added 25 aspen logs (each 2.5 m length × 0.5 m diameter) to 100‐m reaches of each of three forested headwater streams in the Upper Peninsula of Michigan, U.S.A. These wood‐poor streams drain forests that were completely harvested of timber over a century ago and have been selectively logged for the past 50–60 years. An upstream unmanipulateds 100‐m reach in each stream served as the control. 3. We evaluated responses in organic matter processing by measuring red maple leaf decomposition 1 year before and 2 years after wood addition. We also quantified coarse organic matter standing stocks in the main channel and in debris accumulations associated with large wood. In response to wood addition, we predicted both organic matter standing stocks and leaf decomposition rates would increase, thereby enhancing resource availability to higher trophic levels. 4. Leaf decomposition rates did not change following wood addition. Temporal variation in rates among streams was mostly explained by differences in degree days, water velocity, scour/burial and water column inorganic nitrogen concentrations, but not large wood. Variation within streams across years was explained by differences in degree days, water velocity and shredder biomass. 5. Contrary to our prediction, organic matter standing stocks did not increase significantly at the reach scale. However, the experimentally added wood retained c. 4% of total annual coarse benthic organic matter (CBOM) in the first year and an additional c. 15% in the second year, suggesting accumulation over time in the manipulated reaches. The CBOM held by the new logs may be more biologically available because it is less susceptible to burial and transport than material in the streambed. 6. Some shredding macroinvertebrates responded to changes caused by the wood additions. In particular, the common caddisfly shredder, Lepidostoma sp., increased in abundance in leaf bags following wood addition, whereas the biomass of the winter stoneflies, Capniidae, declined in the first year. 7. Considerable funds are spent to restore in‐stream habitat, but few restorations are monitored, particularly over long periods (>5 years). Our results show that longer‐term monitoring is needed to determine the efficacy of these restorations on ecosystem function; organic matter decomposition in our low‐gradient streams did not respond to a substantial increase in large wood after 2 years.