Inhibition of mycorrhizal symbiosis in Leucaena leucocephala by chlorothalonil

The effect of the fungicide, chlorothalonil, on vesicular-arbuscular mycorrhizal (VAM) symbiosis was studied in a greenhouse using Leucaena leucocephala as test plant. Chlorothalonil was applied to soil at 0, 50, 100 and 200 μg g⁻¹. The initial soil solution P levels were 0.003 μg mL⁻¹ (sub-optimal) and 0.026 μg mL⁻¹ (optimal). After 4 weeks, the sub-optimal P level was raised to 0.6 μg mL⁻¹ (high). The soil was either uninoculated or inoculated with the VAM fungus, Glomus aggregatum. The fungicide reduced mycorrhizal colonization of roots, development of mycorrhizal effectiveness, shoot P concentration and uptake and dry matter yields at all concentrations tested, although the highest inhibitory effect was noted as the concentration of the fungicide was increased from 50 to 100 μg g⁻¹. Phosphorus applied after four weeks tended to partially offset the deleterious effects of chlorothalonil in plants grown in the inoculated and uninoculated soil which suggests that the fungicide was interfering with plant P uptake. The results suggest that the use of chlorothalonil should be restricted to levels below 50 μg g⁻¹ if the benefits of mycorrhizal symbiosis are to be expected. Contribution from Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No. 3464. Contribution from Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No. 3464.     

Effects of heavy-metal additions on ammonification and nitrification in soils contaminated with cadmium,lead and zinc

Summary The sensitivity of the mineralization of nitrogen by a range of soils contaminated with heavy metals (up to 340 μg Cd g⁻¹, 7500 μg Pb g⁻¹ and 34000 μg Zn g⁻¹) to the addition of heavy metals in solution were studied using pot incubations (ammonification) and a soil perfusion technique (nitrification). The ammonification of peptone showed little correlation between treatments with Cd, Zn (1000 and 5000 μg g⁻¹) and Pb (10000 and 20000 μg g⁻¹) and origin of the soil. Nitrification was considerably more sensitive to heavy metals than ammonification. All the soils had active, often large, populations of ammonifying and nitrifying organisms which showed substantial similarities between the soils. The rate of nitrifying activity (NO₃−N production) was logrithmic in most cases. The presence of tolerant populations of nitrifying organisms in the contaminated soils was demonstrated. Tolerance was also eventually acquired after a longer lag phase, by the non-contaminated soil populations although the rate of activity was often reduced. Metals added in solution were adsorbed by the soil within 4 hours. Differences in toxicity between metal salts (chlorides, sulphates and acetate) were attributed to the amount left in solution. However, in many instances, acetate was found to stimulate all the stages in the mineralisation of nitrogen.     

Phosphorus absorption and utilization efficiency of pigeon pea (Cajanus cajan (L) Millsp.) in relation to dry matter production and dinitrogen fixation

The effect of P supply on absorption and utilization efficiency of P in relation to dry matter production and dinitrogen fixation was examined in 8 pigeon pea cultivars with different growth duration and a soybean cultivar under field conditions. In all the pigeon pea cultivars, the maximum whole plant dry weight was obtained in a P-deficient soil at 100 kg P ha⁻¹ application. The short duration cultivars had smaller whole plant dry weights at low P rates (5 and 25 kg P ha⁻¹) and poor response to P application compared with the medium and long duration cultivars. Increasing the P application rate significantly increased dinitrogen fixation in all the cultivars. At the low P rates, the total nodule activity (TNA) was lower in the short than in the medium and the long duration cultivars. However, at 200 kg P ha⁻¹ application, dinitrogen fixation did not vary among these cultivars except for one short duration cultivar whichregistered very low values. Dry matter production and dinitrogen fixation are strongly controlled by P absorption ability rather than P utilization efficiency. The low absorption ability of the short duration cultivars is mainly due to poor root development. The high P concentrations in the nodules of all the cultivars suggest that nodules have advantage over host plant interms of P distribution under P deficient conditions. Our results suggest that P plays an important role in dinitrogen fixation through an effective translocation of P to the leaf. Thus when P supply is limited, efficient cultivars obtained reasonably high yield through an effective translocation of the absorbed P to the leaf.     

Screening of diazotrophic bacteria Azopirillum spp. for nitrogen fixation and auxin production in multiple field sites in southern Brazil

Isolation of microorganisms, screening for desirable characters and selection of efficient strains are important steps to optimize high crop yields and improve the sustainability of the ecosystem. The objective of this study was isolate and identify Azopirillum spp. with enhanced potential to promote plant growth among the natural bacterial population associated with rhizosphere soil, roots and stem of maize collected from five maize-growing regions within the southern state of Rio Grande do Sul in Brazil. Diazotrophic microorganisms were isolated using semi-solid N-free and solid selective media NFb. In order to select the most efficient isolates as candidates for plant growth promotion, the purified bacterial strains were studied for cell morphology, and Gram staining, streptomycin resistance, as well as screened for their potential for nitrogen fixation and auxin production under sterile conditions. Among 224 isolates obtained 121 were able to fix nitrogen and produce auxin. The 30 most promising isolates produced indole-3-acetic acid (IAA) ranging in concentration from 3.51 μg to 246.69 μg IAA mg⁻¹. Nitrogen fixation ranged from 15.43 μg to 95.21 μg of N mg protein⁻¹ day⁻¹ From the 30 most productive isolates, chromosomal DNA was extracted and a portion of the nifH gene was amplified and sequenced. Twenty-nine isolates were found to be similar to the Azospirillum genus and one isolate was found to be similar to Herbaspirillum seropedicae. These bacterial isolates revealed potential to increase crop productivity, however field crop experiments in Rio Grande do Sul climatic conditions should be done in order to formulate recommendations for their use as inoculants.     

Studies on inositol-mediated expression of MAL gene encoding maltase and phospholipid biosynthesis in Schizosaccharomyces pombe

In this study, the effects of inositol addition on expression of the MAL gene encoding maltase and phosphatidylinositol (PI) biosynthesis in Schizosaccharomyces pombe (a naturally inositol-requiring strain) were examined. We found that specific maltase activity was at its maximum when the concentration of added inositol reached 6 μg ml⁻¹ in a synthetic medium containing 2.0% (w/v) glucose. When the concentration of added inositol was 1 μg ml⁻¹ in the medium, repression of MAL gene expression occurred at glucose concentration higher than 0.2% (w/v). However, when S. pombe was cultured in the synthetic medium containing 6 μg ml⁻¹, repression of maltase gene expression occurred only at initial glucose concentration above 1.0% (w/v). More mRNA encoding maltase was detected in the cells grown in the medium with 6 μg ml⁻¹ inositol than in those grown in the same medium with 1 μg ml⁻¹ inositol. These results demonstrate that higher inositol concentrations in the synthetic medium could derepress MAL gene expression in S. pombe. PI content of the yeast cells grown in the synthetic medium with 6 μg ml⁻¹ of inositol was higher than that of the yeast cells grown in the same medium with 1 μg ml⁻¹ of inositol. This means that PI may be involved in the derepression of MAL gene expression in S. pombe.     

Urea decomposing activity of fractionated brackish phytoplankton in Lake Nakaumi

The influence of brackish phytoplankton cell classes upon the response of urea decomposition was investigated in Lake Nakaumi. The urea decomposition rate was 5 to 350 μmol urea m⁻³ h⁻¹ in the light and 3 to 137 μmol urea m⁻³ h⁻¹ in the dark. The urea decomposition rates in the light were obviously higher than in the dark. An extremely high rate (350 μmol urea m⁻³ h⁻¹) was observed in Yonago Bay. The rate in the smaller fraction (<5 μm) exceeded that in the middle (5–25 μm) and larger fractions (>25 μm). The chlorophyll- and photosynthesis-specific rates for urea decomposition in the light were 0.5 to 3.9 μmol urea mg chl.a ⁻¹ h⁻¹ and 0.3 to 1.3 μmol urea mg photo.C⁻¹. The specific urea decomposing activities were higher in the smaller fraction than in the other two fractions. The present results suggest that in brackish waters urea decomposition occurred with coupling to the standing crop and photosynthetic activity of phytoplankton. Received: May 22, 1999 / Accepted: August 15, 1999     

Water stress and photoinhibition in acclimatization ofRosa hybrida plantlets

Summary MicropropagatedRosa hybrida plantlets were simultaneously rooted and acclimatized under 100 and 200 μmol m⁻² s⁻¹ light for 2 wk. At the end of the first week of acclimatization, the plantlets were transferred onto a low water potential medium (from −0.06 MPa to −0.3 MPa). Dry weight was decreased by increased hight and low water potential. Photoinhibition of photosynthesis, expressed as a decrease in Fv/Fm ratio and ΦPSII and an increase in 1 −qp, occurred in plants grown under 200 μmol m⁻² s⁻¹. When high light (200 μmol m⁻² s⁻¹) and water stress were applied simultaneously, their effects on chlorophyll fluorescence parameters depended on stress duration; after 1 d of water stress, photoinhibition was more pronounced; after 7 d of stress, Fv/Fm ratio and ΦPSII were higher than after 1 d of stress; photoinhibition was reduced. This suggests that after a 1-d stress, the effect of water stress alone included a superimposed effect of photoinhibition to which the water-stressed plants were sensitized; after 7 d, plantlets had adapted to water stress. The photoprotective effects under high light might result in energy dissipative mechanisms linked to photochemical and nonphotochemical quenching other than CO₂ fixation.     

Effect of salinity and inoculation on growth,nitrogen fixation and nutrient uptake ofVigna radiata (L.) Wilczek

This study reports the effect of salinity and inoculation on growth, ion uptake and nitrogen fixation byVigna radiata. A soil EC_e level of 7.5 dS m⁻¹ was quite detrimental causing about 60% decline in dry matter and grain yield of mungbean plants whereas a soil EC_e level of 10.0 dS m⁻¹ was almost toxic. In contrast most of the studied strains of Rhizobium were salt tolerant. Nevertheless, nodulation, nitrogen fixation and total nitrogen concentration in the plant was drastically affected at high salt concentration. A noticeable decline in acetylene reduction activity occurred when salinity level increased to 7.5 dS m⁻¹.     

Rapid biodegradation of metsulfuron-methyl by a soil fungus in pure cultures and soil

Summary Four strains of bacteria, 9 strains of fungi and 20 strains of actinomycetes capable of utilizing metsulfuron-methyl as sole carbon and energy source were isolated from a metsulfuron-methyl-treated soil by the enrichment culture method. A fungus named DS11F was selected as the most highly effective one according to the maximum tolerance concentration of 1,200 mg l⁻¹ and metsulfuron-methyl-degrading rate of 0.0716 g g⁻¹ cells h⁻¹, and was identified as an unknown strain of Penicillium sp. on the basis of colony growth, morphology and biochemical characteristics.␣Through liquid pure culture, the optimal metsulfuron-methyl-degrading conditions of DS11F were determined to be metsulfuron-methyl concentration 22.6 mg l⁻¹, inoculum concentration 12.25 mg l⁻¹, pH 7.0 and temperature 30°C. As additional C sources, supernatant of soaked compost could increase metsulfuron-methyl degradation by 8%, but glucose was ineffective. DS11F inoculation was found to significantly enhance the degradation of metsulfuron-methyl in soil, with the reduction of the concentration reaching 50% in 6 days. Admixture of compost could promote metsulfuron-methyl degradation to some extent. The growth of the inocula in the soils remained dominant and degradation resumed immediately when metsulfuron-methyl was applied again. The results show that addition of the isolated Penicillium sp. enhances the degradation of metsulfuron-methyl in water and soil.     

Diel variation in urea decomposing activity in the euphotic zone of brackish Lake Nakaumi

Diel variations in urea decomposing activity in the euphotic zone of brackish Lake Nakaumi were measured under fixed light intensity. The decomposition rate of urea was 17 to 44 μ mol urea m⁻³ h⁻¹ in the light and 10 to 27 μ mol urea m⁻³ h⁻¹ in the dark. Higher decomposition rates were obtained in the upper euphotic zone. A clear diel periodicity in the urea decomposition rate was observed, with high rates from 1200 to 1600 and low rates from 0000 to 0400. Chlorophyll a specific decomposing activity ranged from 12 to 21 μg urea C mg chl.a ⁻¹ h⁻¹ in the light and 7 to 13 μg urea C mg chl.a ⁻¹ h⁻¹ in the dark. In the light, high values were obtained from 1600 to 2000 and low values from 0400 to 0800. The diel change in specific decomposing activity exhibited a similar pattern to that of the photosynthetic assimilation number, following the diel change in photosynthetic activity. Received: March 10, 1999 / Accepted: October 22, 1999     

Soil O2 and CO2 effects on root respiration of cacti

Through use of a recently developed technique that can measure CO₂ exchange by individual attached roots, the influences of soil O₂ and CO₂ concentrations on root respiration were determined for two species of shallow-rooted cacti that typically occur in porous, well-drained soils. Although soil O₂ concentrations in the rooting zone in the field were indistinguishable from that in the ambient air (21% by volume), the CO₂ concentrations 10 cm below the soil surface averaged 540 μLL⁻¹ for the barrel cactusFerocactus acanthodes under dry conditions and 2400 μLL⁻¹ under wet conditions in a loamy sand. For the widely cultivated platyopuntiaOpuntia ficus-indica in a sandy clay loam, the CO₂ concentration at 10 cm averaged 1080 μLL⁻¹ under dry conditions and 4170 μLL⁻¹ under wet conditions. For both species, the respiration rate in the laboratory was zero at 0% O₂ and increased to its maximum value at 5% O₂ for rain roots (roots induced by watering) and 16% O₂ for established roots. Established roots ofO. ficus-indica were slightly more tolerant of elevated CO₂ than were those ofF. acanthodes, 5000 μLL⁻¹ inhibiting respiration by 35% and 46%, respectively. For both species, root respiration was reduced to zero at 20,000 μLL⁻¹ (2%) CO₂. In contrast to the reversible effects of 0% O₂, inhibition by 2% CO₂ was irreversible and led to the death of cortical cells in established roots in 6 h. Although the restriction of various cacti and other CAM plants to porous soils has generally been attributed to their requirement for high O₂ concentrations, the present results indicate that susceptibility of root respiration to elevated soil CO₂ concentrations may be more important.     

Effect of irradiation intensity on cell growth and kalata B1 accumulation in <Emphasis Type="Italic">Oldenlandia affinis</Emphasis> cultures

Light irradiation had remarkable effects on callus growth of Oldenlandia affinis with an optimum intensity of 35 μmol m⁻² s⁻¹. Biosynthesis of kalata B1, the main cyclic peptide in O. affinis, was induced and triggered with rising irradiation intensities. The highest concentration of kalata B1, 0.49 mg g⁻¹ DW characterised by the maximum productivity of 3.88 μg per litre and day was analysed at 120 μmol m⁻² s⁻¹, although callus growth was repressed. The light saturation point was established to be 35 μmol m⁻² s⁻¹, where kalata B1 productivity was in a similar order (3.41 μg per day) due to the higher growth index. O. affinis suspension cultures were shown to accumulate comparable specific kalata B1 concentrations in a delayed growth associated production pattern. These were dependent on irradiation intensity (0.16 mg g⁻¹ at 2 μmol m⁻² s⁻¹; 0.28 mg g⁻¹ at 35 μmol m⁻² s⁻¹). The batch cultivation process resulted in a maximum productivity of 27.30 μg per litre and day with culture doubling times of 1.16 d⁻¹. Submers operation represented a 8-fold product enhancement compared to callus cultivation.     

Light-Dependency of Growth and Secondary Metabolite Production in the Captive Zooxanthellate Soft Coral Sinularia flexibilis

The branching zooxanthellate soft coral Sinularia flexibillis releases antimicrobial and toxic compounds with potential pharmaceutical importance. As photosynthesis by the symbiotic algae is vital to the host, the light-dependency of the coral, including its specific growth rate (μ day⁻¹) and the physiological response to a range of light intensities (10–1,000 μmol quanta m⁻² s⁻¹) was studied for 12 weeks. Although a range of irradiances from 100 to 400 μmol quanta m⁻² s⁻¹ was favorable for S. flexibilis, based on chlorophyll content, a light intensity around 100 μmol quanta m⁻² s⁻¹ was found to be optimal. The contents of both zooxanthellae and chlorophyll a were highest at 100 μmol quanta m⁻² s⁻¹. The specific budding rate showed almost the same pattern as the specific growth rate. The concentration of the terpene flexibilide, produced by this species, increased at high light intensities (200–600 μmol quanta m⁻² s⁻¹).     

Linkages between N turnover and plant community structure in a tundra landscape

The spatial distribution of organic soil nitrogen (N) in alpine tundra was studied along a natural environmental gradient, covering five plant communities, at the Latnjajaure Field Station, northern Swedish Lapland. The five communities (mesic meadow, meadow snowbed, dry heath, mesic heath, and heath snowbed) are the dominant types in this region and are differentiated by soil pH. Net N mineralization, net ammonification, and net nitrification were measured using 40-day laboratory incubations based on extractable NH₄⁺ and NO₃⁻. Nitrification enzyme activity (NEA), denitrification enzyme activity (DEA), amino acid concentrations, and microbial respiration were measured for soils from each plant community. The results show that net N mineralization rates were more than three times higher in the meadow ecosystems (mesic meadow 0.7 μg N g⁻¹ OM day⁻¹ and meadow snowbed 0.6 μg N g⁻¹ OM day⁻¹) than the heath ecosystems (dry heath 0.2 μg N g⁻¹ OM day⁻¹, mesic heath 0.1 μg N g⁻¹ OM day⁻¹ and heath snowbed 0.2 μg N g⁻¹ OM day⁻¹). The net N mineralization rates were negatively correlated to organic soil C/N ratio (r = −0.652, P < 0.001) and positively correlated to soil pH (r = 0.701, P < 0.001). Net nitrification, inorganic N concentrations, and NEA rates also differed between plant communities; the values for the mesic meadow were at least four times higher than the other plant communities, and the snowbeds formed an intermediate group. Moreover, the results show a different pattern of distribution for individual amino acids across the plant communities, with snowbeds tending to have the highest amino acid N concentrations. The differences between plant communities along this natural gradient also illustrate variations between the dominant mycorrhizal associations in facilitating N capture by the characteristic functional groups of plants. Responsible Editor: Bernard Nicolardot     

Plant regeneration from phyllode explants of Acacia crassicarpa via organogenesis

In this paper we report the establishment of Acacia crassicarpa regeneration through organogenesis. We used phyllode (leaf) explants excised from 60-day-old in vitro seedlings for green compact nodule induction and, tested Murashige and Skoog (MS) media supplemented with various concentrations of 1-phenyl-3-(thiadiazol-5-yl) urea (thidiazuron) (TDZ) and α-naphthaleneacetic acid (NAA). Under the optimized condition, green compact nodules and adventitious shoots were induced in 10 and 40 days, respectively, on the medium containing a combination of 0.5 mg l⁻¹ TDZ and 0.5 mg l⁻¹ NAA. This medium also yielded the highest rate (56%) of adventitious shoots forming from the nodules. Efficient shoot elongation was achieved by transferring the clusters of adventitious shoots to medium containing 0.1 mg l⁻¹ TDZ within 2 months. The elongated adventitious shoots were rooted at a rate of 96.5% on half-strength MS medium with 0.5 mg l⁻¹ 3-indolebutyric acid (IBA) in 1 month. Rooted plantlets were hardened and successfully established in soil with an 80% survival rate. To our knowledge, this is the first report describing a detailed protocol for regeneration through organogenesis using phyllodes as explants for A. crassicarpa.     

Tissue culture and plant regeneration of blue grama grass, Bouteloua gracilis (H.B.K.) Lag. Ex Steud

Summary As a first step towards applying biotechnology to blue grama, Bouteloua gracilis (H. B. K.) Lag. ex Steud., we have developed a regenerable tissue culture system for this grass. Shoot apices were isolated from 3-d-old seedlings and cultured in 15 different growth regulator formulations combining 2,4-dichlorophenoxyacetic acid (2,4-D), Picloram (4-amino-3, 5,6-trichloropicolinic acid), N⁶-benzyladenine (BA) or adenine (6-aminopurine). The highest induction of organogenic callus was obtained with formulations containing 1 mg l⁻¹ (4.52 μM) 2,4-D plus 0.5 mg l⁻¹ (2.22 μM) BA. and 2 mg l⁻¹ (8.88 μM) BA plus 1 mg l⁻¹ (4.14 μM) Picloram with or without 40 mg l⁻¹ (296.08 μM) adenine. Lower frequencies of induction were obtained for embryogenic as compared to organogenic callus. The most efficient treatments for induction of embryogenic callus contained 2 mg l⁻¹ (9.05 μM) 2,4-D combined with 0.25 (1.11 μM) or 0.50 mg l⁻¹ (2.22 μM) BA, or 1 mg l⁻¹ (4.52 μM) 2,4-D with 0.50 mg l⁻¹ (2.22 μM) BA. Regeneration was achieved in hormonefree Murashige anmd Skoog (MS) medium, half-strength MS medium or MS medium plus 1 mg l⁻¹ (1.44 μM) gibberellic acid. The number of plantlets regenerated per 500 mg callus fresh weight on MS medium ranged from 9 for 2 mg l⁻¹ (9.05 μM) 2,4-D to 62.2 for induction medium containing 2 mg l⁻¹ (8,28 μM) Picloram, 1 mg l⁻¹ (4.44 μM) BA and 40 mg l⁻¹ (296.08 μM) adenine. Regnerated plants grown in soil under greenhouse conditions reached maturity and produced seeds.     

Bioremediation of Fungicides by Spent Mushroom Substrate and Its Associated Microflora

Experiments were conducted both under in vitro and in situ conditions to determine the biodegradation potential of button mushroom spent substrate (SMS) and its dominating microbes (fungi and bacteria) for carbendazim and mancozeb, the commonly used agricultural fungicides. During 6 days of incubation at 30 ± 2°C under broth culture conditions, highest degradation of carbendazim (17.45%) was recorded with B-1 bacterial isolate, while highest degradation of mancozeb (18.05%) was recorded with Trichoderma sp. In fungicide pre-mixed sterilized SMS, highest degradation of carbendazim (100.00–66.50 μg g⁻¹) was recorded with mixed inoculum of Trichoderma sp. and Aspergillus sp., whereas highest degradation of mancozeb (100.00–50.50 μg g⁻¹) was with mixed inoculum of Trichoderma sp., Aspergillus sp. and B–I bacterial isolate in 15 days of incubation at 30 ± 2°C. All these microbes both individually as well as in different combinations grew well and produced extracellular lignolytic enzymes on SMS, which helped in fungicides degradation. Under in situ conditions, among three different proportions of SMS (10, 20 and 30%, w/w) mixed with fungicide pre-mixed soil (100 μg g⁻¹ of soil), the degradation of carbendazim was highest in 30% SMS treatment, while for mancozeb it was in 20% SMS treatment. The residue levels of both fungicides decreased to half of their initial concentration after 1 month of SMS mixing.     

Nitrogen gas (N2+N2O) flux from urea applied to lowland rice as affected by green manure

A field study was conducted on a clay soil (Andaqueptic Haplaquoll) in the Philippines to directly measure the evolution of (N₂+N₂O)−¹⁵N from 98 atom %¹⁵N-labeled urea broadcast at 29 kg N ha⁻¹ into 0.05-m-deep floodwater at 15 days after transplanting (DT) rice. The flux of (N₂+N₂O)−¹⁵N during the 19 days following urea application never exceeded 28 g N ha⁻¹ day⁻¹. The total recovery of (N₂+N₂O)−¹⁵N evolved from the field was only 0.51% of the applied N, whereas total gaseous¹⁵N loss estimated from unrecovered¹⁵N in the¹⁵N balance was 41% of the applied N. Floodwater (nitrate+nitrite)−N in the 5 days following urea application never exceeded 0.14 g N m⁻³ or 0.3% of the applied N. Prior cropping of cowpea [Vigna unguiculata (L.) Walp.] to flowering with subsequent incorporation of the green manure (dry matter=2.5 Mg ha⁻¹, C/N=15) at 15 days before rice transplanting had no effect on fate of urea applied to rice at 15 DT. The recovery of (N₂+N₂O)−¹⁵N and total¹⁵N loss during the 19 days following urea application were 0.46 and 40%, respectively. Direct recovery of evolved (N₂+N₂O)−¹⁵N and total¹⁵N loss from 27 kg applied nitrate-N ha⁻¹ were 20% and 53% during the same 19-day period. The failure of directly-recovered (N₂+N₂O)−¹⁵N to match total¹⁵N loss from added nitrate-¹⁵N might be due to entrapment of denitrification end products in soil or transport of gaseous end products to the atmosphere through rice plants. The rapid conversion of added nitrate-N to (N₂+N₂O)−N, the apparently sufficient water soluble soil organic C for denitrification (101 μg C g⁻¹ in the top 0.15-m soil layer), and the low floodwater nitrate following urea application suggested that denitrification loss from urea was controlled by supply of nitrate rather than by availability of organic C.     

Antimicrobial activity of an endophytic <Emphasis Type="Italic">Xylaria</Emphasis> sp.YX-28 and identification of its antimicrobial compound 7-amino-4-methylcoumarin

An endophytic Xylaria sp., having broad antimicrobial activity, was isolated and characterized from Ginkgo biloba L. From the culture extracts of this fungus, a bioactive compound P3 was isolated by bioactivity-guided fractionation and identified as 7-amino-4-methylcoumarin by nuclear magnetic resonance, infrared, and mass spectrometry spectral data. The compound showed strong antibacterial and antifungal activities in vitro against Staphylococcus aureus [minimal inhibitory concentrations (MIC) 16 μg·ml⁻¹], Escherichia coli (MIC, 10 μg·ml⁻¹), Salmonella typhia (MIC, 20 μg·ml⁻¹), Salmonella typhimurium (MIC, 15 μg·ml⁻¹), Salmonella enteritidis (MIC, 8.5 μg·ml⁻¹), Aeromonas hydrophila (MIC, 4 μg·ml⁻¹), Yersinia sp. (MIC, 12.5 μg·ml⁻¹), Vibrio anguillarum (MIC, 25 μg·ml⁻¹), Shigella sp. (MIC, 6.3 μg·ml⁻¹), Vibrio parahaemolyticus (MIC, 12.5 μg·ml⁻¹), Candida albicans (MIC, 15 μg·ml⁻¹), Penicillium expansum (MIC, 40 μg·ml⁻¹), and Aspergillus niger (MIC, 25 μg·ml⁻¹). This is the first report of 7-amino-4-methylcoumarin in fungus and of the antimicrobial activity of this metabolite. The obtained results provide promising baseline information for the potential use of this unusual endophytic fungus and its components in the control of food spoilage and food-borne diseases.     

A reliable protocol for plant regeneration from callus culture of <Emphasis Type="Italic">Phalaenopsis</Emphasis>

Summary Callus of Phalaenopsis Nebula was induced from seed-derived protocorms on 1/2 Murashige and Skoog (MS) basal medium plus 0–1.0 mg l⁻¹ (0–4.52 μM) N-phenyl-N′-1,2,3,-thiadiazol-5-yl urea (TDZ) and/or 0–10 mg l⁻¹ (0–45.24 μ M) 2,4-dichlorophenoxyacetic acid (2,4-D). Protocorms 2 mo. old performed better than 1-mo.-old protocorms for callus induction. More calluses formed on 1/2 MS basal medium supplemented with 0.1–1.0 mg l⁻¹ (0.45–4.52 μM) TDZ. These calluses could be maintained by subculturing every month with basal medium supplemented with 0.5 mg l⁻¹ (2.27 μM) TDZ and 0.5 mg l⁻¹ (2.26 μM) 2,4-D. Protocorm-like bodies were formed, and plants regenerated from these calluses on 1/2 MS basal medium alone or supplemented with 0.1–1.0 mg l⁻¹ (0.45–4.52 μM) TDZ. Plantlets were then potted on sphagnum moss in the greenhouse and grew well. No chromosomal abnormalities were found among the root-tip samples of 21 of the regenerated plantlets that were successfully acclimatized.