Phenanthrene stimulates the degradation of pyrene and fluoranthene by <Emphasis Type="Italic">Burkholderia</Emphasis> sp. VUN10013

Pyrene and fluoranthene, when supplied as the sole carbon source, were not degraded by Burkholderia sp. VUN10013. However, when added in a mixture with phenanthrene, both pyrene and fluoranthene were degraded in liquid broth and soil. The amounts of pyrene and fluoranthene in liquid media (initial concentrations of 50 mg l⁻¹ each) decreased to 42.1% and 41.1%, respectively, after 21 days. The amounts of pyrene and fluoranthene in soil (initial concentrations of 75 mg kg⁻¹ dry soil each) decreased to 25.8% and 12.1%, respectively, after 60 days. None of the high molecular weight (HMW) polycylic aromatic hydrocarbons (PAHs) tested adversely affected phenanthrene degradation by this bacterial strain and the amount of phenanthrene decreased rapidly within 3 and 15 days of incubation in liquid broth and soil, respectively. Anthracene also stimulated the degradation of pyrene or fluoranthene by Burkholderia sp. VUN10013, but to a lesser extent than phenanthrene. The extent of anthracene degradation decreased in the presence of these HMW PAHs.     

Role of autophagy in triacylglycerol biosynthesis in <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis> revealed by chemical inducer and inhibitors

Autophagy mediates degradation and recycling of cellular components and plays an important role in senescence and adaptive responses to biotic and abiotic stresses. Nutrient deprivation has been shown to trigger triacylglycerol (TAG) accumulation and also induces autophagy in various green algae. However, the functional relationship between TAG metabolism and autophagy remains unclear. To gain preliminary evidence supporting a role of autophagy in TAG synthesis, Chlamydomonas reinhardtii CC-2686 was grown in Tris-acetate phosphate medium with or without nitrogen and treated with an autophagy inducer (rapamycin) or inhibitors (wortmannin, 3-methyladenine, and bafilomycin A1). Fluorescence microscopic analysis of Nile red-stained cells following 72-h treatments showed that rapamycin induced accumulation of subcellular lipid droplets which are storage sites of TAG. Rapamycin treatment in combination with nitrogen starvation led to a greater abundance of lipid droplets. Wortmannin and bafilomycin A1, but not 3-methyladenine, inhibited lipid droplet accumulation in rapamycin-treated cells and to a less extent in nitrogen-depleted cells. These results suggested that autophagy contributes to TAG synthesis in C. reinhardtii, but is not a necessary process. Autophagy induction may also be used to further enhance TAG accumulation in microalgae under nutrient deprivation.     

Quantitative detection of the oil-degrading bacterium Acinetobacter sp. strain MUB1 by hybridization probe based real-time PCR

Quantitative detection of the oil-degrading bacterium Acinetobacter sp. strain MUB1 was performed using the SoilMaster^™ DNA Extraction Kit (Epicentre, Madison, Wisconsin) and hybridization probe based real-time PCR. The detection target was the alkane hydroxylase gene (alkM). Standard curve construction showed a linear relation between log values of cell concentrations and real-time PCR threshold cycles over five orders of magnitude between 5.4±3.0×10⁶ and 5.4±3.0×10² CFU ml⁻¹ cell suspension. The detection limit was about 540 CFU ml⁻¹, which was ten times more sensitive than conventional PCR. The quantification of Acinetobacter sp. strain MUB1 cells in soil samples resulted in 46.67%, 82.41%, and 87.59% DNA recovery with a detection limit of 5.4±3.0×10⁴ CFU g⁻¹ dry soil. In this study, a method was developed for the specific, sensitive, and rapid quantification of the Acinetobacter sp. strain MUB1 in soil samples.     

Survey of aflatoxin-producing fungi in certain fermented foods and beverages in Thailand

Aflatoxin-producing fungi were found in fermented foods and beverages: fermented rice (kaomak), soybean sauce (taotjo), peanut butter, soy sauce (shoyu), Thai red and white wine, and rice sugar wine. These foods were extracted directly and tested for aflatoxins by thin-layer chromatography (TLC) and high pressure liquid chromatography (HPLC).Four strains of aflatoxin-producing fungi were isolated from peanut butter, taotjo, and shoyu. Direct extracts of 10% of the peanut butters tested and 5% of the kaomak tested contained large amounts of aflatoxins. The HPLC procedure used in this experiment utilized chloroform-ethyl acetate (31)     

Enhanced Biodegradation of Anthracene in Acidic Soil by Inoculated <Emphasis Type="Italic">Burkholderia</Emphasis> sp. VUN10013

The ability of Burkholderia sp. VUN10013 to degrade anthracene in microcosms of two acidic Thai soils was studied. The addition of Burkholderia sp. VUN10013 (initial concentration of 10(5) cells g(-1) dry soil) to autoclaved soil collected from the Plew District, Chanthaburi Province, Thailand, supplemented with anthracene (50 mg kg(-1) dry soil) resulted in complete degradation of the added anthracene within 20 days. In contrast, under the same test conditions but using autoclaved soil collected from the Kitchagude District, Chanthaburi Province, Thailand, only approximately 46.3% of the added anthracene was degraded after 60 days of incubation. In nonautoclaved soils, without adding the VUN10013 inocula, 22.8 and 19.1% of the anthracene in Plew and Kitchagude soils, respectively, were degraded by indigenous bacteria after 60 days. In nonautoclaved soil inoculated with Burkholderia sp. VUN10013, the rate and extent of anthracene degradation were considerably better than those seen in autoclaved soils or in uninoculated nonautoclaved soils in that only 8.2 and 9.1% of anthracene remained in nonautoclaved Plew and Kitchagude soils, respectively, after 10 days of incubation. The results showed that the indigenous microorganisms in the pristine acidic soils have limited ability to degrade anthracene. Inoculation with the anthracene-degrading Burkholderia sp. VUN10013 significantly enhanced anthracene degradation in such acidic soils. The indigenous microorganisms greatly assisted the VUN10013 inoculum in anthracene degradation, especially in the more acidic Kitchagude soil.     

Potential of Sonchus Arvensis for the Phytoremediation of Lead-Contaminated Soil

Sonchus arvensis is one of the pioneer plant species that were found in the abandoned Bo Ngam Pb mine in Thailand. S. arvensis was collected from three sites. The highest Pb shoot concentration was 9317 mg kg^?1 and the highest translocation factor (TF) and bioaccumulation factor (BF) values were 2.5 and 6.0, respectively. To investigate Pb uptake capacity of S. arvensis, a hydroponic experiment was performed for 15 d. S. arvensis exposed to 5 mg L^?1 Pb solution had the highest Pb shoot accumulation (849 mg kg^?1). In a pot study, S. arvensis was grown in Pb mine soils amended with organic and inorganic fertilizers for 2 mo. The addition of organic fertilizer to the soil increased plant dry biomass sharply. All treatments with ethylene-diamine-tetra-acetic acid (EDTA) had Pb accumulation in shoots greater than 1000 mg kg^?1 and the highest Pb shoot accumulation was found in S. arvensis grown in soil amended with organic fertilizer and EDTA (1397 mg kg^?1). In a field trial study, S. arvensis was grown at three sites in the mine area for 6 mo. S. arvensis could tolerate a total Pb of 100,000 mg kg^?1 in the soil and accumulated Pb in the shoots up to 3664 mg kg^?1 with high TF (2.19) and BF (2.38) values. These results suggest that S. arvensis is a good candidate for Pb phytoremediation.     

Biodegradation of crude oil by soil microorganisms in the tropic

Five microorganisms, three bacteria and two yeasts, capable of degrading Tapis light crude oil were isolated from oil-contaminated soil in Bangkok, Thailand. Soil enrichment culture was done by inoculating the soil in mineral salt medium with 0.5% v/v Tapis crude oil as the sole carbon source. Crude oil biodegradation was measured by gas chromatography method. Five strains of pure microorganisms with petroleum degrading ability were isolated: three were bacteria and the other two were yeasts. Candida tropicalis strains 7Y and 15Y were identified as efficient oil degraders. Strain 15Y was more efficient, it was able to reduce 87.3% of the total petroleum or 99.6% of n-alkanes within the 7-day incubation period at room temperature of 25 ± 2 °C.     

LEAD TOLERANCE AND ACCUMULATION IN PTERIS VITTATA AND PITYROGRAMMA CALOMELANOS,AND THEIR POTENTIAL FOR PHYTOREMEDIATION OF LEAD-CONTAMINATED SOIL

A field survey was conducted to search for Pb accumulation in fern species at Bo Ngam Pb mine, Thailand. Eleven fern species including Pteris vittata accumulated Pb in the range of 23.3–295.6 mg kg^?1 in the aboveground parts. Hydroponic, pot, and field trial experiments were carried out to investigate Pb-accumulation ability in ferns; including P. vittata and the ornamental species, Pityrogramma calomelanos, Nephrolepis exaltata cv. Gracillimum, and N. exaltata cv. Smirha. In hydroponic experiment, Pi. calomelanos accumulated the highest concentration of Pb (root 14161.1 mg kg^?1, frond 402.7 mg kg^?1). The pot study showed that P. vittata, Pi. calomelanos, and N. exaltata cv. Gracillimum grew well when grown in soil Pb at 92900 mg kg^?1. N. exaltata cv. Gracillimum accumulated the highest Pb concentration in the frond (5074 mg kg^?1) and P. vittata accumulated the highest Pb concentration in the root (16257.5 mg kg^?1). All fern species exhibited TF values less than 1 in both hydroponic and pot experiments. When P. vittata and Pi. calomelanos were grown at mine soils for 6 months, P. vittata tolerated higher soil Pb (94584–101405 mg kg^?1) and accumulated more Pb in frond (4829.6 mg kg^?1) and showed TF > 1 after 2 months of growth. These results indicated that P. vittata can be potentially useful for phytoremediation of Pb-contaminated soil.     

Genome-wide identification of genes involved in tolerance to various environmental stresses inSaccharomyces cerevisiae

During fermentation, yeast cells are exposed to a number of stresses — such as high alcohol concentration, high osmotic pressure, and temperature fluctuation — so some overlap of mechanisms involved in the response to these stresses has been suggested. To identify the genes required for tolerance to alcohol (ethanol, methanol, and 1-propanol), heat, osmotic stress, and oxidative stress, we performed genome-wide screening by using 4828 yeast deletion mutants. Our screens identified 95, 54, 125, 178, 42, and 30 deletion mutants sensitive to ethanol, methanol, 1-propanol, heat, NaCl, and H₂O₂, respectively. These deleted genes were then classified based on their cellular functions, and cross-sensitivities between stresses were determined. A large number of genes involved in vacuolar H⁺-ATPase (V-ATPase) function, cytoskeleton biogenesis, and cell wall integrity, were required for tolerance to alcohol, suggesting their protective role against alcohol stress. Our results revealed a partial overlap between genes required for alcohol tolerance and those required for thermotolerance. Genes involved in cell wall integrity and the actin cytoskeleton are required for both alcohol tolerance and thermotolerance, whereas the RNA polymerase II mediator complex seems to be specific to heat tolerance. However, no significant overlap of genes required for osmotic stress and oxidative stress with those required for other stresses was observed. Interestingly, although mitochondrial function is likely involved in tolerance to several stresses, it was found to be less important for thermotolerance. The genes identified in this study should be helpful for future research into the molecular mechanisms of stress response.     

Effect of corn plant on survival and phenanthrene degradation capacity of Pseudomonas sp. UG14LR in two soils

A study was undertaken to assess if corn (Zea mays L.) can enhance phenanthrene degradation in two soils inoculated with Pseudomonas sp. UG14Lr. Corn increased the number of UG14Lr cells in both soils, especially in the acidic soiL Phenanthrene was degraded to a greater extent in UG14Lr-inoculated or corn-planted soils than uninoculated and unplanted soils. The spiked phenanthrene was completely removed within 70 days in all the treatments in slightly alkaline soil. However, in acidic soil, complete phenanthrene removal was found only in the corn-planted treatments. The shoot and root lengths of corn grown in UG14Lr-inoculated soils were not different from those in non-inoculated soil between the treatments. The results showed that in unplanted soil, low pH adversely affected the survival and phenanthrene degradation ability of UG14Lr. Planting of corn significantly enhanced the survival of UG14Lr cells in both the bulk and rhizospheric soil, and this in turn significantly improved phenanthrene degradation in acidic soil. Re-inoculation of UG14Lr in the acidic soil increased the number of UG14Lr cells and enhanced phenanthrene degradation in unplanted soil. However, in corn-planted acidic soils, re-inoculation of UG14Lr did not further enhance the already active phenanthrene degradation occurring in both the bulk or rhizospheric soils.