Isolation and chemical analysis of the cell wall of Morchella sp.

Cell walls from Morchella sp. were isolated after mechanical breakage of the fungus. Analysis of the material revealed that the most prominent compound was protein, phosphate being present in lesser amounts. Non-nitrogenous polysaccharides accounted for 17%. They were mostly soluble in hot HCl. The following monosaccharides were identified by chromatography of hydrolyzed samples: glucose, mannose and galactose. No lipids were detected in the purified cell walls. Chitin was present in a concentration of 16%. It was identified by cytochemical reactions, infrared spectra, and X-ray diffraction analysis. The cell wall of Morchella did not contain chitosan nor cellulose.     

Isolation and expression of the catA gene encoding the major vegetative catalase in Streptomyces coelicolor Müller.

We isolated the catA gene for the major vegetative catalase from Streptomyces coelicolor Müller. It encodes a polypeptide of 488 residues (55,440 Da) that is highly homologous to typical monofunctional catalases. We investigated catA expression by analyzing both catA mRNA and catalase activity. catA expression was increased by H2O2 treatment but did not increase during stationary phase. A putative catalase (CatB) cross-reactive with anti-CatA antibody appeared during stationary phase and in the aerial mycelium.     

cis-acting control elements for Krüppel expression in the Drosophila embryo.

Krüppel (Kr), a gap gene of Drosophila, shows complex spatial patterns of expression during the different stages of embryogenesis. In order to identify cis-acting sequences required for normal Kr gene expression, we analysed the expression patterns of fusion gene constructs in transgenic embryos. In these constructs, bacterial lacZ expression was placed under the control of Kr sequences in front of a basal promoter. We identified cis-acting Kr control units which drive beta-galactosidase expression in 10 known locations of Kr expression in early and late embryos. More than one cis-regulatory element drives the expression in the anterior domain at the blastoderm stage, in the nervous system, the midline precursor cells and in the amino-serosa. In addition, two cis-acting elements direct the first zygotic expression of Kr in a striped subpattern within the central region of the blastoderm embryo. Both elements respond to alterations in the activities of maternal organizer genes known to be required for Kr expression in establishing the thoracic and anterior abdominal segments in the wild-type embryo.     

早春首次降雨时间及降雨量对古尔班通古特沙漠两种短命植物形态特征与叶绿素荧光的影响

早春降雨事件是影响荒漠短命植物生长的重要因子, 为了解古尔班通古特沙漠积雪完全融化后早春首次降雨时间及降雨量对短命植物光合生长的影响。该研究选取两种优势短命植物尖喙牻牛儿苗(Erodium oxyrhinchum)和琉苞菊(Centaurea pulchella)为研究对象, 设置3个首次降雨时间(积雪完全融化后第10、20和30天)和3个降雨水平(5、10和15 mm), 探究两物种的形态、生物量及光化学效率的响应差异。结果显示: 积雪完全融化后第10天进行首次降雨处理时, 随着首次降雨量的增加, 尖喙牻牛儿苗的叶面积和总生物量呈显著增加趋势, 最大光化学效率呈下降趋势。在第20和30天进行首次降雨处理时, 尖喙牻牛儿苗和琉苞菊的叶面积均呈显著增加趋势。在积雪完全融化后5 mm降雨处理时, 随着首次降雨时间的延迟, 尖喙牻牛儿苗和琉苞菊的最大光化学效率呈先下降后上升趋势; 10 mm降雨处理时, 尖喙牻牛儿苗的冠根比降低, 根长和总生物量呈先增加后下降趋势, 琉苞菊的根长呈显著增加趋势; 15 mm降雨处理时, 尖喙牻牛儿苗的叶面积、冠根比和最大光化学效率呈显著增加趋势; 琉苞菊的冠根比和最大光化学效率呈下降趋势, 总生物量呈显著增加趋势。尖喙牻牛儿苗和琉苞菊的根长分别与0-5 cm和5-10 cm土壤含水量呈显著正相关关系。这说明早春首次降雨量比首次降雨时间更能影响植株生长, 但不同物种对降雨格局变化的响应不一致, 这可能通过影响早春短命植物的形态特征, 进而影响早春荒漠生态系统的稳定性。     

Estimating the transfer rates of bacterial plasmids with an adapted Luria–Delbrück fluctuation analysis

To increase our basic understanding of the ecology and evolution of conjugative plasmids, we need reliable estimates of their rate of transfer between bacterial cells. Current assays to measure transfer rate are based on deterministic modeling frameworks. However, some cell numbers in these assays can be very small, making estimates that rely on these numbers prone to noise. Here, we take a different approach to estimate plasmid transfer rate, which explicitly embraces this noise. Inspired by the classic fluctuation analysis of Luria and Delbrück, our method is grounded in a stochastic modeling framework. In addition to capturing the random nature of plasmid conjugation, our new methodology, the Luria–Delbrück method (“LDM”), can be used on a diverse set of bacterial systems, including cases for which current approaches are inaccurate. A notable example involves plasmid transfer between different strains or species where the rate that one type of cell donates the plasmid is not equal to the rate at which the other cell type donates. Asymmetry in these rates has the potential to bias or constrain current transfer estimates, thereby limiting our capabilities for estimating transfer in microbial communities. In contrast, the LDM overcomes obstacles of traditional methods by avoiding restrictive assumptions about growth and transfer rates for each population within the assay. Using stochastic simulations and experiments, we show that the LDM has high accuracy and precision for estimation of transfer rates compared to the most widely used methods, which can produce estimates that differ from the LDM estimate by orders of magnitude.

Plasmid transfer can often spread resistance between important clinical pathogens. This study shows that widely used methods can lead to biased estimates of plasmid transfer rate by several orders of magnitude, and presents a new approach, inspired by the classic Luria-Delbrück approach, for accurately assessing this fundamental rate parameter     

Effluxes of nitrous oxide,methane and carbon dioxide and their responses to increasing nitrogen deposition in the Gurbantünggüt Desert of Xinjiang,China

Aims Desert soils play an important role in the exchange of major greenhouse gas (GHG) between atmosphere and soil. However, many uncertainties existed in understanding of desert soil role, especially in efflux evaluation under a changing environment. Methods We conducted plot-based field study in center of the Gurbantünggüt Desert, Xinjiang, and applied six rates of simulated nitrogen (N) deposition on the plots, i.e. 0 (N0), 0.5 (N0.5), 1.0 (N1), 3.0 (N3), 6.0 (N6) and 24.0 (N24) g·m^-2·a^-1. The exchange rates of N₂O, CH₄ and CO₂ during two growing seasons were measured for two years after N applications. Important findings The average efflux of two growing seasons from control plots (N0) were 4.8 μg·m^-2·h^-1, -30.5 μg·m^-2·h^-1 and 46.7 mg·m^-2·h^-1 for N₂O, CH₄ and CO_2, respectively. The effluxes varied significantly among seasons. N0, N0.5 and N1 showed similar exchange of N₂O in spring and summer, which was relatively higher than in autumn, while the rates of N₂O in N6 and N24 were controled by time points of N applications. The uptake of CH₄ was relatively higher in both spring and summer, and lower in autumn. Emission of CO₂ changed minor from spring to summer, and greatly decreased in autumn in the first measured year. In the second year, the emission patterns were changed by rates of N added. N additions generally stimulated the emission of N₂O, while the effects varied in different seasons and years. In addition, no obvious trends were found in the emission factor of N₂O. The uptake of CH₄ was not significantly affected by N additions. N additions did not change CO₂ emissions in the first year, while high N significantly reduced the CO₂ emissions in spring and summer of the second year, without affected in autumn. Structure equation model analysis on the factors suggested that N₂O, CH₄ and CO₂ were dominantly affected by the N application rates, soil temperature or moisture and plant density, respectively. Over the growing seasons, both the net efflux and the global warming potential caused by N additions were small.     

Isolation and identification of pathogenic Naegleria from Florida lakes.

Five cases of primary amoebic meningoencephalitis associated with swimming in freshwater lakes have been recorded in Florida over the past 14 years. The present study demonstrated that pathogenic Naegleria, the causative agent, is relatively widespread. Twelve of 26 lakes sampled only once yielded the amoeba. Populations in three of five lakes sampled routinely reached levels of one amoeba per 25 ml of water tested during the hot summer months. Overwintering in freshwater lake bottom sediments was demonstrated, showing that thermal-discharge pollution of waters plays a miniscule, if any, role in the maintenance of pathogenic Naegleria in nature in this semitropical area.     

Cloning of an avilamycin biosynthetic gene cluster from Streptomyces viridochromogenes Tü57.

A 65-kb region of DNA from Streptomyces viridochromogenes Tü57, containing genes encoding proteins involved in the biosynthesis of avilamycins, was isolated. The DNA sequence of a 6.4-kb fragment from this region revealed four open reading frames (ORF1 to ORF4), three of which are fully contained within the sequenced fragment. The deduced amino acid sequence of AviM, encoded by ORF2, shows 37% identity to a 6-methylsalicylic acid synthase from Penicillium patulum. Cultures of S. lividans TK24 and S. coelicolor CH999 containing plasmids with ORF2 on a 5.5-kb PstI fragment were able to produce orsellinic acid, an unreduced version of 6-methylsalicylic acid. The amino acid sequence encoded by ORF3 (AviD) is 62% identical to that of StrD, a dTDP-glucose synthase from S. griseus. The deduced amino acid sequence of AviE, encoded by ORF4, shows 55% identity to a dTDP-glucose dehydratase (StrE) from S. griseus. Gene insertional inactivation experiments of aviE abolished avilamycin production, indicating the involvement of aviE in the biosynthesis of avilamycins.     

Molecular cloning and high-level expression of a bromoperoxidase gene from Streptomyces aureofaciens Tü24.

A bromoperoxidase gene was cloned from Streptomyces aureofaciens Tü24 into Streptomyces lividans TK64 by using the promoter-probe vector pIJ486. Subcloning of DNA from the original, unstable clone allowed the gene to be localized to a 1.7-kilobase (kb) fragment of DNA. Southern blotting showed that the cloned 1.7-kb insert hybridized to a 4.3-kb fragment in an SstI digest of S. aureofaciens Tü24 total DNA. The 1.7-kb insert was shown to code for a protein with the electrophoretic properties of the subunits of the nonheme bromoperoxidase isolated from S. aureofaciens Tü24. The protein produced by S. lividans TK64 transformed with pHM621, which contained an 8.0-kb insert, was shown to be identical to the S. aureofaciens Tü24 bromoperoxidase in terms of its electrophoretic mobility on denaturing and nondenaturing polyacrylamide gels and its NH2-terminal amino acid sequence. The bromoperoxidase was overproduced (up to 180 times) by S. lividans TK64 containing pHM621. Based on the heat stability of the S. aureofaciens Tü24 bromoperoxidase, a new and simple purification procedure with very high yields was developed.     

The chemical composition of the cyst wall of the potato cyst-nematode, Heterodera rostochiensis

1. Cyst walls of the potato cyst-nematode (Heterodera rostochiensis Woll.) were isolated by sieving a suspension of crushed cysts. About 12mg. of dried cyst walls was obtained from 1000 cysts. 2. The cyst walls contained mainly protein (72%, calculated from nitrogen content). On acid hydrolysis about 77% of the cyst wall went into solution. Of 19 amino acids present, proline, glycine, and alanine were the most abundant, and made up about 50% by weight of the total amino acids. The amino acid composition suggested that collagen-like proteins predominated in the cyst wall and larval cuticle. 3. A small amount of glucosamine (1.5%) was present in the hydrolysates, but chitin was not detected in the cyst walls. 4. Other components of the cyst walls were lipid (2%), carbohydrate (0.5%) and a small amount of inorganic matter (ash, 5%). Polyphenols (2% by wt. of the cyst walls) occurred in the acid hydrolysates. The dark pigments of the cyst wall were not indole-containing melanins.     

Resistance of pathogenic Naegleria to some common physical and chemical agents.

Resistance of pathogenic Naegleria to drying, low and high temperature, and two halogens was studied. Dying made trophozoites nonviable instantaneously and cysts nonviable in less than 5 min. Trophozoites degenerated in hours at temperatures below 10 degrees C and in minutes when frozen; cysts survived according to the equation th - t0/theta 1,440/1.122T (t0 is survival at 0 degrees C; Tis temperature between 0 and 10 degrees C), but 1.5 h at --10 degrees C to 1 h at --30 degrees C. At 51, 55, 58, 63, and 65 degrees C, trophozoites survived about 30, 10, 5, 1 and less than 0.5 min, respectively, cysts survived three to four times longer at 51 degrees C and six to seven times longer at 55 to 65 degrees C. Cyst destruction rates by heat indicated first-order kinetics with 25,400 cal/1 degree C for energy of activation. Cyst destruction rates by free chlorine and I2 also conformed to first-order kinetics. Concentration-contact time curves yielded concentration coefficient values of 1.05 for free chlorine and 1.4 for I2 and point to superchlorination as an effective means of destroying the cysts if free residuals are used as a guide and allowance is provided for low temperature and/or high pH waters.