Dual Labeling of Pseudomonas putida with Fluorescent Proteins for In Situ Monitoring of Conjugal Transfer of the TOL Plasmid

We describe here a dual-labeling technique involving the green fluorescent protein (GFP) and the red fluorescent protein (DsRed) for in situ monitoring of horizontal gene transfer via conjugation. A GFPmut3b-tagged derivative of narrow-host-range TOL plasmid (pWWO) was delivered to Pseudomonas putida KT2442, which was chromosomally labeled with dsRed by transposon insertion via biparental mating. Green and red fluorescent proteins were coexpressed in donor P. putida cells. Cells expressing both fluorescent proteins were smaller in size than cells expressing GFP alone. Donors and transconjugants in mixed culture or sludge samples were discriminated on the basis of their fluorescence by using confocal laser scanning microscopy. Conjugal plasmid transfer frequencies on agar surfaces and in sludge microcosms were determined microscopically without cultivation. This method worked well for in situ monitoring of horizontal gene transfer in addition to tracking the fate of microorganisms released into complex environments. To the best of our knowledge, this is the first study that discusses the coexpression of GFP and DsRed for conjugal gene transfer studies.     

昆虫病原细菌“010”的鉴定

1970年初,我们发现从蜜蜂幼虫中分离的芽孢杆菌“010”在生产性能和对昆虫的致病方面,均显示了良好的特性。该年我组曾与南苑公社化工厂共同对此菌的生产及毒效进行了初步摸索。继之,考察了它的营养要求及某些生产工艺条件,并对不同害虫进行了室内及大田的毒力测定。1971—1972年,通过培养基试验和上罐考察,选出两组综合利用较为理想的培养基。其中一组配比是:豆饼粉2.4％、玉米浆2％、麦芽粉4％、碳酸钙0.1％。菌数曾达48亿/毫升。“010”杀虫范     

Plasmid-mediated bioaugmentation of activated sludge bacteria in a sequencing batch moving bed reactor using pNB2

AIMS: The applicability of plasmid pNB2 for bioaugmentation of bacteria in model wastewater treatment reactors receiving 3-chloroaniline (3-CA) was investigated. METHODS AND RESULTS: A setup of three biofilm reactors was studied, all initially inoculated with bacteria from activated sludge. Reactor PB received a Pseudomonas putida pNB2 donor strain not able to degrade 3-CA. Positive control reactor P received a 3-CA degrading Comamonas testosteroni pNB2-transconjugant. The negative control reactor N remained unchanged. Reactor P showed 3-CA degradation from the beginning of the experiment whereas in reactor PB, degradation started after an initial lag period. No degradation was observed in reactor N. PCR analysis showed that the P. putida donor abundance dropped in reactor PB, whereas the plasmid abundance did not, indicating transfer to other bacteria. A number of different 3-CA degrading C. testosteroni strains carrying pNB2 could be isolated from reactor PB. CONCLUSIONS: A successful plasmid-mediated bioaugmentation was achieved with C. testosteroni being the dominant 3-CA degrading pNB2 transconjugant species active in reactor PB. SIGNIFICANCE AND IMPACT OF THE STUDY: The study underlines the potential of gene transfer to contribute to establishment and spread of genetic information in general, particularly emphasizing the spread of xenobiotic degrading potential by dissemination of catabolic genes.     

Comparing global models of terrestrial net primary productivity (NPP): analysis of differences in light absorption and light-use efficiency

Twelve global net primary productivity (NPP) models were compared: BIOME3, CASA, CARAIB, FBM, GLO-PEM, HYBRID, KGBM, PLAI, SDBM, SIB2, SILVAN and TURC. These models all use solar radiation as an input, and compute either absorbed solar radiation directly, or the amount of leaves used to absorb solar radiation, represented by the leaf area index (LAI). For all models, we obtained or estimated photosynthetically active radiation absorbed by the canopy (APAR). We then computed the light use efficiency for NPP (LUE) on an annual basis as the ratio of NPP to APAR. We analysed the relative importance for NPP of APAR and LUE. The analyses consider the global values of these factors, their spatial patterns represented by latitudinal variations, and the overall grid cell by grid cell variability. Spatial variability in NPP within a model proved to be determined by APAR, and differences among models by LUE. There was a compensation between APAR and LUE, so that global NPP values fell within the range of ‘generally accepted values’. Overall, APAR was lower for satellite driven models than for the other models. Most computed values of LUE were within the range of published values, except for one model.     

我国几种土壤甲螨包括一新种及二新亚种记述（蜱螨亚纲：甲螨亚目）

本文报道了新近采得的我国土壤甲螨六种,其中新种和新亚种定名为:中华弯步甲螨Gibbicepheus chinensis sp. nov.,长新领甲螨帽尔山亚种Caenosamerus spatiosus maoershanensis subsp. nov.,克氏蕾甲螨帽尔山亚种Gemmazetes crosby maoershanensis subsp. nov.,并记述了我国新纪录甲螨三种:大全罗甲螨Perlohm(?) gigantea(Aoki,1960)、水沢,德之甲螨乳Tolamocecheus mizusouai Aoki,1966及沼沢梁甲螨Lamellobates palustris Hammer,1958。     

Comparative sequence analysis of the internal transcribed spacer 1 of Ochrobactrum species

The internal 16S/23S rDNA (rrs/rrl) internal spacer region 1 (ITS1) of 54 Ochrobactrum strains and close relatives was analysed. Separation of ITS1 containing PCR products by gel-electrophoresis, DGGE, cloning and sequencing revealed ITS1 length and sequence heterogeneity. We found up to 5 different allelic ITS1 stretches within a single strain (Ochrobactrum intermedium LMG 3301T), and 2-3 different ITS1 alleles in O. tritici. Within ITS1, ITS1c, being part of the conserved double-stranded rrn processing stem dsPS1, produced the most reliable segment tree. The overall ITS1, ITS1c and rrs phylogenetic tree topologies were generally consistent, but there was evidence for horizontal rrn (segment) transfer in O. tritici LMG 2134 (formerly O. anthropi). Good correlations were found between ITS1, ITS1c and rrs sequence similarity and DNA-DNA hybridization values indicating that phylogenetic analysis of ITS1 and ITS1c both can be used to preliminarily deduce the phylogenetic affiliation if HGT was excluded. Strains sharing > 96.19% ITS1c (> 95.11% ITS1) similarity fell within a species, and < or = 68.42% ITS1c (< or = 70.33% ITS1) similarity outside a genus. Both ITS1 and ITS1c analysis resolved microdiversity more profoundly than rrs analysis and revealed clades (genomovars) within O. anthropi that were also produced in rep cluster analysis. There was no evidence for habitat-specific ITS1 genomovars within Ochrobactrum species. Diversity of Ochrobactrum was higher in soil than at the rhizoplane below and at the species level. Isolates from soil contained only 1 rrn type whereas isolates from human clinical, animal and rhizoplane specimens could contain more.     

Antifibrotic properties of c-Ski and its regulation of cardiac myofibroblast phenotype and contractility

Cardiac myofibroblasts are key players in chronic remodeling of the cardiac extracellular matrix, which is mediated in part by elevated transforming growth factor-β? (TGF-β?). The c-Ski proto-oncoprotein has been shown to modify TGF-β? post-receptor signaling through receptor-activated Smads (R-Smads); however, little is known about how c-Ski regulates fibroblast phenotype and function. We sought to elucidate the function of c-Ski in primary cardiac myofibroblasts using a c-Ski overexpression system. Cardiac myofibroblasts expressed three forms of c-Ski with the predominant band at 105 kDa, and adenoviral c-Ski treatment resulted in overexpression of 95-kDa c-Ski in cellular nuclei. Exogenous c-Ski led to significant inhibition of type I collagen secretion and myofibroblast contractility using two-dimensional semifloating gel contraction assay in both basal and with TGF-β? (10 ng/ml for 24 h) stimulation. Overexpressed c-Ski did not inhibit nuclear translocation of phosphorylated R-Smad2, despite their binding, as demonstrated by immunoprecipitation. Acute treatment of primary myofibroblasts with TGF-β? in vitro revealed a marked nuclear shuttling of c-Ski at 24 and 48 h following stimulation. Remarkably, overexpression of c-Ski led to a stepwise reduction of the myofibroblast marker α-smooth muscle actin with increasing multiplicity of infection, and these results indicate that 95-kDa c-Ski overexpression may effect a loss of the myofibroblastic phenotype. Furthermore, adenovirus (Ad) for hemagglutinin-tagged c-Ski infection led to a reduction in the number of myofibroblasts versus Ad-LacZ-infected and uninfected controls, due to induction of apoptosis. Finally, we observed a significant increase in 105-kDa c-Ski in the cytosolic fraction of cells of the infarct scar and adjacent remnant myocardium vs. noninfarcted controls.