Complete Sequence of Plasmid pMP1 from the Marine Environmental Vibrio vulnificus and Location of Its Replication Origin

A novel cryptic plasmid, pMP1, from an environmental Vibrio vulnificus MP-4 isolated from Mai Po Nature Reserve in Hong Kong, has been characterized. The 7.6-kb plasmid had guanine–cytosine content of 40.03% and encoded four open reading frames (ORFs) with >100 amino acids. The predicted protein of ORF1 contained 478 amino acids showing 29% identity and 50% similarity over 309 amino acids to the integrase of Vibrio cholerae phage VP2. ORF2 encoded a putative protein of 596 amino acids, which were 23% identity and 42% similarity over 455 amino acids to the tail tape measure protein TP901 of Chromohalobacter salexigens phage. ORF3 and ORF4 encoded putative proteins of 103 and 287 amino acids, respectively, but showed no homologies to any known proteins. Further experiments indicated that a 3.2-kb fragment from EcoRI digestion could self-replicate. Analysis indicated that a sequence upstream of ORF4 had the features characteristic of theta-type replicons: AT-rich region, six potential direct repeats (iterons) spaced approximately two DNA helical turn apart (about 23 bp), two copies of 9 bp dnaA boxes, three Dam methylation sites, and five inverted repeats. Complementation experiments confirmed that the protein encoded by ORF4 was required for plasmid replication. We propose that ORF4 encode a new type of Rep protein and pMP1 is a new type of theta plasmid.     

Macrobenthos of a tidal impoundment at the Mai Po Marshes Nature Reserve,Hong Kong

The composition and spatial distribution patterns of the macrobenthic faunal assemblages of an 8-ha tidal impoundment operated as a traditional shrimp pond at the Mai Po Marshes Nature Reserve, Hong Kong, were studied in relation to temporal changes in local environmental conditions. Species richness, abundance and biomass of macrobenthos across 5 different sub-habitats (seaward, middle, and landward parts of open water unvegetated areas, and Phragmites- and Kandelia-dominated, vegetated areas) within the pond were examined bimonthly between January 1997 and January 1998. Grab samples were collected randomly within the sub-habitats. Key physical environmental parameters of the sampling sites were also measured. A total of 46 species of macrobenthos was recorded: 11 polychaetes, 11 molluscs, 13 crustaceans and 11 insects. Mean species density in the five sub-habitats ranged from 0 to 3907 indm^–2, with mean biomass ranging from 0 to 96.9 gm^–2. The macrobenthos showed spatial and temporal differences among the sub-habitats and across sampling times. Species abundances of Polychaeta, Mollusca and Crustacea were significantly higher in the three open water areas than in the two vegetated (Phragmites- and Kandelia-dominated) areas, with an inverse pattern for Insecta. There were no clear temporal patterns although abundance and biomass generally increased in the cooler months. Results of a canonical correspondence analysis suggest that macrobenthic species richness, abundance and biomass in the open areas were positively correlated with salinity, while water depth, dissolved oxygen and sediment organic matter content had little relationship with the macrobenthic assemblage parameters. Ordination by multi-dimensional scaling suggests that different habitats supported distinct macrobenthic assemblages. The macrobenthic assemblage in the tidal pond was less species rich but denser than those in the neighboring tidal mangrove and mudflat, suggesting that conversion of these areas into extensively managed tidal aquaculture ponds results in reduced species richness in tropical mangrove habitats.     

Laticola dae n. sp. (Monogenea: Diplectanidae) from Epinephelus maculatus (Perciformes: Serranidae) off New Caledonia

Laticola dae n. sp. is described from specimens collected from the gill-filaments of the highfin grouper Epinephelus maculatus, a coral reef fish caught off Nouméa, New Caledonia, South Pacific. The species is characterised by a spoon-shaped sclerotised male copulatory organ, with four thin walls and 73–m in outer length, and a sclerotised vagina in form of a disc, 16–m in diameter, with a smaller hemisphere on one side. Laticola Yang et al., 2006 was described to accommodate diplectanids from Lates calcarifer (Centropomidae); this is the first Laticola described from a serranid. Other diplectanids, including several species of Pseudorhabdosynochus Yamaguti, 1958, were also found on the same species of fish; specimens of L. dae represented about half of the diplectanids collected; all other species were rare.

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Résumé Laticola dae n. sp. est décrit à partir de spécimens collectés sur les filaments branchiaux de la loche grisette, Epinephelus maculatus, un poisson de récif corallien pêché au large de Nouméa, Nouvelle-Calédonie, Pacifique Sud. L’espèce est caractérisée par un organe copulateur mâle sclérifié en forme de cuiller, avec quatre parois fines, long de 73–m, et un vagin sclérifié en forme de disque, de diamètre 16–m, avec un hémisphère plus petit d’un côté. Laticola Yang et al., 2006 a été décrit pour rassembler des Diplectanidae de Lates calcarifer (Centropomidae) ; ceci est le premier Laticola décrit d’un Serranidae. D’autres Diplectanidae, y compris plusieurs espèces de Pseudorhabdosynochus, ont aussi été trouvés chez ce poisson ; les spécimens de L. dae représentaient environ la moitié des Diplectanidae récoltés, toutes les autres espèces étaient rares.

     

The Herbivore Assemblage, Herbivory and Leaf Chemistry of the Mangrove Kandelia obovata in Two Contrasting Forests in Hong Kong

The herbivore assemblage, intensity of herbivory and factors determining herbivory levels on the mangrove Kandelia obovata (previously K. candel, Rhizophoraceae) were studied over a 13-month period at two forests with contrasting growing conditions in Hong Kong. Mai Po was part of an eutrophic embayment in the Pearl River estuary and generally offered more favourable conditions for mangrove growth, whereas Ting Kok had a rocky substratum and oceanic salinity. Twenty-four insect herbivore species were recorded on K. obovata, with lepidopteran larvae that consume leaf lamina being the dominant species. While leaf litter production was similar at the two forests, herbivory level at Mai Po (mean = 3.9% in terms of leaf area loss) was more severe than that at Ting Kok (mean = 2.3%). Peak herbivory levels were found in summer at both locations (6.5% for Mai Po and 3.8% for Ting Kok). Young leaves of K. obovata at both locations were generally preferred by the herbivores from the period of late spring to summer. Concentrations of most feeding deterrents (ash, crude fibre, and total soluble tannins) were significantly higher in both young and mature leaves at Ting Kok, whereas leaf nutrients (total nitrogen and water) were the same at the two sites. Young leaves at Ting Kok contained about 30% more tannins than their counterparts at Mai Po. Significant differences in leaf chemistry also existed between young and mature leaves at either site. The differences were concomitant with the observed patterns of leaf herbivory on K. obovata, and suggest a potential relationship between environmental quality and plant defence against herbivory.     

Production of Trichoderma strains with pesticide-polyresistance by mutagenesis and protoplast fusion

The sensitivity of two cold-tolerant Trichoderma strains belonging to the species T. harzianum and T.␣atroviride was determined to a series of pesticides widely used in agriculture. From the 16 pesticides tested, seven fungicides: copper sulfate, carbendazim, mancozeb, tebuconazole, imazalil, captan and thiram inhibited colony growth of the test strains significantly with minimal inhibitory concentrations of 300, 0.4, 50, 100, 100, 100 and 50 g/ml, respectively. Mutants resistant to carbendazim and tebuconazole were produced from both wild type strains by means of UV-mutagenesis. The cross-resistance capabilities and in␣vitro antagonistic properties of the mutants were determined. Carbendazim-resistant mutants showed total cross-resistance to benomyl and thiabendazole at a concentration of 20 g/ml. Intraspecific protoplast fusion was carried out between carbendazim- and tebuconazole-resistant mutants of both parental strains, and putative haploid recombinants with stable resistance to both pesticides were produced in the case of T.␣atroviride. These pesticide-polyresistant progenies are potential candidates for application in an integrated pest management system.This work was presented as an oral lecture in section ‘Agriculture, Soil, Forest Microbiology’ at the BioMicroWorld2005 conference.     

Effect of Sodium Salicylate on the Population Dynamics of the Rhizobacterium Pseudomonas aureofaciens in the Wheat Rhizoplane and Adjacent Soil

The effect of sodium salicylate on the population dynamics of the rhizobacterium Pseudomonas aureofaciens BS1393 and its variant bearing the naphthalene biodegradation plasmid pBS216 was studied in the wheat rhizoplane and adjacent soil. Optimum salicylate concentration for the maintenance of the plasmid-bearing strain and for the normal growth of wheat was found to be 250 g/g soil. When the soil was supplemented with salicylate, the population of P. aureofaciens BS1393(pBS216) in the wheat rhizoplane and adjacent soil was, respectively, 4- and 20-fold higher than that of the parent strain lacking the plasmid.     

Seasonal abundance and distribution of <Emphasis Type="Italic">Vibrio</Emphasis> species in the treated effluent of wastewater treatment facilities in suburban and urban communities of Eastern Cape Province,South Africa

We assessed the seasonal abundance and distribution of Vibrio species as well as some selected environmental parameters in the treated effluents of two wastewater treatment plants (WWTP), one each located in a suburban and urban community of Eastern Cape Province, South Africa. Vibrio population density ranged from 2.1×10⁵ to 4.36×10⁴ CFU/ml in the suburban community and from 2.80×10⁵ to 1.80×10⁵ CFU/ml in the urban community. Vibrio species associated with 180 μ, 60 μ, and 20 μ plankton sizes were observed at densities of 0–136×10³ CFU/ml, 0–8.40×10² CFU/ml, and 0–6.80×10² CFU/ml, respectively at the suburban community’s WWTP. In the urban community, observed densities of culturable Vibrio were 0–2.80×10² CFU/ml (180 μ), 0–6.60×10² CFU/ml (60 μm), and 0–1.80× 10³ CFU/ml (20 μm). The abundance of free-living Vibrio species ranged from 0 to 1.0×10² and 1.0×10³ CFU/ml in the suburban and urban communities’ WWTPs, respectively. Molecular confirmation of the presumptive Vibrio isolates revealed the presence of V. fluvialis (41.38%), V. vulnificus (34.48%), and V. parahaemolyticus (24.14%) in the suburban community effluents. In the urban community molecular confirmation revealed that the same species were present at slightly different percentages, V. fluvialis (40%), V. vulnificus (36%), and V. parahaemolyticus (24%). There was no significant correlation between Vibrio abundance and season, either as free-living or plankton-associated entities, but Vibrio species abundance was positively correlated with temperature (r=0.565; p<0.01), salinity, and dissolved oxygen (p<0.05). Turbidity and pH showed significant seasonal variation (p<0.05) across the seasons in both locations. This study underscores the potential of WWTPs to be sources of Vibrio pathogens in the watershed of suburban and urban communities in South Africa.     

Diversity ofFrankia strains isolated from a single alder stand

One hundredFrankia strains isolated from variousAlnus species in a single alder stand were tested for plasmid presence. Plasmid DNA was observed in five of the frankiae strains and was analyzed. We found that plasmids with a similar molecular weight exhibited in fact minor divergences in restriction patterns. The genetic diversity among the five isolates which contained plasmids and seven isolates which contained no plasmid DNA were examined by using restriction endonucleas digestions, Southern hybridization ofnifHDK,nifAB genes, andFrankia cryptic DNA fragments determined at random. Results indicate that genomic DNA digestion patterns and Southern hybridizations to anifHDK probe were not able to discriminate between closely related frankiae. On the other hand, plasmid presence, Southern hybridization to anifAB proble or to a crypticFrankia probe allowed us to delineate groupings of these isolates.     

Site-specific recombination promotes linkage between trimethoprim- and sulfonamide resistance genes. Sequence characterization of dhfrV and sulI and a recombination active locus of Tn21

Summary A new gene for trimethoprim resistance, dhfrV, found in several plasmid isolates with different characteristics, was sequenced and found to correspond to a peptide of 157 amino acids showing 75% similarity with the previously characterized, drug resistant dihydrofolate reductase of type I. The sequenced surroundings of dhfrV in plasmid pLMO20, were found to be almost identical with genetic areas surrounding resistance genes in transposon Tn21 and in R plasmid R388. The trimethoprim resistance genes of pLMO20 and R388 and the spectinomycin resistance gene of Tn21 could be regarded as having been inserted, by recombination, into an evolutionary older structure containg the sulfonamide resistance gene, sulI. The latter gene was sequenced and found to correspond to a peptide of 279 amino acids and with a molecular weight of 30126 daltons. The inserted genes were found to be governed by a promoter situated in the highly conserved structure and also controlling expression of sulI. The insertion points of the different resistance genes were precisely defined, and at the 3 ends of the inserted genes inverted repeats allowing the formation of stem and loop structures were found. Similar structures were found at the 3 ends of the antibiotic resistance genes in Tn7, which could indicate similar recombination mechanisms to be effective in the evolutionary construction of all these different resistance elements.     

Photosynthetic induction in leaves of co-occurring <Emphasis Type="Italic">Fagus lucida</Emphasis> and <Emphasis Type="Italic">Castanopsis lamontii</Emphasis> saplings grown in contrasting light environments

Photosynthetic induction times and photoinhibition in relation to simulated sunflecks (sudden increase of irradiance from 20 to 1,500 μmol m⁻² s⁻¹) were examined in leaves of co-occurring Fagus lucida (a deciduous tree) and Castanopsis lamontii (an evergreen tree) saplings grown either in a beech forest understory or in an adjacent open site during a late rainy season. Two hypotheses were tested: (1) understory leaves would display faster photosynthetic induction times and greater photoinhibition than open-grown leaves; and (2) evergreen species would have slower photosynthetic induction times and lighter photoinhibition than deciduous species. Times to reach 90% of maximal CO₂ assimilation rate (t _90%A ) and stomatal conductance did not differ between species, but showed faster by 3–5 min in open-grown leaves than understory leaves due to higher initial stomatal conductance (g _{s initial}) and induction state 1 min into simulated sunflecks (IS_1min) in the former. Our analysis across the published data on photosynthetic induction of 48 broad-leaved woody species again revealed the negative correlations between t _90%A and either g _{s initial} or IS_1min, and the similarity of t _90%A and between evergreen and deciduous species. Measurements of maximum PSII photochemical efficiency (F _v/F _m) indicated that photoinhibition occurred in saplings in any of the growth habitats during sunfleck-induced photosynthetic induction. Despite no interspecific differences in the degree of photoinhibition, understory leaves of both species suffered heavier photoinhibition than open-grown leaves, as indicated by a stronger decrease of F _v/F _m in the former. Dynamic changes in the quantum yields of PSII photochemistry and ΔpH- and xanthophyll-regulated thermal dissipation and adjustments in the partitioning of electron flow between assimilative and non-assimilative processes were functional to resist photoinhibition. However, such photoinhibition, together with stomatal and biochemical limitations, would decrease carbon gain during simulated sunflecks, particularly in understory leaves.     

Tolerance of pesticides and antibiotic resistance in bacteria isolated from wastewater-irrigated soil

A total of 64 bacterial isolates (40 Pseudomonas spp., 12 Azotobacter and 12 Rhizobium spp.) were characterized on the basis of morphological, cultural and biochemical characteristics. All the isolates were tested for their tolerance to the pesticides endosulfan, carbofuran, and malathion. 12.5% of the Pseudomonas isolates from soil tolerated concentrations of 1600 g malathion ml whereas 7.5% of isolates tolerated the same concentration of carbofuran. However, Pseudomonas isolates demonstrated a tolerance limit to endosulfan at a concentration of 800 g/ml. Asymbiotic N₂-fixers (Azotobacter) and symbiotic N₂-fixers (Rhizobium spp.) were also able to tolerate concentrations of pesticides up to 1600 g/ml. All the isolates were further tested for their antibiotic susceptibility against seven different antibiotics, nalidixic acid, cloxacillin, chloramphenicol, tetracycline, amoxycillin, methicillin, doxycycline. 100% of the Pseudomonas isolates were resistant to cloxacillin and 57.5% were resistant to methicillin. 7.5% of the isolates exhibited multiple resistance to five different antibiotics in three different combinations whereas 25% of the isolates showed multiple resistance to four different antibiotics in seven different combinations. Some of the resistant isolates were also screened for plasmid DNA and found to harbour a single plasmid.     

Rapid Phenotypic Characterization of <Emphasis Type="Italic">Vibrio</Emphasis> Isolates by Pyrolysis Metastable Atom Bombardment Mass Spectrometry

Pyrolysis mass spectrometry was investigated for rapid characterization of food-borne bacterial pathogens. Nine isolates of Vibrio parahaemolyticus and one isolate each of Vibrio fluvialis, Vibrio hollisae, and Vibrio vulnificus were analyzed. Pyrolysis mass spectra, generated via an alternative ionization method, metastable atom bombardment, were subject to principal component-discriminant analysis. The spectral patterns were used to distinguish Vibrio isolates differing in species, serotype and expression of the thermostable direct hemolysin gene. The patterns of similarity and dissimilarity amongst spectra in the Vibrio test set generally reflected those associated with species, serotype or hemolysin-producing genes, though the combined influence of these and other variables in the multi-dimensional data did not produce a simple clustering with respect to any one of these characteristics. These results suggested that with enough examples to model the most common combinations, the method should be able to characterize Vibrio isolates according to their phenotypic characteristics. Pyrolysis-mass spectrometry with metastable atom bombardment and pattern recognition appeared suitable for rapid infraspecific comparison of Vibrio isolates. This integrated analytical, pattern-recognition system should be examined further for potential utility in clinical and public health diagnostic contexts.     

Genetic variation in nine Shorea species (Dipterocarpaceae) in Indonesia revealed by AFLPs

Shorea is the largest and most important genus of the Dipterocarpaceae. The genetic diversity and structure of nine Shorea species from two different locations, namely Nanjak Makmur in Sumatra and Sumalindo in Borneo, were evaluated using amplified fragment length polymorphism (AFLP) markers. A total of 274 trees were investigated at 85 polymorphic AFLP loci. Levels of genetic diversity of these species ranged from  = 0.100 for S. acuminata to  = 0.165 for S. blumutensis. The population of rare species S. blumutensis possessed the highest genetic diversity suggesting that geographically restricted species can have levels of genetic variation comparable to closely related widespread common congeners. Analyses of molecular variance revealed that the genetic variation was mainly found among species in both locations (57.7% in Sumatra; 56.3% in Borneo). The unweighted pairgroup method using arithmetic averages dendrogram of all samples revealed an almost complete separation of species. Thus, AFLP markers proved appropriate for phylogenetic studies of Shorea species. Specific markers have been detected showing high-frequency differences among species and between regions within species. Sequence information of these markers can be used to develop specific polymerase chain reaction markers for wood identification. The possibility of interspecific hybridization was discussed.     

Determination of the chemical composition of ulvan, a cell wall polysaccharide from Ulva spp. (Ulvales, Chlorophyta) by FT-IR and chemometrics

The water-soluble cell wall polysaccharides from green seaweeds of Ulva spp. (Ulvales, Chlorophyta), referred to as ulvan, demonstrate composition- and structure-related functional properties. Mid-infrared spectroscopy combined with chemometric techniques was investigated as a means to rapidly predict the chemical composition of ulvan extracts. A calibration was realized with 41 ulvan extracts from two Ulva species. The variables studied included the constituent sugars (rhamnose, xylose, glucose, galactose, glucuronic acid, iduronic acid), protein, and sulfate contents. The correlation between Fourier transform infrared and chemical data was developed using partial least squares (PLS) regression with full cross-validation (leave one out). The coefficients of determination in cross-validation () and the standard error in cross-validation were determined for each variable. The PLS model validation resulted in a coefficient of determination () and a standard error in prediction. Good predictions were obtained for rhamnose ( = 0.9244), xylose ( = 0.8758), glucuronic acid ( = 0.9415), and sulfate ( = 0.9218), which are the main ulvan constituents. However, minor components such as proteins, glucose, galactose, and iduronic acid were not correctly predicted. This study showed that mid-infrared spectroscopy combined with PLS regression is a reliable and fast method for the quantification of the main chemical constituents of ulvan extracts.     

Occurrence ofVibrio and other bacteria on the sea nettle,Chrysaora quinquecirrha

Sixty-two specimens of the sea nettle,Chrysaora quinquecirrha, were caught in the lower Chesapeake Bay, homogenized, and samples plated on a yeast extract-Bay water agar. Bacterial colonies were selected randomly, purified, and tested for 180 characteristics. Computer analysis permitted clustering of the 208 isolates into 15 groups (comprised of 133 strains) plus 75 nongrouped strains which failed to associate with any group at the 70% similarity level. The majority of the isolates (68.8%) wereVibrio species. These included 110 of the grouped strains (forming 12 of the 15 groups) and 33 of the nongrouped strains. The remainder of the isolates were distributed as follows:Pseudomonas (11.6%),Bacillus (8.2%),Flavobacterium (2.4%),Acinetobacter (2.4%),Moraxella (1.9%),Cytophaga (1.9%), Gram-positive cocci (1.4%), and miscellaneous (1.4%). All theBacillus were isolated from a group of moribund nettles and reflect an abnormal condition.Vibrio species predominated in the five catches of healthy nettles, but were distinctly different for each catch.     

Acclimation to light in seedlings of <Emphasis Type="Italic">Quercus petraea</Emphasis> (Mattuschka) Liebl. and <Emphasis Type="Italic">Quercus pyrenaica</Emphasis> Willd. planted along a forest-edge gradient

Photosynthetic acclimation of two co-occurring deciduous oaks (Quercus petraea and Quercus pyrenaica) to a natural light gradient was studied during one growing season. In the spring of 2003, 90 seedlings per species were planted along a transect resulting from a dense Pinus sylvestris stand, an adjacent thinned area and a 10-m-wide firebreak (16.5–60.9% Global Site Factor (GSF)). In two dates of the following summer, we measured leaf gas exchange, carboxylation efficiency (CE), chlorophyll and nitrogen content, light–response curves of chlorophyll a fluorescence parameters, and leaf mass per area (LMA). Summer was mild, as evidenced by leaf predawn water potential (Ψ_pd), which reduced the interactive effect of water stress on the response of seedlings to light. Q. pyrenaica had higher LMA, CE, stomatal conductance (g _{s max}) and photosynthesis per unit area than Q. petraea at all growth irradiances. , LMA, g _{s max} and electron transport rate (ETR) all increased with light availability (GSF) in a similar fashion in both species. Light had also a clear effect on the organization of Photosystem II (PS II), as deduced by chlorophyll a fluorescence curves. Chlorophyll concentration (Chl_m) decreased with increasing light availability in Q. pyrenaica but it did not in Q. petraea. Seedlings of Q. petraea acclimated to higher irradiances showed a greater non-photochemical quenching (NPQ) than those of Q. pyrenaica. This suggests a higher susceptibility to high light in Q. petraea, which would be consistent with a better adaptation to shade, inferred from the lower LMA or the lower rate of photosynthesis.     

The effect of temperature on post-feeding ammonia excretion and oxygen consumption in the southern catfish

The post-prandial rates of ammonia excretion (TAN) and oxygen consumption in the southern catfish (Silurus meridionalis) were assessed at 2 h intervals post-feeding until the rates returned to those of the fasting rates, at 17.5, 22.5, 27.5, and 32.5°C, respectively. Both fasting TAN and increased with temperature, and were lower than those previously reported for many fish species. The relationship between fasting TAN (mmol NH₃–N kg⁻¹ h⁻¹) and temperature (T, °C) was described as: fasting TAN = 0.144e ^0.0266T (r = 0.526, n = 27, P < 0.05). The magnitude of ammonia excretion and its ratio to total N intake during the specific dynamic action (SDA) tended to increase initially, and then decrease with increasing temperature. The ammonia quotient (AQ), calculated as mol NH₃–N/mol O₂, following feeding decreased as temperature increased. The relationship between AQ during SDA and temperature was described as: AQ_{during SDA} = 0.303e ^−0.0143T (r = 0.739, n = 21, P < 0.05). Our results suggest that ammonia excretion and oxygen consumption post-feeding are operating independently of each other. Furthermore, it appears that the importance of protein as a metabolic substrate in postprandial fish decreases with temperature.     

Restriction enzyme digestion patterns ofFrankia plasmids

Summary After the initial screening of more than 200Frankia strains, the plasmid DNA observed in eight Frankiae was analyzed.In situ lysis was performed to obtain an estimate of their copy number and molecular weight. Four plasmid classes were distinguished, 7–9, 18–20, 30–35 and 50–55 kb. Twelve plasmids were thus analysed with restriction enzymes to determine their plasmid restriction patterns.While someFrankia plasmids with comparable molecular weights were found to be heterologous in their restriction enzyme pattern, an 8 kb plasmid found in bothFrankia sp. ArI3, isolated fromAlnus rubra andFrankia sp. CpI1 isolated fromComptonia peregrina showed undistinguishable fingerprints. Furthermore, an 18 kb plasmid found in the same two strains, also showed homologous restriction enzyme patterns. However, the copy numbers of the two ArI3 plasmids were higher than those of the CpI1 plasmids.Similarly, strains ACN1^AG, , isolated fromAlnus crispa all contained a 50 kb plasmid, and the three plasmids were found upon restriction analysis to be undistinguishable.In one strain, ARgX17c isolated fromAlnus rugosa, it was found through restriction enzyme analysis that two plasmids of a similar molecular weight were in fact heterologous.The possible origin of the homologous plasmids and their potential as specificFrankia markers to be used in ecological studies are discussed.     

Biological functions of antioxidants in plant transformation

Browning and necrosis of transformed cells/tissues, and difficulty to regenerate transgenic plants from the transformed cells/tissues (recalcitrance) are common in Agrobacterium-mediated transformation process in many plant species. In addition, most crop transformation methods that use NPTII selection produce a significant number of nontransgenic shoots, called “shoot escapes” even under stringent selection conditions. These common problems of plant transformation, (browning and necrosis of transformed cells/tissues, recalcitrance, and the occurrence of shoot escapes) severely reduces transformation efficiency. Recent research indicates that reactive oxygen species (ROS) such as superoxide radical , the hydrogen peroxide (H₂O₂), the hydroxyl radical (OH′), and the peroxyl radical () may be playing an important role in tissue browning and necrosis during transformation. This review examines the role of ROS in in vitro recalcitrance and genetic transformation and the opportunities to improve transformation efficiency using antioxidants.     

In vitro susceptibility ofTrichophyton rubrum isolates to griseofulvin and tioconazole. Induction and isolation of a resistant mutant to both antimycotic drugs

The in vitro susceptibility of three clinicalTrichophyton rubrum isolates to griseofulvin and tioconazole, determined by the minimal inhibitory concentration (MIC), was 2 and 0.5 to 1.0g/ml, respectively. One mutant (gril) obtained after mutagenic treatment of one of these isolates was selected and showed simultaneous resistance to griseofulvin (MIC > 2000g/ml) and tioconazole (MIC=1.0g/ml). The clinical importance and the possibility of a multidrug resistance (MDR)-type mechanism being involved in this event is discussed.