Medium-chain-length polyhydroxyalkanoate production by newly isolated Pseudomonas sp. TN301 from a wide range of polyaromatic and monoaromatic hydrocarbons.

AIMS: The aim of this study was to convert numerous polyaromatic and monoaromatic hydrocarbons into biodegradable polymer medium-chain-length polyhydroxyalkanoate (mcl-PHA). METHODS AND RESULTS: Using naphthalene enrichment cultivation method, we have isolated seven bacterial strains from the river sediment exposed to petrochemical industry effluents. In addition to naphthalene, all seven strains could utilize between 12 and 17 different aromatic substrates, including toluene, benzene and biphenyl. Only one isolate that was identified as Pseudomonas sp. TN301 could accumulate mcl-PHA from naphthalene to 23% of cell dry weight. Owing to poor solubility, a method of supplying highly hydrophobic polyaromatic hydrocarbons to a culture medium was developed. The best biomass and mcl-PHA yields were achieved with the addition of synthetic surfactant Tween 80 (0.5 g l(-1)). We have shown that Pseudomonas sp. TN301 can accumulate mcl-PHA from a wide range of polyaromatic and monoaromatic hydrocarbons, and mixtures thereof, while it could also accumulate polyphosphates and was tolerant to the presence of heavy metal (100 mmol l(-1) cadmium and 20 mmol l(-1) nickel). CONCLUSIONS: A new Pseudomonas strain was isolated and identified with the ability to accumulate mcl-PHA from a variety of aromatic hydrocarbons. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is the first report on the ability of a bacterial strain to convert a range of polyaromatic hydrocarbon compounds to the biodegradable polymer (mcl-PHA). Mcl-PHA is gaining importance as a promising biodegradable thermoelastomer, and therefore, isolation of new producing strains is highly significant. Furthermore, this strain has the ability to utilize a range of hydrocarbons, which often occur as mixtures and could potentially be employed in the recently described efforts to convert waste materials to PHA.

Structure and inheritance of ribosomal DNA variants in cultivated and wild hop, Humulus lupulus L.

Genetic variation was assessed among cultivated and wild hop, Humulus lupulus, by restriction fragment length polymorphisms (RFLPs) of the ribosomal RNA genes (rDNA). Two rDNA length variants of 10.3 and 9.3 kbp represented by three phenotypes designated A, B and C were detected with XhoI. Restriction-site mapping showed that hop rDNA is structurally similar to those of most higher plants. A high level of homogeneity existed in rDNA repeat lengths among the diverse hop genotypes. Generally, phenotype A was predominant in wild and cultivated European and Asian genotypes; phenotype B in North American cultivars; while phenotype C was present only in native North American hop, providing a potential molecular marker for the identification of this germ plasm. The rDNA data provided genetic evidence for the separation of native and cultivated American genotypes and supports the hypothesis that North American hop cultivars are of hybrid origin from European and native American genotypes. The segregation of rDNA phenotypes in four F1 families suggests that a single locus with two co-dominant alleles controls genetic variability for rDNA variants in hop.

Random amplified polymorphic DNA (RAPD) markers in hop, Humulus lupulus: level of genetic variability and segregation in F1 progeny.

The level of genetic variation in 24 hop genotypes was studied using the recently developed technique for producing random amplified polymorphic DNAs (RAPDs). Of the 60 primers screened, eight produced polymorphic RAPD bands, 38 produced bands that were monomorphic for all genotypes and 19 did not produce any amplification product. It appeared that the level of polymorphism among the genotypes was generally low. Three of the primers, A11, A17 and C9, were used to determine the stability and segregation of RAPD markers in five families with a total of 182 F1 progeny. The segregation ratios of these markers in the f1 progeny suggested that they were inherited in a Mendelian manner. RAPD markers were stable and may be useful for the construction of linkage maps in hop.

Chloroplast DNA differences between cultivated hop, Humulus lupulus and the related species H. japonicus.

Chloroplast DNA (cpDNA) of Humulus Lupulus and H. japonicus was examined by restriction endonuclease analysis with BamHI, BanI, BclI, BstEII, DraI, EcoRI, EcoRV, HindIII, KpnI, PaeR7I, PstI, PvuII, SalI and XhoI. The restriction fragment patterns showed that the cpDNAs shared a large number of restriction sites. However, the chloroplast genomes of the two species could be distinguished by differences in restriction site and restriction fragment patterns in the PstI, PvuII, BclI, EcoRV, DraI and HindIII digests. On the basis of the complexity of restriction enzyme patterns, the enzymes PstI, PvuII, SalI, KpnI and XhoI were selected for mapping the chloroplast genomes. Single and double restriction enzyme digests of cpDNA from the two species were hybridized to cpDNA probes of barley and tobacco. The data obtained from molecular hybridization experiments were used to construct the cleavage site maps. Except for the PstI digest, the arrangement of cpDNA restriction sites was found to be the same for both species. An extra PstI site was present in H. lupulus. Three small insertions/deletions of about 0.8 kbp each were detected in the chloroplast genomes of the two species. Two of these insertions/deletions were present in the large and one in the small singlecopy region of the chloroplast genome. The cpDNA of Humulus was found to be a circular molecule of approximately 148 kbp that contains two inverted repeat regions of 23 kbp each, a small and a large single -copy region of approximately 20 kbp and 81 kbp, respectively. The chloroplast genome of hop has the same physical and structural organization as that found in most angiosperms.