Rhizosphere and endophytic fungal communities are considered critically important for plant health and soil fertility. In response to continuous cropping, Panax notoginseng becomes vulnerable to attack by fungal pathogens. In the present study, culture-independent Illumina MiSeq was used to investigate the rhizospheric and root endophytic fungi in response to continuous Panax notoginseng cropping practices. The results demonstrated that fungal diversity is increased inside the roots and in rhizospheric. Ascomycota, Zygomycota, Basidiomycota and Chytridiomycota were the dominant phyla detected during the continuous cropping of Panax notoginseng. The fungal diversity in the rhizospheric soil and roots of root-rot P. notoginseng plants are less than that of healthy plants in the same cultivating year, thus showing that root-rot disease also affects the community structure and diversity of rhizospheric and root endophytic fungi. Similarities in the major fungal components show that endophytic fungal communities are similar to rhizospheric soil fungal community based on a specialized subset of organisms. Canonical correspondence analysis on the fungal communities in root-rot rhizospheric from both healthy plants and rotation soils reveals that the soil pH and organic matter have the greatest impact upon the microbial community composition during continuous cropping, whereas soil nutrition status does not significantly affect the fungal community composition in response to continuous cropping practices. In addition, the results suggest that the unclassified genera Leotiomycetes, Cylindrocarpon, Fusarium and Mycocentrospora are shown as the potential pathogens which are responsible for the obstacles in continuous cropping of P. notoginseng. Further exploration of these potential pathogens might be useful for the biological control of continuous cropping of P. notoginseng.
Rhizobacteria and endophytic bacteria play important roles in protecting host plants from infection by phytopathogens, which cause soil-borne diseases and severely impair plant health. Panax notoginseng is negatively affected by continuous cropping and becomes vulnerable to attack by microbial pathogens. In the present study, culture-independent Illumina MiSeq was used to investigate root-endophytic and rhizospheric bacteria in response to continuous cropping of P. notoginseng. Numbers of rhizospheric bacteria decreased with continuous P. notoginseng cropping, while the effects of continuous cropping on endophytic bacteria were not statistically significant. Bacterial diversity was higher in healthy P. notoginseng rhizospheric soils and roots than in those of diseased P. notoginseng. The most dominant phyla detected during continuous cropping were Proteobacteria, Cyanobacteria, Actinobacteria, and Acidobacteria. The genera Pseudomonas, Rhodoplanes, Candidatus Solibacter, and Streptomyces were dominant in P. notoginseng rhizospheric soils and roots. Erwinia, Stenotrophomonas, Pseudomonas, and Sphingobium were specifically detected in relatively high proportions among root-rot rhizospheric bacteria and endogenous root bacteria in plants under continuous cropping, suggesting that they may be the pathogens responsible for the negative effects of continuous cropping on P. notoginseng. Based on canonical correspondence analysis of the bacterial communities that were identified from healthy plants and fallow soils, total phosphorus (P), pH, and organic soil matter exerted the greatest impacts upon bacterial community composition during continuous cropping. In general, continuous cropping practices for P. notoginseng and severe root-rot disease notably affected the community structure and the diversity of rhizospheric and root endophytic bacteria. Our study elucidated the ecological significance of microbial communities in healthy plant maintenance, and our results may inform biological control strategies during the continuous cropping of P. notoginseng.
Thermus strains are regarded as models to investigate the mechanism of thermostability of thermophiles, and phages from Thermus are particularly interesting because of their way to regulate gene expression. In this research, a Thermus bacteriophage named TSP4 (Thermus Siphoviridae phage) was isolated from Tengchong hot springs in China, and characteristics of morphology, temperature for phage production, pH and organic solvent sensitivity, DNA restriction endonuclease digestion and protein composition of TSP4 were further studied. TSP4 belonged to the Siphoviridae family and had a hexagonal head of 73 nm in diameter, an extremely long and flexible tail of 785 nm in length and 10 nm in width. TSP4 was very stable at 65 °C and pH 7.6. The capsid was apparently devoid of lipid. By SDS-PAGE, six protein bands were found in purified virions. Despite their exceptional habitats separated by thousands of kilometers, the characteristics of this thermophilic phage showed high similarity to Thermus siphoviruses P23-45 and P74-26 isolated from Kamchatka peninsula hot springs in the Far East, Russia.
A recombinant protein expression system working at low temperatures is expected to be useful for the production of thermolabile proteins. We constructed a low-temperature expression system using an Antarctic cold-adapted bacterium, Shewanella sp. strain Ac10, as the host. We evaluated the promoters for proteins abundantly produced at 4°C in this bacterium to express foreign proteins. We used 27 promoters and a broad-host-range vector, pJRD215, to produce -lactamase in Shewanella sp. strain Ac10. The maximum yield was obtained when the promoter for putative alkyl hydroperoxide reductase (AhpC) was used and the recombinant cells were grown to late stationary phase. The yield was 91 mg/liter of culture at 4°C and 139 mg/liter of culture at 18°C. We used this system to produce putative peptidases, PepF, LAP, and PepQ, and a putative glucosidase, BglA, from a psychrophilic bacterium, Desulfotalea psychrophila DSM12343. We obtained 48, 7.1, 28, and 5.4 mg/liter of culture of these proteins, respectively, in a soluble fraction. The amounts of PepF and PepQ produced by this system were greater than those produced by the Escherichia coli T7 promoter system.
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