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Malfanova N.,Leiden University | Malfanova N.,All Russian Research Institute for Agricultural Microbiology ARRIAM | Franzil L.,University of Liege | Lugtenberg B.,Leiden University | And 2 more authors.
Archives of Microbiology

In a previous study (Malfanova et al. in Microbial Biotech 4:523-532, 2011), we described the isolation and partial characterization of the biocontrol endophytic bacterium B. subtilis HC8. Using thin-layer chromatography, we have detected several bioactive antifungal compounds in the methanolic extract from the acidprecipitated supernatant of HC8. In the present study, we have further analyzed this methanolic extract using liquid chromatography-mass spectrometry. Based on the comparison of retention times and molecular masses with those of known antifungal compounds, we identified three families of lipopeptide antibiotics. These include four iturins A having fatty acyl chain lengths of C14 to C17, eight fengycins A (from C14 to C18 and from C15 to C17 containing a double bond in the acyl chain), four fengycins B (C15 to C18), and five surfactins (C12 to C16). Evaluation of the antifungal activity of the isolated lipopeptides showed that fengycins are the most active ones. To our knowledge, this is the first report of an endophytic Bacillus subtilis producing all three major families of lipopeptide antibiotics containing a very heterogeneous mixture of homologues. The questions remain open which of these lipopeptides (1) are being produced during interaction with the plant and (2) are contributing to the biocontrol activity of HC8. © The Author(s) 2012. Source

Malfanova N.,Leiden University | Malfanova N.,All Russian Research Institute for Agricultural Microbiology ARRIAM | Kamilova F.,Leiden University | Validov S.,Leiden University | And 2 more authors.
Archives of Microbiology

Twenty endophytic bacteria were isolated from surface-sterilized stems and roots of cucumber plants. After removal of potential siblings and human pathogens, the remaining seven strains were identified based on their 16S rDNA as Pseudomonas fluorescens (2 strains) and P. putida (5 strains). Three strains, namely P. fluorescens CS1, P. fluorescens CR2 and P. putida CR3, were able to suppress tomato foot and root rot (TFRR). Special attention was paid to the characterization of the BIOLOG carbon oxidation profiles of the isolated pseudomonads in order to identify nutrients which might be important for their endophytic lifestyle. Comparative analysis of the profiles of these seven strains with those of seven rhizospheric Pseudomonas spp. revealed that endophytes were able to oxidize L-arabinose and 2,3-butanediol significantly more often than the rhizospheric group. An independent growth experiment performed in tubes using L-arabinose and 2,3-butanediol as sole carbon sources showed the same results as seen using BIOLOG for L-arabinose, but not for 2,3-butanediol. Since L-arabinose is one of the most abundant sugars in xylem of cucumber plants and was not detected in their rhizosphere, our data suggest that utilization of L-arabinose might be a trait contributing to the endophytic lifestyle of the isolated Pseudomonas endophytes. © Springer-Verlag 2012. Source

Egamberdieva D.,National University of Uzbekistan | Egamberdieva D.,Leiden University | Kucharova Z.,National University of Uzbekistan | Davranov K.,National University of Uzbekistan | And 8 more authors.
Biology and Fertility of Soils

The aim of the present work was to test known bacterial plant growth-promoting strains for their ability to promote cucumber plant growth in salinated soil and to improve cucumber fruit yield by protecting these plants against soil-borne pathogens. Fifty-two plant-beneficial bacterial strains were evaluated for their ability to protect plants against cucumber foot and root rot after bacterization of the seeds and infestation of salinated soil with the isolated Fusarium solani pathogen. Based on the results of initial screenings, five efficient strains were selected, namely Serratia plymuthica RR-2-5-10, Stenotrophomonas rhizophila e-p10, Pseudomonas fluorescens SPB2145, Pseudomonas extremorientalis TSAU20, and P. fluorescens PCL1751. All five strains are salt tolerant since they grow well in a medium to which 3% NaCl was added. Infestation of the soil with F. solani resulted in an increase of the percentage of diseased plants from 17 to 54. Priming of seedlings with the five selected bacterial strains reduced this proportion to as low as 10%. In addition, in the absence of an added pathogen, all five strains showed a significant stimulatory effect on cucumber plant growth, increasing the dry weight of whole cucumber plants up to 62% in comparison to the non-bacterized control. The strains also increased cucumber fruit yield in greenhouse varying from 9% to 32%. We conclude that seed priming with the selected microbes is a very promising approach for improving horticulture in salinated soils. Moreover, allochthonous strains isolated from non-salinated soil, from a moderate or even cold climate, and from other plants than cucumber, functioned as well as autochthonous strains as cucumber-beneficial bacteria in salinated Uzbek soils. These results show that these plant-beneficial strains are robust and they strongly suggest they can also be used successfully in case the climate gets warmer and the soils will become more salinated. Finally, the mechanisms by which they may exert their plant-beneficial action are discussed. © 2010 Springer-Verlag. Source

Malfanova N.,Leiden University | Malfanova N.,All Russian Research Institute for Agricultural Microbiology ARRIAM | Kamilova F.,Leiden University | Validov S.,Leiden University | And 4 more authors.
Microbial Biotechnology

Thirty endophytic bacteria were isolated from various plant species growing near Saint-Petersburg, Russia. Based on a screening for various traits, including plant-beneficial properties and DNA fragment patterns, potential siblings were removed. The remaining isolates were taxonomically identified using 16S rDNA sequences and potential human and plant pathogens were removed. The remaining strains were tested for their ability to promote radish root growth and to protect tomato plants against tomato foot and root rot. One strain, Bacillus subtilis HC8, isolated from the giant hogweed Heracleum sosnowskyi Manden, significantly promoted plant growth and protected tomato against tomato foot and root rot. Metabolites possibly responsible for these plant-beneficial properties were identified as the hormone gibberellin and (lipo)peptide antibiotics respectively. The antibiotic properties of strain HC8 are similar to those of the commercially available plant-beneficial strain Bacillus amyloliquefaciens FZB42. However, thin layer chromatography profiles of the two strains differ. It is speculated that endophytes such as B. subtilis HC8 contribute to the fast growth of giant hogweed. © 2011 Leiden University, Institute of Biology. Journal compilation © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd. Source

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