Tomasz Płociniczak

Metabolic potential and community structure of endophytic and rhizosphere bacteria associated with the roots of the halophyte Aster tripolium L.

The submitted work assumes that the abundance and diversity of endophytic and rhizosphere microorganisms co-existing with the halophytic plant Aster tripolium L. growing in a salty meadow in the vicinity of a soda factory (central Poland) represent unique populations of cultivable bacterial strains. Endophytic and rhizosphere bacteria were (i) isolated and identified based on 16S rDNA sequences; (ii) screened for nifH and acdS genes; and (iii) analyzed based on selected metabolic properties. Moreover, total microbial biomass and community structures of the roots (endophytes), rhizosphere and soil were evaluated using a cultivation-independent technique (PLFA) to characterize plant-microbial interactions under natural salt conditions. The identification of the isolated strains showed domination by Gram-positive bacteria (mostly Bacillus spp.) both in the rhizosphere (90.9%) and roots (72.7%) of A. tripolium. Rhizosphere bacterial strains exhibited broader metabolic capacities, while endophytes exhibited higher specificities for metabolic activity. The PLFA analysis showed that the total bacterial biomass decreased in the following order (rhizosphere<soil<endophytes) and confirmed the dominance of Gram-positive endophytic bacteria in the roots of the halophyte. The described strain collection provides a valuable basis for a subsequent applications of bacteria in improvement of site adaptation of plants in saline soils.

Isolation of hydrocarbon-degrading and biosurfactant-producing bacteria and assessment their plant growth-promoting traits

Forty-two hydrocarbon-degrading bacterial strains were isolated from the soil heavily contaminated with petroleum hydrocarbons. Forty-one strains were identified based on their whole-cell fatty acid profiles using the MIDI-MIS method. Thirty-three of them belong to species Rhodococcus erythropolis, while the others to the genera Rahnella (4), Serratia (3) and Proteus (1). Isolates were screened for their ability to produce biosurfactants/bioemulsifiers. For all of them the activity of several mechanisms characteristic for plant growth-promoting bacteria was also determined. In order to investigate surface active and emulsifying abilities of isolates following methods: oil-spreading, blood agar, methylene blue agar and determination of emulsification index, were used. Among studied bacteria 12 strains (CD 112, CD 126, CD 131, CD 132, CD 135, CD 147, CD 154, CD 155, CD 158, CD 161, CD 166 and CD 167) have been chosen as promising candidates for the production of biosurfactants and/or bioemulsifiers. Among them 2 strains (R. erythropolis CD 126 and Rahnella aquatilis CD 132) had the highest potential to be used in the bioaugmentation of PH-contaminated soil. Moreover, 15 of tested strains (CD 105, CD 106, CD 108, CD 111, CD 116, CD 120, CD 124, CD 125, CD 130, CD 132, CD 134, CD 154, CD 156, CD 161 and CD 170) showed the activity of four mechanisms (ACC deaminase activity, IAA and siderophore production, phosphate solubilization) considered to be characteristic for plant growth-promoting bacteria. Two of them (R. erythropolis CD 106 and R. erythropolis CD 111) showed the highest activity of above-mentioned mechanisms and thus are considered as promising agents in microbe assisted phytoremediation.

Endophytic and rhizosphere bacteria associated with the roots of the halophyte Salicornia europaea L. - community structure and metabolic potential.

The main objective of our study was to assess density and diversity of rhizosphere (R) and endophytic (E) microorganisms associated with the halophyte S. europaea. Microorganisms were isolated from two saline sites (S1: 55dSm(-1), anthropogenic origin; S2: 112dSm(-1), natural salinity) located in central Poland and compared with microbial populations in the soil (S) using culture-independent (phospholipid fatty acids analysis, PLFA) and culture-dependent techniques. The endophytic and rhizosphere bacteria were identified and screened for nifH and acdS genes, and their metabolic properties were assessed. Strains with the potential to promote plant growth were selected for further study. PLFA analysis revealed that Gram-negative bacteria were dominant at both saline test sites; the total microbial biomass depended on the site (S1E>S). In contrast, culture-dependent techniques revealed that Gram-positive bacteria (Actinobacteria and Firmicutes) were dominant (S1: E-77.3%, R-86.3% and S2: E-59.1%, R-87.5%). Proteobacteria were observed in the rhizosphere at the lowest frequency (S1: 13.7% and S2: 12.5%). Greater salinity decreased the range and specificity of metabolic activity among the endophytes. These conditions also resulted in a broader spectrum of metabolic abilities in rhizobacteria; however, these metabolic processes were present at lower levels.

Rhizospheric Bacterial Strain Brevibacterium casei MH8a Colonizes Plant Tissues and Enhances Cd, Zn, Cu Phytoextraction by White Mustard

Environmental pollution by heavy metals has become a serious problem in the world. Phytoextraction, which is one of the plant-based technologies, has attracted the most attention for the bioremediation of soils polluted with these contaminants. The aim of this study was to determine whether the multiple-tolerant bacterium, Brevibacterium casei MH8a isolated from the heavy metal-contaminated rhizosphere soil of Sinapis alba L., is able to promote plant growth and enhance Cd, Zn, and Cu uptake by white mustard under laboratory conditions. Additionally, the ability of the rifampicin-resistant spontaneous mutant of MH8a to colonize plant tissues and its mechanisms of plant growth promotion were also examined. In order to assess the ecological consequences of bioaugmentation on autochthonous bacteria, the phospholipid fatty acid (PLFA) analysis was used. The MH8a strain exhibited the ability to produce ammonia, 1-amino-cyclopropane-1-carboxylic acid deaminase, indole 3-acetic acid and HCN but was not able to solubilize inorganic phosphate and produce siderophores. Introduction of MH8a into soil significantly increased S. alba biomass and the accumulation of Cd (208%), Zn (86%), and Cu (39%) in plant shoots in comparison with those grown in non-inoculated soil. Introduced into the soil, MH8a was able to enter the plant and was found in the roots and leaves of inoculated plants thus indicating its endophytic features. PLFA analysis revealed that the MH8a that was introduced into soil had a temporary influence on the structure of the autochthonous bacterial communities. The plant growth-promoting features of the MH8a strain and its ability to enhance the metal uptake by white mustard and its long-term survival in soil as well as its temporary impact on autochthonous microorganisms make the strain a suitable candidate for the promotion of plant growth and the efficiency of phytoextraction.

Mutation in HvCBP20 (Cap Binding Protein 20) Adapts Barley to Drought Stress at Phenotypic and Transcriptomic Levels

CBP20 (Cap-Binding Protein 20) encodes a small subunit of the cap-binding complex (CBC), which is involved in the conserved cell processes related to RNA metabolism in plants and, simultaneously, engaged in the signaling network of drought response, which is dependent on ABA. Here, we report the enhanced tolerance to drought stress of barley mutant in the HvCBP20 gene manifested at the morphological, physiological, and transcriptomic levels. Physiological analyses revealed differences between the hvcbp20.ab mutant and its WT in response to a water deficiency. The mutant exhibited a higher relative water content (RWC), a lower stomatal conductance and changed epidermal pattern compared to the WT after drought stress. Transcriptome analysis using the Agilent Barley Microarray integrated with observed phenotypic traits allowed to conclude that the hvcbp20.ab mutant exhibited better fitness to stress conditions by its much more efficient and earlier activation of stress-preventing mechanisms. The network hubs involved in the adjustment of hvcbp20.ab mutant to the drought conditions were proposed. These results enabled to make a significant progress in understanding the role of CBP20 in the drought stress response.

Improvement of phytoremediation of an aged petroleum hydrocarbon-contaminated soil by Rhodococcus erythropolis CD 106 strain.

ABSTRACT The aim of this study was to assess the impact of soil inoculation with the Rhodococcus erythropolis CD 106 strain on the effectiveness of the phytoremediation of an aged hydrocarbon-contaminated [approx. 1% total petroleum hydrocarbon (TPH)] soil using ryegrass (Lolium perenne). The introduction of CD 106 into the soil significantly increased the biomass of ryegrass and the removal of hydrocarbons in planted soil. The fresh weight of the shoots and roots of plants inoculated with CD 106 increased by 49% and 30%, respectively. After 210 days of the experiment, the concentration of TPH was reduced by 31.2%, whereas in the planted, non-inoculated soil, it was reduced by 16.8%. By contrast, the concentration of petroleum hydrocarbon decreased by 18.7% in non-planted soil bioaugmented with the CD 106 strain. The rifampicin-resistant CD 106 strain survived after inoculation into soil and was detected in the soil during the entire experimental period, but the number of CD 106 cells decreased constantly during the enhanced phytoremediation and bioaugmentation experiments. The plant growth-promoting and hydrocarbon-degrading properties of CD 106, which are connected with its long-term survival and limited impact on autochthonous microflora, make this strain a good candidate for improving the phytoremediation efficiency of soil contaminated with hydrocarbons.