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1

Selekcja bakterii ryzosferowych usprawniających procesy fitoremediacji gleb zawierających związki metali ciężkich

100%

Dąbrowska G. , Hrynkiewicz K. , Kłosowska K. , Goc A.

Ochrona Środowiska

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2011

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tom Vol. 33, nr 2

53-58

PL

Wiele spośród bakterii ryzosferowych charakteryzuje się wysoką opornością na jony metali ciężkich i odgrywa ważną rolę w promowaniu wzrostu roślin w ich obecności. W badaniach przeanalizowano szczepy bakterii Bacillus sp. (ML 1-2), Bacteroidetes bacterium (RI 116-1), Flavobacterium sp. (RI 111-11), Pseudomonas fluorescens (LI C1), Serratia entomophila (RI 111-21) i Variovorax sp. (ML 3-12), wyizolowane z gleb skażonych związkami metali ciężkich lub zdegradowanych antropogenicznie. W warunkach in vitro wyselekcjonowano bakterie oporne na jony wybranych metali ciężkich (Cd, Cu, Zn i Pb). Wykazano, że S. entomophila (RI 111-21) i P. fluorescens (LI C1) należą do bakterii o najwyższym stopniu oporności na obecność jonów metali ciężkich w podłożu, zwłaszcza cynku i ołowiu. Szczepami o najwyższej wrażliwości na przeanalizowane jony metali cięż-kich okazały się B. bacterium (RI 116-1) i Flavobacterium sp. (RI 111-11). Stwierdzono, że na wzrost i rozwój siewek rzepaku negatywnie wpływały jony metali ciężkich uszeregowane w kolejności: Pb>Zn>Cu>Cd. Inokulacja rzepaku ozimego (Brassica napus L. odmiana Kronos) wykazała, że niektóre bakterie promowały wzrost i rozwój siewek w obecności jonów metali ciężkich, zwłaszcza miedzi i kadmu. Szczepy Bacillus sp. (ML 1-2) i B. bacterium (RI 116-1) najsilniej promowały wzrost siewek rzepaku w środowisku zawierającym jony metali ciężkich, stąd też wydają się mieć największy potencjał w procesach fitoremediacji gleb.

EN

Many soil bacteria show a high tolerance to heavy metals and play a vital role in promoting plant growth in a heavy-metal-contaminated environment. In the study reported on in this paper the bacterial strains being analyzed included Bacillus sp. (ML 1-2), Bacteroidetes bacterium (RI 116-1), Flavobacterium sp. (RI 111-11), Pseudomonas fluorescens (LI C1), Serratia entomophila (RI 111-21), and Variovorax sp. (ML 3-12), which had been isolated from soils contaminated by heavy metal compounds, or degraded due to anthropogenic activity. Under in-vitro conditions selected were bacteria tolerant to Cd, Cu, Zn and Pb. It has been demonstrated that among the strains examined S. entomophila (RI 111-21) and P. fluorescens (LI C1) are endowed with the highest tolerance to the heavy metals in the substrate, specifically to Zn and Pb, whereas B.?bacterium (RI 116-1) and Flavobacterium sp. (RI 111-11) display the highest vulnerability to heavy metals. Taking into account their negative influence on the growth and development of rape seedlings, the four heavy metals have been listed in the following order: Pb>Zn>Cu>Cd. Inoculation of winter rape (Brassica napus L. var. Kronos) has shown that some of the bacterial strains promote the growth and development of seedlings in the presence of heavy metals, especially copper and cadmium. The strains Bacillus sp. (ML 1-2) and B. bacterium (RI 116-1), which were parti-cularly active in promoting rape seedling growth in an environment containing heavy metals, seem to have the greatest potential for the phytoremediation of soils.

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Zastosowanie zmodyfikowanych bakterii glebowych do poprawy skuteczności fitoremediacji środowiska zanieczyszczonego jonami metali śladowych

100%

Dąbrowska G. , Hrynkiewicz K. , Janczak K. , Żurańska M.

Ochrona Środowiska

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2014

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tom Vol. 36, nr 1

21--26

EN

Eight bacterial strains from areas contaminated with trace metals were selected for the experiments: Bacillus sp., Bacteroidetes bacterium, Flavobacterium sp., Luteibactor rhizovicina, Pseudomonas fluorescens, Pseudomonas sp., Serratia entomophila and Variovorax sp. Bacteria were transformed with pET21a expression plasmid encoding Brassica napus L. metallothionein 2 (BnMT2). Transformant colonies were examined for growth on solid culture medium containing trace metal ions (Cd and Pb). It was found that most of the analyzed bacterial strains were resistant to ampicillin, though the antibiotic inhibited their growth. It was demonstrated that growth of bacterial strains on a medium containing Pb ions was not inhibited, while Cd ion content limited or inhibited growth of some bacterial strains. No significant differences were noted in growth between transformed and untransformed bacteria of a given strain on the medium containing Pb ions. The BnMT2 transgene presence abolished adverse effects of Cd ions on Flavobacterium sp., Variovorax sp. and L.rhizovicina. Modified bacterial strains were demonstrated for potential use in treatment of soil contaminated with trace metals, especially Cd ions.

PL

W przeprowadzonych badaniach wykorzystano osiem szczepów bakterii: Bacillus sp., Bacteroidetes bacterium, Flavobacterium sp., Luteibactor rhizovicina, Pseudomonas fluorescens, Pseudomonas sp., Serratia entomophila, Variovorax sp. wyizolowanych z terenów zanieczyszczonych jonami metali śladowych. Bakterie stransformowano plazmidem ekspresyjnym pET21a zawierającym fragment genu kodujący metalotioneinę typu 2 Brassica napus L. (BnMT2). Po uzyskaniu kolonii bakteryjnych (transformantów) sprawdzono ich wzrost na podłożu stałym zawierających jony metali śladowych (Cd i Pb). Stwierdzono, że większość analizowanych szczepów bakterii wykazuje oporność na ampicylinę, chociaż antybiotyk ogranicza ich wzrost. Wykazano, iż wzrost badanych szczepów na podłożu zawierającym jony ołowiu nie był hamowany, natomiast jony kadmu ograniczały lub hamowały wzrost niektórych szczepów bakterii. Nie stwierdzono znacznych różnic we wzroście pomiędzy bakteriami transformowanymi i nietransformowanymi danego szczepu na podłożu zawierającym jony ołowiu. Obecność transgenu BnMT2 niwelowała szkodliwe działanie jonów kadmu w przypadku Flavobacterium sp., Variovorax sp. i L. rhizovicina. Wykazano, że zmodyfikowane szczepy bakterii mogą znaleźć zastosowanie w procesie oczyszczania gleby zawierającej jony metali śladowych, a zwłaszcza kadmu.

3

Analysis of microbiologically stimulated biomass of Salix viminalis L. in the presence of CD2plus under in vitro conditions - implications for phytoremediation

100%

Zloch M. , Tyburski J. , Hrynkiewicz K.

Acta Biologica Cracoviensia. Series Botanica

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2015

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tom 57

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nr 2

4

Wpływ szczepienia bakteryjnego na wzrost miskanta i liczebność bakterii i grzybów w glebie zawierającej polimery, Cz. 2. Polimery niebiodegradowalne

100%

Janczak K. , Dąbrowska G. , Znajewska Z. , Hrynkiewicz K.

Przemysł Chemiczny

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2014

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tom T. 93, nr 12

2222-2225

PL

Porównano wpływ inokulacji bakteriami glebowymi miskanta olbrzymiego (Miscanthus x giganteus) na liczebność populacji grzybów i bakterii w glebie zawierającej niebiodegradowalne materiały polimerowe (polietylen, poli(tereftalan etylenu)). Uzyskane wyniki wskazują, że szczep Serratia sp. może być wykorzystywany w procesie biodegradacji tworzyw sztucznych z udziałem miskanta olbrzymiego, gdyż w największym stopniu stymulował wzrost roślin i wpływał na wzrost ogólnej liczebności autochtonicznych mikroorganizmów w glebie zawierającej niebiodegradowalne materiały polimerowe.

EN

Polyethylene and polyethylene terephthalate films (thickness 0.083 and 0.087 mm, resp.) were prepd. by extrusion, cuy and added to soil used then for growing Miscanthus x giganteus inoculated with bacteria. The strain Serratia sp. strongestly promoted the plant growth and an increase in the no. of bacteria anf fungi in the soil.

5

Metabolic potential of microorganisms associated with the halophyte Aster tripolium L. in saline soils

100%

Szymanska S. , Piernik A. , Hrynkiewicz K.

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tom 18

EN

Increased soil salinization may be caused by a natural (e.g. climate change) and anthropogenic (e.g. improper fertilization and irrigation of agricultural land) factors. The submitted work assumes that microorganisms associated with plant halophytes have a unique metabolic properties that can stimulate plant growth under salt stress. The aim of the study was to determine the abundance and metabolic biodiversity of endophytic and rhizosphere microorganisms co-existing with Aster tripolium L. and compare them with the properties of soil microorganisms not affected by plant roots at a salty meadows in the vicinity of a soda factory (central Poland). In order to select halotolerant and halophilic microorganisms culture medium was enriched with various concentrations of NaCl (0, 100, 200, 400, 600 mM NaCl). Total metabolic activity of endophytic, rhizosphere and soil populations was measured to compare the community-level physiological profiles. Results of our study revealed that bacterial and fungal density increased in the following order: endophytes < soil < rhizosphere. Only the highest concentration (600 mM) of NaCl decreased the number of microorganisms. The highest total microbial metabolic activity was observed for the rhizosphere, while the activity of endophytes was higher compare to soil populations. To carbon sources which significantly differentiated zones belonged: D-lactose, 4-hydroxybenzoic acid and L-asparagine. The results are preliminary studies leading to the development of inoculum based on selected microbial halotolerant and halophilic strains which can be used in agriculture and/or recultivation of saline soils.

6

Response of birch and alder root endophytes as well as rhizosphere and bulk soil microorganisms to heavy metal pollution

100%

Zloch M. , Kowalkowski T. , Szymanska S. , Hrynkiewicz K.

Polish Journal of Ecology

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2014

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tom 62

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nr 1

EN

Research on soil microorganisms associated with plant roots (endophytes) and their outside habitats (rhizosphere, soil) under exposure to high concentrations of heavy metals in the environment may contribute to an increased knowledge about the effects of unfavorable pollutants in the natural ecosystem and improve the efficiency of remediation processes of degraded soils. The main goal of our study was to evaluate properties of microbial populations inhabiting different but spatially closed habitats: interior of the tree roots – endophytes (Betula pendula L. and Alnus glutinosa L.), rhizosphere and soil not influenced by plant roots at two heavy metal polluted areas in southern Poland where the content of cadmium, lead and zinc ranged 9.2–14.9, 26.2–75.1 and 1080–1559 mg∙kg–1 of dried soil, respectively. In our experiment, we analyzed their density as well as determined the total metabolic activity and ability for utilization of different carbon sources. The density of endophytes (5.76–7.16 log10 c.f.u. × g dw–1) was higher than the density of bacteria inhabiting outside habitats – the rhizosphere and soil (5.43–5.91 and 5.08–5.57 log10 c.f.u. × g dw–1, respectively) and was not affected by heavy metal content. However, the total metabolic activity of endophytic populations was strongly influenced by the physicochemical properties of the soil and was significantly lower at more contaminated site. In general, the metabolic activity of endophytes was ~15 times lower than the activity of rhizobacteria and at the same level or lower than the activity of soil microorganisms. Discriminant analysis revealed a higher utilization of α-Cyclodextrin and D-Xylose by microorganisms isolated from more polluted test site. The use of an artificial neural network (ANN) followed by principal component analysis (PCA) allowed the distinguishing of preferences in the utilization of carbon sources by microorganisms due to tree species but not the zone of isolation – endophytes, rhizosphere, soil. Our study revealed no significant impact of the metal concentrations present in the soil on the density of microorganisms (bacteria and fungi) inhabiting three analyzed zones (interior of the roots, rhziosphere, soil); however, their total metabolic activity was decreased in more contaminated site. Since the density of endophytes was higher compared to the number of bacteria present in the outside habitats (rhizosphere and soil), and their total metabolic activity was in general lower, we suggest a high specialization of microorganisms associated with the cellular spaces of plant roots compared to other microbial populations.

7

Wrażliwość odmian jarych i ozimych rzepaku na metale ciężkie i bakterie glebowe

100%

Dabrowska G. , Hrynkiewicz K. , Mierek-Adamska A.

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tom 33

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nr 2

8

Do arbuscular mycorrhizal fungi affect metallothionein MT2 expression in Brassica napus L. roots?

100%

Dabrowska G. , Hrynkiewicz K. , Trejgell A.

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nr 1

EN

Arbuscular mycorrhizal fungi are the most widespread root fungal symbionts, forming associations with the vast majority of plant species. Ectomycorrhizal development alters gene expression in plant symbionts. In this work we examined the effect of arbuscular mycorrhizal fungi spores on the growth and development of Brassica and on the expression of BnMT2 in winter rape. In a pot experiment, rape seedlings growing on different types of sterile and nonsterile soils were inoculated simultaneously with mycorrhizal fungi spores of Acaulospora longula, Glomus geosporum, Glomus mosseae and Scutellospora calospora. As compared with control plants growing in the absence of spores, ten-week-old seedlings of Brassica napus L. in sterile soil inoculated with arbuscular spores had longer shoots and higher fresh biomass of above-ground plant parts. In other types of substrates enriched with mycorrhizal fungi spores, the plants were smaller than non-inoculated plants. The presence of AMF spores stimulated the elongation growth of hypocotyls in both analyzed substrates. BnMT2 expression was highest in plants growing on the sterile substrate. Generally, the presence of mycorrhizal fungi spores appeared to have an adverse effect on the growth of rape plants.

9

Interactive physiological response of potato (Solanum tuberosum L.) plants to fungal colonization and Potato virus Y (PVY) infection

75%

Thiem D. , Szmidt-Jaworska A. , Baum C. , Muders K. , Niedojadlo K. , Hrynkiewicz K.

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nr 2

EN

Potato plants can be colonized by various viruses and by symbiotic, saprophytic and pathogenic fungi. However, the significance of interactions of viral infection and fungal colonization is hardly known. This work presents a model experiment in which the influence of three different types of fungal associations on the growth and physiology of the potato variety Pirol was tested individually or in combination with infection by PVY. It was hypothesized that simultaneous viral and fungal infections increase the biotic stress of the host plant, but mutualistic plant-fungal associations can mask the impact of viral infection. In the present study, a symbiotic arbsucular mycorrhizal fungus, Glomus intraradices, significantly stimulated the growth of plants infected with PVY. In contrast, two saprophytic Trichoderma spp. strains either did not influence or even inhibited the growth of PVY-infected plants. Also, inoculation of PVY-infected potato plants with a pathogenic strain of Colletotrichum coccodes did not inhibit the plant growth. Growth of the PVY-free potato plants was not promoted by the symbiotic fungus, whereas T. viride, T. harzianum and C. coccodes had an evident inhibitory effect. The strongest growth inhibition and highest concentration of H2O2, as an indicator of biotic stress, was observed in PVY-free potato plants inoculated with T. harzianum and C. coccodes strains. Surprisingly, ultrastructural analysis of PVY-infected plant roots colonized by G. intraradices showed virus-like structures in the arbuscules. This pointed to the possibility of mycorrhizal-mediated transmission of virus particles and has to be further biotic stress; Potato virus Y (PVY); Glomus intraradices; Trichoderma viride; T. harzianum; Colletotrichum coccodesexamined by testing with immunoassays and real transmission to uninfected plants. In conclusion, although mycorrhiza formation might decrease the impact of PVY infection on plants, a possible role of mycorrhizal fungi as virus vectors is discussed.