Marek Siwulski

Species diversity of Trichoderma in Poland

In the present study, we reinvestigate the diversity of Trichoderma in Poland utilizing a combination of morphological and molecular/phylogenetic methods. A total of 170 isolates were collected from six different substrata at 49 sites in Poland. These were divided among 14 taxa as follows: 110 of 170 Trichoderma isolates were identified to the species level by the analysis of their ITS1, ITS2 rDNA sequences as: T. harzianum (43 isolates), T. aggressivum (35), T. citrinoviride (11), T. hamatum (9), T. virens (6), T. longibrachiatum (4), T. polysporum (1), and T. tomentosum (1); 60 isolates belonging to the Viride clade were identified based on a fragment of the translation-elongation factor 1-alpha (tef1) gene as: T. atroviride (20 isolates), T. gamsii (2), T. koningii (17), T. viridescens (13), T. viride (7), and T. koningiopsis (1). Identifications were made using the BLAST interface in TrichOKEY and TrichoBLAST (http://www.isth.info). The most diverse substrata were soil (nine species per 22 isolates) and decaying wood (nine species per 75 isolates). The most abundant species (25%) isolated from all substrata was T. harzianum.

Selected arsenic species: As(III), As(V) and dimethylarsenic acid (DMAA) in Xerocomus badius fruiting bodies

The aim of the study was to determine the content of As(III), As(V) and DMAA (dimethylarsinic acid) in Xerocomus badius fruiting bodies collected from selected Polish forests from areas subjected to very low or high anthropopressure and some commercially available samples obtained from the Polish Sanitary Inspectorate. The arsenic species determination was provided by two independent HPLC-HG-AAS hyphenated systems. The results show high levels (up to 27.1, 40.5 and 88.3 mg kg(-1) for As(III), As(V) and DMAA, respectively) of arsenic and occurrence of different species in mushrooms collected from areas subjected to high anthropopressure and two commercially available samples. For mushroom samples collected from areas not subjected to high anthropopressure and two commercially available samples the arsenic species level was below 0.5 mg kg(-1) for each arsenic form. Therefore, the accumulation of arsenic by mushrooms may lead to high (toxic for humans) arsenic concentrations, and arsenic species levels should be monitored in mushroom foodstuffs.

Content of selected elements in Boletus badius fruiting bodies growing in extremely polluted wastes

The aim of the study was to analyse levels of 17 trace elements and 5 major minerals in 11 Boletus badius fruiting bodies able to grow in extremely polluted waste (flotation tailings) and polluted soil in southern Poland. The presented data widen the limited literature data about the abilities of wild-growing mushroom species to grow on heavily contaminated substrates. Content of elements in waste, soil and mushrooms was analysed by flame atomic absorption spectrometry (FAAS) and cold vapour atomic absorption spectrometry (CVAAS - Hg). The industrial areas differed greatly as regards the content of elements in flotation tailings and soil; therefore differences in Ag, Ba, Cd, Co, Fe, Mo, Ni, Pb, Ca, K, Mg, Na and P accumulation in mushrooms were observed. The highest contents of elements in mushrooms were observed for: As, Al, Cu and Zn (86 ± 28, 549 ± 116, 341 ± 59 and 506 ± 40 mg kg−1 dry matter, respectively). Calculated bioconcentration factor (BCF) values were higher than 1 for Al (15.1–16.9), Fe (10.6–24.4) and Hg (10.2–16.4) only. The main value of the presented results is the fact that one of the common wild-growing mushroom species was able to grow on flotation tailings containing over 22 g kg−1 of As and, additionally, effective accumulation of other elements was observed. In view of the high content of the majority of analysed elements in fruiting bodies, edible mushrooms from such polluted areas are nonconsumable.

Accumulation of elements by edible mushroom species: Part I. Problem of trace element toxicity in mushrooms

The aim of this study was to evaluate Cd, Co, Cu, Hg, Ni, Pb, Sr and Zn accumulation in six edible mushroom species and to assess their risk and benefits to human consumers. Mushrooms (Leccinium aurantiacum, Xerocomus badius, Lactarius deliciosus, Boletus edulis, Cantharellus cibarius and Suillus luteus) were collected from selected regions of Poland during 1990–2010. The highest diversity between studied mushroom species was observed in terms of Cu and Zn accumulation. Significant differences in the accumulation efficiency were found among the six mushroom species examined. The most efficient were Boletus edulis (Cd and Hg), Suillus luteus (Cu and Sr), and Lactarius deliciosus (Pb and Zn). In the case of Co and Ni, the most effective were Xerocomus badius and Leccinium aurantiacum, respectively. The calculated bioconcentration factor (BCF) values of Cd, Cu, Hg, Sr and Zn were > 1 for all species in this study while Co, Ni and Pb usually were bioexcluded (BCF < 1). Additionally, based on the calculated daily intake rates of trace elements determined it can be concluded that occasional consumption of fruiting bodies of L. aurantiacum, X. badius, L. deliciosus, B. edulis, C. cibarius and S. luteus collected in Poland is safe and this finding largely agrees with results from recent studies by other authors.

Potential of Cultivated Ganoderma lucidum Mushrooms for the Production of Supplements Enriched with Essential Elements

Ganoderma lucidum is an important medicinal mushroom species and there is continuous interest in its bioactive properties. This study evaluated whether it may additionally serve as a nutritional supplement for the trace elements: selenium (Se), copper (Cu), and zinc (Zn). Mushrooms were cultivated on substrates enriched with 0.1 to 0.8 mM of inorganic Se alone or in combination with Zn and/or Cu. Supplementation increased accumulation of the elements in fruiting bodies regardless of the applied cultivation model. G. lucidum demonstrated the ability to accumulate significant amounts of organic Se, maximally amounting to (i) over 44 mg/kg when the substrate was supplemented only with Se, (ii) over 20 mg/kg in the Se+Cu model, (iii) over 25 mg/kg in the Se+Zn model, and (iv) 15 mg/kg in the Se+Cu+Zn model. The accumulation of Cu and Zn steadily increased with their initial substrate concentrations. Maximum concentrations found after supplementation with 0.8 mM amounted to over 55 mg/kg (Se+Zn) and 52 mg/kg (Se+Cu+Zn) of Zn, and 29 mg/kg (Se+Cu) and over 31 mg/kg (Se+Cu+Zn) of Cu. The greater the supplemented concentration and number of supplemented elements, the lower the biomass of G. lucidum fruiting bodies. Nevertheless, it still remained high when the substrate was supplemented up to 0.4 mM with each element. These results highlight that G. lucidum can easily incorporate elements from the substrate and that, when biofortified, its dried fruiting bodies may serve as a nutritional source of these essential elements.

Bioaccumulation of elements in three selected mushroom species from southwest Poland

Abstract The contents of 16 minerals and trace elements (Ag, As, Ca, Cd, Co, Cr, Cu, Fe, Hg, Mg, Mn, Ni, Pb, Pt, Ti and Zn) were analyzed in edible mushrooms (Leccinum scabrum, Boletus edulis and Boletus badius) collected in southwest Poland. Content of Co, Ni and Pb was similar in all tested mushroom species, while content of Ag, Ca, Cd, Hg and Ti was significantly higher in B. edulis than in L. scabrum and B. badius. The largest differences between these species were observed for Fe and Zn accumulation. The highest contents of these elements were noted in B. badius bodies (202 ± 88 and 137 ± 24 mg kg−1 dry matter, respectively), lower in B. edulis (131 ± 99 and 89 ± 26 mg kg−1 dry matter, respectively) and lowest in L. scabrum. Differences in As, Cu and Cr content between tested species were observed mainly between L. scabrum and B. badius fruiting bodies. Content of Pt was below 0.01 mg kg−1 dry matter). In the case of Mg and Mn accumulation, differences between B. edulis and B. badius were not observed (478 and 440 mg kg−1 dry matter for Mg and 23 and 19 mg kg−1 dry matter for Mn), and the results showed significantly higher content of these elements than in L. scabrum bodies (312 and 10 mg kg−1 dry matter, respectively). It is worth underlining that clear accumulation shown by the bioconcentration factor (BCF>1) observed for all three mushroom species was noted in the case of elements Ag, Cd, Co, Cu, Hg, Ni and Zn only.

Efficacy of supplementation of selected medicinal mushrooms with inorganic selenium salts

The aim of the study was to evaluate the possibility of supplementation with inorganic forms of selenium (Na2SeO4 and Na2SeO3) in concentrations of 0, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0 and 1.5 mM of three medicinal mushroom species: Agrocybe aegerita, Hericium erinaceus and Ganoderma lucidum. Tested mushroom species grew in Se additions of 0–0.6 mM (A. aegerita and H. erinaceus), while growth of G. lucidum bodies was observed for 0–0.8 mM. For the latter mushroom species, the total Se content was the highest. Content of Seorg was diverse; for control bodies it was the highest for G. lucidum (only organic forms were present), lower for A. aegerita (84% organic forms) and the lowest for H. erinaceus (56% organic forms). Accumulation of Se(IV) was generally significantly higher than Se(VI) for all tested mushroom species. There was no significant decrease of A. aegerita or G. lucidum biomass with the exception of G. lucidum bodies growing under 0.8 mM of Se species addition (15.51 ± 6.53 g). Biomass of H. erinaceus bodies was the highest under 0.2 (197.04 ± 8.73 g), control (191.80 ± 6.06 g) and 0.1 mM (185.04 ± 8.73 g) of both inorganic salts. The addition to the medium of Se salts brought about macroscopic changes in the fruiting bodies of the examined mushrooms. Concentrations exceeding 0.4 mM caused diminution of carpophores or even their total absence. In addition, colour changes of fruiting bodies were also recorded. At Se concentrations of 0.4 and 0.6 mM, A. aegerita fruiting bodies were distinctly lighter and those of H. erinaceus changed colour from purely white to white-pink.

Laccase production and metabolic diversity among Flammulina velutipes strains

Twelve Flammulina velutipes strains originating from Poland were identified using internal transcribed spacer (ITS) region sequencing. Based on the sequences obtained, the genomic relationship of the analyzed strains was determined. All F. velutipes strains were also characterized using Biolog FF MicroPlates to obtain data on C-substrate utilization and mitochondrial activity. The ability to decompose various substrates differed among the F. velutipes strains up to five times. The highest catabolic activities were characteristic for only two strains with capabilities to decompose up to 22 carbon sources. The correlation between carbon repression and laccase production by F. velutipes was analyzed based on glucose assimilation by these strains. Moreover, the influence of metal ions (Cu2+, Cd2+), veratric and ferulic acids, and temperature on laccase activities in the analyzed strains was determined. The results obtained proved that all the inducers influenced laccase expression in almost all the analyzed strains. However, the degree of induction depended not only on the strain used but also on the day of the induction.

Cultivation of Agaricus bisporus enriched with selenium, zinc and copper

BACKGROUND Agaricus bisporus (white button mushroom) is an important culinary and medicinal species of worldwide importance. The present study investigated for the first time whether it may be grown on substrates supplemented with Se alone or in combination with Cu and/or Zn (0.1-0.8 mmol L-1 ) to produce fruiting bodies of increased nutritional value. RESULTS As found, substrate supplementation did not affect yielded biomass up to 0.6 mmol L-1 element concentrations regardless of the cultivation model. At 0.8 mmol L-1 Se + Cu and Se + Zn supplementation biomass comparable with controls still developed. The accumulation of trace elements in the fruiting bodies generally increased over the concentration gradient reaching its maximum at 0.6 mmol L-1 (for Se + Zn and Se + Cu + Zn) and 0.8 mmol L-1 (for Se and Se + Cu). The organic Se constituted the greatest share in total Se quota. As calculated, each 10 g of dried fruiting bodies of A. bisporus obtained from 0.6 or 0.8 mmol L-1 supplementation would represent 342-469% of the Recommended Daily Allowance (RDA) for Se, 43.4-48.5% for Cu and 5.2-5.8% for Zn. CONCLUSION Considering inexpensive methods of A. bisporus cultivation, global popularity and use of this mushroom, its biofortification with Se, Cu and Zn could have a practical application in deficiency prevention and assisted treatment.

The relationship between metal composition, phenolic acid and flavonoid content in Imleria badia from non-polluted and polluted areas

ABSTRACT The aim of this study was to determine the elemental composition, phenolic content and composition and antioxidant properties of Imleria badia (Fr.) Vizzini (former names Boletus badius (Fr.) Fr., and Xerocomus badius (Fr.) E.-J. Gilbert) fruiting bodies collected from sites with different levels of pollution. Imleria badia was relatively tolerant to soil contamination with toxic elements and was able to grow in As, Cd, Hg and Pb concentrations exceeding 15, 2.9, 0.4 and 77 mg kg−1, respectively. The concentration of elements in soil was reflected in the element content in I. badia. The fruiting bodies from polluted sites exhibited significantly higher content of all the analyzed elements. Among 21 individual phenolic compounds only protocatechiuc and caffeic acids, and quercetin were determined in fruiting bodies of I. badia. The differences between the concentration of the quantified phenolic compounds and the total flavonoid content in fruiting bodies of I. badia from unpolluted and polluted sites were not significant. However, the greatest total phenolic content was found in fruiting bodies from the polluted areas. The antioxidative capacity of mushrooms collected from heavily polluted sites was lower than those growing in unpolluted areas. The concentrations of some metals in soil and fruiting soil were positively correlated with phenolic content and IC50.