CC BY
46
UDC 579.222: 547.562.4: 582.284
Veligodska A.K., Fedotov O.V. SCREENING OF CONTENT AND DYNAMIC OF ACCUMULATION OF POLYPHENOLS IN SOME BASIDIOMYCETES SPECIES
Donetsk National University, Vinnitsa, Ukraine e-mail: o.fedotov@donnu.edu.ua
The aim of the study was to investigate the total content of polyphenolic substances in Basidiomycetes carpophores from 50 species, of which 27 belong to the order Polyporales and 23 to the order Agaricales. Introduced 23 strains of 8 species of Basidiomycetes. Methods. Gathered wild carpophores dried and crushed to a particle size of 0,1 till 0,01 mm and searching strains were cultured in Erlenmeyyers flasks by surface method on standard glucose-peptone culture medium. Determination of total content of polyphenolic compounds was carried out in ethanol extracts of mycological material by a modified method of Folin-Chokalteu. Completely dry biomass of carpophores and mycelium was determined gravimetrically. Results. There was identified the species of polyporal fungi Ganoderma applanatum, Ganoderma lucidum, Laetiporus sulphureus and Fomes fomentarius and types of agarical mushrooms Stropharia rugosoannulata, Agrocybe cylindracea, Tricholoma flavovirens, Flammulina velutipes, Pleurotus ostreatus and Fistulina hepatica high in polyphenolic compounds. It was determined the content of polyphenols ranging from more than 60 mg / g completely dry biomass. For introduced strains established dynamics of growth and accumulation of polyphenolic compounds in the mycelium and culture filtrate during fermentation on glucose-peptone medium. All cultures reach a maximum accumulation of biomass on the 12th day of growth. Shizophyllum commune Sc-1101 and 10 and F. velutipes F-202 have been identified as the most productive strains. The lowest accumulation of absolutely dry biomass was recorded for strain P. ostreatus P-192 and strain F. fomentarius Ff-09. Cultures have investigated individual value growth such as biomass accumulation in the applied cultivation conditions, which probably reflects the suitability of the medium for their growth and genotypic characteristics. Strains are overwhelmingly able to accumulate polyphenolic compounds in both mycelium and culture fluid during the whole period of cultivation. Maximum content of polyphenols in the mycelium to 96%, and in the culture fluid - for 91% of strains coincided with the end of their term cultivation. Calculated correlation coefficient between the content of polyphenols in the mycelium and culture fluid showed that there is a very high positive correlation of 73.2%, a positive high at 17.4% and the average 4.5% of experiment data. Conclusion. The strains of species Shizophyllum commune, Pleurotus ostreatus, Fistulina hepatica and Laetiporus sulphureus were selected for further research in order to obtain polyphenols mycelial and extracellular origin.
Key words: polyphenols, Basidiomycetes, carpophores, mycelium, cultural filtrate
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А.К. Велигодська, О.В. Федотов СКРИН1НГ ВМ1СТУ ТА ДИНАМ1КА НАКОПИЧЕННЯ ПОЛ1ФЕНОЛЬНИХ РЕЧОВИН У ДЕЯКИХ ВИД1В БАЗИД1ОМ1ЦЕТ1В
Донецький нацюнальний утверситет, м. Втниця, Украта, e-mail: o.fedotov@donnu.edu.ua
Метою роботи було вивчення загального вм^ту полiфенольних речовин у карпофорах 50 видiв базидюмщепв з яких 27 належать до порядку Polyporales та 23 -порядку Agaricales. 1нтродуковано 23 штами 8 видiв базидiальних грибiв. Методи. Зiбранi дикороcлi карпофори висушували та подрiбнювали до розмiру часток 0,1+0,01 мм, а доcлiднi штами культивували поверхнево в колбах Ерленмейера на стандартному глюкозо-пептонному живильному cередовищi. Визначення загального вмicту полiфенольних речовин проводили у ширтових витяжках мiкологiчного матерiалу за модифiкованою методикою Фолiна-Чокальтеу. Абгалютно cуху бiомаcу карпофорiв та мщелш визначали ваговим методом. Результаты. Виявлеш види трутових грибiв -Ganoderma applanatum, Ganoderma lucidum, Laetiporus sulphureus та Fomes fomentarius i види агарикових грибiв - Stropharia rugosoannulata, Agrocybe cylindracea, Tricholoma flavovirens, Flammulina velutipes, Pleurotus ostreatus та Fistulina hepatica з вигаким вметом полiфенольних речовин - понад 60 мг/г абгалютно cухоi бюмаот. Для штродукованих штамiв, вcтановленi динамiка роcту та накопичення полiфенольних речовин в мщелп та культуральному фiльтратi при ферментаци на глюкозо-пептонному cередовищi. Вci культури доcягають макcимуму накопичення бiомаcи на 12-ту добу росту. Найпродуктивнiшими тут е штами Schizophyllum commune Sc-1101 i Sc-10 та штами F. velutipes F-202. Найнижчi значення накопичення абгалютно cухоi бiомаcи зафiкcовано для штаму P. ostreatus P-192 та штаму F. fomentarius Ff-09. Встановлено, що доcлiдженi культури мають iндивiдуальнi значення росту - накопичення бюмаот в заcтоcованих умовах культивування, що, ймовiрно, вiдображае придатнicть цих умов для Ъ. роcту та генотишчш оcобливоcтi. Штами в переважнiй бiльшоcтi здатнi до накопичення полiфенольних речовин як в мщелп, так i в культуральнш рЦиш протягом ваого термiну культивування. Макcимум вмicту полiфенолiв у мщели для 96 %, та у культуральнш рЦиш - для 91% вЦ загально'1 кiлькоcтi штамiв cпiвпадав iз закiнченням дванадцятидобового термiну \х культивування. Обчиcлення коефщенту кореляцп мiж вмicтом полiфенолiв у мщелп та культуральнш рЦиш одновшових культур показало, що cпоcтерiгаетьcя дуже вигака позитивна кореляцiя у 73,2%, витка позитивна - у 17,4% та cередня - у 4,5% дом^в. Висновок. Вiдiбрано штами видiв Schizophyllum commune, Pleurotus ostreatus, Fistulina hepatica та Laetiporus sulphureus - першективш для подальших доcлiджень з метою отримання полiфенолiв мiцелiального та позаклиинного походження.
Ключовi слова: полiфеноли, базидюмщети, карпофори, мщелш, культуральний ф1хьтрат
А.К. Велигодская, О.В. Федотов СКРИНИНГ СОДЕРЖАНИЯ И ДИНАМИКА НАКОПЛЕНИЯ ПОЛИФЕНОЛЬНЫХ ВЕЩЕСТВ НЕКОТОРЫХ ВИДОВ БАЗИДИОМИЦЕТОВ
Донецкий национальный университет, г. Винница, Украина e-mail: o.fedotov@donnu.edu.ua
Целью работы было изучение общего содержания полифенольных веществ в плодовых телах 50 видов базидиомицетов из которых 27 принадлежат к порядку Polyporales и 23 - порядка Agaricales. Интродуцировано 23 штаммы 8 видов базидиальных грибов. Методы. Собранные дикорастущие карпофоры высушивали и измельчали до размера частиц 0,1±0,01 мм, а исследуемые штаммы культивировали поверхностно в колбах Ерленмейера на стандартной глюкозо-пептонной питательной среде. Определение общего содержания полифенольных веществ проводили в спиртовых вытяжках микологического материала по модифицированной методике Фолина-Чокальтеу. Абсолютно сухую биомассу плодовых тел и мицелия определяли весовым методом. Результаты. Обнаруженные виды трутовых грибов - Ganoderma applanatum, Ganoderma lucidum, Laetiporus sulphureus и Fomes fomentarius и виды агариковых грибов -Stropharia rugosoannulata, Agrocybe cylindracea, Tricholoma flavovirens, Flammulina velutipes, Pleurotus ostreatus и Fistulina hepatica с высоким содержанием полифенольных веществ -более 60 мг / г абсолютно сухой биомассы. Для интродуцированных штаммов установлена динамика роста и накопления полифенольных веществ в мицелии и культуральном фильтрате при ферментации на глюкозо-пептонной среде. Все культуры достигают максимума накопления биомассы на 12-е сутки роста. Здесь самые продуктивные - штаммы Sohizophyttum commune Sc-1101 и Sc-10 и штаммы F. velutipes F-202. Самые низкие значения накопления абсолютно сухой биомассы зафиксированы для штамма P. ostreatus P-192 и штамма F. fomentarius Ff-09. Установлено, что исследованные культуры имеют индивидуальные значения роста - накопление биомассы в примененных условиях культивирования, что, вероятно, отражает пригодность этих условий для их роста и генотипические особенности. Штаммы в подавляющем большинстве способны к накоплению полифенольных веществ как в мицелии, так и в культуральной жидкости в течение всего срока культивирования. Максимум содержания полифенолов в мицелии для 96%, и в культуральной жидкости - для 91% от общего количества штаммов совпадал с окончанием дванадцятидобового срока их культивирования. Вычисления коэффициента корреляции между содержанием полифенолов в мицелии и культуральной жидкости одновозрастных культур показало, что наблюдается очень высокая положительная корреляция в 73,2%, высокая положительная - в 17,4% и средняя - в 4,5% опытов. Вывод. Отобраны штаммы видов Sсhizophyllum commune, Pleurotus ostreatus, Fistulina hepatica и Laetiporus sulphureus -перспективные для дальнейших исследований с целью получения полифенолов мицелиальными и внеклеточного происхождения.
Ключевые слова: полифенолы, базидиомицеты, карпофоры, мицелий, культуральный фильтрат
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In the last decades an actual problem is searching for new biologically active substances (BAS's) and their producers for the purpose of development and manufacturing application of modern drug and therapeutic products (Zaprometov, 1993; Nikitina, 2007).
In particular, polyphenol compounds, which are natural antioxidants preventing development of different pathogenic effects in a cell and as a result of different diseases, are the desired substances in different branches of industry and medicine (Zaprometov, 1993; Fedotov et al., 2012 Asatiani at al., 2010). These include phenolic acids and aldehyde derivatives, substances of polyphenoloxycarbon complex, carotenoids, flavonoids, melanins, tannins, etc. (Asatiani et al., 2010; Wasser, 2010).
It is established that these substances are synthesised by almost all plant and fungi organisms (Nikitina, 2007; Li Fu et al., 2011). The traditional sources of polyphenols are plant raw materials - Camellia sinensis and Humulus lupulus, as well as the fruits of Vitis vinifera (Li Fu et al., 2011, Halvorsen et al., 2011).
A number of studies are devoted to investigation of polyphenols in fungi, especially to investigation of total polyphenols in the fruit bodies of 49 species of edible fungi which belong to genera Boletus, Suillus, Volvariella, Pleurotus et al. (Guthalu Puttaraju Nethravathi et al., 2006; Guo Ya-Jun et al., 2012). However, these studies give a vague idea about the qualitative and quantitative content of polyphenols in higher basidial fungi and mycological material when cultivating them that makes it necessary to carry out further screening operations in this field.
The interest for basidiomycetes, including wood-destroying ones, is firstly associated with their ability to synthesise numerous BAS's. Especially while destructing lignin-cellulose complex, they produce antioxidant substances -oxidoreductases, vitamins, polyphenols, free radical blockers et al., which provide adaptive mechanisms for antioxidant protection of xylotrophs (Peyrat-Maillard et al., 2000; Fedotov, 2007; Wasser, 2010).
Secondly, mycelium cultures of these organisms are not demanding for a nutrient media composition, most of them are edible and non-poisonous, and can be used in the microbiological production of BAS's.
The aim of this study was to determine total polyphenols in the carpophores and mycelium and in the culture filtrate of some basidiomycetes species.
MATERIALS AND METHODS
The carpophores, mycelium and culture filtrate of 50 macromycetes species, 27 of which belong to order Polyporales and 23 - to order Agaricales, division Basidiomycetes were used as the materials for this investigation. The general information about the species of basidial fungi investigated is set forth in the published work (Fedotov et al., 2012) and relevant sections of the article.
Also 23 strains from a pileated fungi culture collection of the Physiology of
Plants Department, Donetsk National University: Fomes fomentarius (L. ex Fr.) Gill. -T-10, Ff-09, Ff-1201; Laetiporus sulphureus (Bull.) Murrill. - Ls-08, Ls-09; Fistulina hepatica Schff. ex Fr. - Fh-08, Fh-18; Flammulina velutipes (Curt.: Fr.) Sing. - F-03, F-06, F-1, F-202; Pleurotus ostreatus (Jacq.: Fr.) P. Kumm. - Hk-35, P-004, P-01, P-039, P-107, P-192, P-208; Schizophyllum commune Fr.:Fr. - Sc-10, Sc-1101, Sc-1102; Trametes hirsuta (Wulf.:Fr.) Pil. - Th-11 and Trichaptum biforme (Fr.) Ryv. - Tb-11.
Most of the introduced strains are isolated into a pure culture from the wild fruit bodies (FBs) of basidiomycetes collected in different areas of Donetsk oblast, the taxonomic position of which was established according to the modern literature (Kirk et al., 2001).
To determine total polyphenols, the collected FBs were dried and ground to the particle size of 0.1±0.01 mm, and the strains under investigation were cultivated superficially in Erlenmeyer flasks of 250 ml in a glucose-peptone growth medium (GPM, рН0 6.5±0.2) of 50 ml with the following composition (g/l): glucose - 10.0; peptone - 3.0; КН2РО4 - 0.6; К2НРО4 - 0.4; MgSO4 ■ 7H2O - 0.5; CaCl2 - 0.05; ZnSO4 ■ 7H2O - 0.001. 10-day mycelia strain cultures on wort agar were used as inoculums. The incubation temperature was 27.5°С.
The cultivation period was 6, 9 and 12 days. Upon completion of the cultivation period, a mycelium was separated from the culture broth by filtration at 5±1°С. The mycelium obtained was additionally dried a little on filtration paper and cooled down to 1±0.5°С
The prepared mycelium was homogenized by grinding in a chilled mortar. The ground carpophores (GCs), homogenized mycelium (HM) and culture filtrate (CF) were used for further tests. Absolutely dry biomass (ADB) of the GCs and mycelium was determined by gravimetric method (State..., 1987).
The determination of total (W) polyphenols (PPs) was carried out in the alcoholic extracts of the mycological material using modified Folin-Ciocalteu method (Мусиенко et al., 2001) and calculated by the standard formula. The tests were performed in three replications.
Statistical manipulation was done using the programs for statistical manipulation of biological test results. A correlation analysis was performed to determine correlation level between PP concentration in the mycelium and CF of one-year cultures. A difference was considered as significant at confidence level Р>0.95 (Prysedskiy, 1999).
RESULTS AND DISCUSSION
Total polyphenols in 225 carpophores of 27 species of Polyporales fungi and in 220 ones of 23 species of Agaricales fungi were evaluated at the first step of investigation (Table 1).
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Table 1. Total content of polyphenols in fruit bodies of some species of Basidiomycetes
Number of the
Species samples Polyphenols, mg/g
Order Polyporales
Auricularia auricula-judae * 12 32,53 ± 3,52
Laeticorticium roseum * 3 20,02 ± 0,58
Chaetoporus ambiquus * 6 20,66 ± 0,95
Sparassis crispa * 9 10,54 ± 0,19
Fibuloporia mollusca * 6 10,54 ± 0,35
Tyromyces lacteus * 9 16,07 ± 0,76
Tyromyces revolutus * 3 12,09 ± 0,16
Tyromyces undosus * 6 10,33 ± 0,13
Irpex lacteus * 9 26,75 ± 0,43
Amyloporia lenis * 3 15,05 ± 0,21
Hydnum ochraceum * 3 10,02 ± 0,24
Trametes squalens * 6 15,07 ± 0,28
Trametes campestris * 6 20,14 ± 0,41
Trametes versicolor * 15 14,13 ± 0,71
Trametes zonatus * 9 15,06 ± 0,52
Fomes fomentarius * 12 248,29± 5,84
Heterobasidion annosum * 12 13,33 ± 0,64
Fomitopsis pinicola * 6 39,19 ± 0,58
Daedalea quercina * 6 9,02 ± 0,13
Piptoporus betulinus * 12 15,10 ± 0,10
Polyporus squamosus * 9 23,20 ± 0,37
Laetiporus sulphureus * 9 117,04 ± 0,56
Ganoderma applanatum * 9 161,08 ± 0,19
Ganoderma lucidum * 15 89,06 ± 1,5
Inonotus obliquus * 12 20,55 ± 0,31
Phellinus igniarius * 9 34,53± 0,55
Phellinus pomaceus * 9 19,04 ± 0,59
Order Agaricales
Agaricus arvensis * 5 24,57 ± 4,07
Agaricus bisporus ** 9 35,44± 0,63
Agaricus campestris * 5 23,46 ± 0,10
Agrocybe cylindracea ** 9 75,85 ± 1,22
Coprinus comatus * 15 25,04 ± 0,58
Coprinus micaceus * 15 25,03± 0,15
Fistulina hepatica * 9 172,25 ± 0,20
Number of the
Species samples Polyphenols, mg/g
Flammulina velutipes * 27 81,25 ± 7,75
Flammulina velutipes ** 3 65,06 ± 0,92
Lentinus edodes ** 9 35,47 ± 0,42
Marasmius oreades * 3 37,08 ± 0,65
Pleurotus citrinopileatus ** 3 37,55 ± 0,11
Pleurotus eryngii ** 6 15,03 ± 0,42
Pleurotus ostreatus * 34 100,56 ± 3,15
Pleurotus ostreatus var. Florida ** 3 53,07 ± 2,01
Kuehneromyces mutabilis * 9 32,28 ± 0,83
Pholiota aurivella * 3 18,02 ± 0,35
Pholiota squarrosa * 3 12,04 ± 0,65
Schizophyllum commune * 21 19,29 ± 0,27
Stropharia aeruginosa * 3 32,53 ± 0,54
Stropharia rugosoannulata ** 6 59,56 ± 1,85
Lyophyllum loricatum * 5 21,37 ± 0,63
Lyophyllum connatum * 5 20,42 ± 0,12
Tricholoma flavovirens * 5 79,08 ± 0,20
Tricholoma sejunctum * 5 31,48 ± 0,52
- wild in nature CF, "**" - commercial CF
The analysis of polyphenols in the carpophores of Polyporales fungi showed the following. Most of their FBs (85%) contain little polyphenols being within the range of 9 mg/g (D. quercina) to 39 mg/g (F. pinicola). The other group comprises 3 species of Polyporales fungi (G. lucidum, L. sulphureus and G. applanatum) containing polyphenols in their FBs in the amount of 89 mg/g to 161 mg/g (ADB). The fruit bodies of tinder fungus F. fomentarius have the highest concentration of poliphenols -more than 248 mg/g (ADB). To compare the results obtained, it should be noted that the method of reactant extraction from the carpophores of shelf fungus Inonotus obliquus (Ach. ex Pers.) Pil. with the maximum PP concentration of 140 mg/g (ADB) by water extraction has been patented (Sysoeva et al., 2005; Patent 2448721).
The analysis of polyphenols in the carpophores of Agaricales fungi showed that most of them (74%) contain little polyphenols in their FBs but it is less compared to Polyporales fungi. Here they are within the range of 12 (P. squarrosa) to 37 mg/g (P. citrinopileatus). 5 species of Agaricales fungi (P. ostreatus, S. rugosoannulata, T. flavovirens, F. velutipes and A. cylindracea) can be put into a group with the medium polyphenol concentration of 53 to 101 mg/g in the fruit bodies. The highest polyphenol concentration of about 172 mg/g was registered in the wild fruit bodies ISSN 2225-5486 (Print), ISSN 2226-9010 (Online). Eiоnогiцниu eicHUKMffnY. 2015. №3
of F. hepatica. However, this parameter is more than 1.5 times lower than the concentration of phenolic substances in the fruit bodies of tinder fungus
F. fomentarius. In contrast, it should be noted that the mean concentration of polyphenols amounts to 450 mg/g in the plant raw material of Camellia sinensis, and 70 mg/g in the mycological material (fruit bodies of P. ostreatus) (Li Fu et al., 2011; Halvorsen et al., 2011).
The analysis of polyphenols in the alcohol extracts from the fruit bodies of 50 basidiomycetes species made it possible to isolate some species of tinder fungi:
G. lucidum, L. sulphureus, G. applanatum and F. fomentarius, and some species of Agaricales fungi: S. rugosoannulata, A. cylindracea, T. flavovirens, F. velutipes, P. ostreatus and F. hepatica having the high concentration of these substances of more than 60 mg/g (ADB).
The next step of the investigation was to isolate pure cultures from the carpophores as well as to study the growth and synthesis rate of polyphenols of some of them when cultivating in the GPM.
The results of ADB accumulation by the strains while their growing (on the 6th, 9th and 12th day of cultivation) are shown on Figure 1. As can be seen, all cultures reach the maximum of this parameter on the 12th day of their growth. The most productive strains here are those of S. commune Sc-1101 and Sc-10 and that of F. velutipes F-202. The lowest values of ADB accumulation were registered for P. ostreatus strain P-192 and F. fomentarius strain Ff-09. Thus, the investigated cultures have their individual growth values - biomass accumulation in the cultivation conditions used that probably reflects suitability of these conditions for their growth.
tg m
8 7 6 5 4 3 2 1 0
□ 6 DC
□ 9 DC
□ 12 DC
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PhPH1"^
, .i-LO
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<10cnC2M>i/510<^(N r ooooomooc^
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Strain
Fig. 1. Accumulation of absolute dry biomass of strains of Basidiomycetes on Day 6, 9, and 12 of cultivation (DC - day of cultivation)
The investigation results of total polyphenols in the mycelium and culture filtrate while growing some basidiomycetes strains are shown on Figure 2 and 3.
□ 6 DC
tg tg
140 120 100 80 60 40 20 0
□ 9 DC
□ 12 DC
PHF fi,j(_LQAAHH P^PhPHK PHPHPH
<4H
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c Strains
Fig. 2. The total content of polyphenols of mycelium of strains of Basidiomycetes on Day 6, 9, 12 of cultivation (DC - day of cultivation)
8 n
0'-H0'-H00'-H00'-H'-Hr 1 OOOOOCOOOCOOn
^ ¿HH Ph c Strain
□ 6 DC
□ 9 DC
□ 12 DC
Fig. 3. The total content of polyphenols of culture filtrate of strains of Basidiomycetes on Day 6, 9, and 12 of cultivation (DC - day of cultivation)
It was established that most of strains were able to accumulate polyphenols both in the mycelium and CF during the whole cultivation period. The maximum of PP concentration in the mycelium for 96% and in the CF for 91% of the total strains coincided with the termination of their cultivation period.
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The trend of polyphenols in the mycelium of the strains under investigation has the following characteristics. The highest concentration of these substances within the range of 107.9 to 129.4 mg/g was registered for P. ostreatus strains P-039 and P-208, and S. commune strain Sc-1102 on the 12th day of their growth. The lowest values of PP concentration of 29.9 to 50.1 mg/g were registered for F. velutipes strains F-1, F-202 and F-03, and P. ostreatus strain P-192 at the end of their cultivation period.
The trend analysis of polyphenols in the culture filtrate of the strains under investigation showed the following. The highest PP concentration was registered on the 12th day of their growth within the range of 5.4 to 6.8 mg/ml for the strains of F hepatica Fh-18, L. sulphureus Ls-08 and P. ostreatus P-01, and the lowest (0.9 to 1.7 mg/ml) - for the strains of F. hepatica Fh-08, P. ostreatus P-192 and F. velutipes F-1.
In all cases the concentration of polyphenols in the mycelium was much higher than the concentration of these substances in the culture filtrate and varied on the 12th day of cultivation from 11.4 times for L. sulphureus strain Ls-08 to 52.9 times for F hepatica strain Fh-08. A considerable difference between ability of strains to synthesise and accumulate PPs in the mycelium and CF may be most likely explained by their genotype realization under test conditions.
The calculation of a correlation coefficient between PP concentration in the mycelium and CF of one-year cultures showed the following. A very high positive correlation is observed in 73.2%, high positive one - in 17.4% and medium one - in 4.5% of tests.
Thus, the test results of total polyphenols in some basidiomycetes species enable us to make the following conclusions. The species of tinder fungi - Ganoderma applanatum, Ganoderma lucidum, Laetiporus sulphureus and Fomes fomentarius, and the species of Agaricales fungi - Stropharia rugosoannulata, Agrocybe cylindracea, Tricholoma flavovirens, Flammulina velutipes, Pleurotus ostreatus and Fistulina hepatica are characterized by the highest concentration of polyphenols in their carpophores. Most of introduced strains can accumulate polyphenols both in the mycelium and CF during the whole period of their cultivation.
The strains of P. ostreatus P-01, F. hepatica Fh-18 and L. sulphureus Ls-08 are perspective for further investigations aimed to obtain polyphenols of extracellular origin and the strains of S. commune Sc-1102, and P. ostreatus P-039 and P-208 - those of mycelial one.
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Поступила в редакцию 18.10.2015
Как цитировать:
Veligodska, A.K., Fedotov, O.V. (2015). Screening of content and dynamic of accumulation of polyphenols in some Basidiomycetes species. Биологический вестник Мелитопольского государственного педагогического университета имени Богдана Хмельницкого, 5 (3), 42-54. cross«* http://dx.doi.org/10.7905/bbmspu.v5i3.984
© Veligodska, Fedotov, 2015
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