Mycorrhization of oak and hazel trees with different species of the genus Tuber

The root system of oak (Quercus cerris L., Q. robur L., Q. petrea, Lieblein and Q. borealis L.) and hazel trees (Corylus avellana L.) raised in greenhouse conditions was inoculated with a suspension of ascospores of: Tuber aestivum Vitt, T. melanosporum Vitt., T. macrosporum Vitt. (black truffles) and T. magnatum Pico ex Vitt. (white truffle). Roots were examined after 4, 8, 10 and 36 months. After 4 months symbiosis could be observed at ultrastructural level. Mycelium from isolated mycoclenes was applied to oak tree root systems with an aim of proving its identity and viability. Mycorrhiza was obtained and the mycelium was reisolated.Korenov sistem sadnica hrasta (Quercus cerris L., Q. robur L., Q. petrea, Lieblein i Q. borealis L.) i leske (Corylus avellana L.) odgajanih u uslovima staklare inokulisan je ca suspenzijom askospora Tuber aestivum Vitt., T. melanosporum Vitt., T. macrosporum Vitt. (crni tartufi) i T. magnatum Pico ex Vitt. (beli tartuf). Korenovi su posmatrani posle 4, 8, 10 i 36 meseci. Nakon 4 meseca simbioza je dokazana na ultrastrukturnom nivou. Micelija izoloaana sa mikoklena aplikovana je na korenov sistem sadnica hrasta sa ciljem dokazivanja njenog identiteta i vijabiliteta. Mikoriza je dobijena i micelija je reizolovana.nul

Mycorrhization of oak and hazel trees with different species of the genus Tuber

The root system of oak (Quercus cerris L., Q. robur L., Q. petrea, Lieblein and Q. borealis L.) and hazel trees (Corylus avellana L.) raised in greenhouse conditions was inoculated with a suspension of ascospores of: Tuber aestivum Vitt, T. melanosporum Vitt., T. macrosporum Vitt. (black truffles) and T. magnatum Pico ex Vitt. (white truffle). Roots were examined after 4, 8, 10 and 36 months. After 4 months symbiosis could be observed at ultrastructural level. Mycelium from isolated mycoclenes was applied to oak tree root systems with an aim of proving its identity and viability. Mycorrhiza was obtained and the mycelium was reisolated.Korenov sistem sadnica hrasta (Quercus cerris L., Q. robur L., Q. petrea, Lieblein i Q. borealis L.) i leske (Corylus avellana L.) odgajanih u uslovima staklare inokulisan je ca suspenzijom askospora Tuber aestivum Vitt., T. melanosporum Vitt., T. macrosporum Vitt. (crni tartufi) i T. magnatum Pico ex Vitt. (beli tartuf). Korenovi su posmatrani posle 4, 8, 10 i 36 meseci. Nakon 4 meseca simbioza je dokazana na ultrastrukturnom nivou. Micelija izoloaana sa mikoklena aplikovana je na korenov sistem sadnica hrasta sa ciljem dokazivanja njenog identiteta i vijabiliteta. Mikoriza je dobijena i micelija je reizolovana.nul

Effect of cultivation conditions on ligninolytic enzyme production by Ganoderma carnosum

Ganoderma carnosum has been found in Europe only, at coniferous trees and it is difficult to distinguish it morphologically from G. lucidum. Since G. carnosum has not been studied biochemically yet, the aim of this study was to analyse its ability to produce Mn-dependent peroxidase (MnP), versatile peroxidase (VP) and laccase (Lac) under conditions of solid-state fermentation of selected plant raw materials as carbon sources, in the presence of different nitrogen concentrations in the medium. Wheat straw, corn stem, oak and grapevine sawdust were the analysed plant raw materials. Nitrogen source in synthetic medium was NH4NO3 and its concentrations were: 10mM N and 20 mM N. Enzyme activity was determined spectrophotometrically, using ABTS and phenol red, as the substrates for Lac and Mn-oxidizing peroxidases, respectively. Maximum level of MnP activity (56.82 U/l) was obtained in the medium with wheat straw and nitrogen concentration of 10 mM. Best carbon source for VP production was grapevine sawdust at nitrogen concentration of 10 mM (80.80 U/l). The obtained Lac activity was very low in the medium with wheat straw (1.80 U/l), while it was not detected in the presence of other three analyzed plant raw materials. Maximum of total protein content (0.06 mgml-1) was noted in the medium where oak sawdust was carbon source and nitrogen concentration was 20 mM.Vrste roda Ganoderma su producenti mnogih biološki aktivnih supstanci pa su objekti proučavanja mnogih medicinskih i farmaceutskih studija. U novije vreme se sve više pažnje poklanja upoznavanju njihovog ligninolitičkog enzimskog sistema, u cilju uspešne primene u različitim biotehnološkim procesima. Za razliku od G. lucidum, čiji je ligninolitički sistem intenzivno proučavan, ostale vrste ovog roda su retko bile objekti istraživanja. G. carnosum se teško morfološki razlikuje od G. lucidum. i još uvek nije biohemijski proučena, pa je cilj našeg istraživanja bio analiza produkcije Mn-oksidujućih peroksidaza i lakaza u uslovima čvrste kultivacije, na različitim biljnim ostacima (pšenična slama, stabljike kukuruza, piljevina hrasta i vinove loze) u prisustvu azota u obliku NH4NO3 i u koncentracijama od 10 mM, odnosno 20 mM. Maksimum produkcije Mn-zavisne peroksidaze dobijen je na pšeničnoj slami pri koncentraciji azota od 10 mM (56.82 U/l). Piljevina vinove loze i koncentracija azota od 10 mM bili su optimalni za sintezu verzatil peroksidaze (80.80 U/l). Dobijena aktivnost lakaza je bila izuzetno niska u medijumu sa pšeničnom slamom (1.80 U/l), dok na ostalim biljnim ostacima nije zabeležena. Maksimalni sadržaj ukupnih proteina je bio najveći u medijumu sa piljevinom hrasta kao izvorom ugljenika i koncentracijom azota od 10 mM.Projekat ministarstva br. 14304

Produktivnost ozime raži u organskom i konvencionalnom sistemu gajenja

Rye is a cereal which is very much demanded at the market, for making a special kind of bread, but it is little grown in Serbia. The aim of this paper is to investigate possibilities of organic growing of winter rye, comparing with the conventional one, in agroecological conditions of Valjevo hilly region during 2008/2009 and 2009/2010 cropping seasons. The trial was set up in a village of Kotešica, on soil which had not been used for agriculture for 7 years. In organic cropping system three combinations of microbiological fertiliser baktofil with zeolite and hydrogel were used prior to sowing. Half of each plot was treated with foliar microbiological fertiliser Slavol during crop growing period. In conventional cropping system three variants with mineral fertilisers were included: NPK, NPK+zeolite, NPK+hydrogel. Results of the yield obtained in the experiment showed a significant difference between two seasons, 2008/2009 and 2009/2010. In comparison with the control, the treatments in an organic cropping system resulted with statistically insignificant differences for mean values in both years, while the mean in conventional cropping system has significantly higher yield of winter rye. Organic cropping system under conditions of Valjevo hilly region did not give significantly lower rye yield compared with the conventional one in a moderate growing season such was 2008/2009. The combination of soil microbiological fertiliser (Baktofil) with foliar fertiliser (Slavol) and zeolite gave the highest winter rye grain yield in all other treatments in the second year of investigation. In a very wet season (2009/2010) mineral fertiliser NPK showed an advantage, especially in combinations with zeolite, and this treatment can be recommended.Raž je žito, koje je veoma traženo, naročito za spravljanje specijalnih vrsta hleba, ali se malo gaji u Srbiji. Cilj ovog rada je da se ispita mogućnost organskog gajenja ove biljne vrste, u odnosu na konvencionalni u konkretnim agroekološkim uslovima valjevskog pobrđa 2008/2009. i 2009/2010. godine. Ogled je postavljen u selu Kotešica, na zemljištu koje nije korišćeno u konvencionalnoj proizvodnji 7 godina. U organskom sistemu poljoprivredne proizvodnje, korišćene su kombinacije mikrobiološkog đubriva baktofila sa dva poboljšivača zemljišta zeolita i hidrogela, kojima je tretirano zemljište neposredno pred setvu. Polovina svake elementarne parcele je prihranjena folijarno, takođe mikrobiološkim đubrivom, slavolom u toku vegetacionog perioda biljaka. U konvencionalnom sistemu poljoprivredne proizvodnje uključene su tri varijante sa kompleksnim mineralnim NPK đubrivom i kombinacija sa zeolitom i hidrogelom, kojima je tretirano zemljište neposredno pred setvu. Dobijeni rezultati prosečnih prinosa ozime raži pokazuju značajne razlike između vegetacionih sezona 2008/2009. i 2009/2010. U poređenju sa kontrolom, tretmani u organskom sistemu gajenja nisu dali značajne razlike prosečnih vrednosti prinosa u obe godine istraživanja, dok je prosečan prinos dobijen u konvencionalnom sistemu gajenja bio značajno veći. U uslovima umerene vegetacione sezone 2008/2009. godine u regionu valjevskog pobrđa u organskom sistemu gajenja nije ustanovljeno smanjenje prinosa u odnosu na konvencionalni. Kombinacija zemljišnog sa folijarnim mikrobiološkim đubrivom i zeolitom je dala najbolji rezultat i najveći prinos ozime raži u drugoj godini istraživanja, pa se preporučuje kao najbolja kombinacija u organskom sistemu gajenja. U veoma vlažnoj godini, kao što je bila 2009/2010. mineralna đubriva su pokazala prednost, naročito u kombinaciji sa zeolitom i ovaj tretman se može preporučiti u sličnim uslovima

Antimicrobial activity of secondary metabolites isolated from Centaurea spruneri Boiss. & Heldr.

Two coumarins, scopoletin (1) and isoscopoletin (2), two simple phenolic acids, protocatechuic acid (3) and isovanillic acid (4) and one flavonoid, eriodictyol (5) were isolated from the aerial parts of Centaurea spruneri. The structure of the compounds was established by spectroscopic methods. The in vitro antimicrobial activity of the isolated compounds was tested against eight bacteria and eight fungal species, using a microdilution method. All compounds tested showed moderate antibacterial and antifungal activities. Their minimum inhibitory concentrations were in the range 0.655-2.38 µmol ml-1 and their minimal bactericidal concentrations ranged from 0.694 to 4.15 µmol ml-1 against the tested bacterial species. All compounds showed fungistatic activity at 0.259-2.38 µmol ml-1 and fungicidal at 0.69-2.6 µmol ml-1 against all fungi tested.Iz nadzemnog dela Centaurea spruneri izolovana su dva kumarina skopoletin (1) i izoskopoletin (2), dve proste fenolske kiseline protokatehuinska kiselina (3) i izovanilinska kiselina (4) i flavonoid eriodiktiol. Hemijske strukture poznatih jedinjenja su utvrđene korišćenjem 1D i 2D NMR, MS i UV spektroskopskih analiza. U in vitro testu za određivanje antimikrobne aktivnosti izolovanih jedinjenja korišćena je mikrodiluciona metoda. Dobijena jedinjenja testirana su na po osam bakterijskih i gljivičnih vrsta. Sva jedinjenja su imala umerenu antibakterijsku i antifungalnu aktivnost. Minimalna inhibitorna koncentracija testiranih jedinjenja se kretala od 0,655 do 2,38 µmol ml-1, a minimalna baktericidna koncentracija 0,694-4,15 µmol ml-1. Testirana jedinjenja su pokazala fungicidnu aktivnost od 0,259-2,38 µmol ml-1 i fungistatičku 0,694-2,60 µmol ml-1, na sve testirane gljive.Projekat ministarstva br. 14304

Osetljivost referentnih sojeva i oralnih kliničkih izolata Candida spp. - na odabrana etarska ulja familije Apiaceae

The aim of this study was to investigate the chemical composition and antifungal activity of four essential oils from Apiaceae family, Pastinaca sativa, Foeniculum vulgare, Pimpinella anisum and Anethum graveolens, against Candida spp. reference strains and clinical isolates from human oral cavities. Chemical analysis of the essential oils (GCFID and GC/MS) showed that predominating chemical classes in the essential oil of A. graveolens was monoterpene hydrocarbons (49.4 %) and the major constituent of the oil was carvone (42.2 %): while in the oils of P. anisum, F. vulgare and P. sativa majority of compounds belong to phenylpropenes (80.9 %, 65.9 % and 49.4 %, respectively): and the major oil constituents were trans-anethole (65.1 % and 80.8 %) and myristicine (49.0 %): respectively. Minimum inhibitory (MIC) and minimum fungicidal (MFC) concentrations were determined using microdilution method. All the tested oils inhibited growth of Candida spp. The strongest antifungal activity was shown by A. graveolens essential oil (MIC 0.63-2.5 mg mL-1; MFC 1.25-5 mg mL-1). In addition to the common use of parsnip, fennel, anise and dill in traditional medicine, present study validates use of their essential oils in treatments intended for reduction and elimination of Candida spp. population in human patients with fungal infection.Cilj ovog istraživanja bio je da se ispita hemijski sastav i antifungalna aktivnost četiri etarska ulja familije Apiaceae, Pastinaca sativa, Foeniculum vulgare, Pimpinella anisum i Anethum graveolens, na referentne sojeve i kliničke izolate Candida spp. iz usne duplje ljudi. Hemijska analiza ispitivanih etarskih ulja (GC-FID i GC/MS) je pokazala da u uljima dominiraju sledeće hemijske klase: u ulju A. graveolens - monoterpenski ugljovodonici (49,4%) i glavni sastojak je karvon (42,2 %): dok u uljima P. anisum F. vulgare P. sativa većina jedinjenja pripada fenilpropenima (80,9 %, 65,9 % i 49,4 %, respektivno): pri čemu su glavne komponente prva dva ulja trans-anetol (65,1 % i 80.8 %): a trećeg, miristicin (49.0 %). Minimalne inhibitorne (MIC) i minimalne fungicidne koncentracije ulja (MFC) su određene primenom mikrodilucione metode. Ispitivana tri ulja su inhibirala rast svih Candida spp.; najjaču antifungalnu aktivnost je pokazalo ulje A. graveolens (MIC 0.63-2.5 mg ml-1; MFC 1.25-5 mg ml-1). Pored česte primene paškanata, morača, anisa i mirođije u tradicionalnoj medicini, ova studija potvrđuje upotrebu njihovih etarskih ulja u tretmanima redukcije i eliminisanju populacije Candida spp. kod humanih pacijenata sa gljivičnim infekcijama

Effect of cultivation conditions on ligninolytic enzyme production by Ganoderma carnosum

Ganoderma carnosum has been found in Europe only, at coniferous trees and it is difficult to distinguish it morphologically from G. lucidum. Since G. carnosum has not been studied biochemically yet, the aim of this study was to analyse its ability to produce Mn-dependent peroxidase (MnP), versatile peroxidase (VP) and laccase (Lac) under conditions of solid-state fermentation of selected plant raw materials as carbon sources, in the presence of different nitrogen concentrations in the medium. Wheat straw, corn stem, oak and grapevine sawdust were the analysed plant raw materials. Nitrogen source in synthetic medium was NH4NO3 and its concentrations were: 10mM N and 20 mM N. Enzyme activity was determined spectrophotometrically, using ABTS and phenol red, as the substrates for Lac and Mn-oxidizing peroxidases, respectively. Maximum level of MnP activity (56.82 U/l) was obtained in the medium with wheat straw and nitrogen concentration of 10 mM. Best carbon source for VP production was grapevine sawdust at nitrogen concentration of 10 mM (80.80 U/l). The obtained Lac activity was very low in the medium with wheat straw (1.80 U/l), while it was not detected in the presence of other three analyzed plant raw materials. Maximum of total protein content (0.06 mgml-1) was noted in the medium where oak sawdust was carbon source and nitrogen concentration was 20 mM.Vrste roda Ganoderma su producenti mnogih biološki aktivnih supstanci pa su objekti proučavanja mnogih medicinskih i farmaceutskih studija. U novije vreme se sve više pažnje poklanja upoznavanju njihovog ligninolitičkog enzimskog sistema, u cilju uspešne primene u različitim biotehnološkim procesima. Za razliku od G. lucidum, čiji je ligninolitički sistem intenzivno proučavan, ostale vrste ovog roda su retko bile objekti istraživanja. G. carnosum se teško morfološki razlikuje od G. lucidum. i još uvek nije biohemijski proučena, pa je cilj našeg istraživanja bio analiza produkcije Mn-oksidujućih peroksidaza i lakaza u uslovima čvrste kultivacije, na različitim biljnim ostacima (pšenična slama, stabljike kukuruza, piljevina hrasta i vinove loze) u prisustvu azota u obliku NH4NO3 i u koncentracijama od 10 mM, odnosno 20 mM. Maksimum produkcije Mn-zavisne peroksidaze dobijen je na pšeničnoj slami pri koncentraciji azota od 10 mM (56.82 U/l). Piljevina vinove loze i koncentracija azota od 10 mM bili su optimalni za sintezu verzatil peroksidaze (80.80 U/l). Dobijena aktivnost lakaza je bila izuzetno niska u medijumu sa pšeničnom slamom (1.80 U/l), dok na ostalim biljnim ostacima nije zabeležena. Maksimalni sadržaj ukupnih proteina je bio najveći u medijumu sa piljevinom hrasta kao izvorom ugljenika i koncentracijom azota od 10 mM.Projekat ministarstva br. 14304

Sensitivity of clinical isolates of candida to essential oils from burseraceae family

The aim of this study was to investigate the chemical composition and antifungal activity of four commercial essential oils from the Burseraceae family - two Boswellia carterii Flueck oils, Canarium luzonicum (Blume) A. Gray oil, and Commiphora myrrha (Nees) Engl oil, against most common Candida spp. recovered from the human oral cavity. The essential oil samples were analyzed by GC- FID and GC/MS. The analysis showed that major essential oils' components were. - pinene (23.04 % and 31.84 %), limonene (45.62 %) and curzerene (34.65 %), respectively. Minimum inhibitory (MIC) and minimum fungicidal (MFC) concentrations were determined using a microdilution standardized technique. All tested Candida spp. clinical isolates and ATCC strains showed susceptibility to tested essential oils in a dose dependent manner. The strongest antifungal activity was shown by essential oil of B. carterii, sample 2; the average MIC values ranged from 1.25 to 1.34 mg/ml, and MFC values ranged from 2.50 to 3.75 mg/ml, depending on the fungus. This study supports the possible use of essential oils from the Bursecaceae family in reduction and elimination of Candida spp. populations in patients with oral cavity fungal infections

Investigation on antibacterial synergism of Origanum vulgare and Thymus vulgaris essential oils

Essential oils are well known as strong antimicrobial agents of plant origin. In spite of this, the antimicrobial synergism of essential oils isolated from different plant species is poorly investigated. The following study examines the synergism of the essential oils of Origanum vulgare L. and Thymus vulgaris L against pathogenic bacteria, Staphylococcus aureus and Salmonella typhimurium. First, the antibacterial effect of the oils was tested, and the minimal inhibitory concentrations (MIC) of both oils were determined using the microdilution method. To test whether the oils act synergistically, every possible combination of essential oil concentrations was used in a dynamic checkerboard method. The results indicated that the oils indeed acted synergistically with fractional inhibitory concentration indexes of 0.45 and 0.50. [Projekat Ministarstva nauke Republike Srbije, br. 173032