Endophytic fungi isolated from nettle lady´s mantle and broadleaf plantain

Töö eesmärgiks oli ravimtaimedest endofüütsete seente eraldamine puhaskultuuri, nende kirjeldamine ja määramine. Uuritavad taimed olid suur teeleht (Plantago major), kõrvenõges (Urtica dioica) ja harilik kortsleht (Alchemilla vulgaris). Endofüütide vähese uurituse tõttu on võimalik leida uusi mikroorganisme elusate taimede seesmistest kudedest. Endofüüte on järjest rohkem hakatud uurima, seoses nende suure osatähtsusega nii looduslikes kooslustes, põllumajanduses kui meditsiinis. Käesolevas töös loodeti leida uusi seenetüvesid ravimtaimedest, mida edaspidises töös saaks kasutada haigustõrjeks. Uurimusliku töö tulemusena eraldati 85 puhaskultuuri, millede identifitseerimiseks mõõdeti nende morfoloogilisi tunnuseid (sh koniidi ja hüüfi suurused, kasvukiirused, kultuuri värvus) ning eraldati sekveneerimiseks ITS rDNA. Selle alusel määrati 12 kultuuri: Lewia infectoria, Hypoxylon howeanum, Fusarium culmorum, Apodus deciduus, Leptosphaeria sp, Pleosporales sp, Colletotrichum clavatum, Hypoxylon fuscum, Colletotrichum clavatum, Phoma multirostrata (2), Phoma sp (2). Edaspidi tahetakse uurida, kumb toodab inimesele kasulikku bioaktiivset ainet, kas seen või ravimtaim ning kas erinevatel taimedel on erinevad endofüütsed seened.The aim of this research was to isolate endophytic fungi to pure culture, to describe and determine them. The studied plants were broadleaf plantain (Plantago major), stinging nettle (Urtica dioica) and lady's mantle (Alchemilla vulgaris). Due to the fact that endophyte-related research is quite new, it is possible to discover new microorganisms in inner tissues of plants. The research on endophytes has been growing over time, because of their relative importance to natural community, agriculture and medicine. This research is looking for fungal strains from herbal plants, which could be used for disease control in further work. As a result, 85 pure cultures were isolated from plants, which were identified by measuring their morphological features (including conidia and hypha sizes, growth rate, pure culture colour) and separating ITS rDNA for sequencing. On this basis the 12 culture: Lewia infectoria, Hypoxylon howeanum, Fusarium culmorum, Apodus deciduus, Leptosphaeria sp, Pleosporales sp, Colletotrichum clavatum, Hypoxylon fuscum, Colletotrichum clavatum, Phoma multirostrata (2), Phoma sp (2). In the future, researchers want to find out whether fungi or herbs produce benefitial bioactive substance for people, and do different plants have different endophytic fungi

Antimicrobial properties of endophytic fungi

Maailmas on üha suuremaks probleemiks inimese haigustekitajate ravimiresistentsus. Samuti on kliimamuutusi seostatud suureneva metsapatogeenide levikuga. Selles vallas omavad potentsiaali endofüütsed seened – seened, kes elavad taimekudede vahel ja ei põhjusta peremeestaimedel enamasti haigusnähtusid. Nende ainevahetusproduktidest võib leida uusi aineid, mis on antibiootiliste või antifungaalsete omadustega ning omavad potentsiaali bioloogilises tõrjes. Töö eesmärgiks oli uurida valitud endofüütsete seente antimikroobseid omadusi bakterite ja puitu kahjustavatate seente suhtes. Bakteritest kasutati Staphylococcus aureus, Escherichia coli ja Proteus vulgaris´t. Lisaks uuriti mõju pärmseenele Candida albicans. Metsapatogeenidest tehti katseid Heterobasidion annosum (nii männilt kui kuuselt pärit viljakehade puhaskultuurid), Diplodia sapinea, Fomes fomentarius, Fomitopsis pinicola, Piptoporus betulinus ja Lophodermium seditiosum´iga. Uuritavad endofüüdid on pärit Eestist, Nigeeriast ja Prantsuse Gujaanast. Kokku kasutati töös 56 endofüüti. Töös antakse ülevaade teemaga seonduvast kirjandusest ja patogeenidest. Uurimustöös kasutati kahte eri metoodikat: endofüütide vedelfiltraate kasutati S. aureus, E. coli, C. albicans vastase mõju uurimisel ning kaksikkultuurimeetodi kaudu tuvastati endofüütide inhibeeriv mõju P. vulgaris ja puitu lagundavate seente puhul. Uurimusliku töö tulemusena leiti üks endofüüt, mis inimpatogeenide uuringus inhibeeris E. coli kasvu. P. vulgaris katses oli patogeeni kasvu pidurdavaid endofüüte 15. Puitu lagundavate seente uuringus andsid Eesti taimedest isoleeritud endofüüdid troopika omadest paremaid tulemusi. Kõige enam inhibeeriti kasvu Lophodermium seditiosumi puhul, antagonismi protsent oli erinevate endofüütide puhul 19,4-44,1%. Antud uuringust võib järeldada, et endofüütsed seened olid efektiivsemad P. vulgaris ja puitu lagundavate seente tõrjes. Tulevikus saaks antud tulemusi rakendada bioloogilises tõrjes, kuid esmalt tuleks tuvastada spetsiifilised ained, mis omavad inhibeerivat efekti. Seejärel saaks looduses nakatada puid kõige efektiivsemate endofüütidega.Drug resistance of the human pathogens is an increasing problem in the world. Also, increasing spread of forest pathogens have been accociated with the changes in the climate. Endophytic fungi - fungi that exist between plant tissues and do not cause disease to the host, have great potential in this area. Among their metabolic products there could be new substances that have antibiotic or antifungal properties and have potential for biological control. The aim of this research was to investigate antimicrobial properties of selected endophytic fungi in regards to bacteria and wood decaying fungi. Bacteria used in this research were Staphylococcus aureus, Escherichia coli and Proteus vulgaris. Effect on yeast fungus Candida albicans was also investigated. Heterobasidion annosum (fruiting bodies pure cultures from spruce and pine), Diplodia sapinea, Fomes fomentarius, Fomitopsis pinicola, Piptoporus betulinus ja Lophodermium seditiosum were the used forest pathogens. Endophytes used are from Estonia, Nigeria and French Guyana. All together 56 endophytes were used in this reasearch. An overview of literature used and pathogens is given. There were two methods used: liquid filtrates were used to investigate the impact against S. aureus, E. coli, C. albicans and dual culture method to detect inhibitory efect of endophytes on P.vulgaris and wood decaying fungi. As a result of this research, one endophyte that inhibited E.coli in human pathogen survey was found. In the survey of P.vulgaris, 15 endophytes inhibited the pathogen. In the study with wood decaying fungi, endophytes inhibited from Estonia showed better results. The most inhibited pathogen was Lophodermium seditiosum, antagonism percent with different endophytes was 19,4 – 44,1 %. In conclusion of this study, endophytic fungi were more efficient against P.vulgaris and wood decaying fungi. In the future these results could be used in biological control, but first specific substances with inhibitory effect should be established. After that trees should be infected with the most effective endophytes

Synthesis and characterization of comb-shaped polyesters based on 2-octadecyl-1,3-propanediol and phthalic acids

Three isomeric poly(2-octadecyl-1,3-trimethylene phthalate)s were prepared by melt transesterification of the corresponding diphenyl ester and 2-octadecyl-1,3-propanediol. Intrinsic viscosity, size exclusion chromatography and light scattering determinations yielded weight-average molecular masses of 110 000 and higher. High-resolution, solution C-13 n.m.r. spectroscopy analysis of both the diphenyl ester precursors and polyesters identified the main-chain units, the isomeric aromatic acid residue and glycol carbons, the linking and the terminal ends of the octadecyl side chain, and finally allowed detection of phenyl ester end groups at 1-2% level. No absorption band due to hydroxyl end groups was monitored in the i.r. polyester spectra. I.r. spectroscopy, d.s.c. and powder X-ray diffractometry provided a clear indication of the crystallinity of the prepared polyesters. The relevant melting enthalpy values and X-ray diffraction patterns were typical of comb-shaped polymers in which the paraffinic side chains crystallize independently with their characteristic hexagonal packing