Provider: - Institution: - Data provided by Europeana Collections- Mycorrhizas are symbiotic associations formed between the group of specialized soil fungi and roots of higher plants. Mycorrhizas are present in all terrestrial ecosystems and play a central role in the nutrient uptake. In addition, mycorrhizal plants are often more resistant to diseases caused by microbial soil borne pathogens and to the effects of drought. Ectomycorhiza is being established with roots of trees, especially the families Betulaceae, Pinaceae, Fagaceae and Salicaceae, representing ecologically and economically most important tree species being dominated in boreal, Mediterranean forests and forests in temperate region. Analysis of diversity and distribution of ectomyccorhizas is important for understanding of their ecological function. According to published data, 50 ectomycorrhizal fungal genera have been recorded in Montenegro until now: 12 Genera of Ascomycota (50 taxa) and 38 Genera of Basidiomicota (267 taxa), representing c.ca one third of the total number of macromycetes’ records here. Pinus heldrechii Christ, being tertiar relict and subendemic, is one of the most interesting elements of Balkan’s dendroflorae. Fungal communities in P. heldreichii forests are only slightly examined, while their ectomycorrhizal communities have not been described or even confirmed. Due to evident ecological adaptibility, P. heldreichii could be used for afforestation of extreme terrains. In that sense, ectomycorrhizal fungi surviving on these soils should receive a special scientific attention. The overall aim of this doctorial dissertation was to introduce the ectomycorrhizal communities of Pinus heldreichii, also as to examine the possibilities of application of ectomycorrhizal fungi in nurrsery production of conifers. The collection of c.ca 40 of fungal culture’s isolates of putative ectomycorrhizal fungi from Montenegro area was established and used in further researches. Using different methods of inoculation under laboratory conditions, ectomycorrhiza were achieved between P. heldreichii and 18 fungal species. Genetic characterisation of fungal cultures was performed using molecular methods (ITS sequencing), and 22 fungal ITS sequences (20 ectomycorrhizal) were deposed in NCBI GenBank. This way, fungi from 10 ectomycorrhizal genera were characterised: Amanita, Boletus, Hebeloma, Lactarius, Russula, Suillus,Tricholoma, Pisolithus, Scleroderma, also as Chalciporus and Hypholoma. Using RFLP analysis with Hinf I, Alu I, Mbo I, Bsur, EcoRI i Rsa I restrictive enzymes, RFLP patterns for 25 taxa were obtained, which could serve as an efficient tool for their further monitoring. Characterization of ectomycorrhizal fungal communities of P. heldreichii, done by molecular analysis of plant roots on 2 localities with different soil development, shows their differences in composition and structure. In pioneer forest community of P. heldreichii on calcareous soils in initial phase, 11 morphotypes were identified on root samples, from which 29 ectomycorrhizal taxa were extracted. Species from genera Suillus, Rhizopogon, Amphinema, Tomentela, Willcoxina, Sebacine, Pseudotomentella, Lactarius, Tricholoma, Inocybe were predominant in ectomycorrhizal community on this locality. In climatogenous P. heldreichii forest on calcareous brown forest soil, 7 morphotypes and 21 ectomycorrhizal taxa were identified. On this locality, Amphinema taxa were predominant, while Tomentella, Sebacine, Pseudotomentella, Suillus, Russula and Lactarius followed. Examination of physiological characteristics of Chalciporus ammarelus (Quel.) Bataile, Lactarius deliciosus (L.) Gray, Russula sanguinaria (Schumach.) Rauschert, Suillus granulatus (L.) Rousell, Suillus collinitus (Fr) Kuntze, Tricholoma batchii Gulden, Tricholoma imbricatum (Fr.) Kumm, Pisolithus arhizus (Scop.) Rauschert, Scleroderma sp. in vitro, displays differences in appearances of cultures of same fungus developed on different temperatures, nitrogen (NH4 +, NO3 -, protein) and carbon sources (glucose, sucrose, dextrin, arabinose, xylose, starch). Examined isolates from P. heldreichii forests showed the best growth at 22°, while the temperature of 25°C suited better P. arhizus and Scleroderma. Examined isolates could have used ammonium, nitrate and protein as nitrogen sources, and, practically, all of them were able to use the different carbohydrates as the carbon sources, although those characteristics were variable between the species. All examined isolates exhibited good growth on pH between 4 and 7,5. Seedling mycorrhization acts as an efficient tool for improving the quality of seedlings. The effectiveness of different autochthonous ectomycorrhizal fungi to produce containerized ectomycorrhizal seedlings of Pinus nigra in open field conditions was investigated. Spore inoculation with P. arhizus, S. granulatus, S. collinitus, Xerocomus rubellus (106, 107, 108), Boletus luridus, Boletus fechtneri and Boletus aestivalis (107) and vegetative inoculation with P. arhizus, S. granulatus, Scleroderma sp. (1:16, 1:8, 1:4) on ectomycorrhizal formation and seedling growth were tested and proved to be an effective method of obtaining containerized ectomycorrhizal P. nigra seedlings under open field conditions after 11 months. Seedling growth also as ectomycorrhizal formation and development differs according to applied fungal species and inoculation method. According to obtained results, and having in mind the simplicity of application and low costs, it would be feasible to use the spore inocula of P. arhizus with 106 - 107spores per plant, also as S. collinitus and S. granulatus, with 106 spores per plant, to produce ectomycorrhizal P. nigra plants on a large scale. Controlled mycorrhizal inoculation of seedlings is not a common practice in Montenegrin and Serbian nurseries; as such, the obtained results will contribute to the enhancement of nursery production of Pinus nigra and other conifers.- Mikoriza predstavlja zajednicu između grupe gljiva koje žive u zemljištu i korena viših biljaka. Mikoriza se javlja u svim terestričnim ekosistemima i ima važnu ulogu u usvajanju hranljivih materija. Osim toga, biljke sa mikorizom su otpornije na bolesti i sušu. Ekrtomikoriza je tip mikorizne simbioze karakterističan za drveće iz familija Betulaceae, Pinaceae, Fagaceae i Salicaceae, koje predstavlja ekološki i ekonomski najznačajnije drvenaste vrste, dominantne u borealnim, mediteranskim šumama i šumama umerenog pojasa. Analiza diverziteta i rasprostranjenja ektomikorize je značajna za razumevanje njene ekološke uloge. Na osnovu objavljenih podataka, dosadašnjim istraživanjima je u Crnoj Gori zabeleženo 48 rodova ektomikoriznih gljiva: 12 rodova Ascomycota (50 taksona) i 38 rodova Basidiomycota (267 taksona), što predstavlja oko trećinu makromiceta zabeleženih u Crnoj Gori. Munika (Pinus heldrechii Christ.), tercijalni relikt i subendemit, predstavlja jedan od najinteresantnijih elemenata dendroflore Balkana. Zajednice gljiva u šumama munike su slabo istražene, dok ektomikorizne zajednice munike nisu do sada opisivane, niti potvrđene. Zbog svoje ekološke plastičnosti munika se može koristiti za pošumljavanje ekstremnih terena. U tom smislu pažnju zavređuju i ektomikorizne gljive koje opstaju baš na ovakvim staništima. Cilj ove doktorske disetacije bio je upoznavanje sa ektomikoriznim zajednicama munike, kao i ispitivanje mogućnosti primene i korišćenja autohtonih ektomikoriznih gljiva u rasadničkoj proizvodnji četinara. Formirana je kolekcija koja je sadržala oko 40 kultura pretpostavljeno ektomikoriznih gljiva sa prostora Crne Gore, koje su korišćene u daljim istraživanjima. Upotrebom različitih metoda inokulacije u laboratorijskim uslovima ostvarena je ektomikoriza između munike i 18 vrsta gljiva. Upotrebom molekularnih metoda izvršena je genetička karakterizacija pojedinih kultura ektomikoriznih gljiva. U Banku Gena NCBI deponavane su 22 sekvence gljiva od kojih 20 pripada mikoriznim vrstama. Na ovaj način su genetički okarakterisane gljive iz 10 ektomikoriznih rodova : Amanita, Boletus, Hebeloma, Lactarius, Russula, Suillus, Tricholoma, Pisolithus, Scleroderma, kao i Chalciporus i Hypholoma. Upotrebom restrikcionih enzima Hinf I, Alu I, Mbo I, Bsur, EcoR I i Rsa I dobijeni su restrikcioni šabloni za 25 vrste ektomikoriznih gljiva koji mogu poslužiti za njihovo praćenje. Karakterizacija ektomikoriznih zajednica munike, obavljena molekularnom analizom korena, na dva lokaliteta sa različito razvijenim zemljištem, pokazuje da se one razlikuju po sastavu i strukturi. U pionirskoj zajednici munike sa gotovo potpuno nerazvijenim zemljištem, na uzorcima korena identifikovano je 11 različitih morfotipova iz kojih je izolovano je 29 ektomikoriznih taksona. Ektomikoriznom zajednicom munike na ovom staništu dominiraju vrste iz rodova Suillus, Rhizopogon, Amphinema, Tomentela, Willcoxina, Sebacine, Pseudotomentella, Lactarius, Tricholoma, Inocybe. U klimatogenoj zajednici munike na razvijenom smeđem zemljištu na krečnjacima, u uzorcima korena identifikovano je 7 različitih morfotipova, odakle je izolovan 21 mikorizni takson. Ovde dominiraju Amphinema taksoni, a zatim Tomentella, Sebacine, Pseudotomentella, Suillus, Russula, Lactarius. Ispitivanja fizioloških karakteristika Chalciporus ammarelus (Quel) Bataile, Lactarius deliciosus (L.) Gray, Russula sanguinaria (Schumach.) Rauschert, Suillus granulatus (L.) Rousell, Suillus collinitus (Fr) Kuntze, Tricholoma batchii Gulden, Tricholoma imbricatum (Fr.) Kumm, Pisolithus arhizus (Scop.) Rauschert, Scleroderma sp. u laboratorijskim uslovima pokazuju da postoje razlike u izgledu kultura iste gljive kada se ona razvija na različitim temperaturama, različitim izvorima azota i ugljenka. Ispitivani izolati poreklom iz šume munike najbolje rastu na temperaturi od 22°C, dok izolati P. arhizus i Scleroderma bolje rastu na 25°C. Pokazalo se da su svi ispitivani izolati mogli da koriste amonijačni