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Projects / Programmes source: ARIS

Biotic diversity of fungal endophytes in biochemically defined spruces, their ecophysiological characteristics and antagonistic activity against pests

Research activity

Code Science Field Subfield
4.01.01  Biotechnical sciences  Forestry, wood and paper technology  Forest - forestry 

Code Science Field
B230  Biomedical sciences  Microbiology, bacteriology, virology, mycology 
B250  Biomedical sciences  Entomology, plant parasitology 
Keywords
endophytic fungi, biotic diversity, biology of endophytes, ecophysiology of endophytes, antagonistic activity, pests, mutualistic symbiosis, genetics of Norway spruce, Pice abies (L.) Karst., growing site
Evaluation (rules)
source: COBISS
Researchers (6)
no. Code Name and surname Research area Role Period No. of publicationsNo. of publications
1.  11253  PhD Jurij Diaci  Forestry, wood and paper technology  Researcher  1997 - 1999 
2.  02491  PhD Maja Jurc  Forestry, wood and paper technology  Head  1997 - 1999 
3.  17335  Nina Rotar    Researcher  1997 - 1999 
4.  17336  Matej Rupel    Researcher  1997 - 1999 
5.  10264  PhD Primož Simončič  Forestry, wood and paper technology  Researcher  1999 
6.  00989  PhD Mitja Zupančič  Biology  Researcher  1998 - 1999 
Organisations (2)
no. Code Research organisation City Registration number No. of publicationsNo. of publications
1.  0404  Slovenian Forestry Institute  Ljubljana  5051673000 
2.  0618  Research Centre of the Slovenian Academy of Sciences and Arts  Ljubljana  5105498000 
Abstract
In proposed research endophytic fungal population in the needles of Norway spruce are investigated, the influence of the host and growing sites on this population will be determined and also the interaction between some dominant fungi and insects, feeding on needles. -The research is performed at supposed natural growing sites of Norway spruce and on its secondary growing sites. The growing sites were characterised by the standard central-European phytocenological method. The samples were taken at following plant associations: Sphagno-Piceetum, Rhytidiadelpho lorei-Piceetum, Adenostylo glabrae-Piceetum, Aposeri-Piceetum and in some planted spruce stands on beech growing sites. -Analysis of soil samples from all sampling locations were performed. -For the genetic definition of spruces, from which samples were taken the results of the ongoing research, started in 1995 in Slovenian Forestry Institute will be used. In this research the method of isoenzyme electrophoresis of needle extracts was introduced, tested regarding extraction buffers, gel systems and enzyme systems. Samples of needles for endophyte research were taken from the same trees from which samples were taken for the above mentioned research. -The nutrition of sampled trees (N, P, K, Ca, Mg), analyses of total sulphur (S) content and the heavy metals content (Pb, Zn, Cd) were analysed. -The gathered needles (1-8 years old) were sorted in the laboratory by their age, ten healthy and not damaged needles were selected from each yearly segment. They were rinsed under running tap water for about 30 minutes, 3mm long segments were cut from them, their surface was sterilised and they were placed in petri dishes on diferent growing media. 15 segments were placed in each petri dish and incubated at room temperature. During the first three weeks the growth of fungi were controlled daily and weekly for the following three months. After three months the fungal cultures were stored in refrigirator and controlled occasionally. Identification of fungal taxa was done on the basis of culture characteristics and on the basis of the morphological characteristics of fruitbodies and spores. All isolated fungi were stored as dry or living cultures in Herbarium of SFI (LJU). Reference material will be deposited in herbarium and in culture collection of IMI, Egham, Surrey, GB and in Microbiological collection of Chemistry Institute in Ljubljana as well. -The growth increment of five most important (most frequent) endophytes was measured on malt agar at different temperetures, at various amounts of available water (aw) in the growing medium (aw1 = 0,81, aw2 = 0,90, aw3 = 0,94), at different acidity of the growing medium and with combinations of the mentioned factors. All experiments were done in three replicates. We used BIOLOG tests (GH MicroPlates, BIOLOG, Inc, CA) in analyses of the ecological niche (NOI) of dominat endophytic species. -Insect species colonising spruce needles will be determined on all trees on sampling locations. The abundance of gathered species, their developmental phase and degree of survival will be detected. It is expected to follow the development of most frequent insects in vitro feeding them with needles of sampled trees with the intention to analyse the effect of consuming needles, infected with different endophytes. -The degree of alliance and homogenity among morphologicaly and biologicaly different genotypes (different degree of mycelium growth of the same species in different conditions, presumed different ergosterol and other sterols content) of 2-3 endophytic fungi will be established with molecular techniques. The method of random amplified polymorphic DNA (RAPD), one of the applications of basic PCR techniques, will be used (Williams et all. 1990).
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