Rezumatul tezei de doctorat - USAMV Cluj-Napoca

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2010 CONTENTS CHAPTER I ........................................................................................................................................... 4 EXPERIMENTAL HYPOTHESIS, RESEARCH OBJECTIVES, EXPERIMENTAL DISPOSAL, MATERIAL AND METHOD........................................................... 4 I.1. EXPERIMENTAL HYPOTHESIS AND RESEARCH OBJECTIVES ..................................... 4 I.2. EXPERIMENTAL DISPOSAL................................................................................................... 5 I.3. MATERIAL AND METHOD ..................................................................................................... 7 I.3.1. Sample collecting.................................................................................................................. 7 I.3.2. The initiation of Saprolegnia culture in laboratory conditions............................................. 7 I.3.3. Culture media used ............................................................................................................... 8 I.3.3.1. Solid culture media used in the experiment ................................................................... 9 I.3.3.2. Liquid culture media used in the experiment............................................................... 50 I.3.3.3. Antibiotics used in the experiment ............................................................................... 51 I.3.4. Morphological characterization of Saprolegnia strains...................................................... 51 I.3.5. Fungal DNA extraction and <strong>de</strong>tection ................................................................................ 52 I.3.5.1. Fungal DNA extraction using extraction kits (QIAGEN) ............................................ 52 I.3.5.2. Fungal DNA extraction using solutions....................................................................... 52 I.3.6. DNA quantification using direct method of DNA purity and concentration with Nanodrop ND-1000 spectrophotometer ....................................................................................... 53 I.3.7. ITS region from fungi and primers used............................................................................. 53 I.3.8. Molecular techniques used in the experiment..................................................................... 55 I.3.8.1. PCR amplification of Saprolegnia samples ................................................................. 55 I.3.8.2. PCR-RFLP techique - Restriction Fragment Length Polymorphism .......................... 56 I.3.9. Methods used in establishing the phylogenetic diversity and relationship of Saprolegniaceae family fungi species........................................................... 56 I.3.9.1. Automatic sequencing .................................................................................................. 56 CHAPTER II........................................................................................................................................ 56 PERSONAL RESEARCH RESULTS................................................................................................. 56 II.1. THE RESULTS OF SAPROLEGNIA CULTURE IN LABORATORY CONDITIONS ..................................................................................................... 57 II.2. RESULTS REGARDING SAPROLEGNIA’S GROWTH AND DEVELOPMENT ON THE CULTURE MEDIA ........................................................................... 58 II.2.1. Comparative results regarding the growth of Saprolegnia colonies on solid culture media ........................................................................... 58 II.2.2. Comparative results regarding the <strong>de</strong>velopment of Saprolegnia colonies on the liquid culture media.................................................................... 19 II.3. THE MORPHOLOGICAL DESCRIPTION OF SAPROLEGNIA STRAINS......................... 19 II.4. RESULTS REGARDING SAPROLEGNIA’S DNA EXTRACTION AND QUANTIFICATION.................................................................................... 61 II.4.1. DNA extraction.................................................................................................................. 61 II.5. THE DNA AMPLIFICATION RESULTS OF FUNGAL SPECIES FROM SAPROLEGNIACEAE FAMILY ......................................................................................... 62 II.5.1. DNA amplification using PCR .......................................................................................... 62 II.6. THE DNA ENZYMATIC RESTRICTION RESULTS OF FUNGAL 42
SPECIES FROM SAPROLEGNIACEAE FAMILY USING PCR-RFLP METHOD...................... 63 II.7. RESULTS REGARDING THE GENETIC PHYLOGENY STUDY OF SAPROLEGNIACEAE FAMILY FUNGI.................................................................... 67 II.8. SEQUENCING RESULTS OF THE SAPROLEGNIACEAE FAMILY FUNGAL SPECIES FROM THE STUDIED LOCATIONS .......................................................... 28 II.9. THE INCIDENCE OF SAPROLEGNIASIS IN THE CENTRAL AND NORTH-WESTERN ROMANIAN AREAL......................................................................... 28 CHAPTER III....................................................................................................................................... 72 CONCLUSIONS AND RECOMMENDATIONS............................................................................... 72 SELECTIVE BIBLIOGRAPHY.......................................................................................................... 76 43
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