Rezumatul tezei de doctorat - USAMV Cluj-Napoca

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 detection ................................................................................ 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 development
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