Rezumatul tezei de doctorat - USAMV Cluj-Napoca

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The dendrogram interpretation, performed after the reading of DNA fragment lengths, from the 69 analyzed fishponds, allows us to conclude the following aspects regarding the Saprolegniaceae family fungi phylogeny (fig.4). All the identified fungal types have an common ancestral origin, the first speciation being realized at the ancestral variety, at a linkage distance of 58 cM (centiMorgans). One of the speciations evolved for a longer period of time, in a part of the analyzed fishponds, when caused by a point mutation, placed on a distance of 20 cM, appeared another 2 speciations. One of them evolves until today, being localized in the fishponds 1-5, 8,9 from Ariniş fishery, in the 7 fishponds from Motiş fishery (var.1), in the fishponds 1-3, 5-6 from Ciurila, Daia fishponds 1-4, Iernut fishponds 1-3 and 5, Cefa fishponds 1-2, 4-7 and 9, and all the 7 fishponds from Ineu fishery (var.7). The other speciation, which evolved from a point mutation, at the level of 20 cM, for a period of time, suffered another point mutation, at the linkage distance of 10 cM, generating another two speciations. One of them (var.3) still evolves and was identified in the fishponds 1-4, 6-8 and 10 from Adrian fishery and in all the 5 fishponds from łaga fishery (var.6). The second branch which broked from the ancestral form, at the distance of 58 cM evolved until a point mutation caused the creation of two new speciations, at the linkage distance of 42cM. One of the branches evolved for a shorter period of time, when at the linkage distance of 35 cM realized a point mutation, which caused the appearance of another two new speciations. One of them was identified in the fishponds 6 and 7 from Ariniş fishery (var.2). The other speciation, after a point mutation, at a linkage distance of 30 cM, caused the evolution of two taxonomical units, identified today. One of them evolves and it is very active in the fishpond 4 from Ciurila fishery (var.8), and in the fishpond 4 from Chiochiş fishery (var.10). The other broked branch is identified and present today in the fishpond 4 from Iernut fishery and fishpond 5 from Daia (var.11). The second branch, broked at a linkage distance of 42 cM, evolved for a longer period of time, in a part of the analyzed fishponds, then, at a distance of 10 cM, after another mutation, resulted 3 speciations, two of them genetic related (var.4 and 9), identified in the fishpond 4 from Adrian fishery and in fishponds 1-3 and 5 from Chiochiş fishery. At a 68
higher genetic distance, the individuals broked from the same branch, are evolving today in the fishpond 9 from Adrian fishery (var.5) and in the fishponds 8 and 10, from Cefa fishery (var.12). II.8. SEQUENCING RESULTS OF THE SAPROLEGNIACEAE FAMILY FUNGAL SPECIES FROM THE STUDIED LOCATIONS After the automatic sequencing, performed with ABI Prism sequencer, by Mycrosinth (Switzerland), there were identified two aquatic fungal species, in the studied fishponds, represented by Saprolegnia ferax and Achlya bisexualis. In the case of Saprolegnia ferax species sequencing, the fragment sequenced with the primers ITS1(forward) and ITS4 (reverse), has a length of 743 bp, a identity of 99% and has two point mutations compared to the matrix species from GeneBank. At 31 position, there is observed a deletion between A-C nucleotides, of a C nucleotide, and at 368 position, there is a substitution of A with C. After sequencing Achlya bisexualis species, the fragment sequenced with the primers ITS1(forward) and ITS4 (reverse), has a length of 761 bp, a identity of 99% and has 7 point mutations compared to the matrix species from GeneBank. At 112 position, there is rematked a substitution of C with T; at 119 position, a substitution of G with A; at 194 position, a substitution of T with G; at 370 and 371 positions, two substitutions of C with A; at 416 position, a substitution of A with T, and at 633 position, a substitution of A with G. II.9. THE INCIDENCE OF SAPROLEGNIASIS IN THE CENTRAL AND NORTH-WESTERN ROMANIAN AREAL Saprolegniasis is the ”silent killer” of the aquatic species, the disease producing extremely important losses in the fisheries, particularly in the cyprinid ones. Saprolegniasis is one of the most important causes of economic losses in aquaculture, fungal infections are second only to bacterial diseases in economic importance. Fungal infections are generally restricted to chronic, steady losses. In Japan, there is an annual mortality rate of 50% in coho salmon (Oncorhynchus kisutch) due to Saprolegnia parasitica 69
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UNIVERSITATEA DE ŞTIINłE AGRICOLE
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II.6. REZULTATELE RESTRICłIEI ENZI
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diverse specii piscicole. De aici s
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Fig.1. LocaŃiile în care s-au efe
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A fost constituit un număr de 207
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Sabouraud Dextrose Broth (SDB-bulio
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Echipament, materiale şi reactivi
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Primerii folosiŃi de către noi î
Page 17 and 18: CAPITOLUL II REZULTATELE CERCETĂRI
Page 19 and 20: Mediile diametrului coloniilor de S
Page 21 and 22: laborator specifice metodologiei de
Page 23 and 24: Deoarece am utilizat în experiment
Page 25 and 26: fragmente: primul la aproximativ 45
Page 27 and 28: II.7. REZULTATE PRIVIND STUDIUL ÎN
Page 29 and 30: mai mare genetică, indivizii despr
Page 31 and 32: MenŃionăm faptul că în toate ba
Page 33 and 34: CAPITOLUL III CONCLUZII ŞI RECOMAN
Page 35 and 36: isexualis (761 pb, o identitate de
Page 37 and 38: BIBLIOGRAFIE SELECTIVĂ 1. Colao Ma
Page 39 and 40: 19. Oroian, T.E., 2006, SelecŃia a
Page 41 and 42: UNIVERSITY OF AGRICULTURAL SCIENCES
Page 43 and 44: SPECIES FROM SAPROLEGNIACEAE FAMILY
Page 45 and 46: The specific literature presents up
Page 47 and 48: Fig.1. The locations where the rese
Page 49 and 50: estimation of the spawns infestatio
Page 51 and 52: pellets were filtred, washed with d
Page 53 and 54: The DNA rapid extraction using PBS
Page 55 and 56: We used a comparatively mixture of
Page 57 and 58: II.1. THE RESULTS OF SAPROLEGNIA CU
Page 59 and 60: Reading time 24 48 72 n Culture med
Page 61 and 62: asexual one. We can precisely concl
Page 63 and 64: 1 2 3 4 5 6 7 8 9 700pb- Fig.3. Ele
Page 65 and 66: pairs length. The sequencing result
Page 67: II.7. RESULTS REGARDING THE GENETIC
Page 71 and 72: We mention the fact that in al the
Page 73 and 74: extraction have values between 1,10
Page 75 and 76: 13. In researches on the aquatic fu
Page 77 and 78: 40. Frisvad, J.C., Bridge, P.D., Ar
Page 79: 61. *** http://www.sigmaaldrich.com
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