UPORABA MEDGENSKE REGIJE 16S-23S rDNA ZA IDENTIFIKACIJO OCETNOKISLINSKIH BAKTERIJ, IZOLIRANIH IZ RAZLIČNIH HABITATOV

Copot, Darija

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Title:	UPORABA MEDGENSKE REGIJE 16S-23S rDNA ZA IDENTIFIKACIJO OCETNOKISLINSKIH BAKTERIJ, IZOLIRANIH IZ RAZLIČNIH HABITATOV
Authors:	ID Copot, Darija (Author) ID Trček, Janja (Mentor) More about this mentor...
Files:	MAG_Copot_Darija_2014.pdf (1,44 MB) MD5: 8CA5D1EF1A11D7A1660F4B6A92D79181
Language:	Slovenian
Work type:	Master's thesis/paper
Typology:	2.09 - Master's Thesis
Organization:	FNM - Faculty of Natural Sciences and Mathematics
Abstract:	Namen magistrske naloge je bil pridobiti čiste seve ocetnokislinskih bakterij iz industrijskih bioreaktorjev za proizvodnjo vinskega kisa, domačega kisa, grozdnih jagod in cvetov izbranih rastlin. V ta namen smo bakterije izolirali na različnih selektivnih gojiščih v aerobni atmosferi ter jih identificirali s fenotipskimi in z molekularno-biološkimi pristopi, t. j. s preiskovanjem restrikcijskih vzorcev v reakciji PCR pomnoženih regij 16S–23S rDNA. Restrikcijske vzorce smo primerjali z restrikcijskimi vzorci homolognih regij referenčnih sevov ocetnokislinkih bakterij in tako določili njihovo najbolj sorodno bakterijsko skupino oz. vrsto. Iz starter kulture za proizvodnjo rdečega vinskega kisa in sedmih vzorčnih točk enega oksidacijskega ciklusa proizvodnje kisa smo pridobili 84 izolatov. Po en naključno izbran izolat iz vsake vzorčne točke smo nadalje na osnovi restrikcijskih vzorcev 16S‒23S rDNA uvrstili v skupino Gluconacetobacter intermedius/Gluconacetobacter oboediens. Nadaljnja določitev nukleotidnega zaporedja je potrdila, da gre za vrsto Gluconacetobacter oboediens. Ekstremni pogoji rasti v industrijskih bioreaktorjih za proizvodnjo kisa omogočajo preživetje le visoko specializiranim bakterijskim vrstam, sposobnim rasti v prisotnosti več kot 6 % ocetne kisline, med katere spada tudi vrsta, ki smo jo identificirali v tem delu. Pri vseh preiskanih izolatih smo z analizo RAPD-PCR določili enake RAPD-vzorce, kar kaže na visoko genetsko homogenost mikrobiote za industrijsko proizvodnjo rdečega vinskega kisa. Iz domačega kisa, kjer koncentracija ocetne kisline ne preseže 6 %, smo izolirali vrsto Acetobacter aceti. Iz površine grozdnih jagod in cvetov izbranih rastlin nismo izolirali ocetnokislinskih bakterij, smo pa iz cvetov izolirali bakterije, ki smo jih na osnovi preiskanih nukleotidnih zaporedij odseka gena za 16S rRNA uvrstili v Rahnella sp., Serratia sp. in Pantoea agglomerans. Te bakterije so bile že v preteklosti opisane za te ekološke habitate.
Keywords:	ocetnokislinske bakterije, 16S–23S rDNA, restrikcija, identifikacija, habitat
Place of publishing:	Maribor
Publisher:	[D. Copot]
Year of publishing:	2014
PID:	20.500.12556/DKUM-45779
UDC:	579.6(043.2)
COBISS.SI-ID:	20819208
NUK URN:	URN:SI:UM:DK:Y9BYNCND
Publication date in DKUM:	25.09.2014
Views:	2651
Downloads:	243
Metadata:
Categories:	FNM
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Secondary language

Language:	English
Title:	APPLICATION OF THE 16S-23S rDNA SPACER REGION FOR IDENTIFICATION OF ACETIC ACID BACTERIA ISOLATED FROM DIFFERENT HABITATS
Abstract:	The purpose of the master's thesis was to obtain pure strains of acetic acid bacteria from industrial bioreactors for production of wine vinegar, home made vinegar, grape berries and flowers of selected plants. To this end, we have isolated the bacteria on different selective media in an aerobic atmosphere and identified them by the phenotypic and molecular biological approaches, i.e. by investigating the restriction patterns of the PCR amplified 16S‒23S rDNA regions. Restriction patterns were compared with the restriction patterns of the homologous regions of the reference strains of acetic acid bacteria. From the samples of wine vinegar we isolated strains from the starter culture for vinegar production and from the seven sampling points of one oxidizing cycle for vinegar production, all together 84 isolates. From each of the sampling point one randomly selected isolate was further identified on the basis of the 16S‒23S rDNA restriction profiles as belonging to the group Gluconacetobacer intermedius/Gluconacetobacter oboediens. Further sequence analysis confimed that the strains belong to the species Gluconacetobacter oboediens. Extreme growing conditions in industrial bioreactors for the production of vinegar allow the survival of highly specialized bacterial species, to which belongs also the species identified in this work. Further RAPD analysis could not identify genotypic differences among the analyzed strains, suggesting a high genotypic homogeneity among the strains of red wine vinegar-producing microbiota. From home made vinegar, wherein the concentrations of acetic acid does not exceed 6 %, Acetobacter aceti was isolated. From the surface of the berries and flowers of selected plants we did not succeed to isolate acetic acid bacteria. However, we isolated from flowers the strains, that were on the basis of partial 16S rDNA sequences identified as Rahnella sp., Serratia sp. and Pantoea agglomerans. The species of these genera were already before isolated from this habitat.
Keywords:	acetic acid bacteria, 16S–23S rDNA restriction, identification, habitat

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