Author/Editor		Ambrožič-Dolinšek, Jana; Ravnikar, Maja; Žel, Jana; Demšar, Tina; Camloh, Marjana; Cankar, Katarina; Dreo, Tanja
Title		Tissue culture of Pyrethrum (Tanacetum cinerariifolium) and associated microbial contamination
Translated title		Tkivna kultura bolhača (Tanacetum cinerariifolium) in z njo povezana okužba z mikroorganizmi
Type		članek
Source		Acta Biol Slov
Vol. and No.		Letnik 53, št. 1
Publication year		2010
Volume		str. 63-68
Language		eng
Abstract		Microbial contamination was observed on several subcultures of Pyrethrum (Tanacetum cinerariifolium) (Trevir.) Schultz-bip. callus lines. The presence of microorganisms was detected by isolation of contaminants in pure culture from 7 out of 34 callus lines and direct amplification of eubacterial 16S rDNA in the pyrethrum callus and plants and isolated bacteria. Altogether 16 contaminants were further analyzed, observing their morphology on several media and restriction of amplified 16S rDNA. Analysis revealed presence and persistence of morphologically and genetically diverse bacteria in pyrethrum tissue culture. Due to cross reactivity of 16S rDNA primers with DNA of plant origin, no conclusions could be drawn on the origin of contaminants.
Summary		Po vzpostavitvi kalusnih linij bolhača (Tanacetum cinerariifolium) (Trevir.) Schultz-bip. smo v kulturah opazili okužbo z mikroorganizmi. Z različnimi izolacijskimi postopki in gojitvenimi metodami smo iz 7 od 34 kalusnih linij izolirali posamezne seve mikroorganizmov. Nobeden od izolatov ni bil izoliran iz več kot dveh kalusnih linij. S pomnoževanjem eubakterijske 16S rDNA in nadaljnjo restrikcijsko analizo z encimom Mn/I smo analizirali kalusno tkivo, rastline bolhača in izolirane bakterije. Analiza je pokazala morfološko in genetsko raznolikost bakterij prisotnih v tkivni kulturi bolhača. Zaradi navzkrižne reaktivnosti uporabljenih 16S rDNA oligonukleotidnih začetnikov, na prisotnost podobnih bakterijskih sevov v kalusnem tkivu in steblih bolhača, ni bilo mogoče sklepati na izvor okužb.