Author/Editor		Demšar, Tina; Toplak, Nataša; Gruden, Kristina; Ravnikar, Maja
Title		Uporaba molekularnih metod za določanje bakterije povzročiteljice obročkaste gnilobe krompirja
Translated title		The use of molecular biology methods for detection of potato ring rot disease
Type		članek
Source		In: Maček J, editor. Zbornik predavanj in referatov 6. slovenskega posvetovanja o varstvu rastlin; 2003 mar 4-6; Zreče. Ljubljana: Društvo za varstvo rastlin Slovenije,
Publication year		2003
Volume		str. 118-22
Language		slo
Abstract		Clavibacter michiganensis (Smith) Davis et aL ssp. sepedonicus (Spieckermamm et Kotthoff) Davis et aL is the causal agent of bacterial ring rot disease of potato. Bacteria is listed on LA.II. List of harmful organisms in Slovenia. Diagnostic of bacteria in our laboratory is conducted as survey in the frame of official plant protection service. It was not yet found in Slovenia, but we had identified bacterium in imported ware potato for the first time in year 2001. A testing procedure according to EU directive (93/85/ECC) is based on screening serological immunofluoresce or ELISA test. If serological test on potato extract is positive, inoculation of test plants Solanum melongena cv. Black Beauty is pertormed. From inoculated test plants bacteria is then isolated on medium and confirmed with different biochemical tests and molecular methods. In our laboratory different molecular methods for detection of ring rot disease in potato extracts and confirmation of bacterial isolates are used: PCR, Real Time PCR and FISH (fluorescent in situ hybridization).
Summary		Obročkasto gnilobo krompirja povzroča bakterija Clavibacter michiganensis (Smith) Davis et al. ssp. sepedonicus (Spieckermamm et Kotthoff) Davis et al. Bakterija je v Sloveniji uvrščena na seznam I.A.II. škodljivih organizmov. Diagnostika bakterije v našem laboratoriju poteka v okviru posebnega nadzora, ki ga vrši uradna služba za varstvo rastlin. V Sloveniji bakterije še nismo našli, prvič pa smo bakterijo identificirali v pošiljkah jedilnega krompirja iz uvoza leta 2001. Postopek laboratorijskega testiranja, ki ga predpisuje direktiva EU (93/85/ECC), temelji na presevnem serološkem imunofluorescenčnem ali ELISA testu. Če je serološki test krompirjevega ekstrakta pozitiven, je potrebna inokulacija testnih rastlin jajčevcev Solanum melongena cv. Black Beauty. Bakterijo iz inokuliranih testnih rastlin izoliramo na gojišča in potrdimo z ustreznimi biokemijskimi testi in molekularnimi metodami. V našem laboratoriju smo za določanje obročkaste gnilobe krompirja uvedli naslednje molekularne metode: PCR, PCR v realnem času ( Real Time PCR) in metodo FISH (fluorescent in situ hybridization), ki omogočajo določanje te bakterije v ekstraktih krompirja in potrditev izolirane bakterijske kulture.
Descriptors		POTATOES PLANT DISEASES BACTERIA ENZYME-LINKED IMMUNOSORBENT ASSAY POLYMERASE CHAIN REACTION IN SITU HYBRIDIZATION, FLUORESCENCE