Avtor/Urednik		Štebih, Dejan; Dreo, Tanja; Demšar, Tina; Blatnik, Aleš; Petrovič, Nataša; Ravnikar, Maja
Naslov		Določanje in identifikacija bakterije Erwinia amylovora (Burrill) Winslow et al. v okuženi hruški
Prevedeni naslov		Detection and identification of Erwinia amylovora (Burrill) Winslow et al. in infected pear tree
Tip		članek
Vir		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,
Leto izdaje		2003
Obseg		str. 467-71
Jezik		slo
Abstrakt		In the frame of systematic survey, conducted by official plant protection service in Slovenia, National Institute of Biology performs laboratory diagnosis of bacteria Erwinia amylovora (Burrill) Winslow et al., the causal agent of fire blight. Frequently, samples with characteristic symptoms arrive to the laboratory. However, similar symptoms can be caused by other plant pathogens such as Pseudomonas syringae pv. syringae which has already been demonstrated in the past. In 2001, Erwinia amylovora was detected in a pear sample with symptoms of fire blight for the first time in Slovenia. Different methods that can be used for detection of Erwinia amylovora were used on infected material with and without symptoms. Isolation on media (KB, NSA), pathogenicity test on pear slices, indirect immunofluorescence (IIF), agglutination test and PCR with different sets of primers were performed. Isolated bacteria were sent for confirmation with analysis of fatty acids profile to Plant Protection Service in Wageningen, the Netherlands. As expected, Erwinia amylovora was detected with IIF test in all samples with symptoms, where concentration of bacteria is high but also in asymptomatic samples with lower concentration of bacteria that have been collected from the same pear tree. Other tests were less reliable particularly in case of latent infection. We established, that the DNA extraction using DNeasyTM Tissue Kit reduces inhibition in PCR reaction compared to extraction by boiling of the sample. Avoiding inhibition is essential, particularly when the concentration of bacteria in sample is low. For reliable and accurate laboratory determination of Erwinia amylovora, the use of several properly combined detection methods is recommended.
Izvleček		V okviru sistematičnega nadzora rastlin, ki ga vodi Uprava RS za varstvo rastlin in semenarstvo, izvajamo na Nacionalnem inštitutu za biologijo laboratorijsko diagnostiko povzročiteljice bakterijskega hruševega ožiga, bakterije Erwinia amylovora (Burrill) Winsiow et aL Pogosto prispejo vzorci z značilnimi bolezenskimi znamenji, ki pa jih lahko povzročijo drugi povzročitelji bolezni, kot na primer bakterija Pseudomonas syringae pv. syringae, ki smo jo v preteklosti že nekajkrat identificirali. V letu 2001 smo v vzorcu hruške z bolezenskimi znamenji, značilnimi za bakterijski hrušev ožig, določili bakterijo Envinia amylovora. To je bil prvi primer najdbe hruševega ožiga v Sloveniji. Na okuženem materialu smo uporabili različne metode, ki jih je možno uporabljati za določanje bakterije Envinia amylovora in sicer na poganjkih z bolezenskimi znamenji in na poganjkih brez njih. Za določanje bakterije Erwinia amylovora smo uporabili uveljavljene metode določanja te bakterije: izolacija na gojišču (KB, NSA), test patogenosti na rezinah hrušk. Od seroloških metod smo izvedli test indirektne imunofluorescence (IIF), od molekularnih pa test PCR z raziičnimi pari začetnih oligonukleotidov. Izolirano bakterijo smo poslali v potrditev z analizo profila maščobnih kislin v Piant Protection Service v Wageningen na Nizozemskem. (Izvleček skrajšan na 2000 znakov)
Deskriptorji		FRUIT PLANT DISEASES ERWINIA DNA, BACTERIAL POLYMERASE CHAIN REACTION