Prvi rezultati uporabe novih laboratorijskih metod za ugotavljanje ZASTOPAnosti virusa, ki povzroča razbrazdanje lesa na vinski trti v Sloveniji  

Nataša PETROVIČ¹, Petra ŠOSTER², Zora KOROŠEC - KORUZA³, Maja RAVNIKAR⁴, Bao Zhong MENG⁵ in Dennis GONSALVES ⁶

^1,2,4, Nacionalni inštitut za biologijo, Oddelek za rastlinsko fiziologijo in biotehnologijo, SI-1000 Ljubljana, Slovenija,
^5,6 Department of  Plant Pathology, Cornell University, New York State Agricultural Experiment Station (NYSAES), Geneva, NY-14456, ZDA,  
³ Univerza v Ljubljani, Biotehniška fakulteta, Oddelek za agronomijo, SI-1000 Ljubljana, Slovenija

Zdravstvena selekcija je nepogrešljiv del klonske selekcije vinske trte, ki jo v Sloveniji izvajajo že od leta 1956, od leta 1990 pa z uporabo ELISA testiranja. Testiranje na zastopanost virusov in potrjevalna shema ustrezajo mednarodnim in evropskim priporočilom in standardom in poteka v Sloveniji v dveh trsničarskih centrih, koordinirata in nadzorujeta pa ga Kmetijski inštitut Slovenije in Oddelek za agronomijo Biotehniške fakultete Univerze v Ljubljani.

Zanimivo in plodno sodelovanje se je pričelo med Nacionalnim inštitutom za biologijo, Oddelkom za agronomijo BF Univerze v Ljubljani in Oddelkom za rastlinsko patologijo Univerze Cornell v ZDA. Skupaj uvajamo alternativne hitre laboratorijske metode za ugotavljanje zastopanosti povzročitelja bolezni razbrazdanja pri vinski trti (Rupestris stem pitting associated virus 1, RSPaV-1). Nove detekcijske metode smo prenesli iz ameriškega  v slovenski laboratorij za ovrednotenje njihovega delovanja v smislu rutinskih analiz. Metode predstavljajo: serološke metode (imunski pivnik in ELISA) in molekularne metode (RT-PCR). Uvajanje tehnik je rodilo že prve zanimive rezultate analize zastopanosti RSPaV-1 virusa v indikatorski vrsti Vitis rupestris cv. Saint George ter Vitis vinifera cv. Refošk, ki smo ju testirali zaradi njunega posebnega statusa: trsi V. rupestris St. George služijo odvzemu cepičev za indikatorje pri indeksiranju, izbrani trsi Refoška pa že nekaj let kažejo znake razbrazdanja lesa bodisi na podlagi ali na žlahtnem cepljenem delu.

Preliminarne analize kažejo okuženost vseh šestih od šestih analiziranih trsov V. rupestris cv. St. George in okuženost štirih od petih analiziranih trsov V. vinifera cv. Refošk. Zanimivo je dejstvo, da dva od štirih okuženih trsov Refoška ne kažeta nobenih znakov razbrazdanja, eden kaže hude znake razbrazdanja na podlagi in eden na žlahtnem cepljenem delu. Ti rezultati nakazujejo velik pomen laboratorijskih metod za ugotavljanje zastopanosti RSPaV-1, tako za preverjanje zdravstvenega stanja materiala, ki se uporablja za indikatorje pri indeksiranju, kakor tudi za ugotavljanje latentnih okužb s tem virusom. 

ABSTRACT

First results on the use of laboratory methods for detection of rupestris stem pitting associated virus 1 in grapevines in Slovenia

Sanitary selection is an essential part of the grapevine clonal selection, which has been in Slovenia conducted since 1956, and since 1990 by extensively using ELISA testing. Virus testing and certification follow international and European recommendations and standards. The routine testing is conducted at the two Selection stations - nuclei, and co-ordinated and supervised by the Agricultural Institute of Slovenia and by Department of Agronomy at the Biotechnical Faculty of the University of Ljubljana.

An interesting and fruitful cooperation has started between the National Institute of Biology, Biotechnical Faculty of the University of Ljubljana in Slovenia, and Department of Plant Pathology at the Cornell University in the USA in introducing alternative quick laboratory methods for a detection of the causal agent of rupestris stem pitting disease, a rupestris stem pitting associated virus 1 (RSPaV-1). New potential detection techniques and tools have been transferred from the US to Slovenian laboratory as part of the evaluation of their performance for the routine use. They represent serological techniques (ELISA and Western blot) and RT-PCR. The techniques have already generated some interesting preliminary results on the presence of the RSPaV-1 in V. rupestris cv. St. George, and in V. vinifera cv. Refošk, which have been both analysed because of their special status: tested vines of St. George are stock material for  indexing, and selected cv. Refošk was known from the previous research for its severe decline of vines due to all forms of rugose wood disease.

Our preliminary analyses show the infection of six (out of six analysed) St. George plants, and the infection of four (out of five analysed) Refošk vines. Interestingly, two of the positively analysed Refošk vines show no symptoms, one is showing severe symptoms of rugose wood on the rootstock, and one on the upper grafted part. The preliminary results indicate the great importance of the laboratory methods for detection of RSP disease agents, for the purposes of  re-evaluation of the rupestris indexing material, and for detection of latent infections.