IMPROVED DETECTION OF PVS BY MOLECULAR HYBRIDIZATION AND BIOLISTIC INOCULATION

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F60109807%3A_____%2F07%3A8P001891" target="_blank" >RIV/60109807:_____/07:8P001891 - isvavai.cz</a>

Result on the web

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DOI - Digital Object Identifier

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Result language

angličtina

Original language name

IMPROVED DETECTION OF PVS BY MOLECULAR HYBRIDIZATION AND BIOLISTIC INOCULATION

Original language description

New extraction procedure was developed to detect potato viruses S and Y by a dot-blot hybridization. While a classical AMESS extraction is ineffective for these potato viruses an improved extraction buffer that contains 5% SDS enabled a sensitive detection without need to purify RNA with an expensive procedures. An alternative biolistic inoculation method was developed for potato virus S (PVS). Biolistic inoculation with CsCl-purified PVS led to strong pathogenic effect on tomato cv. Nevske and to highlevels of PVS accumulation. As detected by the dot-blot hybridization with a "bypass" PVS probe, an infection was achieved even after biolistic inoculation with total RNA isolated from infected plants i.e. without any need to purify virus RNA. This suggests much simpler preparation of biolistic inocula for special handling of plant viruses and their mixtures.

Czech name

ZLEPŚENÁ DETEKCE VIRU S BRAMBORU (PVS) MOLEKULÁRNÍ HYBRIDIZACÍ A BIOLOSTICKÁ INOKULACE.

Czech description

Byla vyvinuta nová extrakční procedura pro detekci viru S a viru Y bramboru pomocí dot-blot hybridizace. Zatímco klasické AMESS extrakční postupy byly pro tyto viry neefektivní, citlivá detekce byla dosažena pomocí pufru s 5% SDS též bez nutnosti purifikovat RNA. Pro PVS byla rovněž vyvinuta alternativní, biolistická metoda inokulace. Touto metodou byl purifikovaným PVS dosaženo silného patogenního účinku u hostitelské rostlin rajčete cv. Něvské a rovněž byla prokázána vysoká úroveň akumulace tohoto viru v rostlině. Dot-blot hybridizace se sondami překlenující vybrané úseky génomu PVS ("bypass sondy) citlivě detekovala infikované rostliny, bez nutnosti purifikovat celkovou RNA.

Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

GF - Diseases, pests, weeds and plant protection

OECD FORD branch

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Project

<a href="/en/project/QF3109" target="_blank" >QF3109: The differentiation of Potato Virus S (PVS) and the identification of isolates of quarantine significance by means of molecular diagnostic techniques.</a><br>

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Publication year

2007

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Vědecké práce

ISSN

1802-940X

e-ISSN

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Volume of the periodical

15

Issue of the periodical within the volume

prosinec

Country of publishing house

CZ - CZECH REPUBLIC

Number of pages

8

Pages from-to

141-148

UT code for WoS article

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EID of the result in the Scopus database

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