Cloves crop is a routinely practice often used in procedures for detecting allexiviruses in garlic bulbs to obtainment foliar tissue to be analyzed by serological and/or molecular tests. The availability of the plants under greenhouse implies is expenses with the maintenance and requires abaut 30 days. In areas free of these viruses, there is also the risk of their introduction and dissemination. This study presents an adjustment of protocol aiming a fast detection of allexiviruses, using primordial leaves. Cloves of consumption-garlic, from Rio Grande do Sul, Brazil, and imported from Argentina were dissected for obtaining primordial leaves and total RNA extraction, using 0,1g of tissue of each sample. RT-PCR reactions were performed with a pair of primers able to amplify a 500 bp fragment, corresponding to the internal region of the coat protein gene from several species of Allexivirus genus. A band in the expected height (500 pb) was visualized in agarose gels and further confirmed using Southern Blot test and by sequencing as Garlic virus C (GarV-C, AY170322.1). Total RNA obtained from foliar primordia of cloves and its use in RT-PCR analysis is an economic, fast and secure methodology for allexivirus detection in garlic bulbs.
Allium sativum; allexiviruses; Southern Blot; RT-PCR
