A rapid and effective RNA release procedure for virus detection in woody plants by reverse transcription-polymerase chain reaction.
A rapid and effective RNA release procedure (RNA-RP) for detection of the Apple stem pitting virus (ASPV) and the Apple stem grooving virus (ASGV) in woody plants by reverse transcription-polymerase chain reaction (RT-PCR) was developed. RNA-RP released RNA into crude homogenate. RNA-RP was compared with classical phenol/chlorophorm extraction of RNA. The RT-PCR detection of ASPV and ASGV was shown to be similar by both the RNA preparation procedures. RNA-RP in contrast to the classical phenol/chloroform procedure represents a reliable and easy-to hand protocol convenient for routine virus detection. Leaf tissues, especially dormant bud leaves and leaves during blossom, were shown to be the most optimal material for detection of these viruses, irrespective of the used RNA preparation procedure.
Kundu
J K
JK
Department of Virology, Research Institute of Crop Production, Drnovská 507, 161 06 Prague 6, Czech Republic. jiban@vurv.cz
D027845:Malus / Q000821:virology*; D010935:Plant Diseases / Q000821:virology*; D018515:Plant Leaves / Q000821:virology; D010942:Plant Viruses / Q000235:genetics / Q000302:isolation & purification*; D012328:RNA Viruses / Q000235:genetics / Q000302:isolation & purification*; D012367:RNA, Viral / Q000302:isolation & purification*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors
D012367:RNA, Viral