Antiserum prodction and reverse transcription polymeraes chain reaction (RT-PCR) for detection of bean leaf roll virus

摘要

Bean leaf roll virus (BLRV) is a recently isolated virus from?Vicia fabae?beans which was collected from eight different locations at Ismailia governorate, Egypt.?This virus is belonging to the genus?luteovirus?and resembles its members in mode of transmission, symptoms and morphological appearance.?In this study, BLRV was identified using multi techniques, direct enzyme linked immunosorbent assay (ELISA), antiserum with purified antigen, insect transmission and reverse transcription-polymerase chain reaction (RT-PCR). All plants collected from El- Tall El-Caber farms gave positive reactions with specific BLRV antibodies. Moreover, typical symptoms of BLRV, stunting, leaf roll, mosaic, chlorosis and yellowing, appeared on all plant cultivars under test (Giza 714, Giza 429, Giza461, Giza717 and Giza3). Six aphid species,?Acyrthosiphon pisum,?Aphis craccivora,?Aphis fabae,?Aphis gossypii,?Macrosiphum euphorbiae?and?Myzus persicae, have obligatory transmitted BLRV in a circulative persistent manner. The percentages of transmission, acquisition and inoculation feeding period have been determined. RT-PCR technique was used for the detection and identification of the isolated virus particles from the infected faba bean plants. Using specific oligonucleotide primers, the coat protein gene,?cp, was amplified from BLRV genome producing 400 bp fragments.