Articles
DETECTION OF BEAN YELLOW MOSAIC VIRUS IN GLADIOLI CORMS
Article number
377_23
Pages
209 – 220
Language
Abstract
Bean yellow mosaic virus (BYMV) could not be detected in corms of infected gladiolus plants by either ELISA or by RNA hybridization.
The virus could be detected by both procedures in crude extracts from parts of the corm which were cut (wounded) 2–5 weeks before testing.
However, the virus concentration in intact corms was below the detection limit of these methods.
Using the polymerase chain reaction (PCR), BYMV RNA could be detected in intact corms.
In a few corms the virus could not be detected by the standard PCR, but was detected by a procedure in which the PCR products from the viral RNA was further amplified by transcription (PCR-TAS). Quantitation of the virus concentration in corms using PCR is discussed.
The mechanism(s) which causes BYMV titer to be low in the corm tissue, and the induction of virus multiplication by wounding or healing effects is not yet understood.
Proteolysis was shown to interfere with serological detection of BYMV in corms.
Proteolytic activity was detected in corm extracts but not from leaves, when purified BYMV was used as a substrate.
Analysis of the reactions of a series of monoclonal antibodies with coat protein from cut corms showed that cleavage of the N-terminus occurred in corm extracts.
Applying a rapid extraction procedure yielded intact coat protein, implying that cleavage occurs during extraction rather than in vivo.
The virus could be detected by both procedures in crude extracts from parts of the corm which were cut (wounded) 2–5 weeks before testing.
However, the virus concentration in intact corms was below the detection limit of these methods.
Using the polymerase chain reaction (PCR), BYMV RNA could be detected in intact corms.
In a few corms the virus could not be detected by the standard PCR, but was detected by a procedure in which the PCR products from the viral RNA was further amplified by transcription (PCR-TAS). Quantitation of the virus concentration in corms using PCR is discussed.
The mechanism(s) which causes BYMV titer to be low in the corm tissue, and the induction of virus multiplication by wounding or healing effects is not yet understood.
Proteolysis was shown to interfere with serological detection of BYMV in corms.
Proteolytic activity was detected in corm extracts but not from leaves, when purified BYMV was used as a substrate.
Analysis of the reactions of a series of monoclonal antibodies with coat protein from cut corms showed that cleavage of the N-terminus occurred in corm extracts.
Applying a rapid extraction procedure yielded intact coat protein, implying that cleavage occurs during extraction rather than in vivo.
Authors
A. Stein, A. Rosner, J. Hammond
Keywords
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