Articles
DETECTION OF PELARGONIUM FLOWER BREAK CARMOVIRUS USING ELISA AND A TRANSCRIBED RNA PROBE
Article number
432_41
Pages
338 – 345
Language
Abstract
Pelargonium, probably the best-selling pot plant in the world, is propagated mainly vegetatively, with viral diseases consequently being a major threat to the production and quality of the crop.
Pelargonium flower break carmovirus (PFBV) which is the most common virus affecting the plant in Israel and Europe, was purified recently and used for the production of a polyclonal antiserum.
An ELISA system enabling the detection of PFBV in plant tissue, was used in surveys of propagation nurseries, where PFBV was found to be quite widespread, especially in propagation material imported from Europe.
During ELISA indexing of commercial Israeli nurseries for PFBV, plants initially indexed as negative were found to be positive in subsequent tests.
An in vitro transcribed RNA probe (riboprobe) system was developed to detect PFBV in tissue extracts.
RNA transcripts of a 1500 bP complementary DNA (cDNA) fragment inserted into plasmid pGEM-7Zf(+) were obtained, using T7 RNA polymerase.
Dot blot hybridization using 32P-labeled T7 transcripts was compared to ELISA for the detection of PFBV in pelargonium tissues.
The virus could be detected efficiently in high dilutions of sap from infected plants and at low concentrations of purified virus.
The riboprobe and ELISA are both powerful tools for the detection of PFBV, the limits of detection by cRNA hybridization being similar to ELISA. The relatively low sensitivity of the riboprobe may probably be due to a recombination of fragments.
Pelargonium flower break carmovirus (PFBV) which is the most common virus affecting the plant in Israel and Europe, was purified recently and used for the production of a polyclonal antiserum.
An ELISA system enabling the detection of PFBV in plant tissue, was used in surveys of propagation nurseries, where PFBV was found to be quite widespread, especially in propagation material imported from Europe.
During ELISA indexing of commercial Israeli nurseries for PFBV, plants initially indexed as negative were found to be positive in subsequent tests.
An in vitro transcribed RNA probe (riboprobe) system was developed to detect PFBV in tissue extracts.
RNA transcripts of a 1500 bP complementary DNA (cDNA) fragment inserted into plasmid pGEM-7Zf(+) were obtained, using T7 RNA polymerase.
Dot blot hybridization using 32P-labeled T7 transcripts was compared to ELISA for the detection of PFBV in pelargonium tissues.
The virus could be detected efficiently in high dilutions of sap from infected plants and at low concentrations of purified virus.
The riboprobe and ELISA are both powerful tools for the detection of PFBV, the limits of detection by cRNA hybridization being similar to ELISA. The relatively low sensitivity of the riboprobe may probably be due to a recombination of fragments.
Authors
A. Franck, A. Gera, Y. Antignus, G. Loebenstein
Keywords
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