Articles
NEW POLYMERASE CHAIN REACTION APPROACH FOR IDENTIFICATION OF DASHEEN MOSAIC VIRUS IN ANTHURIUM ANDRAEANUM AND OTHER RNA PLANT VIRUSES
Article number
1072_19
Pages
157 – 164
Language
English
Abstract
Correct identification of plant diseases caused by viruses, including those of ornamental plants can be accomplished by several methods.
Although serology constitutes the most used method for plant virus identification on a large scale, the use of molecular techniques for plant virus identification and characterization is increasing all over the world.
Several molecular techniques have been developed and the reverse transcription polymerase chain reaction (RT-PCR) has been demonstrated to be a suitable method for research with RNA plant viruses.
In the present study a variation of RT-PCR involving previous virus immune precipitation (IP-RT-PCR) was used for identification of Dasheen mosaic virus (DMV), genus Potyvirus, in anthurium (Anthurium andraeanum), a promising ornamental produced in the State of Ceará, Brazil.
Leaves and flowers from symptomatic plants exhibiting mosaic, chlorotic stripes along the foliar veins were collected in a commercial anthurium plantation in Baturité Mountain in the State of Ceará and taken to the Plant Virus Laboratory at the Federal University of Ceará. Extracts of plant samples were mixed with DMV polyclonal antiserum, and after incubation the virus was concentrated by low centrifugation.
The viral RNA was extracted from the precipitate and analyzed by RT-PCR, using the universal primers NIb2F and NIb3R for detection of virus from the genus Potyvirus. According to the results, the samples from anthurium plants with virus symptoms presented a band corresponding to a DNA fragment of approximately 350 bp which was expected for those primers, confirming the presence of DMV in the symptomatic plants.
The new PCR approach was also efficient for detecting the presence of five other virus species: Cowpea severe mosaic virus (CPSMV); Cucumber mosaic virus (CMV); Squash mosaic virus (SqMV); Zucchini yellow mosaic virus (ZYMV) and Papaya lethal yellowing virus (PLYV).
Although serology constitutes the most used method for plant virus identification on a large scale, the use of molecular techniques for plant virus identification and characterization is increasing all over the world.
Several molecular techniques have been developed and the reverse transcription polymerase chain reaction (RT-PCR) has been demonstrated to be a suitable method for research with RNA plant viruses.
In the present study a variation of RT-PCR involving previous virus immune precipitation (IP-RT-PCR) was used for identification of Dasheen mosaic virus (DMV), genus Potyvirus, in anthurium (Anthurium andraeanum), a promising ornamental produced in the State of Ceará, Brazil.
Leaves and flowers from symptomatic plants exhibiting mosaic, chlorotic stripes along the foliar veins were collected in a commercial anthurium plantation in Baturité Mountain in the State of Ceará and taken to the Plant Virus Laboratory at the Federal University of Ceará. Extracts of plant samples were mixed with DMV polyclonal antiserum, and after incubation the virus was concentrated by low centrifugation.
The viral RNA was extracted from the precipitate and analyzed by RT-PCR, using the universal primers NIb2F and NIb3R for detection of virus from the genus Potyvirus. According to the results, the samples from anthurium plants with virus symptoms presented a band corresponding to a DNA fragment of approximately 350 bp which was expected for those primers, confirming the presence of DMV in the symptomatic plants.
The new PCR approach was also efficient for detecting the presence of five other virus species: Cowpea severe mosaic virus (CPSMV); Cucumber mosaic virus (CMV); Squash mosaic virus (SqMV); Zucchini yellow mosaic virus (ZYMV) and Papaya lethal yellowing virus (PLYV).
Authors
J.A.A. Lima, A.K.Q. Nascimento
Keywords
virus diagnosis, molecular virology, serology, Bromovirus, Comovirus, Potyvirus, Sobemovirus
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