Articles
DETECTION OF IRIS YELLOW SPOT VIRUS IN LISIANTHUS
Article number
568_4
Pages
43 – 49
Language
English
Abstract
Recently, unusual viral symptoms of systemic necrosis, necrotic spots and rings were observed on lisianthus (Eustoma russellianum) leaves.
Preliminary analyses suggested that the disease was caused by a tospovirus.
Crude sap from symptomatic tissue was mechanically transmitted to Nicotiana benthamiana, Chenopodium quinoa, C. amaranticolor, and Gomphrena globosa. On inoculated plants of N. benthamiana, chlorotic spots developed on inoculated leaves followed by systemic necrosis.
The virus produced local spots on inoculated plants of C. quinoa, C. amaranticolor, and G. globosa. Leaf samples of lisianthus and N. benthamiana were analyzed by transmission electron microscopy (TEM) in leaf dip preparations and thin sections of leaf tissues.
Virus particles typical of a Tospovirus were observed only in samples taken from symptomatic leaves.
Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) tests of leaf sap extracted from naturally infected lisianthus and mechanically inoculated indicator plants, gave a strong positive reaction to Iris yellow spot virus (IYSV), demonstrated that this virus was serologically related to IYSV. Polyclonal antibodies prepared against the nucleocapsid of the virus enable specific detection of the virus in crude sap from infected plants.
Primers specific to the nucleocapsid gene of IYSV were used in a reverse transcription-polymerase chain reaction assay (RT-PCR) to verify the presence of IYSV. RT-PCR gave an expected PCR product of approximately 850 bp.
The amplicon was cloned in pGEM-T vector and the recombinant clone was sequenced.
The sequence of the cloned nucleocapsid gene confirmed the identity of IYSV, thus verifying IYSV infection of lisianthus.
Preliminary analyses suggested that the disease was caused by a tospovirus.
Crude sap from symptomatic tissue was mechanically transmitted to Nicotiana benthamiana, Chenopodium quinoa, C. amaranticolor, and Gomphrena globosa. On inoculated plants of N. benthamiana, chlorotic spots developed on inoculated leaves followed by systemic necrosis.
The virus produced local spots on inoculated plants of C. quinoa, C. amaranticolor, and G. globosa. Leaf samples of lisianthus and N. benthamiana were analyzed by transmission electron microscopy (TEM) in leaf dip preparations and thin sections of leaf tissues.
Virus particles typical of a Tospovirus were observed only in samples taken from symptomatic leaves.
Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) tests of leaf sap extracted from naturally infected lisianthus and mechanically inoculated indicator plants, gave a strong positive reaction to Iris yellow spot virus (IYSV), demonstrated that this virus was serologically related to IYSV. Polyclonal antibodies prepared against the nucleocapsid of the virus enable specific detection of the virus in crude sap from infected plants.
Primers specific to the nucleocapsid gene of IYSV were used in a reverse transcription-polymerase chain reaction assay (RT-PCR) to verify the presence of IYSV. RT-PCR gave an expected PCR product of approximately 850 bp.
The amplicon was cloned in pGEM-T vector and the recombinant clone was sequenced.
The sequence of the cloned nucleocapsid gene confirmed the identity of IYSV, thus verifying IYSV infection of lisianthus.
Authors
A. Gera, A. Kritzman, H. Beckelman, J. Cohen, B. Raccah
Keywords
detection, Gentianaceae, nucleocapsid protein, sequence, tospovirus
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