ARS | Publication request: An Immunocapture RT-PCR Procedure Using Apple stem grooving virus Antibodies Facilitates Molecular Genetic Characterization of Citrus tatter leaf virus from the Original Meyer Lemon Host

Submitted to: Plant Disease
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: August 15, 2007
Publication Date: February 1, 2008
Citation: Hilf, M.E. 2008. An Immunocapture RT-PCR Procedure Using Apple stem grooving virus Antibodies Facilitates Molecular Genetic Characterization of Citrus tatter leaf virus from the Original Meyer Lemon Host. Plant Disease. 92:746-750.

Interpretive Summary: This work describes the development of a sensitive laboratory assay for the detection of citrus tatter leaf virus, which causes a graft incompatibility when it infects citrus which is grafted on rootstocks which are commonly used in the US citrus industry. This technique will augment the standard method of plant inoculation to observe symptom development by this virus and make detection of this virus in important citrus budwood sources more accurate and sensitive.

Technical Abstract: A magnetic bead-based immonocapture system using polyclonal antiserum against Apple stem grooving virus (ASGV) successfully facilitated polymerase chain reaction (PCR) amplification of sequences from three Citrus tatter leaf virus (CTLV) isolates originally isolated from the citrus host Meyer lemon. Primers designed from a pairwise alignment of genomic sequences of CTLV isolates from lily and from kumquat amplified two non-overlapping genomic regions of 625 and 1165 bp (~28% of the CTLV genome) which were cloned and sequenced. Despite being propagated separately in the glasshouse for more than forty years, the CTLV sequences from separate Meyer lemon sources were identical, but had only ~80% nucleotide identity with the homologous regions of CTLV genomes of isolates from lily and from kumquat. Neighbor-joining phylogenetic analysis indicated the CTLV isolates from Meyer lemon were distinct from but more closely related to CTLV from kumquat than from lily, and these CTLV sequences showed equivalent genetic distances from two ASGV isolates.