An Immunocapture RT-PCR Procedure Using Apple stem grooving virus Antibodies Facilitates Analysis of Citrus tatter leaf virus from the Original Meyer Lemon Host

摘要

A magnetic bead-based immunocapture 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 nonoverlapping genomic regions of 625 and 1,165 bp (approximately 28% of the CTLV genome) which were cloned and sequenced. Despite being propagated separately in the glasshouse for more than 40 years, the CTLV sequences from separate Meyer lemon sources were identical but had only approximately 80% nucleotide identity with the homologous regions of CTLV genomes of isolates from lily and 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.