Molecular detection of apple chlorotic leaf spot virus in different hosts in Central Anatolia

Abstract: Partial molecular properties of apple chlorotic leaf spot virus (ACLSV) isolates were studied on previously known ACLSV infected pear and quince trees. Isolates were taken from infected fruit trees in Ankara. RT PCR test was used for molecular detection. cDNA Sequence analysis was performed on the coat protein (CP) coding regions for isolates from pear and quince, in order to determine the sequence similarity among ACLSV isolates. Sequences of the investigated isolates were also compared with those of genome fragments of ACLSV isolates published in the GenBank database. The similarity in the percentage figures of the sequence variability of sequences of pear isolates and GenBank ACLSV sequences was between 79 and 92%, respectively. DAS ELISA and biological indexing to bean were used for verification. Results were obtained with total nucleic acid preparations from a set of ACLSV isolates taken leaf samples of pear and quince trees. ACLSV was not detected in symptomless leaves of these trees. RT-PCR amplification of ACLSV isolates gave expected products at 677 bp with the ACLSV primer set. Virus-free trees were differed from virus infected trees by the conventional RT-PCR assay. The nucleotide sequence of a pear isolate was deposited in the GenBank database under accession numbers HM490312. First ACLSV isolates was sequenced and deposited in GenBank and alternative detection method to biological indexing and ELISA was presented in use.