Genome Analyses of Carnation Mottle Virus Iranian Isolate (FUM2)

پژوهش های حفاظت گیاهان ایران

Article 6, Volume 25, Issue 2 - Serial Number 15, June 2011   

Document Type: Research Article

DOI: 10.22067/jpp.v25i2.10102

Authors

B. Jafarpour^* ; Mohammad Ali Sabokkhiz; M. Falahati Rastegar; S. Pakbaz

Abstract

Abstract
During the winter and spring of 2007 and 2008, 450 samples (leaves of carnation plants) showing symptoms of yellowing, mottling, leaf malformation, and stunting were collected from greenhouses in Mashhad and Chenaran regions. The samples were tested by DAS-ELISA for the presence of CarMV. Total RNA was extracted by RNXTM (-Plus) solution from positive infected samples confirmed in DAS-ELISA tests. AccuPowerR RT Pre Mix Kit was used for synthesis of cDNA with reverse specific primer according to two segments of genome. PCR amplification of cDNA was carried out using AccupowerR PCR PreMix. RT-PCR assay amplified two DNA fragments approximately 1037bp and 676 bp using. PCR products directly sequenced. The nucleotide sequence identity was also compared with different isolates of the world. The determined sequences of FUM2 isolate of CarMV were compared which previously reported 23 CarMV isolates, using Bioedit software and ClastalW2. A Neighbor-joining method of MEGA 3.1 was applied to construct unrooted trees for 3 genes (p7, p9 and partial p38). Analysis of the phylogenetic tree showed that our isolate is in group I and subgroup A.

Keywords: Carnation mottle virus (CarMV), FUM2, RT-PCR, Phylogenetical position, Iran