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Molecular Pathogens
MP2011, Vol.2, No.2
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Figure 4 Electron micrographs of ultrathin section sections
from FMV-infected leaf of
F. carica
L. showing profile of long
elongated and flexuous virus-like particles
Note: (C) cytoplasm; (Ch) Chloroplast and (v) virus-like
particles. X=50 000
were used and successfully amplified 969 bp of
NIb
gene of FMV in the infected tissue. This band was
successfully amplified from RNA extracted from
purified virus as well (Figure 5). Sequence and
sequence analysis for the amplified 969 bp revealed
that this DNA fragment was amplified within the
nuclear inclusion body (
NIb
). This sequence was
deposited in GenBank under accession number (Acc#
GQ871933). Phylogenetic analyses, shown in Figure
6a, revealed that the
NIb
, a phylogenetic tree, was
generated from sequence data of 14 virus isolates by
UMPGADA distance analysis with maximum
sequence difference of 0.8. The maximum nucleotide
sequence divergence was exhibited in lineage I.
Meanwhile, the FMV isolates appeared in the other
lineage as monophyletic sister clade, as shown in
Figure 6b. The phylogenetic analysis indicated that the
Egyptian FMV isolate was closely related to Arkansas
(Acc# FJ769161) and Italy (Acc# FM864225) fig
mosaic viruses with similarity 56%. When the
phylogeny was constructed based on the deduced
amino acids sequences, the similarity between the
Italian fig mosaic virus and ours (Egyptian fig mosaic
virus) was increased up to 55%. Meanwhile, the
Arkansas isolate was grouped with other Italian
isolates (FM991954 and FM992851) with similarity
42%.
1.4 Nucleotide sequence and sequence analysis of
CP
gene
RT-PCR using degenerate primer set designed (based
on amino acid conserved region of 67 mosaic virus
Figure 5 PCR products of the
Fig mosaic virus
coat protein
gene and
Fig Mosaic Virus NIB
gene
Note: A: PCR products of the
Fig mosaic virus
coat protein
gene using degenerate primers; B: PCR products of
Fig Mosaic
Virus NIB
gene using universal primers; M: 1 kb Ladder DNA
Marker; 1: Negative control (plant healthy tissues); 2,3: PCR
products amplified from infected fig leaves and from the partial
purified fig virus particles
Figure 6 Phylogenetic analysis based on the nucleotide and the
deduced amino acid sequences
Note: A: Phylogenetic analysis based on the nucleotide
showing the genetic relationship between the
NIb
genes of
FMV with those of selected other viruses; B: Phylogenetic
analysis based on the deduced amino acid sequences showing
the genetic relationship between the
NIb
genes of FMV with
those of selected other viruses