Total genomic DNA of all isolates of Alternaria was isolated
by CTAB method and molecular characterization by ITS sequence. The genetic
identification of different isolates of Alternaria was confirmed by the DNA
sequence analysis, a rapid mean of identification. Sequence similarities were
determined by molecular method using DNA and ITS region. DNA analysis not only
enables the species identification but also permits phylogenetic analysis.
Internally transcribed spacer (ITS) region including 5.8S rRNA coding region in
ribosomal DNA is one of the favorite targets for this purpose. The ITS region
was amplified using the polymerase chain reaction (PCR) and the universal
primers ITSl and ITS4. The focus of present study was on the genetic diversity
analysis of the ITS regions of rRNA gene complex of local isolates of
Alternaria in Pakistan. The genomic DNA of these isolates KI1, KI4, KI5, KI6,
KI, KI8, KI9, and KI10 was amplified using primers ITS1 and ITS4 designed at
the end, and start of conserved 18S and 28S region and between ITS1 and ITS4
respectively. By the comparison of isolates of all species have maximum genetic
diversity only KI7 and KI8 have maximum homology with each other that showed
that both are may be originated from same ancestor. Their phylogenetic
relationships in GenBank were analyzed. Morphological and molecular data
obtained might be useful in determining the taxonomy and diversity of
Alternaria species.
No comments:
Post a Comment