K. Elena & A. Grigoriou
50
digested with the enzymes
Alu
I,
Msp
I and
Taq
I, while the BPIC2514 isolate of
P. syringae
showed a different band pattern (Fig. 5).
Employing the BLAST procedure inNCBI,
the ITS sequence of P1 isolate was found to
share high homology with
P. primulae
strain
CBS620.97 (AF266802.1),
isolated from
Prim-
ula acaulis
with identities 790/791
.
Accord-
ing to a phylogenetic tree based on neigh-
bour-joining analysis [program NCBI/BLAST/
blastin (nucleotide databases)], the species
with the highest degree of similarity to
P.
primulae
was
P. porri
(Fig. 4). After sequenc-
ing, the P1 isolate was deposited to Bena-
ki Phytopathological Institute Collection
(BPIC) with culture number BPIC2584.
Pathogenicity tests
The isolates of the fungus from pars-
ley were pathogenic to parsley, but not to
other plant species. Parsley pants devel-
oped symptoms similar to those observed
in the field and most of them died three
to five weeks after inoculation, while con-
trol plants (non-inoculated) and the oth-
er plant species remained healthy (Table
1).
P. primulae
was reisolated from the in-
fected crowns and roots of the artificially
inoculated plants. The other
Phytophtho-
ra
species tested did not infect the parsley
plants (Table 2).
Apple fruits developed a brown rot from
the wound to the center, while the controls,
the inoculated potato tubers and the onion
bulbs remained healthy (Table 3) (18).
The results of the the repeated experi-
ments and the tests with the other parsley
isolates were similar.
Discussion
Pathogenicity of
P. primulae
was con-
firmed on primula plants, since this plant
species is the main host of
P. primulae
(18).
Tomlinson has identified for the first time
Fig. 4.
Phylogenetic tree for
Phytophthora
species, from PCR amplified products of ribosomal DNA, using the ITS6
and ITS4 primers, based on neighbour-joining analysis by “Macrogen”. The ITS sequence of parsley isolate BPIC2584
(Ic||1_9269) was close to strain CBS620.97 (AF266802) of
P. primulae
. The nearest species, exhibiting the highest de-
gree of similarity to
P. primulae
is
P. porri.
1...,42,43,44,45,46,47,48,49,50,51 53,54,55,56,57,58,59,60