containing
breeding
line
Massachusetts
#2,
for
this
gene
was
found
in
'an
aberrant
plant
of
the
breeding
line
Massachusetts
#2'
.
This
plant
was
resistant
to
race
4,
but
susceptible
to
another
race,
which
later
appeared
to
be
2
.
3
.
4
.
11
(Kerr
&
Patrick,
1977)
.
The
Cf
-
11
line
was
crossed
to
other
(unknown)
lines,
resulting
in
Ontario
7716,
which
was
used
in
later
experiments
.
Upon
inoculation
of
different
plant
genotypes
with
the
systemic
virus
PVX,
expressing
the
avirulence
gene
avr4,
Vantage,
Moneymaker
-
Cf
-
4
(MM
Cf
-
4
),
Ontario
7522
and
Ontario
7716
exhibited
necrosis,
indicating
that
these
lines
all
contain
an
active
Cf
-
4
gene
.
Southern
analysis
with
genomic
DNA
of
these
lines
digested
with
EcoRl,
Bg/ll,
Hindlll
and
EcoRV
and
probed
with
a
Cf
-
4
or
a
Cf
-
9
probe
indicated
that
the
patterns
of
MM
Cf
-
4
,
Ontario
7522
and
Ontario
7715
were
identical
and
different
from
control
lines
.
In
experiments
using
PCR
primers
derived
from
the
DNA
sequence
of
Cf
-
9
no
difference
in
PCR
pattern
between
MM
Cf
-
4
,
Ontario
7522
and
Ontario
7716
was
detected
again
.
Later,
specific
PCR
primers
were
designed,
based
on
the
sequence
of
Cf
-
4
.
When
used
at
high
annealing
temperatures,
no
difference
was
found
between
these
lines,
also
after
digestion
of
the
PCR
products
with
Bg
/ll,
Eco
RI,
Hin
cll,
Hind
lll
and
Eco
RV,
while
they
did
differ
from
banding
patterns
of
other
lines
used
.
The
experiments
mentioned
in
this
paper
suggest
the
presence
of
a
functional
Cf
-
4
gene
in
the
Cf
-
8
and
Cf
-
11
lines
.
The
line
Cf
-
11
seems
to
harbour
another
gene,
Cf
-
11
,
for
which
a
corresponding
avirulence
gene
exists
.
Because
the
Cf
-
4
gene
was
already
present
in
Massachusetts
#2
and
Cf
-
11
was
found
in
a
plant
of
the
same