Document ID: EPA-HQ-OPP-2006-0299-0003
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2006-05-26T04:00Z

Monsanto
Company
04­
CR­
138E­
1
Page
1
of
11
Title
Petition
to
Establish
a
Temporary
Exemption
from
the
Requirement
of
a
Tolerance
for
Bacillus
thuringiensis
Cry2Ab2
Insecticidal
Protein
and
the
Genetic
Material
Necessary
for
Its
Production
in
Corn
When
Used
as
Plant­
incorporated
Protectant
in
Food
and
Feed
Commodities
of
Field
Corn,
Sweet
Corn,
and
Popcorn
Volume
1
of
1
Author
N.
N.
Bogdanova,
D.
V.
M.

Date
June
28,
2005
Submitted
By
Monsanto
Company
1300
I
(
Eye)
Street,
NW
Suite
450
East
Washington,
DC
20005
Monsanto
Reference
No.

04­
CR­
138E­
1
Monsanto
Company
04­
CR­
138E­
1
Page
2
of
11
Table
of
Contents
Page
Introduction
..................................................................................................................
3
Section
A.
The
name,
chemical
identity,
and
composition
of
the
plant­
incorporated
protectant......................................................................
4
Section
B.
The
amount,
frequency,
and
time
of
application
of
the
plant­
incorporated
protectant......................................................................
5
Section
C.
Full
reports
on
investigations
made
with
respect
to
the
safety
of
the
plant­
incorporated
protectant..................................................
6
Section
D.
The
results
of
tests
on
the
amount
of
plant­
incorporated
protectant
remaining,
including
a
description
of
the
analytical
method
used
...............
7
Section
E.
Practical
methods
for
removing
plant­
incorporated
protectant
that
exceeds
any
proposed
tolerance...........................................................
8
Section
F.
Proposed
tolerances
for
the
plant­
incorporated
protectant
if
tolerances
are
proposed...........................................................................
9
Section
G.
Reasonable
grounds
in
support
of
this
petition
..........................................
10
Attachment
I.
Applicant's
notice
of
filing
summary
........................................................
11
Monsanto
Company
04­
CR­
138E­
1
Page
3
of
11
Introduction
On
May
11,
2001,
the
EPA
granted
a
time­
limited
exemption
from
the
requirement
of
a
tolerance
for
the
plant­
incorporated
protectant,
Bacillus
thuringiensis
Cry2Ab2
protein
and
the
genetic
material
necessary
for
its
production
in
food
and
feed
commodities
of
field
corn,
sweet
corn,
and
popcorn
(
corn
event
MON
840
[
vector
ZMBK28]),
and
in
cotton,
cotton
seed,
cotton
oil,
cotton
meal,
cotton
hay,
cotton
hulls,
cotton
forage,
and
cotton
gin
byproducts
when
applied/
used
as
plant
pesticide
(
40
CFR
§
180.1215).
Development
of
MON
840
has
since
been
discontinued
and
that
time­
limited
exemption
expired
on
May
1,
2004.

Subsequently
Monsanto
has
developed
a
second
generation
insect­
protected
corn,
vector
ZMIR245,
that
provides
activity
against
an
increased
spectrum
of
lepidopteran
insect
pests.
Corn
developed
from
vector
ZMIR245
produces
two
insecticidal
proteins,
Cry1A.
105
and
Cry2Ab2.
The
combination
of
Cry1A.
105
and
Cry2Ab2
in
a
single
plant
allows
better
insect
control
and
offers
a
tool
for
management
of
potential
insect
resistance.
The
Cry2Ab2
protein
and
its
expression
cassette
in
ZMIR245
are
identical
to
those
in
MON
840.

By
way
of
this
petition,
Monsanto
is
requesting
that
EPA
establish
a
temporary
exemption
from
the
requirement
of
a
tolerance
for
the
Cry2Ab2
in
corn
as
part
of
an
Experimental
Use
Permit
524­
EUP­
OT.
The
results
of
product
safety
studies
already
submitted
to
and
reviewed
by
EPA
support
this
petition.
A
summary
of
the
results
of
these
studies
has
been
prepared
in
accordance
with
the
formatting
guidelines
delineated
in
PR
Notice
97­
1.
Monsanto
consents
to
publication
of
this
summary
as
part
of
the
Agency's
notice
of
filing
in
the
Federal
Register
and
as
part
of
the
proposed
or
final
regulation.

Petitioner:

__________________________________
Natalia
N.
Bogdanova,
D.
V.
M.
Regulatory
Affairs
Manager
Monsanto
Company
Monsanto
Company
04­
CR­
138E­
1
Page
4
of
11
Section
A:
The
name,
chemical
identity,
and
composition
of
the
plant­
incorporated
protectant
The
Bacillus
thuringiensis
var.
kurstaki
protein
Cry2Ab2
when
used
as
plant­
incorporated
protectant
is
the
subject
of
this
tolerance
exemption
petition.
This
exemption
would
include
the
genetic
material
necessary
for
production
of
Cry2Ab2
in
all
food
and
feed
commodities
of
field
corn,
sweet
corn,
and
popcorn.

The
nomenclature
for
the
Cry2Ab2
protein
has
changed
over
the
past
years.
This
protein
has
been
referred
to
as
Cry2Ab,
Cry2Ab2,
Insect
Protected
Protein
2
(
IPP2),
and
B.
t.
k.
P2.
Monsanto
Company
04­
CR­
138E­
1
Page
5
of
11
Section
B:
The
amount,
frequency,
and
time
of
application
of
the
plant­
incorporated
protectant
Because
the
Cry2Ab2
is
a
plant­
incorporated
protectant,
it
is
not
applied
to
plants.
The
protein
has
been
found
to
be
present
in
corn
plants
throughout
their
development
and
in
all
tissues.
Levels
of
Cry2Ab2
in
corn
tissues
previously
reviewed
by
the
Agency
are
described
in
the
report
listed
below.

MRID
450863­
06
Tonnu,
B­
P.,
J.
C.
Jennings,
and
R.
P.
Lirette.
2000.
Insect
protection
protein
2
and
NPTII
levels
in
samples
collected
from
corn
event
MON
840
in
1998
US
field
trials.

Levels
of
the
Cry2Ab2
protein
in
corn
events
derived
from
the
vector
ZMIR245
are
as
follows:

Tissue
Cry2Ab2
levels
(
µ
g/
g
dry
weight)
Leaf
27
­
93
Pollen
<
LOQ1
Grain
1.4
­
3
Forage
33
­
100
1
Limit
of
Quantitation
for
the
Cry2Ab2
ELISA
is
0.10
µ
g/
g/
dwt.
Monsanto
Company
04­
CR­
138E­
1
Page
6
of
11
Section
C:
Full
reports
of
investigations
made
with
respect
to
the
safety
of
the
plantincorporated
protectant
Monsanto
has
submitted
data
that
establish
the
food
safety
of
Cry2Ab2
protein
when
used
as
a
plant­
incorporated
protectant.
These
data
are
summarized
in
the
attached
Applicant's
Notice
of
Filing
Summary.

The
following
studies
have
been
reviewed
and
found
acceptable
by
the
Agency:

MRID
Toxicity
449666­
02
Bechtel,
C.
L.
1999.
Acute
oral
toxicity
study
of
insect
protection
protein
2
(
IPP2)
in
mice.

Digestive
Fate
449666­
03
Leach,
J.
N.,
R.
E.
Hileman,
J.
W.
Martin,
M.
A.
Nemeth,
and
J.
D.
Astwood.
1999.
Assessment
of
the
in
vitro
digestibility
of
insect
protection
protein
2
(
IPP2)
utilizing
mammalian
digestive
fate
models.

Allergenicity
449666­
04
Hileman,
R.
E.
and
J.
D.
Astwood.
1999.
Bioinformatics
analysis
of
insect
protection
protein
2
(
IPP2)
sequence
utilizing
an
allergen
database.

449666­
05
Hileman,
R.
E.
and
J.
D.
Astwood.
1999.
Bioinformatics
evaluation
of
insect
protection
protein
2
(
IPP2)
sequence
utilizing
toxin
and
public
domain
genetic
databases.
Monsanto
Company
04­
CR­
138E­
1
Page
7
of
11
Section
D:
The
results
of
tests
on
the
amount
of
plant­
incorporated
protectant
remaining,
including
a
description
of
the
analytical
method
used
Another
analytical
method
should
not
be
required
because
this
petition
requests
a
temporary
exemption
from
the
requirement
of
a
tolerance.
In
addition,
an
analytical
method
for
detection
of
the
Cry2Ab2
proteins
is
available
commercially,
2
and
a
Monsanto­
validated
analytical
method
has
also
been
submitted
previously
to
EPA:

MRID
454154­
01
Kolwyck,
D.,
K.
Gustafson,
and
T.
B.
Stone.
2001.
Validated
method
for
extraction
and
direct
ELISA
analysis
of
Cry2Ab2
in
corn
grain.

2
QuickStixTM
Combo
Kit,
Envirologix,
Cat.
#
AS
012
LS
Monsanto
Company
04­
CR­
138E­
1
Page
8
of
11
Section
E:
Practical
methods
for
removing
plant­
incorporated
protectant
that
exceeds
any
proposed
tolerance
Such
a
method
is
not
required
because
a
temporary
exemption
from
the
requirement
of
a
tolerance
is
being
sought.
Monsanto
Company
04­
CR­
138E­
1
Page
9
of
11
Section
F:
Proposed
tolerances
for
the
plant­
incorporated
protectant
if
tolerances
are
proposed
A
temporary
exemption
from
the
requirement
of
a
tolerance
is
being
sought
for
Cry2Ab2
protein
and
the
genetic
material
necessary
for
its
production
in
food
and
feed
commodities
of
field
corn,
sweet
corn,
and
popcorn.

Monsanto
proposes
that
40
CFR
§
180
be
revised
as
follows:

§
180.
XXXX.
Bacillus
thuringiensis
Cry2Ab2
protein
and
the
genetic
material
for
its
production
in
corn;
temporary
exemption
from
the
requirement
of
a
tolerance.

Bacillus
thuringiensis
Cry2Ab2
protein
and
the
genetic
material
necessary
for
its
production
in
corn
is
exempt
from
the
requirement
of
a
tolerance
when
used
as
plant­
incorporated
protectant
in
food
and
feed
commodities
of
field
corn,
sweet
corn,
and
popcorn.
"
Genetic
material
necessary
for
its
production"
means
the
genetic
material
encoding
the
Cry2Ab2
protein
and
its
regulatory
regions.
"
Regulatory
regions"
are
the
genetic
material,
such
as
promoters,
terminators,
and
enhancers
that
control
the
expression
of
the
genetic
material
encoding
the
Cry2Ab2
protein.
Monsanto
Company
04­
CR­
138E­
1
Page
10
of
11
Section
G:
Reasonable
grounds
in
support
of
this
petition.

Reasonable
grounds
to
support
this
petition
for
a
temporary
exemption
from
the
requirement
of
a
tolerance
for
the
Bacillus
thuringiensis
Cry2Ab2
protein
and
the
genetic
material
necessary
for
its
production
in
food
and
feed
commodities
of
field
corn,
sweet
corn,
and
popcorn
when
used
as
plant­
incorporated
protectant
are
presented
in
Section
C.
This
temporary
tolerance
exemption
is
required
for
field­
testing
of
ZMIR245
lepidopteranprotected
corn.
Monsanto
Company
04­
CR­
138E­
1
Page
11
of
11
Attachment
I.
Applicant's
Notice
of
Filing
Summary
A
summary
of
this
pesticide
petition
for
a
temporary
exemption
from
the
requirement
of
a
tolerance
for
residues
of
Bacillus
thuringiensis
Cry2Ab2
protein
and
the
genetic
material
necessary
for
its
production
in
food
and
feed
commodities
of
field
corn,
sweet
corn,
and
popcorn
has
been
combined
with
the
petition
requesting
establishment
of
a
temporary
exemption
from
the
requirement
of
a
tolerance
for
Bt
Cry1A.
105
insecticidal
protein,
which
was
submitted
to
EPA
on
January
31,
2005.
Monsanto
consents
to
the
use
of
all
or
part
of
this
summary
by
the
Agency
in
preparing
the
notice
of
filing
and
the
proposed
or
final
regulation.

An
electronic
copy
of
this
petition
summary
in
Word
format
is
also
being
provided
on
a
CDROM
disk.