Document ID: EPA-HQ-OPP-2005-0122-0003
Agency: epa
Document Type: Supporting & Related Material
Title: 
Posted Date: 2005-07-26T04:00Z

UNITED
STATES
ENVIRONMENTAL
PROTECTION
AGENCY
WASHINGTON,
D.
C.
20460
OFFICE
OF
PREVENTION,
PESTICIDES
AND
TOXIC
SUBSTANCES
January
6,
2005
MEMORANDUM
SUBJECT:
Review
of
materials
submitted
by
Dow
in
support
of
proposed
amendments
to
the
Section
G
Experimental
Use
Permit
(
EUP
No:
68467­
EUP­
7,
DP#
310100)
for
Cry34/
35Ab1
corn.
Submission
dated
February
10,
2004
and
revised
December
20,
2004.

TO:
Michael
Mendelsohn
Senior
Regulatory
Action
Leader
Microbial
Pesticides
Branch,
Biopesticides
and
Pollution
Prevention
Division
(
7511C)

FROM:
Tessa
Milofsky,
M.
S.
Agronomist
Microbial
Pesticides
Branch,
Biopesticides
and
Pollution
Prevention
Division
(
7511C)

PEER
REVIEW:
Alan
Reynolds,
M.
S.
Entomologist
Microbial
Pesticides
Branch,
Biopesticides
and
Pollution
Prevention
Division
(
7511C)

CONCLUSION
This
request
to
extend
the
duration
and
increase
the
acreage
of
the
existing
Cry34/
35
EUP
is
acceptable.
It
is
recommended
that
3,096
EUP
acres
should
be
granted
(
4,946
acres
requested).

RECOMMENDATIONS
 
Proposed
acreage
for
the
Cry34/
35Ab1
corn
breeding
and
observation
nursery,
Cry34/
35Ab1
corn
efficacy
trial,
and
insect
resistance
management
studies
is
similar
to
that
approved
for
the
existing
EUP
and
is
considered
acceptable.
It
is
recommended,
however,
that
future
submissions
should
provide
more
detailed
protocol
descriptions
(
e.
g.
states
where
specific
trials
will
be
conducted,
number
of
trials
per
state,
number
of
hybrid/
inbred
lines
to
be
tested,
etc.)
that
clearly
justify
the
need
for
the
trial
and
acreage.
2
 
Given
available
information,
it
is
recommended
that
2,000
acres
should
be
granted
for
agronomic
observation
trials
(
2,401
acres
were
requested).
Information
on
the
number
of
experimental
locations
per
state,
acres
per
location,
and
hybrid/
inbred
lines
tested
should
be
provided
in
the
final
report.

 
The
registrant
requested
2,199
EUP
acres
to
evaluate
glufosinate­
ammonium
herbicide
tolerance
in
Cry34/
35Ab1
corn
lines.
A
variety
of
effective
laboratory,
greenhouse,
and
small­
scale
field
plot
bioassays
are
frequently
used
to
verify
the
presence
of
herbicide
tolerance
genes
in
transgenic
corn.
However,
field
scale
trials
can
be
necessary
to
evaluate
the
effect
that
herbicide
applications
may
have
on
plant
development,
flowering
time,
herbicide
susceptibility/
resistance,
herbicide
injury
scores,
"
staygreen",
and
barren
plant
counts.
Although
fieldbased
assays
can
require
considerable
acreage,
the
submission
does
not
provide
sufficient
information
to
justify
the
2,199
acre
request.
Given
available
information,
it
is
recommended
that
750
acres
should
be
granted
for
this
protocol.

 
The
participant/
collaborator
information
is
sufficient
provided
that
collaborator
names
and
contact
information
are
submitted
prior
to
experiment
initiation.

 
The
final
Section
G
submission
addressed
most
deficiencies
identified
by
the
Agency.
However,
state­
by­
state
breakdowns
of
acreage
and
locations
needed
per
protocol
were
not
included.
It
is
recommended
that
these
details
should
be
provided
in
the
final
report.

BACKGROUND
This
is
a
request
to
extend
the
duration
and
increase
the
allowable
acreage
permitted
under
the
EUP
granted
for
Cry34/
35Ab11
corn,
which
protects
against
western
corn
rootworm
(
WCRW;
Diabrotica
virgifera
virgifera)
and
northern
corn
rootworm
(
NCRW;
Diabrotica
berberi)
.
The
EUP
also
includes
acreage
for
stacked
events,
containing
Cry34/
35Ab1
and
Cry1F
insecticidal
proteins,
which
provide
combined
protection
against
corn
rootworm
and
lepidopteran
pests.
The
present
EUP
(
last
amended
April
29,
2004)
permits
test
plant
cultivation
on
1,177
acres
in
25
different
states
(
including
Puerto
Rico)
and
is
slated
to
expire
April
30,
2005.
Cry34Ab1
and
Cry35Ab1
proteins
are
temporarily
exempted
(
exemption
will
expire
April
30,
2006)
from
the
requirement
of
a
tolerance
when
used
as
plant­
incorporated
protectants
in
the
food
and
feed
commodities
of
field
corn,
sweet
corn,
and
popcorn
(
CFR
2004,
§
180.1242).
Consequently,
non­
crop
destruct
status
has
been
granted,
meaning
that
confinement
and
destruction
of
plant
material
(
e.
g.
pollen,
seeds,
stalks,
etc.)
is
not
required.
It
should
be
noted,
however,
that
1
The
EUP
covers
corn
containing
the
Cry34/
35Ab1
Insecticidal
Crystal
Protein
(
ICP)
produced
by
construct
17662
(
events
DAS­
45216­
6,
DAS­
59122­
7,
DAS­
45214­
4,
DAS­
59110­
4,
DAS­
59132­
8,
DAS­
06317­
5)
and
could
include
corn
produced
by
traditional
breeding
of
the
rootworm­
protected
corn
with
lepidopteran­
protected
Bt
corn
(
e.
g.
Herculex
1
Corn
(
Bt
Cry1F,
event
1507).
3
a
Scientific
Advisory
Panel
is
scheduled
to
meet
on
March
1­
2,
2005
"
to
consider
and
review
human
health
issues
associated
with
the
Cry34Ab1
protein".

PROGRAM
REVIEW
The
submission
requests
that
the
present
EUP
be
extended
until
April
30,
2006,
so
that
the
registrant
can
"
fully
evaluate
this
product
and
its
benefits."
The
registrant
also
requests
that
the
allowable
acreage
be
increased
to
4,946
acres
for
the
2005
growing
season.
The
attached
EUP
spreadsheet
(
Appendix
A)
describes
the
proposed
acreages
for
each
state.
It
should
be
noted
that
the
product's
temporary
exemption
from
tolerance
expires
April
30,
2006
and
thus,
corresponds
with
the
proposed
expiration
date
of
the
existing
EUP.

Participant/
Collaborator
Information
The
names,
telephone
numbers,
and
mailing
addresses
of
participants
who
will
supervise
experimental
work
covered
under
this
EUP
are
provided
in
the
submission.
Information
on
cooperators
was
not
included,
yet
the
registrant
states
that
this
listing
will
be
provided
prior
to
planting
or
initiation
of
the
research
trials.

BPPD
Review
Participant/
collaborator
information
is
sufficient
provided
that
undisclosed
collaborator
names
and
contact
information
are
submitted
prior
to
experiment
initiation.

Acreage
Calculation
Total
EUP
acreage
is
equivalent
to
the
sum
of
the
acreage
comprised
in
each
EUP
test
block.
This
calculation
would
include
all
EUP
corn
plants
(
Cry34/
35
and
stacked
plants),
non­
EUP
corn
plants
(
e.
g.
breeding
plants
or
isoline
control
plots),
and
associated
border
rows
contained
within
the
perimeter
of
the
test
block.
The
section
also
includes
field
plans
that
illustrate
how
blocks
of
EUP
corn
may
be
positioned
in
relation
to
non­
EUP
corn
fields,
providing
a
visual
representation
of
experimental
block
location.

BPPD
Review
This
approach
to
EUP
acreage
calculation
is
acceptable.

Field
Trial
Containment/
Disposition
Procedures
Field
trial
containment
is
said
to
comply
with
the
USDA­
APHIS
performance
standards
for
regulated
corn
trials.
Six
approaches
to
gene
flow
containment
are
listed
(
e.
g.
bagging,
660
ft.
isolation,
temporal
isolation,
etc.),
of
which
one
or
more
will
be
used
in
the
proposed
trials.

Sections
covering
plant
sampling,
harvest
procedures,
final
disposition,
and
volunteer
plant
monitoring,
include
brief
(
from
one
to
several
sentences)
descriptions
of
containment
procedures
that
may
be
implemented
during
these
phases.

BPPD
Review
The
isolation
methods
presented
are
acceptable.
4
Specific
Trial
Protocols
Specific
trial
protocols
implemented
for
the
period
March
2004­
March
2005
and
proposed
for
the
period
March
2005­
April
2006
are
listed
below.
Two
tables
are
presented
for
2005­
2006,
because
the
initial
request
(
submitted
2­
5­
04)
was
replaced
by
a
revised
submission
dated
12­
20­
04.
It
should
be
noted
that
the
revised
Section
G
was
submitted
in
response
the
Agency's
request
for
more
information
on
agronomic
observation
and
demonstration
trial
protocols.

March
2004­
March
2005
Requested/
Granted
EUP
Protocols
EUP
Acres
Lbs.
of
Seed
Breeding
and
observation
nursery
277
4,617
Agronomic
observation
trials
805
13,417
Glufosinate
herbicide
tolerance
study
8
133
Efficacy
trial
45
750
Insect
resistance
management
studies
8
133
Demonstration
trials
34
567
Totals
1,177
19,617
March
2005­
April
2006
Requested
(
2­
5­
04)
EUP
Protocols
EUP
Acres
Lbs.
of
Seed
Breeding
and
observation
nursery
277
4,617
Agronomic
observation
trials
4,070
40,017
Glufosinate
herbicide
tolerance
study
8
36,650
Efficacy
trial
44
733
Insect
resistance
management
studies
8
417
Demonstration
trials
3,280
54,666.7
Totals
7,687
128,117
March
2005­
April
2006
Requested
(
12­
20­
04)
EUP
Protocols
EUP
Acres
Lbs.
of
Seed
Breeding
and
observation
nursery
277
4,617
Agronomic
observation
trials
2,401
40,017
Glufosinate
herbicide
tolerance
study
2,199
36,650
Efficacy
trial
44
733
Insect
resistance
management
studies
25
417
Totals
4,946
82,433
Protocol
descriptions
include
information
on
the
objective,
experimental
design,
plot
treatments,
genotypes
and
vectors,
and
estimated
planting
and
harvest
schedules
for
each
trial.
Protocol
objectives
are
listed
below.
5
Protocol:
Breeding
and
observation
nursery
Objective:
Develop
and
evaluate
a
variety
of
Cry34/
35Ab1
corn
lines.

Protocol:
Agronomic
observation
trials
Objective:
To
assess
agronomic
and
phenotypic
effects
that
may
result
from
inserting
the
Cry34/
35Ab1
protein,
and
the
genetic
material
required
for
its
production,
in
corn
lines.

Protocol:
Glufosinate
herbicide
tolerance
studies
Objective:
To
evaluate
the
effect
of
glufosinate­
ammonium
herbicide
application
on
Cry34/
35Ab1
corn
lines.

Protocol:
Efficacy
trials
Objective:
To
assess
the
efficacy
of
Cry34/
35Ab1
corn
lines
in
controlling
western
corn
rootworm,
northern
corn
rootworm,
and
other
insects.

Protocol:
Insect
resistance
management
trials
Objective:
To
provide
information
needed
to
develop
IRM
strategies
appropriate
for
Cry34/
35Ab1
corn
lines.
Experiments
will
focus
on
identifying
Cry34/
35
dose
for
corn
rootworm
pests.

Requested
acreages
for
breeding
and
observation,
efficacy
trials,
and
insect
resistance
management
protocols
are
generally
in
line
with
past
EUP
acreage
requests
for
insect
protected
corn
products.
However,
the
acreage
requested
for
agronomic
observation
and
demonstration
trials
were
considerably
larger
than
those
requested
and
granted
for
most
EUPs.
Consequently,
the
Agency
requested
that
more
detailed
protocol
descriptions
should
be
submitted
for
these
study
areas.
Additional
information
requested
by
the
Agency
included:
descriptions
of
specific
experimental
trials
included
within
a
protocol,
experimental
objectives
and
expected
outcomes
for
each
trial,
experimental
design,
number
of
hybrids
to
be
used,
number
of
locations
required
per
protocol
and/
or
per
experiment,
acreage
per
experiment,
and
a
table
showing
required
acreage
and
number
of
experimental
locations
per
state.
The
registrant
was
also
notified
that
acreage
for
demonstration
trials
is
generally
not
permitted
under
EUPs.

The
revised
Section
G,
dated
December
20,
2004,
included
an
updated
acreage
request
(
these
changes
were
not
requested
by
the
Agency).
Most
notably,
demonstration
acreage
was
removed,
acreage
for
herbicide
tolerance
was
raised
from
8
acres
to
2,199
acres,
and
acreage
for
agronomic
observation
trials
was
reduced
from
4,070
to
2,401
acres.
The
revised
submission
also
addressed
some
deficiencies
identified
for
the
agronomic
observation
protocol.
Information
presented
for
agronomic
observation
and
herbicide
tolerance
protocols
is
summarized
below.
Other
protocols
are
not
summarized,
because
acreage
requests
for
these
trials
are
similar
to
what
has
been
granted
for
other
EUPs.

Agronomic
observation
protocol
Acreage
requested
rose
from
805
acres
in
2004
to
2,401
(
revised
Section
G)
acres
in
2005.
Examples
of
agronomic
trials
that
may
be
conducted
under
this
protocol
include:
6
1.
Hybrid
yield:
used
to
compare
yield
among
hybrids;
trials
would
have
multiple
replicates
and
contain
as
many
as
150
to
200
hybrid
varieties,
with
a
variety
of
maturities
ranging
from
85
(
zone
2)
to
115
(
zone
8)
RM.
2.
Inbred
yield:
used
to
compare
yields
and
agronomic
traits
among
inbred
lines;
as
many
as
25
inbreds
may
be
evaluated.
3.
Hybrid
and
inbred
observation
plots:
conducted
to
observe
differences
in
agronomic
traits
between
different
hybrids
and
their
isogenic
checks;
as
many
as
200
to
225
hybrids
and
inbreds
could
be
evaluated.
4.
Flowering
trials:
important
for
evaluation
of
pollen
shed,
pollen
viability,
and
the
timing
of
flowering
in
relation
to
silk
emergence
on
female
plants;
trials
could
be
run
in
a
variety
of
environments
and
soil
types.
5.
Grade
out
studies:
conducted
to
evaluate
seed
size
of
female
plants
grown
in
multiple
environments
and
soil
types.
6.
Disease
susceptibility
evaluations:
used
to
evaluate
hybrid
susceptibility
to
grey
leaf
spot
and
other
fungal
diseases.

Herbicide
tolerance
protocol
Requested
acreage
for
glufosinate­
ammonium
herbicide
tolerance
increased
from
8
acres
in
2004
to
2,199
acres
in
2005
(
revised
Section
G).
The
stated
objective
of
this
protocol
is
"
to
evaluate
the
effect
of
glufosinate­
ammonium
herbicide
application
on
Cry34/
35Ab1
corn
lines
and 
confirm
there
will
not
be
any
reduction
of
expression
of
the
PAT
gene
(
which
confers
herbicide
tolerance)."
The
registrant
further
states,
"
A
reduction
in
gene
expression
can
be
difficult
to
detect
unless
and
until
the
PIP
is
planted
widely
in
largescale
environments."

Various
experimental
plot
designs
will
be
used.
Experiments
will
include
inbred
lines,
hybrids,
and
stacked
products
of
varying
genetic
constitutions.

BPPD
Review
Proposed
acreages
for
Cry34/
35Ab1
corn
breeding
and
observation
nurseries,
efficacy
trials,
and
insect
resistance
management
studies
are
acceptable.
However,
it
is
recommended
that
future
submissions
should
include
expanded
discussions
of
all
EUP
protocols
(
see
Future
Submissions
section
below).

Although
trials
included
under
the
agronomic
observation
protocol
are
acceptable,
the
registrant's
request
for
2,401
acres
may
exceed
the
acreage
required
to
complete
necessary
experiments.
Additional
information
on
the
number
of
experimental
locations
per
state,
acres
per
location,
and
hybrid/
inbred
lines
tested
could
provide
justification
for
greater
acreage.
However,
given
available
information,
it
is
recommended
that
2,000
acres
should
be
granted
for
these
trials.

The
registrant
also
requested
2,199
EUP
acres
to
evaluate
glufosinate­
ammonium
herbicide
tolerance
in
Cry34/
35Ab1
corn
lines.
The
protocol
description
states,
"[
Cry34/
35]
plants
may
be
evaluated
for
yield,
and
for
effects
of
the
herbicide
on
plant
development
and
agronomic
traits."
Information
on
specific
experiments
is
not
provided.
7
Field­
scale
trials
can
be
useful
in
evaluating
the
effect
that
herbicide
applications
may
have
on
plant
development,
flowering
time,
herbicide
susceptibility/
resistance,
herbicide
injury
scores,
"
staygreen",
and
barren
plant
counts.
However,
a
variety
of
effective
laboratory
(
ELISA,
PCR,
and
isozyme
electrophoresis),
greenhouse,
and
small­
scale
field
plot
bioassays
are
frequently
used,
by
industry
and
the
research
community
alike,
to
verify
the
presence
of
herbicide
tolerance
genes
in
transgenic
corn.
These
methods
would
require
little,
if
any,
EUP
acreage.

Although
the
registrant's
need
for
field­
based
assays
may
be
justified,
the
submission
did
not
provide
sufficient
information
(
e.
g.
objective,
expected
outcome,
experimental
design,
number
of
hybrids
to
be
tested,
required
acreage
per
location,
etc.)
to
validate
the
2,199
acre
request.
Given
what
is
know
about
the
herbicide
tolerance
protocol,
it
is
recommended
that
750
EUP
acres
should
be
granted
for
these
trials.

March
2005­
April
2006
EUP
Acres
EUP
Protocols
Requested
Agronomically
Justified
Breeding
and
observation
nursery
277
277
Agronomic
observation
trials
2,401
2,000
Glufosinate
herbicide
tolerance
study
2,199
750
Efficacy
trial
44
44
Insect
resistance
management
studies
25
25
Totals
4,946
3,096
Final
Report
The
final
Section
G
submission
addressed
most
deficiencies
identified
by
the
Agency.
However,
state­
by­
state
breakdowns
of
acreage
and
locations
needed
per
protocol
were
not
included.
Consequently,
it
is
recommended
that
these
details
should
be
provided
in
the
final
report.
Specifically,
the
final
report
should
include:
 
A
state­
by­
state
breakdown
of
acreage
per
protocol.
 
A
state­
by­
state
breakdown
of
locations
per
protocol.
 
A
listing
of
experiments
conducted
within
each
protocol.
 
Average
acreage
per
location
for
each
experiment
included
within
each
protocol
(
ideally
this
would
be
presented
on
a
state­
by­
state
basis).

Future
Submissions
If
EUP
requests
are
to
be
objectively
evaluated,
it
is
strongly
recommended
that
future
submissions
should
include
the
following
information:
 
Protocols
to
be
conducted.
 
Experiments
to
be
included
within
each
protocol.
 
Estimated
state­
by­
state
breakdown
of
acreage
per
protocol.
 
Estimated
state­
by­
state
breakdown
of
locations
per
protocol.
 
Estimate
of
acreage
and
locations
required
for
each
experiment
included
within
protocols
(
ideally
this
would
be
presented
on
a
state­
by­
state
basis).
8
 
If
an
acreage
increase
is
requested
for
one
or
more
protocols,
justification
should
be
provided
for
those
increases
(
justification
should
be
provided
on
a
protocol
by
protocol
basis).
 
Objective
and
brief
description
of
each
experiment
included
within
protocols.
Approximate
number
of
hybrids/
inbreds/
isolines
to
be
tested
should
also
be
included
(
may
be
presented
as
average
number
of
lines
per
location).
 
Clear
explanation
of
acreage
calculation,
including
a
breakdown
of
EUP
acreage
designated
for
test
plants,
non­
test
plants,
and
border
rows
included
within
experimental
blocks.
 
It
should
be
noted
that
EUP
acreage
designated
for
demonstration
trials
is
generally
considered
inappropriate.