[Federal Register: January 10, 2006 (Volume 71, Number 6)]
[Notices]
[Page 1506-1507]
From the Federal Register Online via GPO Access [wais.access.gpo.gov]
[DOCID:fr10ja06-14]
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Notices
Federal Register
________________________________________________________________________
This section of the FEDERAL REGISTER contains documents other than rules
or proposed rules that are applicable to the public. Notices of hearings
and investigations, committee meetings, agency decisions and rulings,
delegations of authority, filing of petitions and applications and agency
statements of organization and functions are examples of documents
appearing in this section.
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DEPARTMENT OF AGRICULTURE
Animal and Plant Health Inspection Service
[Docket No. 05-053-2]
University of Wisconsin-Madison; Availability of Environmental
Assessment and a Finding of No Significant Impact for Field Tests of
Genetically Engineered Erwinia carotovora
AGENCY: Animal and Plant Health Inspection Service, USDA.
ACTION: Notice.
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SUMMARY: We are advising the public that the Animal and Plant Health
Inspection Service has prepared an environmental assessment for a field
trial of genetically engineered strains of a bacterium, Erwinia
carotovora, the causal agent of tuber soft rot disease in potato. The
bacteria have been genetically engineered to disrupt the disease
causing pathway. This field trial will allow researchers to better
understand the function of each mutated gene under field conditions.
The environmental assessment provides a basis for our conclusion that
these field tests will not present a risk of introducing or
disseminating a plant pest and will not have a significant impact on
the quality of the human environment. Based on its finding of no
significant impact, the Animal and Plant Health Inspection Service has
determined that an environmental impact statement need not be prepared
for these field tests.
Effective Date: January 10, 2006.
ADDRESSES: You may read the environmental assessment, the finding of no
significant impact, and any comments that we received on Docket No. 05-
053-1 in our reading room. The reading room is located in room 1141 of
the USDA South Building, 14th Street and Independence Avenue SW.,
Washington, DC. Normal reading room hours are 8 a.m. to 4:30 p.m.,
Monday through Friday, except holidays. To be sure someone is there to
help you, please call (202) 690-2817 before coming.
FOR FURTHER INFORMATION CONTACT: Dr. Rudaina Alrefai, Biotechnology
Regulatory Services, APHIS, 4700 River Road Unit 147, Riverdale, MD
20737-1236; (301) 734-4866. To obtain copies of the environmental
assessment (EA), the finding of no significant impact (FONSI), or the
response to comments, contact Ms. Ingrid Berlanger at (301) 734-4885;
e-mail: Ingrid.E.Berlanger@aphis.usda.gov. The EA, FONSI, and response
to comments are also available on the Internet at http://www.aphis.usda.gov/brs/aphisdocs/05_09701r_ea.pdf.
The draft EA is
available at http://www.aphis.usda.gov/brs/aphisdocs/05_09701r_pea.pdf
.
SUPPLEMENTARY INFORMATION:
Background
The regulations in 7 CFR part 340, ``Introduction of Organisms and
Products Altered or Produced Through Genetic Engineering Which Are
Plant Pests or Which There Is Reason to Believe Are Plant Pests,''
regulate, among other things, the introduction (importation, interstate
movement, or release into the environment) of organisms and products
altered or produced through genetic engineering that are plant pests or
that there is reason to believe are plant pests. Such genetically
engineered organisms and products are considered ``regulated
articles.'' A permit must be obtained or a notification acknowledged
before a regulated article may be introduced. The regulations set forth
the permit application requirements and the notification procedures for
the importation, interstate movement, or release into the environment
of a regulated article.
On April 7, 2005, the Animal and Plant Health Inspection Service
(APHIS) received a permit application (APHIS permit number 05-097-01r)
from the University of Wisconsin-Madison, Department of Plant
Pathology, Madison, WI, for a permit for a field trial of Erwinia
carotovora. These bacteria have been genetically engineered not to
express specific hrp/hrc (hypersensitive reaction on non-host plants
and pathogenesis on host plants or conserved among plant and animal
pathogens) genes resulting in the disruption of the disease-causing
mechanism. These mutations are expected to make the bacterial strains
avirulent or non-pathogenic. The application describes four genetically
engineered strains to be used in this field trial.
The E. carotovora ssp. carotovora WPP14 strain was initially
isolated from a diseased potato plant obtained from a commercial farm
in Waushara County, WI. This strain was used to create four new
genetically engineered strains by inserting a marker gene into genes
that may be necessary for E. carotovora infection of potatoes. The four
strains resulting from this mutagenesis that are proposed for use in
this field trial are described below.
Strain WPP40 contains an insertion of a kanamycin
resistance gene (aph) cassette into outD. The outD gene encodes for an
outer membrane porin that is required for a functional type II
secretion system. This mutant is unable to secrete plant cell wall
degrading enzymes and is avirulent. The kanamycin resistance gene
cassette contains aph, which encodes neomycin phosphotransferase which
was originally isolated from Tn5, and two FRT sites derived from
Saccharomyces cerevisiae.
Strain WPP60 has an insertion of a spectromycin resistance
gene (aadA) cassette into hrcC, an outer membrane porin which is
required for a functional type III secretion system. This mutant is
unable to secrete harpins or effector (Avr) proteins. It is
hypersensitive response minus. The spectinomycin resistance gene
cassette is constructed from the aadA gene which encodes
aminoglycoside-3 adenyltransferase, originally derived from Shigella
flexneri, with termination sequences derived from bacterophage T4.
Strain WPP195 has a deletion of hrpN and an insertion of a
chloramphenicol resistance gene (cat) cassette and a modified green
fluorescent protein (GFPmut2) into this locus. This mutant is unable to
produce or secrete the harpin, HrpN. The gfpmut2 gene was originally
cloned from Aequorea victoria and was modified to be brighter. Its
expression is driven by the nptII kan promoter from Tn5. The cat gene
encodes
[[Page 1507]]
cholramphenicol acetyltransferase, which was originally isolated from
Escherichia coli. This construct also contains FRT sites.
Strain WPP198 is an insertion of a similar chloramphenicol
resistance cassette into hrpL, which is a sigma factor required for
expression of the type III secretion system and its secreted
substrates. The mutant is unable to produce or secrete harpins or
effector (Avr) proteins. It is hypersensitive response minus.
The genetically engineered E. carotovora are considered regulated
articles under the regulations in 7 CFR part 340 because they may be
plant pests. The purpose of the field trial is to use genetically
engineered E. carotovora strains with mutations in homologs of the
well-characterized Pseudomonas syringae hrp genes as tools to:
Understand the effects of specific genes on the fitness of
E. carotovora,
Use the results from these experiments to better
understand the function of these genes in plant-bacterial interactions,
and
Compare the results obtained with E. carotovora mutants
with those found for P. syringae to determine if homologous genes play
similar roles in fitness in different environments.
In a notice published in the Federal Register on August 12, 2005
(70 FR 47170-47171, Docket No. 05-053-1), APHIS announced that it had
prepared an environmental assessment (EA) for a field trial of the
genetically engineered strains of E. carotovora and made the EA
available for public review and comment. During the 30 day comment
period for the draft EA, which ended on September 12, 2005, APHIS
received six comments. Responses to the issues raised in the comments
are provided as an attachment to the finding of no significant impact
(FONSI). Three of the comments were from private individuals, one was
from a public interest group, and the remaining two comments were from
the same person, writing first as a private individual and then
representing a public interest group. All six comments opposed the
field test. One individual was generally opposed to field tests of
genetically engineered organisms. However the comment did not provide
support for the opposition. The remaining comments raised two issues.
One issue is the concern that horizontal gene transfer of the
antibiotic resistance gene in these bacterial strains might result in
transfer of this trait to soil or enteric bacteria. This issue was
addressed in the EA and is again addressed in the response to comments.
The second issue is that the experiment is conducted with bacterial
strains that may be as virulent as the native bacteria strains. APHIS
disagrees with the comment that this field trial ``provides high risk
with little or no benefit.'' This issue is also addressed in the
response to comments.
APHIS has edited the EA to include specific contact information in
Appendix I and to add a new section IX, ``Consultations,'' in the final
EA. The changes are not substantive and do not impact the analysis in
the EA. Copies of both the draft EA and the final EA are available (see
FOR FURTHER INFORMATION CONTACT).
Pursuant to its regulations (7 CFR part 340) promulgated under the
Plant Protection Act, APHIS has determined that this field trial will
not pose a risk of the introduction or dissemination of a plant pest
for the following reasons:
(1) Erwinia carotovora is widely spread in the environment and
commonly present on plant roots of numerous species as well as in
lakes, streams, rain, and ground water.
(2) Screening weeds for the past year around potato fields did not
reveal any naturally-occurring ``hypersensitive reaction on non-host
plants and pathogenesis on host plants or conserved among plant and
animal pathogens'' (hrp/hrc) mutants of E. carotovora even though these
mutants have been found on potato. Therefore, it is not likely that the
host range of the bacterium will change because of the modification.
(3) Reversion of the genetically engineered strains would not pose
any additional environmental risk because reverted mutants will be
similar to the other E. carotovora strains that are commonly present on
these plants. The risks associated with the introduction of genetically
engineered organisms generally are the same kind as those associated
with the introduction into the environment of unmodified organisms and
organisms modified by other genetic techniques.
(4) The field trial is a confined release and would have no
significant impact on the environment. The field release conditions and
measures described in the permit should be sufficient to prevent any
unplanned release of the transgenic bacteria or the inoculated plant
material; or the persistence of the transgenic material in the
environment.
(5) This small field test of 0.2 acre should not have any
significant impact on existing agricultural practices because this test
is solely for research purposes. The antibiotic resistance genes
themselves should not cause these mutant strains to have any
competitive advantage in the environment and would not interfere with
current agricultural practices to control the soft rot disease in
potato. Although spraying with streptomycin is used to control Erwinia
amylovora on fruit trees, it is not normally used to control the soft
rot disease in potatoes on this field station.
(6) Resistance to antibiotics is already widely prevalent in
enteric bacteria and soil-borne bacteria. Gene transfer from E.
carotovora to animals and plants is highly unlikely under the
conditions of this field test.
(7) Erwinia species are not known as animal or human pathogens and
there are no references that associate it with human or animal disease
even though farm workers have been exposed to Erwinia spp. for decades.
There should be no risk to university personnel handling the inoculated
potatoes since they hand-inoculate potatoes while wearing gloves and
all diseased plants are removed from the field. No potential impact of
this experiment on people living in the area of the field trial test
plot or any other human population can be identified.
(8) An examination of threatened and endangered species for
Wisconsin listed in the U.S. Fish and Wildlife Service's Threatened and
Endangered Species System \1\ showed that 6 threatened or endangered
plant species and 12 animal species exist or once existed in the State.
Only one plant species has been reported in Waushara County but is not
a host for E. carotovora. None of the listed threatened or endangered
plant and animal species would be impacted by this test.
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\1\ http:// ecos.fws.gov/tess-- public/servlet/gov.doi.tess--
public.servlets. RegionLists?lead--region=3#WI.
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The EA was prepared in accordance with (1) The National
Environmental Policy Act of 1969 (NEPA), as amended (42 U.S.C. 4321 et
seq.), (2) regulations of the Council on Environmental Quality for
implementing the procedural provisions of NEPA (40 CFR parts 1500-
1508), (3) USDA regulations implementing NEPA (7 CFR part 1b), and (4)
APHIS' NEPA Implementing Procedures (7 CFR part 372). Copies of the EA
and FONSI are available from the individual listed under FOR FURTHER
INFORMATION CONTACT.
Authority: 7 U.S.C. 7701-7772 and 7781-7786; 31 U.S.C. 9701; 7
CFR 2.22, 2.80, and 371.3.
Done in Washington, DC, this 3rd day of January 2006.
Kevin Shea,
Acting Administrator, Animal and Plant Health Inspection Service.
[FR Doc. E6-76 Filed 1-9-06; 8:45 am]
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