[Federal Register: August 12, 2005 (Volume 70, Number 155)]
[Notices]
[Page 47170-47171]
From the Federal Register Online via GPO Access [wais.access.gpo.gov]
[DOCID:fr12au05-39]
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DEPARTMENT OF AGRICULTURE
Animal And Plant Health Inspection Service
[Docket No. 05-053-1]
University of Wisconsin-Madison; Availability of Environmental
Assessment 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.
This environmental assessment is available for public review and
comment.
DATES: We will consider all comments we receive on or before September
12, 2005.
ADDRESSES: You may submit comments by either of the following methods:
Postal Mail/Commercial Delivery: Please send four copies
of your comment (an original and three copies) to Docket No. 05-053-1,
Regulatory Analysis and Development, PPD, APHIS, Station 3C71, 4700
River Road Unit 118, Riverdale, MD 20737-1238. Please state that your
comment refers to Docket No. 05-053-1.
EDOCKET: Go to http://www.epa.gov/feddocket to submit or
view public comments. Once you have entered EDOCKET, click on the
``View Open APHIS Dockets'' link to locate Docket No. 05-053-1.
Reading Room: You may read the environmental assessment and any
comments that we receive in our reading room. The reading room is
located in room 1141 of the USDA
[[Page 47171]]
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.
Other Information: You may view APHIS documents published in the
Federal Register and related information on the Internet at http://www.aphis.usda.gov/ppd/rad/webrepor.html
.
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 petition or the
environmental assessment (EA), contact Ms. Ingrid Berlanger at (301)
734-4885; e-mail: ingrid.e.berlanger@aphis.usda.gov. The EA is also
available on the Internet at http://www.aphis.usda.gov/brs/aphisdocs/05_09701r_ea.pdf
.
SUPPLEMENTARY INFORMATION: 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 mutatgenesis 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 and reduced in virulence. The
spectinomycin resistance gene cassette is constructed from the aadD
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 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 and
reduced in virulence.
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.
To provide the public with documentation of APHIS' review and
analysis of any potential environmental impacts and plant pest risk
associated with the proposed field trial of theses strains of
genetically engineered E. carotovora, an environmental assessment (EA)
has been prepared. 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
are available as indicated in the FOR FURTHER INFORMATION CONTACT
section of this notice.
Done in Washington, DC, this 8th day of August 2005.
Kevin Shea,
Acting Administrator, Animal and Plant Health Inspection Service.
[FR Doc. E5-4382 Filed 8-11-05; 8:45 am]
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