[Federal Register Volume 70, Number 155 (Friday, August 12, 2005)]
[Notices]
[Pages 47170-47171]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: E5-4382]


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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: [email protected]. 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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