[Federal Register Volume 71, Number 6 (Tuesday, January 10, 2006)]
[Notices]
[Pages 1506-1507]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: E6-76]


 ========================================================================
 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.
 
 ========================================================================
 

  Federal Register / Vol. 71, No. 6 / Tuesday, January 10, 2006 / 
Notices  

[[Page 1506]]



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.

-----------------------------------------------------------------------

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: [email protected]. 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.
---------------------------------------------------------------------------

    \1\ http://ecos.fws.gov/tess_public/servlet/gov.doi.tess_public.servlets.RegionLists?lead_region=3#WI.
---------------------------------------------------------------------------

    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]
BILLING CODE 3410-34-P