NEWS AND NOTES
USDA'S OFFICE OF AGRICULTURAL BIOTECHNOLOGY TO CLOSE
February 19 marks the closing of the Office of Agricultural Biotechnology (OAB), which was instituted in 1986 to coordinate USDA policies and procedures related to agbiotech. The Department's involvement in biotechnology cuts across many of its research, regulatory, policy, technology transfer, education, and public information programs and agencies. To harmonize biotech initiatives at USDA, OAB created the Biotechnology Council which brought together on a regular basis senior level representatives from eight departmental agencies.
OAB provided administrative support for the Agricultural Biotechnology Research Advisory Committee (ABRAC) which was established to advise the Secretary of Agriculture on agbiotech research funded by USDA, and to provide a biosafety review process of proposed biotechnology experiments not under the jurisdiction of other federal agencies. The ABRAC developed a set of voluntary Guidelines for releasing genetically modified organisms in the environment, and, more recently, Performance Standards for research with genetically modified fish and shellfish. With the closing of the OAB, the ABRAC will also cease operations.
Since its inception, OAB has served as an information source for other federal agencies, the media, the public and the international community. Effective February 19, requests for information should be directed as follows:
Biotechnology Information: Ray Dobert, Director, Biotechnology Information Center, USDA National Agricultural Library; 301-504- 5340, fax: 301-504-7098, email: rdobert@nalusda.gov.
Biotechnology Regulations: John Payne, Acting Director, BBEP, USDA/APHIS; 301-734-7602, fax: 301-734-8669, email: jpayne@aphis.usda.gov.
Technology Transfer and Patenting: Richard Parry, USDA/ARS; 202- 720-3973, fax: 202-720-7549, email: parryr@ars.usda.gov.
Media and Press Information: Maria Bynum, USDA Office of Communications; 202-720-5192, fax: 202-690-3611, email:mbynum@usda.gov.
Anyone with an interest in the scientific, environmental, or methodological techniques underpinning the environmental risk assessment of biotechnology is invited to take part in the 8th Symposium on Environmental Releases of Biotechnology Products: Risk Assessment Methods and Research Progress. The conference, jointly sponsored by Environment Canada, Agriculture and Agri- Food Canada, the USDA and the U.S. EPA, is scheduled for June 23- 26 at the Radisson Hotel in Ottawa, Ontario. There is no registration fee.
The symposium is the latest in a series of conferences designed
to assist risk assessors in the development of effective and
scientifically defensible protocols for risk assessment of
microbial, plant, and animal products having environmental,
agricultural, forestry, mining, and aquaculture applications. The
goals of this year's conference, entitled "Risk Assessment at the
Crossroads", are:
Of particular interest will be the profiling of current inhouse and extramural environmental risk assessment research activities in such areas as:
For information about submitting an abstract, giving a poster presentation, or registration, contact Dr. Morrie Levin (410-234- 8800; fax: 410-234-8896; email: levin@mbisgi.umd.edu) or Dr. Scott Angle (301-405-9041; fax: 301-405-1346; email: ja35@umail.umd.edu).
Biotechnology R&D is conducted primarily in developed countries, but many biotech products, particularly in agriculture, have a global market. Trade issues and the potential for the technology to improve food, feed, and fiber production worldwide are helping to drive international efforts towards harmonization of biotechnology regulations. At the same time, needs and priorities of individual countries will influence how they oversee incountry research, importations, and small and large scale releases of genetically engineered organisms. From time to time, the News Report will take a look at agbiotech developments in other countries. We begin with a report from Australia, with two articles that describe the regulatory system for genetically modified organisms, and the biosafety review of two crops intended for general release as commercial products. The case of Monsanto's Bt cotton is interesting in that it illustrates the necessity of considering local environmental factors when determining the potential impact of a release. The third article, a case study for the release of a virus resistant white clover, systematically identifies potential risks, addresses them, and invites comments on the issues raised.
Pat Traynor, Editor
Information Systems for Biotechnology
The Genetic Manipulation Advisory Committee (GMAC) is a non-statutory body established by the Commonwealth Government to oversee the development and use of novel genetic manipulation techniques in Australia. It assesses whether such work poses potential hazards to the community and the environment and recommends appropriate safety procedures and containment of organisms to researchers, commercial institutions and government regulatory bodies. GMAC has 21 part-time members drawn from disciplines relevant to assessing genetic manipulation proposals. Members are appointed to the Committee by the Minister for Administrative Services and are chosen for their expertise, rather than as representatives of a particular interest group.
Negotiations are continuing regarding the establishment of a new statutory authority to replace GMAC. The Commonwealth-State Consultative Group on Genetic Manipulation is developing the instructions for drafting the proposed legislation and the Department of Industry, Science and Technology has carriage of this legislation.
Proposals for the release of genetically modified organisms (GMOs) into the environment are assessed by GMAC's Scientific and Planned Release Subcommittees which will advise if the proposals raise any safety concerns. The Scientific Subcommittee reviews the molecular aspects of all proposals covered by GMAC's Guidelines, and the Planned Release Subcommittee assesses the hazards associated with the planned release into the environment of genetically manipulated live organisms falling within the Guidelines. (GMAC Guidelines are available free of charge via email request to: gmac.secretariat@das.gov.au)
Advice is typically sent out on planned releases about 12 weeks after the quarterly cut-off date for receipt. This allows time for a summary of the proposal to be published in the Commonwealth of Australia Government Notices Gazette and circulated to interested individuals and organizations registered with GMAC for this purpose. The public has 30 days to comment and these comments are taken into account in GMAC's consideration of the proposal. The proposal is then considered at a Planned Release Subcommittee meeting and advice is provided several weeks after this date.
GMAC produces Public Information Sheets on each planned release proposal it assesses. These include details of the organism, the vector, the genetic modification and procedures for release, together with a summary of the risk assessment and GMAC's recommendations for the planned release. [Public Information Sheets are available at http://binas.unido.or.at/binas/releases/]
Approvals for commercial release are given by regulatory agencies that have statutory power over the end-use of genetically manipulated products, such as the National Registration Authority for Agricultural and Veterinary Chemicals. Permission to import any biological material into Australia must be obtained from the Commonwealth Department of Primary Industries and Energy.
Kirsty McLean, GMAC Secretariat
kirsty.mclean@das.gov.au
The development of some genetically modified organisms is moving towards their commercial release to the Australian market-place. GMAC considers all data on small field and demonstration trials, and if these indicate that the modified organism poses negligible risk (that is, the risk is no greater than that associated with the unmodified organism), the proponent must make a submission to GMAC indicating the intention to proceed to commercial release of the organism. GMAC publishes a notice of this intention, inviting public comment, and makes an assessment of the submission. The proponent must also satisfy the requirements of other agencies that have statutory power over the end-use of genetically manipulated products (e.g. the Therapeutic Goods Administration, the National Registration Authority for Agricultural and Veterinary Chemicals).
GMAC has assessed two applications for general release from a company intending to market genetically modified carnations with an extended vase life, or an altered petal color. GMAC's assessment of the general release of the transgenic plants was that it posed negligible risk to the environment or the community. Carnation has no weedy characteristics and is not closely related to any weed in Australia. The biology of carnation is such that there are no realistic ways for the genetically modified plants to escape from cultivation and become established as populations in the wild, or for gene dispersal from the genetically modified carnation to occur. In the case of the carnations, there is no agency with regulatory authority over ornamental plants and so GMAC advice was sufficient for the general release to proceed.
GMAC has just issued advice on an application from Monsanto Australia for cotton plants expressing the CryIA(c) gene from the bacterium Bacillus thuringiensis (Bt). This gene encodes a protein that is toxic to the major caterpillar pests of cotton. In view of the lack of data on 1) the consequences of transfer of the Bt gene to native Australian Gossypium species, and 2) appropriate resistance management strategies, GMAC's view was that unrestricted commercial release of the Bt cotton should not take place at this stage. However, GMAC recognized that large scale planting of the Bt cotton may provide data that would help in making a decision on whether full commercial release should proceed.
GMAC's specific recommendations for the conduct of the release included: confining the release to the cotton-growing areas of southern Queensland and New South Wales (in view of the diversity of native Gossypium species present in other regions); staging the scale of the release, with gradual increases in the area sown to Bt cotton each season; and limiting the release to five years' duration, with a further review after this period. During this time, further research was required concerning resistance management strategies and the possible ecological consequences of gene transfer to wild Gossypium species.
Kirsty McLean
Editor's Note: Last month's News Report presented highlights from a summary of the USDA/APHIS sponsored workshop on transgenic virus resistant plants. The meeting was held to discuss environmental and agricultural risks that might be associated with the commercialization of these crops. Conclusions and recommendations were necessarily general, due to the diversity of crops being engineered, virus pathogens being targeted, and potential release sites. To look at a specific case, the following article addresses the environmental issues that may be raised by the release in Australia of white clover engineered for resistance to alfalfa mosaic virus (AMV). The author invites your comments.
Two years ago, the Victorian state government's Department of Agriculture (now Agriculture Victoria) relocated its pasture biotechnology research to the Bundoora campus of La Trobe University. I am a plant virologist with interests in plant virus epidemiology and manage a program to introduce resistance to alfalfa mosaic virus into white clover using various methods similar to the coat protein strategy. We have developed a regeneration process which gives us extensive adventitious shoot production on over 80% of the genotypes of all white clover and sub-clover cultivars that we have examined, and on most species of Medicago. We have several transgenic lines of white clover expressing sequences from AMV, some of which are immune and others which show good levels of resistance. We approach now the stage of field trials and it is on this matter I wish to open up direct debate. The following issues are considered with respect to releasing our virus resistant white clover.
Gene Spread
Recombination
Transcapsidation
Synergistic Effects
The list of issues is slowly growing, and growing in complexity. I would therefore welcome any debate on these or other issues, especially those supported by references. Also, subject to agreement with my commercial partners, I would welcome discussion on collaborative work to answer some of these issues, or to evaluate our products.
Ron Garrett, PS&B, Agriculture Victoria
La Trobe University
agrrgg@lube.latrobe.edu.au
LONG TERM STABILITY OF TRANSGENES IN APPLE TREES
Apple trees transformed seven years ago with a reporter gene and a selectable marker have been shown to express the genes and transmit them to progeny. Researchers at Horticultural Research International (West Malling, UK) showed, for the first time in a tree species, long term stability of transgene expression in both fruit and vegetative tissues and Mendelian segregation of both transgenes in the R1 progeny (Bio/Technology, January 1996).
The report presents physiological, biochemical and molecular evidence for stable expression of nopaline synthase (nos) and neomycin phosphotransferase (nptII) genes that were introduced by Agrobacterium-mediated leaf disk transformation in 1986. Transgene inactivation, widely reported in transgenic plants, was not seen in the apple tissues. It remains to be seen whether apples are an exception to the general phenomenon, or whether inactivation would occur in other cultivars, with other genes, or under different environmental conditions.
Transformation of long-lived perennial fruit crops is worthwhile only if introduced genes are stably expressed in the relevant tissues over a long period of time. To be useful in breeding programs, the transgenes must show Mendelian segregation and stable expression in the progeny, as well. Thus the findings reported for transgenic apple trees have significant implications for the use of genetic manipulation in fruit tree improvement programs. Apple trees have already been transformed with a Bacillus thuringiensis endotoxin gene that can control insect predation. Genes for key enzymes in the ripening process have been isolated from apples, cloned and sequenced. It should now be possible to introduce these and other genes with potential to improve apple fruit quality and pest and disease resistance.
- P. T.
Monsanto scientists have added a new selectable marker to the list of agents available for transformation of monocot species. The dual-gene system for herbicide tolerance combines two biochemical mechanisms for tolerance to glyphosate, and has been shown to be effective in wheat and corn, as well as several dicot species. The new marker was reported in a recent publication (Plant Cell Reports, 1995. 15:159).
Previously available selectable markers are limited in their effectiveness with monocots. Kanamycin resistance, widely used to select dicots transformed with the nptII marker gene, is less effective for cereal crops due to their high level of endogenous tolerance to the antibiotic. Transformation with the bar gene, which confers tolerance to glufosinate, allows selection in the presence of the herbicide. In practice, however, the process is not airtight and some nontransformants escape the selection.
The new marker system uses two bacterial genes: CP4, encoding a glyphosate tolerant form of EPSPS, the critical enzyme in aromatic amino acid biosynthesis normally inhibited by the herbicide, and GOX, encoding glyphosate oxidoreductase, an enzyme that degrades the herbicide into a nontoxic compound. The genes are carried on a single plasmid and delivered into immature wheat embryos by particle bombardment.
Actual selection of transformants does not occur until several steps later in the regeneration process. First, bombarded embryos are placed on callus medium for a week, then transferred to selection medium containing 2mM glyphosate. At this stage, non- transformed embryos can grow and form callus even at high levels (up to 4mM) of glyphosate. After 9-12 weeks, the callus is transferred to regenerating medium containing 0.2mM glyphosate. This is the step where selection actually occurs. Transformed callus can regenerate green shoots that form healthy roots and grow into normal-looking, fertile plants. Nontransformed callus cannot develop shoots even when the herbicide concentration is cut in half. Selection appears very effective, as no escape on the regenerating medium has been seen so far.
Genetic engineering of monocots has presented physiological and technical hurdles not seen with broadleaf species. One of the most limiting has been a lack of effective and alternative transformation selection systems. Now, a new selectable marker for wheat lowers one of the barriers to developing a wider range of transgenic cereals and grasses.
- P. T.
PCR FOR SEX DETERMINATION IN CATTLE
A polymerase chain reaction-based method has been developed for sexing of bovine embryos on the farm. The study involved manual biopsy of 24 embryos, with diagnosis of sex based upon restriction fragment length polymorphism (RFLP) analysis of DNA amplified with primers specific to a highly repeated region of the Y chromosome. After 50 cycles, reaction tubes were examined under UV illumination for fluorescence indicative of the presence of amplified Y-chromosomal DNA. Results obtained by this method were in complete agreement with those obtained by the standard method, suggesting that sexing of embryos by PCR, without the traditional electrophoretic detection of product, may provide a useful field test.
J. Glenn Songer
University of Arizona
PCR amplification and RFLP analysis were applied to detection and identification of four pathogenic and ten nonpathogenic avian mycoplasmas. A fragment of the 16S ribosomal RNA gene was amplified using oligonucleotide primers derived from sequences common to all avian mycoplasmas tested. Use of multiple restriction digestions for RFLP analysis allowed differentiation of the fourteen species of mycoplasmas tested. These molecular methods may find wide use in diagnosis of mycoplasma-induced in domestic poultry.
PCR has also found utility in differentiation of Marek's disease virus, a herpesvirus causing a lymphoproliferative disease of chickens, from reticuloendotheliosis virus, a retrovirus which causes bursal and nonbursal lymphomas resembling lesions of Marek's disease. Primers unique for each virus were synthesized, and the assay was standardized with template DNA obtained from chick embryo fibroblasts infected with Marek's disease virus or reticuloendotheliosis virus. Normal and tumor-bearing birds from multiple flocks of chickens and turkeys were studied, examining birds by enzyme immunoassay for antibodies, by virus isolation, and by histopathology. PCR was shown to be a useful tool in differentiating the two infections, far more rapid than other assays.
- J.G.S.
The Mediterranean fruit fly, Ceratitis capitata, is the latest in a very short list of insects to be genetically engineered. In an article in the December 22 issue of Science, a group at the Foundation for Research and Technology in Heraklion, Greece reported the successful introduction of a marker gene into medflies.
Developing techniques for insect transformation has been a challenge since the early 1980s, when another species of fruit fly, the familiar Drosophila, was first transformed. Taking a cue from the earlier work, the medfly group used a transposable element system to shuttle an eye color gene into a mutant strain having white eyes. Almost 4000 medfly embryos were injected with the DNA of the transposable element minos and a wildtype red eye color gene. Insertion of the DNA into cells that would give rise to eggs and sperm would impart the gene to the progeny of the injected embryos. The appearance of descendants with normal red eyes confirmed success of the procedure.
Medflies are an economically important pest of fruit and coffee crops worldwide. This work marks an important milestone in efforts to develop new strategies to genetically disarm them.
- P. T.
Haplosporidium nelsoni is a major pathogen of the eastern oyster in its beds along the Atlantic coast of the US. For diagnostic purposes, it was desirable to develop a DNA probe to allow specific and sensitive detection of the organism. Nucleotide sequences of the small subunit of rDNA from H. nelsoni, other haplosporidians, and various procaryotes were aligned, and an oligonucleotide probe was derived from a sequence unique to H. nelsoni.
The probe, capable of detecting as little as 100 pg of cloned target DNA, was able to detect H. nelsoni-specific sequences in a one microgram sample of genomic DNA from an infected oyster. The probe hybridized well with at least two forms of the pathogen in fixed, paraffin-embedded, infected oyster tissue, but not with non-infected tissue or other, similar oyster pathogens.
- J.G.S.
The accounting and consulting firm Ernst & Young (E&Y) has published an annual primer on the biotechnology industry since 1986. The annual report is thought by many to be a good overview of the state of commercial biotechnology. The 10th anniversary edition was released at the beginning of this year and offers some insights into the biotechnology industry and some specifics on the agricultural biotechnology sector.
The report is entitled "Biotech 96 - Pursuing Sustainability" and focuses on the efforts of biotechnology firms to develop into "sustainable business entities" through traditional means of partnering, merger/acquisition and reshaping strategies in search of longevity. In spite of the perceptions of some that the industry was in distress, a number of industrial indicators were up in 1995. These include product sales, R&D expenditures, and market capitalization. As a benchmark of the industry's growth over the last 10 years, the report compared statistics from 1986 and 1996. Sales increased from $1.1 billion to $9.3 billion, R&D expense grew from $1.7 billion to $7. 7 billion, the number of biotechnology companies expanded from 850 to over 1 ,300, and the number of employees grew from 40,000 to 108,000. Although having grown substantially over the last ten years, the U.S. biotechnology industry is still small in comparison to other high tech sectors. While the U.S. biotechnology industry had 1995 sales of $9.3 billion, the U.S. computer industry, semiconductor industry, and software industry had annual sales of $80 billion, $45 billion, and $97 billion, respectively.
With regard to the agricultural biotechnology sector, the report points out that although the sector has great promise, companies have been slow to get through the regulatory cycle which has hurt profitability. It also notes that the sector's leaders such as Calgene and Mycogen have not been the fast-growing profitable companies a new industry needs to attract investment. In spite of Wall Street's reluctance to pay attention to the ag-sector, E&Y feels that the promise of the technology presents an excellent long-term investment opportunity.
The following table shows a comparison of the average agbiotech company to companies in other sectors of the U.S. biotechnology industry.
Product sales:
Agbiotech......$12.8 million
Therapeutic....$28.2 million
Diagnostic.....$12.6 million
R&D expenses:
Agbiotech......$ 4.4 million
Therapeutic....$21.0 million
Diagnostic.....$ 4.7 million
Net profit (loss):
Agbiotech......($ 6.5 million)
Therapeutic....($10.1 million)
Diagnostic.....($ 3.8 million)
Dr. Jerry Caulder, President and CEO of Mycogen Corporation, describes in the report the challenge that agbiotech can help overcome. "The five billion people who are on earth now are going to expand to ten billion by about 2035. To feed the additional five billion people who are coming to dinner, we have to produce more calories in the next forty years than we have produced since we started farming 10 thousand years ago."
Caulder also points to success drivers of sustainable agbiotech companies. "In the year 2000 and beyond, those companies that succeed will have properly built a technological base that is going to keep their pipelines full. They will also have successfully protected their technology with patents, so that even in comparison to the larger companies, they are perceived as a player, as someone to deal with in a particular business area."
Reference:
Lee, K.B., Jr. and Burrill, G.S., Biotech 96 - Pursuing
Sustainability. Ernst & Young, Palo Alto, CA, 1995.
William O. Bullock
Institute for Biotechnology Information, LLC
Research Triangle Park, NC
April 20-22: Workshop on Transgenic Plants: Biology and Applications, Tuskegee University, Tuskegee, AL. The workshop will provide a forum for students, teachers and researchers to learn first-hand about the latest developments in transgenic plant research from leading scientists, enhance minority participation in plant molecular biology, and promote networking of students with established plant molecular biologists. The program includes lectures by more than 25 eminent scientists on the current status of research on transgenic plants including techniques, strategies for agricultural improvement, commercial applications, ethical issues and social considerations. Also on the agenda are demonstrations of computer databases and internet resources, and panel discussions on "Enhancing Minority Participation in Transgenic Plant Research and Education" and "Funding Opportunities for Research on Transgenic Plants". Contact: Dr. C. S. Prakash, School of Agriculture, Tuskegee University, Tuskegee, AL 36088, Tel: 334-727-8023; FAX: 334- 727-8552; email: prakash@acd.tusk.edu.
May 8-10: International Summit on Phytoremediation, Stouffer Renaissance Hotel, Washington, DC. The conference will address the utilization of plants for environmental clean-up, covering the scope and potential of phytoremediation, regulatory issues, new data and results, and steps to bring the technology to the marketplace. Contact: IBC USA Conferences, Inc. Tel: 508-481-6400, FAX: 508-481-7911
June 11-14: Agbiotech International Conference 1996: Foundation for the Future, Saskatoon, Saskatchewan, Canada. ABIC '96 is intended to put researchers and business people together to explore opportunities for investment and financing; strategic alliances, collaborative partnerships and technology transfer. It will have over 90 sessions covering almost every aspect of agricultural biotechnology. The conference is organized in five concurrent streams: Crop Development, Animal Science, Microbials, Technology Transfer, and Business. Contact: ABIC 96, FAX: 306-664-6615; email: signatur@eagle.wbm.ca or http://www.lights.com/gaba/index.html.
July 1-5: Theoretical Course: Transgenic Organisms: Biological Risk Assessment, Trieste, Italy. Contact: Ms. Elisabetta Lippolis, ICGEB, Padriciano 99, 34012 Trieste, Italy, Tel: 39-40-3757332; FAX: 39-40-226555; email: courses@icgeb.trieste.it.
August 11-24: International Internship Program: Biosafety Aspects of Agricultural Biotechnology, Michigan State University, East Lansing, MI. A two week program provides training in the biosafety review process for environmental release of genetically engineered organisms. The course is targeted to participants from developing countries where transgenic crops are ready to be tested. Contact: ABSP Networking Office, Michigan State University, 414 Plant and Soil Sciences Building, East Lansing, MI 48824-1325, Tel: 517-353-5262; FAX: 517-432-1982; email: kmaredia@msu.edu.
The material in this News Report is compiled by NBIAP's Information Systems for Biotechnology, a joint project of USDA/CSREES and the Virginia Polytechnic Institute and State University. It does not necessarily reflect the views of the U.S. Department of Agriculture or of Virginia Tech. The News Report is distributed free of charge and may be photocopied or otherwise reproduced. P.L. Traynor, Editor.
Information Systems for Biotechnology, 120 Engel Hall, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061-0308, tel: 540-231-2620, fax: 540-231-2614, email: traynor@nbiap.biochem.vt.edu
For internet access to the News Report, textfiles, and databases
use one of the following procedures.
1. Through WWW: http://www.isb.vt.edu/
2. Use telnet or gopher to connect to ftp.nbiap.vt.edu
3. Use ftp to connect to ftp.nbiap.vt.edu. Use "anonymous" as
your user-id, your email address as your password. Type "cd
pub/nbiap".
To have the News Report automatically emailed, send an email message to news@nbiap.biochem.vt.edu and type subscribe newsreport in the message section.