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Genetic transformation
Biolistic-mediated genetic transformation is another effective method to produce transgenics in a wide variety of plant species. Although no transgenic tea plants have been grown in the field using this technique, experimental conditions have been standardized by Australian and Chinese scientists. Genetic characterization
Work on molecular markers in tea began in our laboratory in 1994 and a significant amount of work is continuing worldwide (discussed below). Research in India:
In a separate study, twenty-five diverse tea cultivars were analyzed using the simple sequence repeat anchored polymerase chain reaction (SSR-anchored PCR) or Inter SSR-PCR (ISSR). In both cases, cultivars were analyzed using Shannon's diversity index, which revealed that the China type tea group is more diverse than the Assam group. Additionally, we noted that molecular classification matches conventional classification of tea. A species-specific primer was also developed for distinguishing between the Assam and China type tea cultivars. Amplified fragment length polymorphism (AFLP) markers were also studied in depth to detect diversity and genetic differentiation of several important tea clones, including the famous `Darjeeling tea', mainly to protect cultivars for intellectual property rights purposes. Interestingly, the RFLP technique was also used to detect adulteration with cashew husk in 10 different tea samples4. Research abroad:
In Japan, a wide range of markers has been used with various applications. The markers used for genetic characterization of different green tea cultivars are RAPD, AFLP, SSR, CAPS, and RFLP. Importantly, the RFLP technique was also applied in Japan to prevent adulteration of higher grade with lower grade tea. Several other minor tea-producing countries have used different molecular markers to characterize the tea gene pool of introduced tea cultivars available to that country. Such efforts were made using RAPD in Portugal, ISSR in Taiwan, and RAPD in South Africa. All work focused on the genetic characterization and molecular taxonomy of the introduced variety available in the respective countries. Similarly, South Korea and China tea cultivars were characterized through RAPD or AFLP, and RAPD, respectively. Simple sequence repeats (SSR) were derived from C. japonica, a closely related species of tea in Japan. Using these primer pairs, 53 C. japonica ecotypes were genotyped and population genetic parameters calculated. Later, the same group investigated the spatial genetic structure of C. japonica using four of these microsatellite primers. Spatial distribution of individuals was also assessed to obtain an insight into spatial relationships between individuals and alleles. Gene cloning and expression
References 1. Mondal TK et al. (2004) Plant Cell Tissue Org Cult, Netherlands 76, 195-254. 2. Akula A, Akula C (1999) In: Jain SM, Gupta PK, & Newton RJ (eds.), Somatic embryogenesis in Woody plants, Vol. 5, pp 239-259. Kluwer Academic Publishers: The Netherlands. 3. Mondal TK et al. (2001) Plant Cell Reports 20,712-720. 4. Dhiman & Singh (2003) Planta Med 69(9), 882-4.
Tapan Kumar Mondal
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REGULATING PLANT GENES AND AGBIOTECH PRODUCTS INTERNATIONAL STYLE
Challenges from pathogens, pests, and the climate fuel perpetual efforts to develop new crops with mixed traits. When a fungus decimated maize grown in the southern United States, for example, plant breeders devised a way to defeat the pathogen with a natural resistance trait harbored within a variety of African maize. To combat yellow dwarf disease in U.S. barley varieties, researchers harvested resistance genes from Ethiopian plants, while genes from plants originating in the Caucasus and Spain provided the keys for resisting rust disease in U.S. commercial wheat varieties.
Plant breeders can perform these feats of agricultural innovation because they can select traits from a wealth of genetic resources. And since no single country has the full range of naturally occurring genetic resources, the collection and exchange of germplasm requires international cooperation. Yet not every nation considers this exchange equitable. Accusations of biopiracy often arise from countries in tropical and subtropical regions, which possess the majority of the world's agricultural genetic diversity. At the same time, changing land use practices within these countries lead to the loss of uncollected genetic resources. The Food and Agriculture Organization (FAO) of the United Nations estimates that about three quarters of the genetic diversity found in agricultural crops has vanished over the last century.
These concerns inspired the creation of the UN's International Treaty on Plant Genetic Resources for Food and Agriculture, an agreement adopted after seven years of negotiations by delegates from 116 nations. Although drafted in November 2001, the Treaty only came into force on June 29, ninety days after forty governments ratified it. José Esquinas-Alcázar, secretary of FAO's Commission on Genetic Resources for Food and Agriculture, told BBC News Online that the "treaty will ensure the conservation and availability of raw material for agriculture."
The agreement requires each contracting party to explore and conserve its plant genetic resources for food and agriculture. Parties can work toward this objective by surveying their genetic resources and assessing any threats, and by promoting both in situ conservation and the compilation of genetic resources for preservation in public collections.
The treaty also mandates that contracting parties develop and maintain measures to advance the sustainable use of plant genetic resources. Examples of such measures include extending the genetic base of crops available to farmers and supporting plant breeding efforts that strengthen the capacity to develop varieties adapted to particular ecological conditions.
Under the Treaty, countries agree to establish a Multilateral System to facilitate access to plant genetic material and to share the benefits. The Multilateral System applies to plant genetic resources for food and agriculture listed in the first annex of the Treaty that are under the control of the contracting parties and in the public domain. The 35 itemized food crops and 29 forage crops "represent most of the important food crops on which countries rely," says Esquinas-Alcázar. According to one estimate, the Treaty's annex lists crops representing 80 percent of the world's calorie intake. The 600,000 sample gene bank collection held by the Consultative Group on International Agricultural Research will also be administered under the Treaty.
Restrictions apply to the use of these genetic resources. The material must be used to promote conservation, research, breeding, and training for food and agriculture. Any use of genetic material for chemical, pharmaceutical, and other industrial applications falls outside the scope of the Treaty. If an entity incorporates material accessed from the Multilateral System into a commercial food or agricultural product and does not permit others to use the product without restriction for research and breeding, then the Treaty requires payment of an equitable share of any resulting monetary benefits.
The FAO asserts that the Treaty benefits agricultural research because the Multilateral System will reduce transaction costs for the exchange of plant genetic material between countries. That is, researchers will no longer have to negotiate bilateral agreements with each donor country to obtain germplasm.
But before the Treaty can facilitate any transfer of plant genetic material from its germplasm clearinghouse, the countries that ratified the agreement must decide about conditions for access and benefit-sharing, details that will be embodied in a standard material transfer agreement. Conditions recited in this key document will also determine whether the United States will ratify the Treaty.
At least one mystery resides within the Treaty. Article 12 states that "recipients shall not claim any intellectual property or other rights that limit the facilitated access to the plant genetic resources for food and agriculture, or their genetic parts or components, in the form received from the Multilateral System." Even delegates involved in the drafting of the document could not agree about the meaning of this statement.
It might have been the eruption of biopiracy complaints that motivated the inclusion of the ambiguous passage. During the drafting of the Treaty, protests from India provoked the revocation of a U.S. patent covering a use of turmeric and a European patent on a compound derived from the Neem tree, while South American activists prompted the withdrawal of a U.S. patent on an ayahuasca vine variety. In 2001 alone, India, Pakistan, and Thailand voiced
grievances about a U.S. patent on a type of basmati rice, and Mexico protested a U.S. patent on yellow beans.
A copy of the Treaty is available from the FAO
website (http://www.fao.org/ag/cgrfa/itpgr.htm).
Labeling Food as Genetically Modified and Living Modified Organisms as Pests
Production labeling was met with less enthusiasm. Certain delegates, including the U.S. delegation, asserted that consumers would perceive labeling based solely on the method of production as a safety warning. The Canadian delegation argued that method of production labeling fails to comply with Codex's own rule: only factors accepted worldwide should be taken into account as a basis for decision-making. A copy of the report can be obtained from the website of the Codex Alimentarius Commission (http://www.codexalimentarius.net/).
On June 1, the FAO published new guidelines for determining if a living modified organism (LMO) poses a hazard to plants. About 130 countries have adopted this assessment standard, which focuses on the risk that a foreign gene may transform a plant into a weed. The document also offers advice for determining whether genetically modified insects, fungi, or bacteria could harm plant ecosystems. If authorities deem an LMO a threat, then they can decide whether to prohibit or restrict its import and domestic use. The website of the International Plant Protection Convention offers a copy of the guidelines, entitled "Pest Risk Analysis for Quarantine Pests, Including Analysis of Environmental Risks and Living Modified Organisms" (http://www.ippc.int/).
Selected References
Anonymous. (2004) Plant gene treaty becomes law. BBC News. June 29, 2004. Available at: http://news.bbc.co.uk/2/hi/science/nature/3849489.stm.
Fowler C. (2003) The status of public and proprietary germplasm and information: An assessment of recent developments at FAO. IP Strategy Today, No. 7-2003. Available at: http://www.biodevelopments.org/.
Sullivan SN. (2004) Plant genetic resources and the law. Past, present, and future. Plant Physiology, 135: 10-15.
Phillip B.C. Jones, PhD., J.D.
INDIA PRODUCES INDIGENOUS `GM COTTON'
Insects, disease, and drought present the greatest impediments to realizing expected yields in major crops. In addressing these problems, development of transgenic varieties has assumed significance, primarily through use of durable resistance genes. However, a few multinational companies in developed countries own and patent many of these genes. In developing countries, these novel genes are sometimes available to scientists as `gifts' through personal contacts. Though initial transgenic crop development is dependent on these borrowed genes, the varieties developed from them would not be available for commercial cultivation because of contractual obligations generally underlying the `gift' to investigators (as these genes generally are available for academic and experimental purposes only). At best, GM plants thus developed could be tested only for their efficacy in solving designated problems and not developed for commercial cultivation. Deployment of borrowed genes in transgenic crops might also attract patent problems under new IPR regimes.
Scientists, institutes, and seed companies have therefore decided to convene and begin searching and licensing indigenous genes and technologies for endogenous developments. Apparently this exercise is having an effect, as Monsanto's monopoly on genetically modified cotton in India will soon be broken by Swarna Bharat Biotechnics Private Ltd (SBBPL), Hyderabad, India, a consortium of seven Indian seed companies. SBBPL received licenses for two genes derived from Bacillus thuringiensis (Bt), which protect cotton against bollworm (Helicoverpa armigera) and tobacco caterpillar (Spodoptera litura). The genes are licensed from the National Botanical Research Institute (NBRI), Lucknow, India, for Rs. 7.5 ($ 0.16) million over a three year period and a royalty of 3%. SBBPL is soon likely to get license for a third gene (LecGNA 2) that directs production of lectin, a protein lethal to sucking pests such as aphids, from the publicly funded Centre for Plant Molecular Biology (CPMB), Osmania University, Hyderabad, India.
According to NBRI's Deputy Director, Dr Rakesh Tuli, SBBPL has licensed two genesCry1Ac and another killer gene called Cry1Ec. Of these two genes,
Cry1Ec, used against a tobacco caterpillar, is designed and synthesized at NBRI. Because Cry1Ac, which confers resistance against bollworm, is not protected in India, NBRI's
team altered Cry1Ac promoters to create a version with greater expression and stability. According to some experts, although NBRI has modified the gene, there could be legal implications: GM plants derived from this gene might come under the category of `essentially derived varieties' (EDVs) and cannot be registered under India's Plant Variety Protection Act - 2001. Even more, Monsanto might contest the commercialization of Indian indigenous Bt cotton after January 2005, the date when India has to comply with patents covered under the WTO rules.
Tuli admits that only time might solve the problem. Meanwhile, SBBPL is seeking regulatory approval in India for both Cry1Ac and Cry1Ec, with a view toward introducing
the new Bt cotton by 2006.
The consortium's aim is to enter an era of self-sustaining agribiotech development. Satish Kumar, Managing Director of SBBPL, reiterated that they are ready to source beneficial genes from any publicly funded laboratory where they are available. The consortium opines that the advantages of sourcing indigenous technology are economic and strategic. The profit generated by public sector institutes through licensing helps support reinvestment in developing more agribiotech products to serve local needs. The main benefit for consortium members is economic, as the technology access fee is shared by members of the consortium. In addition, Indian partners help with the regulatory process to obtain product approval. Kumar expects that the price of SBBPL seeds would be two-thirds of Monsanto's.
Through licensing these genes, Rs 10 ($ 0.2) billion spent on chemical pesticides can be saved by SBBPL, as 90% of cotton damage is from bollworm and sap sucking pests. Indian farmers spend about Rs 16 ($ 0.35) billion on chemical pesticides. With the introduction of novel Bt cotton varieties, SBBPL, which has a 30% share of the total Indian cottonseed market, expects to claim some of the Rs 30 ($0.66) billion market per year that is presently monopolized by the joint-venture company Monsanto-Mahyco Biotech, Mumbai, India.
Since scientific knowledge is indigenous in India, and smaller players can access costly technology monopolized by big multinational companies, many eminent scientists, activists, scholars, and institutes welcomed this initiative of SBBPL. The activist groups critical of Monsanto's monopoly are happy, as 42% of India's transgenic research has been based on Monsanto's gene. "Finally, we seem to be getting our act together," said Suman Sahai, Convener of Gene Campaign, New Delhi, India.
However, Monsanto is not daunted by the competitionthey have already co-licensed the Cry 1Ac gene to nine more Indian companies whose products are at different stages. Ranjana Smetacek, the company's spokesperson in India, says Monsanto welcomes the widespread usage of Bt cotton. Meanwhile the Council for Scientific and Industrial Research (CSIR), New Delhi, India, the parent institute of NBRI, is finalizing the list of countries soon to file patents for novel genes. Experts suggest that the consortium may try to exploit those GM technologies on crops for which patents held by multinational companies are now expiring. Prabhakar Rao, Managing Director of Nujiveedu seeds (Hyderabad, India), the largest company in the consortium, said that membership would soon reach 19, as many other countries are willing to partner with them. Hence, it appears that a few multinational seed companies can no longer monopolize agribiotechnologies, and local stakeholders can play a crucial role in technology development and commercialization.
References
1. Nat. Biotechnol. 22, 255-256, 2004.
2. Nat. Biotechnol. 21, 590-591, 2003.
3. Nat. Biotechnol. 19, 895-896, 2001,
P. Janaki Krishna
MOLECULAR MARKER ASSISTED SELECTION DOCUMENT
FAO has published a biotechnology-related summary of an e-mail conference on "Molecular marker assisted selection as a potential tool for genetic improvement of crops, forest trees, livestock and fish in developing countries," which ran from 17 November to 14 December 2003. The document summarizes the main arguments and concerns raised during the moderated e-mail conference. The full summary is available at http://www.fao.org/biotech/logs/C10/summary.htm.
ISB News Report
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. Any opinions, findings, conclusions, or recommendations expressed in this publication are those of the author(s) and do not necessarily reflect the view of the U.S. Department of Agriculture, or Virginia Tech. The News Report may be freely photocopied or otherwise distributed without charge.
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Ruth Irwin, Editor (rirwin@vt.edu)
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