Guatemala and Costa Rica can be considered the most advanced Central American countries in biotechnological research and applications. Both countries are using R&D resources to increase their competitiveness in traditional and non­traditional export commodities, but the route from research to commercial application has many obstacles.

Guatemala and Costa Rica, two small countries where agriculture has considerable socio­economic importance, can not afford to ignore the possible opportunities and risks that biotechnology represents for them. In spite of the difficulties for small countries to assemble professional critical mass, economic as well as social considerations make it advisable to move rapidly into biotechnological R&D. For latecomers there is still room to take advantage of technologies produced abroad, although it is important to bear in mind that successful high­tech purchase can only be based on a substantial local R&D effort.

Importance of biotechnology
The predominance of Guatemala and Costa Rica in agro­biotechnological research was confirmed previously, when the Inter­American Institute for Cooperation on Agriculture (IICA) conducted an assessment of research capabilities in Central America. It appeared that in 1988 nearly 40 per cent of the 29 organizations conducting agro­biotechnological research in Central America were based in Costa Rica and 24 per cent in Guatemala. The remaining organizations were located in other Central American countries, i. e. 3 organizations each in Honduras, Panama, Nicaragua and El Salvador. The concentration of human resources in the region was even higher. Nearly half of the researchers were working for Costa Rican organizations (equivalent of 114 full­time researchers) and 37 per cent (87 researchers) in the Guatemalan ones. Nevertheless, in both countries, the amount of human and financial resources devoted to biotechnological research is still relatively low: the two combined have a critical mass equivalent to the staff of a medium­sized biotechnology company in the USA.
Costa Rica and Guatemala were the first in Central America to set up commissions for planning and linking universities, governmental agencies and enterprises dealing with biotechnology. In addition, since 1986 every national plan for science and technology development in Costa Rica gives priority to biotechnological R&D and technology transfer.
In its efforts to get into biotechnology research, Guatemala has benefitted from serving as the headquarters of a regional organization focusing on manufacturing research, the Central American Research Institute for Industry (ICAITI). Costa Rica has been taking advantage of housing the headquarters of the Tropical Agriculture Research and Training Center (CATIE), another regional organization whose mandate concentrates on agriculture.

The challenge for small economies
Guatemala and Costa Rica constitute small economies. With an area of 50,900 square kilometres and nearly 3 million inhabitants, Costa Rica's gross domestic product (GDP) was about US$ 5 billion in 1991. With nearly twice the area and three times the population, Guatemala had a GDP of US$ 8.6 billion (1991). The smallness of the Costa Rican and Guatemalan economies can be illustrated by a comparison with Sweden, a country with a population of 8.6 million, which had a GDP of about US$ 228 billion in 1990.
Due to their limited size and their relatively incipient industrial activity, Costa Rica trades annually roughly 80 per cent of its GDP (either exported or imported), while Guatemala trades 40 per cent. In contrast, Japan and the USA traded only 16 per cent of their GDP in 1990.
Most of Guatemalan and Costa Rican exports are agriculture related. In 1991, the share of agriculture in exports was roughly 60 per cent in Costa Rica, and 65 per cent in Guatemala. Moreover, despite attempts to diversify their exports, a very limited number of commodities are still extremely important for their foreign exchange revenue. In 1991, coffee, a long­established export product for both countries, still commanded nearly 16 per cent of all foreign exchange revenues in Costa Rica, and 20 per cent in Guatemala. Bananas is the main foreign exchange earner in Costa Rica (25 per cent of exports) and ranks third in Guatemala. Sugar still ranks second in Guatemala. Due to the high contribution of export to the GDP, changes in the export commodity markets can indeed be critical for economic and social stability.

Coffee improvement: Constraints from lab to field
In 1978, IICA started a coffee research programme funded by the United States Agency for International Development (USAID) involving Central American countries, Mexico and the Dominican Republic. One of the first mandates received by this project, officially called the Co­operative Regional Program for Technological Development and Modernization of Coffee Cultivation (PROMECAFE), was related to the control of and research on coffee rust, a new fungus disease in Central America at that time. Catimor cultivars (resistant to the disease) were bought through the project and transferred to the coffee research programmes of PROMECAFE's member countries. Several other issues of agronomic interest were tackled: epidemiology of coffee rust was extensively studied as well as the optimal application of chemicals, nematode control, etc. Biological control of coffee berry borer (Hipothenemus hampei) was also developed and transferred to the countries through this programme.
Mainly through the technical assistance of the French Centre for International Co­operation in Agricultural Research for Development (CIRAD), biotechnology became part of the coffee growing promotion efforts. As early as 1983, micropropagation of coffee was studied, and tissue culture laboratories were installed at CATIE headquarters and at Guatemala's coffee growers' association Asociacion Nacional del Cafe (ANACAFE). Several French biotechnologists are involved in the project, and at the same time local capacities in coffee tissue culture have been upgraded in the framework of PROMECAFE activities.
Despite the huge potential market, at least in Central America, several biological engineering and scaling­up problems of coffee micropropagation must be solved in order to make the application of these technologies economically feasible. The current activity of the laboratories is not geared to the seed market. Sexual propagation of coffee has proven to be much cheaper than micropropagation and still the vast majority, if not all, commercial coffee plantations are sown by the conventional system. Therefore, in the short term micropropagation is unlikely to replace seed in coffee growing.
PROMECAFE's researchers envisage somatic embryogenesis in coffee as an interesting technology that could eventually reduce the costs of micropropagation. Somatic embryogenesis currently faces a major constraint: somaclonal variation caused by mutations. The researchers expect to solve this problem in the coming years.
In spite of the difficulties encountered in developing tissue culture as an economical way to propagate coffee, the technology is considered as very useful in the future for non­propagation purposes. Firstly, the genetic base for coffee is very narrow in the Americas. Its origin can be traced to the Horn of Africa and production is based on a few strains brought to Brazil, Central America and the Caribbean via European botanical gardens around 1725. Therefore, the genetic base of coffee in the Americas has not changed much since the crop was brought to the continent. There is a strong need to widen the genetic pool of coffee, and tissue culture opens the possibility to exchange material safely and cheaply. Several coffee cultivars from its center of origin are in the genebank of CATIE, and at ANACAFE several interesting varieties for improvement are available in the tissue culture laboratory.
Secondly, resistance to nematodes and other coffee diseases could be studied in vitro. Field trials cannot be avoided, but promising in vitro material may be preselected and brought to the field, speeding up improvement, which has proven to be especially valuable in a perennial crop as coffee.
Recently, a researcher from CIRAD has started to use randomly amplified polymorphic DNAs (RAPDs) for the genetic characterization of coffee. The idea is to use RAPDs to analyze coffee germplasm at CATIE in order to speed­up improvement by selecting for sufficient variation. Nearly one thousand accessions at CATIE will be studied using this technology.

In addition to the biological difficulties of coffee micropropagation, another problem facing coffee tissue culture is that of up­scaling, which clearly escapes the domain of biotechnologists. Entrepreneurial and engineering resources should be deployed in order to make this technology available to Central American coffee growers, who have been facing sharp price declines over the last years. Lack of solid biotechnology industry and venture­capital schemes have prevented coffee micropropagation plantlets reaching the fields.

Reaching the market with tissue culture
Drawing on their past experience of producing foundation seed, Instituto de Ciencia y Tecnologia Agropecuaria (ICTA, Guatemala) began to work on the micropropagation of potato. In the past two years they have been providing private companies with foundation seed of potato, which is later propagated and sold to farmers by private seed companies. In this way, it has been estimated that ICTA's tissue culture laboratories annually provided nearly 120,000 plantlets. ICTA trained part of its staff at the University of Valle (Guatemala). Researchers at this university have been working in micropropagation of garlic and cardamom.
ICTA has also developed micropropagation protocols for apple and is negotiating to deliver between 3,500 to 5,000 plantlets per year to a co­operative established by farmers. Additionally, ICTA and a multinational corporation are negotiating the development by ICTA of protocols for seedless watermelon propagation, and the production and delivery of plantlets to the company.
In Costa Rica, micropropagation of bananas has proved to be able to compete with more conventional technologies. During 1985­1991, the area planted with bananas grew rapidly in Costa Rica from about 25,000 hectares to 40,000. The reasons behind the expansion were the prospects of more open European markets, relatively high prices in the mid­1980s, the reallocation of plantations from Colombia to Costa Rica, and private and public stimulation policies. It has been estimated that nearly 20 per cent of the material used during this period was in vitro material. Several Costa Rican biotechnology companies profited from this newly created market. Banana expansion even prompted the creation of some of them. When area expansion halted as quotas were imposed by European markets, some of the micropropagation companies involved in banana micropropagation went bankrupt, while others survived mainly by going into micropropagation of ornamental plants.
Although these examples show that tissue culture can reach the market, it is important to take into account the fact that profitability was not an important motivation for these governmental agencies. Indeed, private efforts to engage in micropropagation technologies are regarded as marginal for several important agricultural commodities in Costa Rica, due to the lack of profitability. The two ICTA laboratories, for instance, have a combined capacity of at least 600,000 plantlets, far more than the actual number they deliver to the market.
An example of a sector in which tissue culture seems to have potential is the ornamental plant, as it typically demands virus­free, homogeneous propagation material. However, it has been estimated that only 2 per cent of propagation material for ornamental plants production comes from in vitro culture. The reasons for this are small­producer predominance and geographical dispersion.
Additionally, increasing competition from India and Malaysia has prevented Costa Rican companies in the tissue­culture business devoting large sums to R&D or purchasing higher yielding (but more expensive) in vitro material. The low profit margin forces Costa Rica to rely more on a low input­low return strategy.
 

Centro de Investigaciones en Biologia Celular y Molecular  The Centro de Investigaciones en Biologia Celular y Molecular (CIBCM) was established at the University of Costa Rica in 1977, as both a research centre and a master's degree training venue for Costa Rican and other Latin American scientists. Since its establishment, 34 students have completed research work for their Master's dissertation at CIBCM.  The Centro's main competitive advantage lies in its long­term involvement with human health and tropical virus diseases in plants. With a current staff of 50 people, 23 of them researchers, the Centro has successfully developed contacts with their counterparts in industrial countries. Most projects are developed jointly with foreign researchers or within international networks.  PCR  For instance, two researchers were sent to the University of California to master the polymerase chain reaction (PCR) technique. As a result CIBCM has been able to use PCR since 1990 to map genetic diseases. Using PCR, researchers are describing the gene sequence that explains the appearance of Deafness of the Monge in nearly 100 Costa Rican individuals from one family (the Monge family). Although genetic therapy to overcome diseases could be difficult to realize, the understanding of the genetic base is the first step to devising future therapies.  Transgenic rice  Drawing on their past experience with virus research, the CIBCM is devoting efforts to genetically transform rice, maize and criollo melon. Maize and rice are staple foods in Costa Rica. The rice consumption per capita, for instance, has been estimated at nearly 45 kilos per year. Even though most domestic rice consumption is still supplied by Costa Rican producers, deregulation is eroding their competitive position vis­a­vis Thai, US, and more recently Vietnamese producers. Besides, rice varieties suitable to Costa Rican agro­ecological conditions are Indicas, instead of Japonicas dominant in the North. Therefore, solutions for technological problems cannot be expected to come from industrialized countries, but must be produced using domestic scientific and technological capacities, or make use of the research conducted at the International Rice Research Institute. Costa Rican maize production has also been confronted with strong competition by US farmers, as a result of higher US yields and subsidies.  Criollo melon is not of the same critical socio­economical importance as rice or maize. Nevertheless, the CIBCM has developed expertise in a virus affecting melons.  Currently, the rice transformation project is most advanced. It is funded by the Rockefeller Foundation, USA, within their rice biotechnology research network. Varieties developed in Costa Rica suffer the hoja blanca virus disease (also called chlorosis or white band) transmitted by the leafhopper Sagatodes oryzicola. The hoja blanca disease is a serious constraint to rice production in the country. Last year, for instance, large­scale producers lost about a third of the crop because of the disease, medium­scale and small­scale farmers lost even more.  In 1990, a project was started by the Rockefeller Foundation to study the molecular biology of the hoja blanca virus. The idea is to transfer the gene of the capside of the virus to the plant, making it resistant to the virus. How this transference confers resistance is not yet entirely known.  At the moment rice regeneration protocols have been completed. Two varieties reached nearly 50 per cent of regeneration in vitro, showing potential for future genetic transformation. At the same time, a CIBCM researcher developed a 'helium pistol` or particle accelerator. This instrument permits the utilization of the so­called biolistic method for the transfer of DNA, by means of a 'bombardment` with particles of gold or tungsten covered by the DNA of interest. Parameters of bombardment are being optimized using common bean (Phaseolus vulgaris) leaves. Simultaneously, electroporation methods for DNA transfer are being studied. With this method, intermittent, continuous electric pulses are applied to plant cells, in which the cell wall has been removed. The cells are suspended in a liquid medium, which allows the electric current to pass. This produces changes in the membrane, facilitating DNA penetration.  At the moment, researchers do not have a deadline for the release of the transgenic rice varieties. Because of long­established contacts with the Rice Improvement Programme of the Ministry of Agriculture, the National Seed Bureau (a governmental regulatory body) and the rice growers themselves, a rapid process of adoption of the new varieties is expected.  Sources  Personal communication with Gabriel Macaya, Ana Mercedes Espinoza and Pedro Leon, CIBCM.

University involvement
Although most promises of biotechnology have not materialized, many developing countries' policy­makers and researchers regard biotechnology as a useful tool. Given that agricultural research is highly site­specific, it is not very likely that researchers in the North will be interested in the development of biotechnological materials for the South.
Strong ties among Northern and Southern universities, however, might enable developing countries to take advantage of the new technologies. Years of budget constraints at Southern universities and a more aggressive entrepreneurial community have intensified the relationship among university researchers and entrepreneurs. Therefore, once university researchers master the techniques, products and services can be transferred to the private sector.
Researchers at the Universidad de San Carlos de Guatemala (USAG) have been involved in biotechnology research since the mid­eighties, and are currently negotiating with sugar cane producers on the development of sugar cane micropropagation protocols. Research at the USAG further includes anther culture in rice and wheat and tissue culture, mainly in common bean. Besides, nearly 700 species of orchids are kept in genebanks through tissue culture, as well as more than half of the national collection of cassava and sweet potato. Tissue culture has proven to be a very cheap way to preserve biodiversity, especially of the tropical fruits that are present in the country. Research in molecular markers for cocoa has recently begun. The creation of a R&D unit for tissue culture is under consideration.
According to Guatemalan researchers, the development of transgenic plants is possible through sending sequentially a group of researchers for doctoral studies abroad. The final objective is to develop new transgenic materials, suited to their own agro­ecological and socio­economical needs. Currently Costa Rican researchers are working on the same construction, mainly at the Centro de Investigacion en Biologia Celular y Molecular, at the University of Costa Rica (see box). In the medium term the results seem to be successful.

Both Guatemala and Costa Rica are to benefit from opportunities that new biotechnologies create. Getting access to research seems to be the first, inevitable step. Transferring these capabilities to the market appears to be a much more difficult task. Nevertheless, relatively less complex biotechnologies, such as tissue culture, have shown the potential of reaching the market. Many other technologies, such as somatic embryogenesis, are still in the laboratories of industrialized countries, but they probably will evolve into commercial technologies. These innovations will reach the markets of those developing countries prepared to take advantage of them: the ones with long­term, strong R&D efforts in biotechnology, capable of researching the advantages and disadvantages of different technologies.
Walter Jaffé/Miguel Rojas (IICA).

Sources
Economic Commission for Latin America and the Caribbean (1994), Statistical Yearbook for Latin America and the Caribbean. Santiago, Chile: ECLAC.

Inter­American Development Bank (1993), Economic and Social Progress in Latin America. CITY: IDB.

IICA (1988), PROMECAFE: 10 años de labores 1978­1988. San Jose: IICA.

IICA and FEDEPRICAP (1989), Oportunidades de las Biotecnologias Agropecuarias en America Central. San Jose, Costa Rica: IICA.

Personal communications with Carlos Humberto Morales, PROMECAFE, IICA Guatemala; Francisco Anzueto, ANACAFE; Edgar Franco, Fernando Rodriguez Bracamonte and Luis Mejia, University of San Carlos the Guatemala; Oscar Arias Moreira, Agribiotecnologias, Costa Rica; Benoit Bertrand, PROMECAFE­CIRAD.