Biotechnology in Developing countries

II AGRICULTURAL BIOTECHNOLOGIES:
ACHIEVEMENTS, COOPERATION, AND PROSPECTS

A. Asia

1. China

Anther culture was being practiced in about 1000 institutions throughout China. Chinese scientists, a world record, obtained a high number of microspore-derived plant species. These species included wheat (Triticum aestivum)] maize, hot pepper (Capsicum annuum), and sugar-beet (Beta Vulgaris). The acreage of haploid paddy rice and wheat was extended to millions of hectares, with outstanding econoic profit [Zhaoxiang and Yongchun, 1990]. The use of embryo rescue technique and in vitro culture of a hybrid between Triticum aestivum and Agropyron elongatum led to a new wheat variety "Xiaoyan no. 6, " which, in 1990, was grown on 38 million ha, with a yield of 16 million tonnes.

It became possible to propagate more than 100 crop species, by in Vitro tissue culture. In vitro micropropagation is also used for the clonal multiplication of banana: in the Guangdong province, 3-4 million banana plantlets are produced annually by tissue culture, and 1 million plantlets have been exported. Meristem culture is also being carried out to produce virus-free seedlings of potato, strawberry, garlic, and asparagus (Asparagus cochinchiensis). Seedlings of virus-free potato have been commercialized and cultivated on about 300,000 ha (i.e., 10% of the total acreage) in the early 1990s. A propagation and extension system for virus-free potato production has been established in several provinces, and yields have been increased by up to 100 or 200% [Guang-Nan Wu, in paper submitted to the workshop on "Assessment of biotechnology for food production in Vietnam, " Hanoi, 9-12 December 1991).

In the Guangxi province, the sugarcane cultivar "Gueitang 11, " with a high content of sucrose, has been mass-propagated and grown on more than 30,000 ha in 4 years, instead of the 10 years generally needed when conventional propagation is used.

At the Shanghai Institute of Biochemistry, plant genetic engineering uses the introduction of alien DNA by the pollen tube after pollination, with a view to developing resistance to diseases in rice, wheat , soybeans, and cotton. This simple technique of genetic transformation is an indication of the unsophisticated approach chosen in some Chinese laboratories to obtain good results in crop breeding. At the National laboratory of Protein Engineering and plant Genetic Engineering of the university of Beijing (Beida), research on transgenic tomato and rice are being carried out with a view to transferring genes coding for resistance to fungal blast and rice rust, respectively [Borry, 1992].

At the Laboratory of Plant Cell Engineering of the Beijing Academy of Agricultural Sciences, research projects are being carried out on.

Anther culture aimed at selecting new winter wheat and rice varieties Culture of protoplasts and cell fusion (a winter wheat variety has been regenerated from protoplasts) Somaclonal variation and embryo cultures in maize and rice Interspecific crosses to breed disease resistance in winter wheat.

Chinese strains of Bacillus thuringiensis and Bacillus sphaericus were applied successfully to vegetables, maize, rice fields, and pine trees, covering over 18 million ha. For more detailed information, see Sasson (1993). The need to increase China's present grain yields can be met by using both conventional breeding and plant biotechnologies. The rice genome project, supported by the National Commission of Science and Technology, is an important, long-term research project, with major potential applications. The Rockefeller Foundation also supports about 15 laboratories for rice biotechnology research.

2. India

It is anticipated that over the 1993-2000 period, about 8 billion rupees would be invested in health care, 2 billion rupees in agriculture, and some 4 billion rupees in other areas, mainly in most conventional biotechnology-derived products, investments in recombinant products being rather small (Ghosh, 1993).

It is anticipated that the consumption of hybrid seeds would rise from 90,000 tonnes in 1992 to 115,000 tonnes in 1995, and about 165,000 tonnes by 2000. Increasing quantities would be produced locally, to exploit the potential of hybrid vigor in paddy, wheat, rapeseed, and mustard (Ghosh, 1993).

Up to 1993, 22 approvals had been granted giving a capacity of 110 million tissue culture-derived plantlets per year for the whole country (Ghosh, 1993).

During 1988-1989, the Department of Biotechnology (DBT) of the Indian Ministry of Science and Technology launched the Oil Palm Demonstration Projects, with a view to reducing edible oil imports, which accounted for 104 million dollars in 1990-1991, and providing a regular source of income to the farmer, as this crop species is perennial. Many state governments, private entrepreneurs, and financial institutions have drawn up ambitious expansion programs for oil palm in Indian (Kumar et al., 1993).

A program for tissue culture of coconut was launched in 1986-1987. Large-scale plantations using in vitro plants are anticipated after evaluating field performance. In 1995-2008, massive-scale production units will be set up to supply high numbers of plantlets of high-yielding coconut varieties to replace traditional low-yielding ones (Kumar et al., 1993).

In 1984-1985, the A. V. Thomas group (AVT) initiated commercial production of cardamom through tissue culture. On average, tissue culture-derived plantlets show an increase of 63% in yield, the mean estimated yield being 360 kg/ha. AVT has also applied tissue culture to tea, for it owns 2000 ha of tea plantations; the company has been successful in increasing tea yield to 2520 kg/ha., compared with the average South Indian tea yield of 2,060 kg/ha (Kumar et at., 1993).

Unicorn Biotek is a recent example of a tissue culture-based company moving modern biotechnologies from laboratory to marketplace. In addition to a project on the large scale production of virus-free banana plantlets, supported by the Department of Biotechnology, Unicorn Biotek, produced and exported 120,000 Spatiphyllus (a foliage ornamental) and 2000 rose plantlets over 12 months at competitive prices in the world market. In addition to selling to clients in Belgium, The Netherlands, Denmark, and the United States, the company is also present on the local market: farmers in Andhra Pradesh, Karnataka, and Maharashtra bought 20,000 disease-free strawberry plantlets from Unicorn Biotek; they can buy higher numbers of high-yielding and disease-free plantlets from the company, which has improved productivity and lowered the price of tissue culture-derived plantlets. Although Unicorn Biotek is a good illustration of efforts by India's more that 40 tissue-culture companies (in 1993) to bring commercially viable products to the market place, it faces competition from major Southeast Asian companies, and also from other Indian companies (Knudsen, 1993).

Together with the Biotech Consortium of India, Ltd., the Department of Biotechnology (DBT) has assisted in setting up two pilot plants for the mass productionof biocontrol agents. The DBT plans to promote small production units in villages by providing training and credits to entrepreneurs. The major focus is to control one of the world's most damaging pests, the cotton bollworm, recorded in 96 crop species and 61 weeds and wild species in India (Kumar, 1992).

Sandoz India, Ltd., produced a Bacillus thuringiensis Berliner-based biopesticide in September 1992. Similarly, Hindustan Lever, Ltd., the Indian subsidiary of Unilever NV, produced a Bacillus thuringiensis isreaelensis-based biopesticide using molasses as a culture medium. It has been successfully tested against insects attacking cabbage, pigeonpea, mazie, safflower, and cotton. The insecticide is also active against black flies and mosquitoes and therefore, can be used in malaria vector control.

The biopesticides extracted from neem (Azadirachta indica) of the Meliaceae family, are attracting increasing attention from scientists and companies throughout the world. Neem extracts reportedly control more than 200 species of insects, mites, and nematodes, and major pests such as locusts, rice, and maize borers, pulse beetles, and rice weevils (Khanna, 1992). In 1993, about 1000 tonnes of Rhizobium inoculants were produced, but it was forecast that, by the year 2000, the consumption of Rhizobium fertilizers may rise to 8,000-10,000 tonnes/yr, covering 50-60% of the 30 million ha of land used for leguminous crops. For the blue-green algae (BGA), used for rice cultivation, more than 400 production ponds have been set up for growing BGA for field trials and nearly 1100 field demonstrations have been organized in different parts of India to popularize the use of BGA fertilizers. The latter increase rice yield by 7-9%, enabling a reduction in chemical fertilizer use of up to 30%. Large-scale use of BGA, which is considerably cheaper than chemical fertilizers, could have a substantial influence, because about 40 million ha of land have been devoted to paddy cultivation (Ghosh, 1993).

Promising results have been obtained in conventional research and development for increased production of sugarcane, wheat, and rice, but more attention has to be directed to oil seeds, pulses, and coarse grain cereals. Despite shortcomings in packaging, storage, and transportation of plantlets, India has the potential of becoming a major international trading partner of horticultural and floricultural products. For more detailed information, see Sasson (1993,1994).

3. Indonesia

The Agency for Agricultural Research and Development-Badan Litbangtan (AARD) - supports the main program on agricultural biotechnologies in Indonesia, and has set up a National Center for Agricultural Biotechnology at the Bogor Central Research Institute for Food Crops (BORIF), nominated as a national center of excellence in biotechnologies by the Ministry for Research and Technology.

Initial research emphasis at the National Center is placed on seed technology, and, as part of the Rockefeller Foundations's International Rice Biotechnology Program, cooperation has been established with the International Rice Research Institute (IRRI, Los Banos, Philippines), concerning the regeneration of plants from calli of javanica rice. This regeneration is a prerequisite for any selection based on somaclonal variation, protoplast fusion, or plant transformation (Dart et al., 1991).

The Australian Center for International Agricultural Research (ACIAR) scheme for collaboration between laboratories in Australia and Indonesia on projects of mutual benefit became highly productive in terms of both scientific achievements and technology transfer. An ACIAR project involving the New South Wales Department of Agriculture concerns the selection of Rhizobium strains for soybean inoculation, the legume species that is grown after rice and in acid soils in newly developed fields in transmigration areas. Another project sponsored by the ACIAAR concerns the improved diagnosis and control of groundnut stripe virus. This project aims to protect groundnut from the virus by transforming the legume species with the viral coat protein gene using Microprojectiles, followed by the regeneration of transformed groundnut calli (Dart et al., 1991).

The Dutch Government supports the setting up of the Lembang Horticulture Research Institute (LEHRI). Biotechnologies are being used at the LEHRI for the major commercial crop species, garlic, potato, and asparagus; the research agenda for garlic and shallot (Allium spp.) includes virus elimination, diagnostic kits for virus identification in plant tissue; cabbage and tomato are also being studied (virus-free seeds, multiplication of elite clones, somaclonal variation). A new research institute in Solok is expected to focus on citrus, papaya, mango, and banana (i.e., on the production of disease-free planting material, varietal improvement, postharvest activities and marketing) (Dart et al., 1991). For more detailed information, see Sasson (1993).

The first oil palm clones derived from somatic embryogenesis were outplanted in the field in 1987. About 1 million plants were sold in 1993. Although clonal plants are sold at five times the price of seedlings, there is a large market in Indonesia trying to catch up with Malaysia in overall palm oil production (Dart et al., 1991).

Some 3 million ha of coconut are under production, with generally low yields. Hybrids. Hybrids are being produced and their cloning by tissue culture has been hindered by difficulties in regenerating plantlets from the cultures. There are commercial incentives to develop clones of the Kopyor mutant coconut tree which produces a nut with special flesh, used in ice cream, confectionery lproducts and preserves, and is worth 10-15 times more than the ordinary nut (Dart et al., 1991). Two coconut projects in the Lampung province of southern Sumatra involve researchers from the French International Cooperation Centre for Agricultural Research for Development-CIRAD's Department of Perennial Crops: one government project for small plantations funded by the World Bank and one private project run with the help of the Multi Agro Corporation on coconut and cocoa.

A facility at the Faculty of Agriculture of the Gadjah Mada University in Yogyakarta produces Rhizobium inoculants (with the trade name Legin) for government-sponsored crop improvement projects. The production capacity is approximately 20 tonnes/yr, an amount sufficient to inoculate about 111,000 ha of soybeans. The full capacity of the facility is probably five times this production. Rhizogin is the trade name of another Rhizobium inoculant prepared and marketed by the private company, Rhizogin-Indonesia, the production capacity of which is sufficient to inoculate approximately 440,000 ha/yr with the recommended dose of 180 kgs. inoculant per 65 kg. of soybean seeds. It is supplying about 70% of the inoculum needed for soybean cultivation in the country. With additional labor and investment in bioreactor capacity, the factory could double or treble its output and unit costs could be reduced markedly (Saono, 1991).

Integrated pest management and biological control of plant pests are likely to have a high return on investment. Between 1986 and 1989, the government phased out pesticide subsidies and banned 57 broad-spectrum formulations of insecticides. A national integrated pest management program was set up, coordinated by the National Development Planning Board; as a result, the spraying frequency per field dropped from 2.2 to 0.8 times per season, resulting in a 50% decline in insecticide consumption, while rice production increased by 12%. The Lembang Horticulture Research Institute 9LEHRI) and the Indonesian Government's extension service teamed up with the Swiss chemical company Ciba-Geigy AG in a pilot project on reducing pesticide consumption in cabbage cultivation; farmers have found that three to six applications of pesticide are adequate, without any loss of yield, compared with the 15 sprays normally applied. The project also relies on the use of natural predators: for cabbage, a parasitoid wasp Diadegma eucerophaga was chosen, which lays eggs in the diamond backmoth larvae, preventing them from reaching maturity. Encouraged by the promising results, ciba-Geigy AG has extended the program to potatoes, then to onion and tomato cultivation (Kumar, 1992). The Dutch Government is supporting an extension program of integrated pest management to complement that of the FAO on rice.

Indonesia gives a high priority to the development of biotechnologies, with direct support from the Ministry of State for Research and Technology and the Departments of Agriculture and Education. Plant tissue culture and micropropagation techniques are well established in several laboratories, and large-scale commercial production of planting material has been achieved for oil palm (Research Institute for Oil Palm at Marihat). Similar developments for other plantation crop species are likely in the mid-1990s. The use of the same technique to eradicate viruses is also likely to support commercial production of planting material of potato and several horticultural crop species, and selected strains of mycorrhizae are expected to become available for commercial inoculant production (Dart et al., 1991).

4. Malaysia

One of the major areas of research and development is the clonal propagation of high-yielding, oil palm varieties, carried out by the Palm Oil Research Institute of Malaysia 9PORIM) and the Federal Land Development Authority. The market is sizeable, as more than 1.6 million ha of plantations existed in the early 1990s. The problem of abnormal flowering has been solved and cloned superior oil palms has been marketed since 1992. All Malaysian laboratories working in this area, including the PORIM, are producing such clones. They were also increasingly involved in the mass propagation of other crop species, with a view to diversifying production and meeting local needs.

The Malaysian Agricultural Research and Development Institute (MARDI) have established a Biotechnology Center devoted to the in vitro culture of the following crop species: coconut (cloning of selected plants, in vitro germination of embryos, callogenesis, and somatic embryogenesis); cocoa (organ culture); papaya (cloning of selected varieties); anther culture of rice hybrids and somaclonal variation of rice; pepper (production of pathogen-free plants to control Phytophthora disease)' strawberry (meristem culture); and pineapple (culture of axillary buds to propagate the crosses of Sarawak and Singapore Spanish varieties, suitable for the canning industry) (Bordier, 1990). A cocoa clone derived from in vitro micropropagation was released at the MARDI Cocoa Research Station near Teluk Intan, Perak (Zakri, 1991). For more detailed information see Sasoon (1193).

The Southeast Asian lowland swamps are home to the sago palm (Metroxylon sagu). The three leading world producers - Malaysia, Indonesia, and Papua New Guinea - are interested in cooperation aimed at developing sago plantations and using sago starch extensively. Japan is also supporting this cooperation through the creation of a Sago Research Fund. In the early 1990s, Sarawak had a total area of 19,720 ha of sago palm, including both semiwild and cultivated stands. Sago flour production increased by about 60% between 1984 and 1990 to 84,991 tonnes. Since 1984, there has been an upturn in the export of sago starch : from 3,406 to 27,502 tonnes, earning the State of Sarawak 11.4 million dollars in revenue (Zulpilip et al., 1991).

A potential area for sago plantation exists in Malaysia, which has 2.4 million ha of peat land, of which 1.66 million ha are in Sarawak. Because of its natural adaptation to peat soils of low nutritional value and high acidity, sago seems to be the only crop species that could grow on these soils without reclamation. Furthermore, sago being little prone to natural disasters, such as drought, pest and disease infestations, and flooding, is considered by farmers a minimal-risk crop species. A langorous cycle makes its cultivation ideally suited to a part-time activity, leaving the farmer time between planting and harvesting to seek other income-earning opportunities (Zulpilip et al., 1991).

With the increased demand from industry for its products and a 9 to 15 year growth cycle, quality varieties or clones of sago palm will need to be planted on a large scale. Although sago palms can be propagated from suckers, the number is limited; therefore, clonal propagation through in vitro techniques is the most suitable means both for producing the vast amount of planting material required for extensive plantations and for improving the quality and vigor of palms (Alang et al., 1993).

Since 1983, research has been conducted at the Department of Biotechnology, Faculty of Food Science and Biotechnology, Universiti Pertanian Malaysia, Serdang, Selangor, towards clonal propagation of sago palm. Several hundred clonal plantlets were produced over a period of 18-24 months from culture initiation; the protocol for inducing embryogenesis from explants and developing embryoids into plantlets was being improved (Alang et al., 1993).