Agriculture and Agricultural Biotechnology: Development Trends Toward the 21st Century

1. Agriculture and Agricultural Biotechnology: Development Trends Toward the 21st Century

2. I. Agricultural Biotechnologies and Breeding a Historical Perspective • Agriculture, has evolved since the dawn of human civilization, • first as a means to guarantee food supply and, • then, as a source of family income and improved profitability. • Food collection then, • Domestication of plants and animals that were found in the wild, combined with gradual, long-term changes in their quality and quantity, were the first signs of what is now collectively termed agriculture.

3. At that time, agriculture replaced the former nomadic habit of food collection for immediate consumption. • A continuous effort to improve plant and animal yield was already documented in the ancient scriptures of many nations of the Old and New World. • The documented history of all cradles of civilizations records of agricultural activities and improvement of plant and animal productivity. • The experiments carried by Jacob, the father, provided strong evidence that he succeeded in selecting for spotted, colored sheep against brown sheep by using classic breeding. The spotted, colored sheep made him rich because he could specify them as his own herd, distinct from that of his father-in-law.

4. Noah's ark was perhaps the first documented breeding institution, on which males and females of all known living creatures were given the chance to survive, breed, and produce the necessary number of offspring for selection of future generations. • Domestication of plants and animals, followed by food storage, coincided, most probably unintentionally, with the growth of microorganisms. • Thus, was born classical food fermentation, the earliest known application of biotechnology, focusing on the use of microorganisms to produce food products. • The technique is well documented for beer brewing, wine production, and bread baking,

5. Cheese, yogurt, vinegar, soy sauce, and bean curd, are additional examples of traditional uses of microorganisms in biotechnology for both food and industrial applications. • Gradual improvements in agricultural techniques, domestication of additional plant and animal species from the wild, step-by-step selection of better-performing and more-adapted genotypes. • Mendel's discoveries and laws, in the period of the 1860s, revolutionized genetics, and led at the beginning of the 20th century, to planned, controlled breeding experiments.

6. Many achievements during this century and toward the end of the 1970s have been successfully implemented since then. • Breeding, or the old biotechnology, was now harnessed as a most successful tool, resulting in better crops and farm animals. • This rapid scientific development has significantly improved agricultural yields and product quality, to better support the increasing human demands for a variety of foods. • More productive genotypes of corn, cereals, rice, and legumes, as well as cattle and poultry, enabled individual farmers and agricultural enterprises alike to provide more food.

7. Soon, this old biotechnology was revolutionized again, when DNA was cloned in 1973. • Toward the end of the 1970s and during the early 1980s, recombinant DNA technologies resulted in the first development of transgenic microorganisms and, subsequently animals and plants. • The novel scientific discoveries and techniques thus paved the way for the new agricultural biotechnology. This historical development is schematically illustrated in Figure 1.

11. II. The Limitation of Traditional Agriculture in Meeting Land, Environmental, and Economic Constraints • Traditional agriculture suffers from several serious limitations in facing current changes in international markets. • The world transition to a global village, and the increased flow of information, are changing the market conditions. • Local pricing policies and growers‘ profitability are no longer effective, except in certain closed communities, and they are increasingly affected by the volume of international trade and international prices of food and other agricultural products. • This is in addition to the limitations of classic breeding (see later).

12. Agriculture, facing reduced land and water availability. • This is aggravated (يتفاقم) by increased soil, water, and air quality deterioration, owing to global climatic changes, desertification, pollution, and industrialization. • Industrialization and urbanization already cause a severe competition over land. • The problem is international and affects Europe, America, the Far East, and the Middle East. • Partial solution is offered by the policy of European countries that issued rules and allocated financing for saving green belts around urban centers.

13. Another way of circumventing some of the problems is technological development of greenhouses, use of solar energy, development of innovative irrigation techniques, and novel applications of water desalination and recycling. • The greenhouse technology (protected agriculture) was developed originally as a means to improve plant quality and to extend crop harvest and marketing beyond the traditional regional seasons. • Similar trends are evident for cattle and poultry husbandry in climate-controlled structures, enabling their productivity in regions that were previously unfavorable. • Classic breeding provided improved plant and animal species suitable for growing under unfavorable environmental conditions, such as saline and arid lands.

14. Advanced greenhouses are fully computerized, and various environmental sensors are capable of sustaining the optimal conditions of humidity, light, temperature, pH, irrigation, and planting. • The resulting production costs endanger their continuous development. In addition, advanced controlled agriculture relies on highly educated growers. • Food production to sustain the expected large increase in world population in the 21st century in developing countries around the globe cannot, however, rely solely on protected agriculture. • Pastures, most cereals and legumes, and forests cannot be maintained as environmentally protected agriculture.

15. III. Combining the New Biotechnology with Classic Breeding is a Means of Promoting Agriculture Beyond the Year 2000 • The term new biotechnology usually applies to the use of recombinant DNA technology. • Biotechnology itself has a broader definition, covering any technique that involves the use of biological species and biomass, or their derivatives, for the generation of beneficial products. • In this respect, classic breeding can be considered synonymous with old biotechnology. • Classic breeding and selection for pest resistance has resulted in better yields. • Selection for genotypes that are more tolerant to extreme climates, water, and soil conditions, has just begun. • Moreover, even classic breeding of plants and animals now relies on the newly developed DNA biotechnologies that permit one to detect successful genotypes, and enable better selection of favorable traits.

16. In fact, molecular biology resulted in the generation of revolutionary powerful and precise breeding tools. • There are many examples of successful integration of these new biotechnologies in classic breeding. • Among these we may list the microscopic visualization of chromosomes in tissue culture, the fluorescent in situ staining by hybridization (FISH) for fluorescent labeling of chromosome markers in cells, restriction fragment length polymorphism (RFLP) for the isolation of DNA markers, development of the simple-sequence repeat (SSR)DNA markers for isolation of important plant traits, and the use of Southern and western blot techniques to validate successful trait isolation. • Contributing successfully to shortening the breeding and selection cycle, and significantly accelerating breeding of plants and animals .

17. The new biotechnology, as discussed in this book, refers to the use of recombinant DNA and in vitro biological techniques in three major areas: • as powerful tools in classic breeding, • as means for generating transgenic plants, animals, and other organisms, and • as a means of integrating microorganisms into various agricultural production systems. • The progress of agricultural research and the use of molecular biology techniques facilitated successful breeding.

18. Agricultural biotechnology (agbiotech) has thus become essential to the success of the growers, who are now more affected by competitors from far away. • Despite many difficulties, the agbiotech industry continues to grow (Table 1). • According to annual reports published by Ernst and Young in 1995 and 1996, the compelling aspect of agbiotech is that agriculture is the world's largest business, and biotechnology can both reduce costs and produce plants with more valuable characteristics. • To improve their product competitive edge, Monsanto Corp. (USA) invested 1 billion US dollars in agbiotech research and development over the last decade, a significant part of which has been directed to Posilac,

19. a version of recombinant bovine growth hormone which is now used for 15% of US dairy cows. • Monsanto also developed a transgenic soybean, resistant to the premier herbicide Roundup, the application of which yields larger crops. Herbicide-resistant corn, cotton, and potato came next. • China is marketing a virus-resistant tomato, while virus resistant potatoes are being tested in Mexico. • Examples of some leading agbiotech companies are given in Table 2. Calgene Inc. (USA), one of the world leaders in agbiotech, was valued at 158 million US dollars by Wall Street, and its revenues reached 117 million US dollars. • Total revenues in North America of publically traded agbiotech companies (see Table 2) reached close to 0.5 billion US dollars in 1995.

20. The Ernst and Young report emphasizes the 25% annual growth in sales of agbiotech-based products, as compared with 21% annual growth of the industry at large. • This also represents a major increase from the mean 18% annual increase calculated for the last decade. • The increase in revenues reflects the impression that investments in research and development-based agbiotech companies are finally paying off. • Public acceptance of agbiotech and engineered organisms is very flexible, and changes continuously. • Thus, for example, the European Council of Ministers agreed to limit the labeling of genetically modified food products. • Britain approved a tomato paste based on modified tomato (Zeneca Corp., UK), rapeseed oil from Plant Genetic System Ltd. (Belgium), and soybean products from Monsanto Corp. (USA) .

21. However, during the summer and fall of 1996, Monsanto Corp. and Unilever Ltd. (UK) faced a strong public resistance, and, there were demonstrations in Germany and The Netherlands against the use of transgenic plants in Europe, based on fears of the unknown and the presumed long-term effects. • Germany is currently blocking the import of transgenic food products. • Several other European countries also have their reservations.

22. A partial solution to some of these difficulties of public acceptance and prices is additional time. • During this time, agbiotech research and development will permit an evaluation of the real consequences of genetically modified organisms, and product prices will eventually decrease. • Along these lines, the European Commission plans to invest large sums of money in programs focusing on the development of public awareness and on promotion of biotechnology in Europe.

23. IV. Biotechnology in Agriculture has Multiple Faces A. Biopharming: Higher Plants and Farm Animals for the Pharmaceutical Industry • A major new direction is biopharming: the use of transgenic technologies to produce human-valuable proteins in genetically modified plants or farm animals. • The biopharming approach is designed to expand the capacity for drug production in a larger repertoire of plants and animals that can act as drug manufacturers, thereby increasing the versatility of production lines and facilities. • Transforming transgenic plants and animals into pharmaceutical production lines may create conditions to optimize production.

24. Cytokines and nutritional proteins can be manufactured in tobacco (a project under development in North Carolina and Israel). • Human serum albumin, a major protein constituent of the blood (which is used to treat emergency blood losses and chronic blood deficiency), and factor VIII (an essential constituent of the body response to repair injuries of blood vessels), can be produced in goat milk to replace the blood-derived products that are currently threatened by risks of acquired immunodeficiency syndrome (AIDS) and other diseases. • In addition, mammals can produce glycosylated proteins that are required for the tertiary structure of most human proteins, which cannot be provided by E. coli.

25. Genzym Transgenic Corp. (USA) uses this technology to develop antithrombin III, recombinant tissue plasminogen activator (tPA). • Human growth hormone is produced in goat milk by Serumtech Ltd. (Israel), and Pharmaceutical Proteins Ltd. (UK) uses this technology to produce anti-a1-transferrin antibody in sheep. • Additional examples are Gene Pharming Ltd. (The Netherlands) that uses cows for production of lactoferrin, collagen, and erythropoietin, and the Red Cross is using swine to develop protein C for blood coagulation.

26. Biopharming combines the advantages of agriculture with those of the pharmaceutical industry, to generate a larger production capacity for novel products to meet the increasing world demand. • With these techniques, the profit margins for the growers of farm animals are expected to increase at least twofold over that for their traditional use for meat and milk. • If successful, this novel trend will add to accelerating agrobusiness and its manufacturing capabilities.

27. B. Plant In Vitro Technologies: Micropropagation, Somatic Cell Genetics, and Transgenic Plants 1. Micropropagation • Plant propagation in tissue culture (micropropagation) is used to develop high-quality clonal standard plants. • These plants are selected for unique horticultural traits, pest resistance, crop quality, or suitability for environmental stress conditions. • Micropropagation has many advantages over traditional plant propagation.

28. The main advantages are attributed to the potential of combining rapid, large-scale propagation of new genotypes, the use of small amount of original germ plasm (particularly at the early breeding stage, when only few plants are available), and generation of pathogen-free propagules (e.g., virus- or bacteria-free). • Further commercialization is dependent on automation, currently under development, and on technological solutions to large-scale micropropagation, combined with novel products (e.g., artificial seeds) and quality control.

29. 2. Somatic Cell Genetics • The contribution of in vitro methods to plant breeding (i.e., somatic cell genetics) is most significant, especially in terms of haploid production and somatic hybridization. • Regeneration of haploid cell lines and plants from microspores is highly important for production of homozygous offspring for further breeding. • This was realized in barley, rice, rapeseed, potato, asparagus, and other plants. • Somatic hybridization by protoplast fusion is an elegant solution to overcome the interspecific crossing barriers.

30. It has already resulted in introgression of useful traits (e.g., cytoplasmic male sterility) in genomes of several cultivated plants, especially Solanaceae and Cruciferae. The embryo rescue technique is another alternative to overcome some crossing barriers.

31. 3. Transgenic Plants • A recent global review of field testing and commercialization of transgenic plants during 1986-1995 was published by James and Krattiger, who have listed more than 3500 field trials of transgenic crops in more than 15,000 individual sites in 34 countries. • This survey gives details of 56 crop plants, mostly in North America and the European Union, that have been engineered for a large number of traits. • Moreover, by the end of 1995, 35 applications had been granted to commercially grow nine transgenic crops involving eight traits in the European Union and additional six countries.

32. The major transgenic crops approved for commercial production (in the United States) include tomato (delayed ripening), cotton (insect- and herbicide-resistance), soybean (herbicide-resistance), corn (herbicide- and insect-resistance, male sterility), canola (modified oil quality), and others. • This reflects the continued impressive increase in using transgenic techniques to breed better plants.

33. C. Germplasm of the Future • Germplasm (i.e., the self-contained units of propagation in plants [seed and vegetative propagules] and reproduction in animals [embryos]) is, in fact, a concentrated package of genes. • This includes biotechnology of hybrid and artificial seeds, germ plasm banks and cryopreservation, in vitro fertilization, and embryo implantation. • Implementation of novel biotechnologies that involve gene manipulation, somatic hybridization, and mutagenesis has led to the identification of the roles of specific mitochondrial, chloroplast, and nuclear DNA elements in cell growth, fertility, and control of blossom.

34. Understanding their roles can lead to the design of tools for regulation of commercially important traits, such as male sterility. • Encapsulation and coating of the somatic embryos results in production of artificial synthetic seeds. • Sales of artificial seeds produced by this technology is estimated to reach 10.8 million US dollars annually. • This is only a fraction of the hugh 37 billion US dollars in sales of hybrid seeds in the world market annually. • The reward for improved seeds is highly profitable. • The market value of 1 kg of elite greenhouse tomato seeds can reach 20,000 US dollars.

35. Similarly important are efforts for improving reproduction of farm animals. • Novel in vitro biotechnologies, including superovulation, in vitro fertilization, and embryo implantation in surrogate mothers, allow breeders to produce multiple embryos with the most desired qualities. • This, together with successful gender selection and embryo manipulation, is a major asset for livestock agriculture. • The results are increased meat and milk production that is currently six to seven times larger than 30 years ago.

36. D. Agbiotech Disease Control, Diagnostics, and Biological Control of Plant Pests • Disease Control: New biotechnology-based vaccines against brucellosis, encephalitis, and hepatitis, for example, became crucial for proper maintenance of farm animals and poultry. • Diagnostics: Diagnostic biotechnology of animals and plants, for early monitoring and detection of diseases, pests, and chemical residues, becomes extremely important, for both improving health care and life expectancy of plants and animals, and for the marketability of the products.

37. Biological control: the global market for chemical pesticides is estimated at 36.8 billion Deutsche mark in 1993. • The market is now saturated, and no market growth was detected in the last 5 years. • On the other hand, the sales of biopesticides, which currently consist of only 10% of the total aforementioned pesticide market, increased 714% annually during the last 5 years. • Fears of biological control are associated with the assumption that resistance to biopesticides may develop with time, and that it may destroy the fragile interbalance of the environment.

38. This fear is, however, counterbalanced by the danger of continuous exposure to organophosphates, bromo- and chloro-derivatives, and others, that have a detrimental environmental effect and pose cumulative health risks. • In addition, the effectiveness of many chemical pesticides has been lost owing to resistance build-up. • The market effects of organic vegetables and fruits, which avoid the use of any chemicals, has not taken a large share of the sales, as their yield is at least 30% lower. • Biopesticides are considered to be more pest-specific, with less negative effects on humans, farm animals, and the environment.

39. Sales of Bacillus thuringiensis (Bt) alone amounted to 100 million US dollars in 1995. • The anticipated global market of baculoviruses for protection of vegetables and cotton is estimated at 2 billion US dollars. • Biosys Inc., is developing beneficial nematodes, especially for garden pest control and citrus protection. • Experiments in Egypt have shown the usefulness of pheromones to destroy pink ballworm in large-scale field experiments. • W. R. Grace Corp. is using azadirachtin, extracted from the oil of the Neem tree seeds, as an insecticide.

40. Ecoscience Inc. developed anticockroach fungi, and Ecogen Inc. is producing biofungicides to protect postharvest spoilage of citrus, apples, and other pome fruits. • Biofertilizers had entered the market as well. • The EPA and USDA approval and clearance for biocontrol products of nematodes, phytopathogenic fungi, and insect pests, as well as bacterial biofertilizers, imply an expected substantial growth in sales.

41. E. Phytoremediation and Bioremediation • Biotechnology holds major promise for control and protection of many environmental problems. • The ability of plants and seaweeds (algae) to absorb heavy metals is considered a useful tool for developing biofilters for effluent detoxification, water desalinization, and sewage treatment. • The technology is referred to as phytoremediation. • Bioremediation, using microorganisms for similar purposes, is currently a favored approach for cleaning water of oil contamination, as tested in the spoilage of water at the Exxon oil company disaster site, or in the Gulf during the Gulf war.

42. The sale of bioremediation products and services in the United States has reached several billion dollars in 1995, of which 500 million dollars were for environmental cleanups. • Another successful trend involves the addition of nutrients, such as nitrogen and phosphorus, to augment (Augmentation) the efficacy of microorganisms in digesting environmental waste. • Fungi are used to degrade DDT, PCB, cyanide, TNT, and other soil pollutants. • The ability of bacteria and fungi to decompose organic material has been successfully employed to clean oil-contaminated shorelines.

43. V. Emerging New Markets for Biotechnology A. Marine Biotechnology: Supply for a Growing Demand • In 1991 the mariculture industry produced 14 million tons of fish, with a market value of 28 billion US dollars. • Since 1996, the supply of fish from open seas and lakes is lagging behind the demand. • The latter is projected to increase by 65% toward 2020. • The shortage in fish supply from mariculture is aggrevated by difficulties in fish egg-laying, egg-spawning, and proper development of young fish during their adaptation to growth in captivity .

44. Recent genetic manipulation elucidated some of the molecular hormonal aspects of egg laying and reproduction, resulting in better understanding and hopes for major progress. • Technologies were developed for growing fish in integrative systems, in which organisms, behaving as a food chain cascade, are placed in a single pond. • The food chain saves costs on food supplements and maintains the system as a balanced ecosystem that controls cleaning of the required water.

45. Other environmentally friendly closed systems for growing fish are supplied with filtered, recycled water. • Fish cultivation in growing cages in deep water in seas and oceans is another recent, novel method. • All together, biotechnology has a repertoire of tools for the success of mariculture, including large-scale enhanced growth of fish, marine invertebrates, and macro and microalgae, both to supplement world food supply and for new, marketable products.

46. B. Cosmoceuticals and Nutroceuticals: Back to Natural Products • The world demand for cosmoceuticals (natural cosmetics) and nutroceuticals (food products enriched with nutrients derived from self-enhanced biochemical production) has increased by 20% annually over the past 7 years . 1. Nutroceuticals • Synthetically designed plant storage proteins can radically improve human and animal nutrition. • Eight of the 20 essential amino acids, not synthesized in humans and farm animals, are made by plants (especially soybean).

47. Storage proteins constitute the bulk of seeds, and production of transgenic plants that are engineered for elevated biosynthesis of specific physiologically stable storage proteins in seeds of cereals and legumes improves their nutritional quality. • Furthermore, chemically produced vitamins are not as efficient as their natural counterparts, and alternative sources are required, suggesting the growth of fresh fruits and vegetables that have been engineered to contain high levels of vitamins. • This may become important, according to some sources, for delaying aging, degenerative neurological diseases, and cancer. • The possible role of antioxidants in improving human

48. health and life expectancy, and for rejuvenating facial skin is slowly revealed. • The demand for antioxidants from natural pigments such as Dunaliela-enriched carotene complex and tomato lycopene, is reflected in a market of about 1 billion US dollars instead of 100 million-dollar only 5 years ago. • The understanding today is that the carotene complex, extracted from sources such as enriched microalgae, is more valuable than pure b-carotene. • Another promising direction is the use of transgenic algae to develop vaccines against gastrointestinal infections. • Algae may also become an important source of novel potent antiviral compounds and pesticides.

49. 2. Cosmoceuticals • Natural cosmetics are in demand by consumers who are frightened by reports of artificial chemicals in cosmetics that are known to have cell transformation activity, that could increase the risk of developing cancer. • Initially, suppliers of cosmoceuticals distributed natural eye and face makeup derived from plant extracts. • These were followed by the use of algal extracts, hyaluronic acid, and liposome technologies to deliver a new generation of effective, longlasting, and safer cosmetics.

50. C. Biochemicals for the Food and Chemical Industries • Traditional uses of plant products in the food industry (e.g., the use of cocoa and coffee beans for chocolate and for coffee production) relies heavily on plant breeding and could thus gain significantly from new plant biotechnologies. • In addition, there is a growing recognition of the importance of natural polysaccharides, for which the market has grown from annual sales of 150 million US dollars in the 1980s to over 500 million dollars in 2000. • These include, for example, carbohydrates with heat hysteresis properties, such as algal (seaweed) extracts for production of agar-agar, carrageenan, and agarose for food and microbiological markets.