Since GM tobacco and potato were first obtained in 1983, the research and development of plant genetic engineering has progressed rapidly in just over ten years. More than 100 plants have been obtained in the world for transgenic plants, including crops such as rice, corn and potatoes; cash crops such as cotton, soybeans, rapeseed, flax and sunflower; tomatoes, cucumbers, mustard, cabbage, cauliflower, carrots, eggplant, and lettuce , celery and other vegetable crops; alfalfa, white clover and other forage; apple, walnut, plum, papaya, melon, strawberry and other fruits; petunia, chrysanthemum, carnation, garland blue flowers and other flowers; to occupy poplar and other afforestation tree species. It should be said that the research on transgenic plants has made encouraging progress.
However, most of the previous work focuses on the easy-to-make model plants, which has led to the rapid development of molecular biology and transgenic technologies for tobacco, potatoes, tomatoes, petunias, and Arabidopsis. In recent years, the number of researches with practical goals has greatly increased. In foreign countries, major seed companies and small companies compete for the development of recombinant DNA technology for the commercial use of important crops. The research institutes and the university’s pioneering principles and technologies are used for development. This led to the advancement of genetic engineering of plants to the practical use of genetic improvement of important foods and legumes. Before 1988, the conversion of important grain and leguminous crops was very difficult. It was only after the successful development of a new biotech tool, the gene gun, that the genetic modification of these crops was not only possible, but often it could be done without Depends on breed or genotype. The gene gun is powered by gunpowder explosion, capacitor discharge, or high-pressure gas, and emits metal particles with a diameter of only about 1 micron. The surface of the microparticles is coated with a preferred gene, and is injected into the plant cells at high speed, and is expressed in the cells to produce active gene products, thereby achieving the purpose of improving the variety.
The original soybean genetic engineering focused on the regeneration of protoplasts and embryogenic suspension cells, but the progress is very slow, and obtaining genetically modified soybeans is a big problem. The emergence of gene guns has made soybean genetic transformation a reality. In fact, soybeans have become a model crop for many difficult-to-transform crops. In just two years from 1988 to 1990, a practical soybean conversion system was established, which is currently the only soybean transformation method that does not rely on genotypes. The genes for the herbicides Bastar and Roundup have also been transferred to soybeans, and field trials have been conducted continuously for the past three years and are expected to be commercialized in the near future. This will be a milestone in the commercialization of genetic engineering of legume crops. Future goals of soybean genetic engineering may include modification of protein and oil components, resistance to insects, resistance to viruses, and other disease resistance.
Rice is the second largest cereal crop in the world, but it is the largest food crop in China with an annual output of 189.92 million tons (China Agriculture Yearbook 1993). Rice production in China and India accounts for 60% of the world's total production. Almost half of the world's population uses rice as the main source of heat. Rice and Xenopus japonica and japonica rice account for 80% of cultivated rice and are consumed by more than 2 billion people in the world.
Rice transgenic plants began in 1988, initially using protoplasts as recipients, using direct DNA transfer methods to regenerate fertile transgenic plants. However, the restriction of protoplast regeneration system is very limited. There are only a few varieties of indica rice, such as Taipei 309, which can be regenerated from protoplasts. The majority of excellent japonica rice varieties and most japonica rice varieties are difficult to be regenerated from protoplasts. Transgenic plants have not yet been obtained from indica rice varieties that can regenerate plants from protoplasts. Since the scutellum of the immature embryo of rice is highly capable of regenerating plants, nearly all rice cultivars can be regenerated from immature embryos. Therefore, some scientists believe that the prospect of application of protoplast transformation in rice is limited. It is better to use the gene gun to transform rice embryos. The callus derived from young embryo and scutellum seedlings recently obtained transgenic plants transformed with Agrobacterium tumefaciens.
From the current research situation, the transgenic efficiency of some important food crops is not high, and it is only successful in a few varieties. How to achieve high efficiency, rapidity, simplicity, and wide applicability of transgenic technology is still an important limiting factor in plant genetic engineering.
What is the breakthrough of plant genes in the reduction of diseases and insects? The development of recombinant DNA technology has allowed the transfer of genes from animals, plants and microorganisms, breaking the natural barriers that are difficult to cross between species. So far, genes of practical value such as antiviral, insect-resistance, herbicide-resistance, altered protein components, male infertility, genes that change flower color and flower shape, and extended shelf life have been transferred to tobacco, potatoes, cotton, tomatoes, respectively. Soybeans, alfalfa, petunia and other crops. Plant genetic engineering will have an inestimable impact on future agriculture.
(1) Anti-virus: The virus is a major enemy of crops, and the yield loss caused by it is extremely high. For potato alone, the loss of potato virus X (PVX) can reach 10%, and the loss of potato virus Y (PVY) can be as high as 80%.
Since the transfer of the coat protein gene of tobacco mosaic virus from tomato to tomato in the United States in 1986 and the development of tomato plants resistant to the tobacco mosaic virus, transgenic plants resistant to the cucumber leaf virus have also been successively successful. Scientists in China have used genetically modified methods to develop antiviral and antiviral tomatoes, and field trials have begun.
(b) Antibacterial and fungi aspects:
Bacteria and fungi are major diseases in agricultural production. History has documented cruel events that have changed the lives of millions of people because of plant diseases. The most striking example is that Ireland (1845-1860) caused 1 million people to suffer from hunger due to the outbreak of potato late blight (fungal disease) and forced another 200 Millions of people immigrated to North America. It is estimated that potato production in the world is reduced by 25% annually due to bacterial diseases, which amounts to about 4 billion U.S. dollars. Conventional breeding breeds have made outstanding contributions to agricultural production, but in some cases, the development of disease-resistant breeding is limited due to the lack of antigens in the crop itself or in wild species. Recombinant DNA technology allows genes of different organisms to transfer to each other, thus opening up a new way to solve problems.
In the haemolymph of the silkworm, silkworm, and pupa pupa, 15 proteins were found after induction. They can be divided into three different bactericidal peptides: cecropin, Attacin, and lysozyme. They have a wide range of gram positive and negative bacteria. Spectral antibacterial activity. Jaynes Laboratories (1991) transferred cecropin B and two artificially synthesized bactericidal peptide genes into tobacco. After inoculation with R. solanacearum, it was found that the incidence of transgenic tobacco was delayed, and the disease index and mortality rate were reduced. Our laboratory collaborated with brothers to transfer the synthetic cecropin B and ShivaA genes into seven potato cultivars in China. Identification of Ralstonia solanacearum in the greenhouse and in the field resulted in the growth of some special genetic strains of organisms than the original cultivars. Level 1-3. Jaynes et al. also used a computer to compare the structure of bactericidal peptides and in vitro bioactivity assays and found that some peptides can kill fungi, Plasmodium, and plant nematodes. They optimistically estimate that in the future it is possible to use a single gene to kill different pathogenic bacteria, fungi and nematodes of plants.
(c) Insect pests:
So far, the most studied and achieved results are two types of genes, namely the insecticidal protein genes of Bacillus thuringiensis and the protein trade-offs and inhibitors genes isolated from crops such as cowpea. When pests feed on this transgenic plant, they kill it.
In 1987, Vaeck et al. demonstrated for the first time that tobacco transformed with the insecticidal protein gene of Bacillus thuringiensis was resistant to Nicotiana halodendron, whose expression level accounted for 0.0001% of the water-soluble protein can completely inhibit Nicotiana. Since then it has also been successful on tomatoes, resistant to tomato fruits and tomato codling moths. The insect-resistant blue flowers have been obtained after transformation of the engineered insecticidal protein gene into cotton according to plant-preferred codons, and field experiments have shown that they are resistant to Brassica juncea, Spodoptera exigua, and Helicoverpa armigera. It is estimated that cotton insecticides cost 645 million U.S. dollars each year, and the breeding of insect-resistant cotton will have a huge effect on reducing the amount of pesticides used and protecting the environment. The second type of insecticidal gene is a protease inhibitor gene isolated from crops such as cowpea and potato. It is known that there are two types of injury-inducing protease inhibitors in solanaceous plants, inhibitor II inhibits chymotrypsin and trypsin, and inhibitor I inhibits chymotrypsin. Transgenic grasses expressing these genes have proven to be broad-spectrum resistant to many insects. In recent years, the laboratory of Mr. Qi Zhengwu of the Shanghai Institute of Biochemistry in China has also isolated the protease inhibitor gene from the Sagittaria and Cucurbitaceae crops, and is being transformed with protein engineering to obtain a more insecticidal and broad-spectrum insect-resistant gene. .
Insect feeding trials have proven that the Su Yunjin insecticidal protein plus protease inhibitors can increase insect killing by 2-20 times. It can be expected that the future development trend will be to transfer both types of genes to plants at the same time in order to increase the resistance to insects and to solve insect resistance problems.
(d) Anti-weedy aspects:
The decrease in crop damage due to weeds in agriculture increased from 8% in the 1940s to 12%. Most of the herbicides currently widely used are non-selective herbicides. Therefore it can only be used before sowing. The development of herbicide-resistant crops through genetic engineering can not only reduce the application of chemical herbicides, reduce environmental pollution, but also give greater flexibility in the selection of crops for crop rotation or intercropping.
Glyphosate is currently the most widely used non-selective herbicide that kills 76 of the 78 species of weeds in the world, is non-toxic to humans and animals, and is easily decomposed by soil microorganisms.
Herbicide-resistant transgenic plants are expected to be one of the earliest commercially engineered plants. However, before application, it is also necessary to consider whether the viability and yield of transgenic plants will be reduced, the potential for crossing with weeds, and the possibility that the crop itself will turn into weeds.
Advances in Practical Phases of Research and Development of Transgenic Plants In order to ensure human health and environmental safety, countries in the field trials of genetically modified plants need to be approved by the relevant government departments. For example, China's State Science and Technology Commission has issued the "Genetic Safety Management Measures." The Ministry of Agriculture is also working out the "Agricultural Biological Genetic Engineering Implementation Regulations." Units and individuals engaged in research and development in this area must provide Details of safety evaluation. From the general trend, the number of field-tested transgenic plants is greatly increased. Since the field trials of each genetically modified plant in 1986, up to 1992, there have been 675 field trials approved worldwide. In 1986, there were only 5 cases. By 1992, it had rapidly increased to 244 cases. Field trials were conducted in 28 countries, including 250 in the United States, 112 in France, 76 in Canada, 54 in Belgium, 42 in the United Kingdom, 33 in the Netherlands, and 108 in the other 22 countries. Of these 675 cases, the vast majority were applied by private companies, and 60 companies accounted for 501 cases. In terms of crops, there were the largest number of potatoes (134 cases), followed by rape (122 cases), tobacco (96 cases), soybean (27 cases), loquat (23 cases), and other crops were less than 10 cases, such as rice 5 example. Among the transgenic plants, herbicide resistance was the most common (247), followed by quality improvement (116 cases), antiviral (104 cases), insect resistance (89 cases), marker gene (81 cases) and disease resistance. (20 cases) and so on. This shows that plant genetic engineering has entered the actual stage of crop improvement. It is predicted that the construction of tobacco, tomatoes, potatoes, rape, cotton, soybeans, etc., can be put on the market within 5-10 years.
Main factors affecting the commercialization of genetically modified crop varieties
1. The question of safety, that is, the effect of certain characteristics transferred on the use of the final product, especially as a food, has no adverse effects on the human body.
2. Mobility of genetically modified DNA, ie whether this DNA will be transferred to other crops or weeds, causing environmental and ecological problems.
3. The post-effects of other agricultural measures, the impact on agriculture and the export of agricultural products from developing countries, etc.
4, the public acceptance, that is, psychological factors.
In short, the research and development of transgenic plants has made great progress, but there are still many issues that deserve further discussion. In spite of this, its commercialization prospect is bright and brilliant.
The 863 Program for the evaluation of the current status of research and development of plant genetic engineering in China has played a key role in promoting the development of plant genetic engineering in China. At present, from the perspective of technological strength, equipment and equipment, it has already had the ability to track the world's advanced level.
However, as a whole, the basic research, development and application of plant biotechnology in China are still two fatal weaknesses that must be strengthened. China's plant genetic engineering
and moisture effectively, hydrating and moisturizing, antioxidant, prevent the
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However, most of the previous work focuses on the easy-to-make model plants, which has led to the rapid development of molecular biology and transgenic technologies for tobacco, potatoes, tomatoes, petunias, and Arabidopsis. In recent years, the number of researches with practical goals has greatly increased. In foreign countries, major seed companies and small companies compete for the development of recombinant DNA technology for the commercial use of important crops. The research institutes and the university’s pioneering principles and technologies are used for development. This led to the advancement of genetic engineering of plants to the practical use of genetic improvement of important foods and legumes. Before 1988, the conversion of important grain and leguminous crops was very difficult. It was only after the successful development of a new biotech tool, the gene gun, that the genetic modification of these crops was not only possible, but often it could be done without Depends on breed or genotype. The gene gun is powered by gunpowder explosion, capacitor discharge, or high-pressure gas, and emits metal particles with a diameter of only about 1 micron. The surface of the microparticles is coated with a preferred gene, and is injected into the plant cells at high speed, and is expressed in the cells to produce active gene products, thereby achieving the purpose of improving the variety.
The original soybean genetic engineering focused on the regeneration of protoplasts and embryogenic suspension cells, but the progress is very slow, and obtaining genetically modified soybeans is a big problem. The emergence of gene guns has made soybean genetic transformation a reality. In fact, soybeans have become a model crop for many difficult-to-transform crops. In just two years from 1988 to 1990, a practical soybean conversion system was established, which is currently the only soybean transformation method that does not rely on genotypes. The genes for the herbicides Bastar and Roundup have also been transferred to soybeans, and field trials have been conducted continuously for the past three years and are expected to be commercialized in the near future. This will be a milestone in the commercialization of genetic engineering of legume crops. Future goals of soybean genetic engineering may include modification of protein and oil components, resistance to insects, resistance to viruses, and other disease resistance.
Rice is the second largest cereal crop in the world, but it is the largest food crop in China with an annual output of 189.92 million tons (China Agriculture Yearbook 1993). Rice production in China and India accounts for 60% of the world's total production. Almost half of the world's population uses rice as the main source of heat. Rice and Xenopus japonica and japonica rice account for 80% of cultivated rice and are consumed by more than 2 billion people in the world.
Rice transgenic plants began in 1988, initially using protoplasts as recipients, using direct DNA transfer methods to regenerate fertile transgenic plants. However, the restriction of protoplast regeneration system is very limited. There are only a few varieties of indica rice, such as Taipei 309, which can be regenerated from protoplasts. The majority of excellent japonica rice varieties and most japonica rice varieties are difficult to be regenerated from protoplasts. Transgenic plants have not yet been obtained from indica rice varieties that can regenerate plants from protoplasts. Since the scutellum of the immature embryo of rice is highly capable of regenerating plants, nearly all rice cultivars can be regenerated from immature embryos. Therefore, some scientists believe that the prospect of application of protoplast transformation in rice is limited. It is better to use the gene gun to transform rice embryos. The callus derived from young embryo and scutellum seedlings recently obtained transgenic plants transformed with Agrobacterium tumefaciens.
From the current research situation, the transgenic efficiency of some important food crops is not high, and it is only successful in a few varieties. How to achieve high efficiency, rapidity, simplicity, and wide applicability of transgenic technology is still an important limiting factor in plant genetic engineering.
What is the breakthrough of plant genes in the reduction of diseases and insects? The development of recombinant DNA technology has allowed the transfer of genes from animals, plants and microorganisms, breaking the natural barriers that are difficult to cross between species. So far, genes of practical value such as antiviral, insect-resistance, herbicide-resistance, altered protein components, male infertility, genes that change flower color and flower shape, and extended shelf life have been transferred to tobacco, potatoes, cotton, tomatoes, respectively. Soybeans, alfalfa, petunia and other crops. Plant genetic engineering will have an inestimable impact on future agriculture.
(1) Anti-virus: The virus is a major enemy of crops, and the yield loss caused by it is extremely high. For potato alone, the loss of potato virus X (PVX) can reach 10%, and the loss of potato virus Y (PVY) can be as high as 80%.
Since the transfer of the coat protein gene of tobacco mosaic virus from tomato to tomato in the United States in 1986 and the development of tomato plants resistant to the tobacco mosaic virus, transgenic plants resistant to the cucumber leaf virus have also been successively successful. Scientists in China have used genetically modified methods to develop antiviral and antiviral tomatoes, and field trials have begun.
(b) Antibacterial and fungi aspects:
Bacteria and fungi are major diseases in agricultural production. History has documented cruel events that have changed the lives of millions of people because of plant diseases. The most striking example is that Ireland (1845-1860) caused 1 million people to suffer from hunger due to the outbreak of potato late blight (fungal disease) and forced another 200 Millions of people immigrated to North America. It is estimated that potato production in the world is reduced by 25% annually due to bacterial diseases, which amounts to about 4 billion U.S. dollars. Conventional breeding breeds have made outstanding contributions to agricultural production, but in some cases, the development of disease-resistant breeding is limited due to the lack of antigens in the crop itself or in wild species. Recombinant DNA technology allows genes of different organisms to transfer to each other, thus opening up a new way to solve problems.
In the haemolymph of the silkworm, silkworm, and pupa pupa, 15 proteins were found after induction. They can be divided into three different bactericidal peptides: cecropin, Attacin, and lysozyme. They have a wide range of gram positive and negative bacteria. Spectral antibacterial activity. Jaynes Laboratories (1991) transferred cecropin B and two artificially synthesized bactericidal peptide genes into tobacco. After inoculation with R. solanacearum, it was found that the incidence of transgenic tobacco was delayed, and the disease index and mortality rate were reduced. Our laboratory collaborated with brothers to transfer the synthetic cecropin B and ShivaA genes into seven potato cultivars in China. Identification of Ralstonia solanacearum in the greenhouse and in the field resulted in the growth of some special genetic strains of organisms than the original cultivars. Level 1-3. Jaynes et al. also used a computer to compare the structure of bactericidal peptides and in vitro bioactivity assays and found that some peptides can kill fungi, Plasmodium, and plant nematodes. They optimistically estimate that in the future it is possible to use a single gene to kill different pathogenic bacteria, fungi and nematodes of plants.
(c) Insect pests:
So far, the most studied and achieved results are two types of genes, namely the insecticidal protein genes of Bacillus thuringiensis and the protein trade-offs and inhibitors genes isolated from crops such as cowpea. When pests feed on this transgenic plant, they kill it.
In 1987, Vaeck et al. demonstrated for the first time that tobacco transformed with the insecticidal protein gene of Bacillus thuringiensis was resistant to Nicotiana halodendron, whose expression level accounted for 0.0001% of the water-soluble protein can completely inhibit Nicotiana. Since then it has also been successful on tomatoes, resistant to tomato fruits and tomato codling moths. The insect-resistant blue flowers have been obtained after transformation of the engineered insecticidal protein gene into cotton according to plant-preferred codons, and field experiments have shown that they are resistant to Brassica juncea, Spodoptera exigua, and Helicoverpa armigera. It is estimated that cotton insecticides cost 645 million U.S. dollars each year, and the breeding of insect-resistant cotton will have a huge effect on reducing the amount of pesticides used and protecting the environment. The second type of insecticidal gene is a protease inhibitor gene isolated from crops such as cowpea and potato. It is known that there are two types of injury-inducing protease inhibitors in solanaceous plants, inhibitor II inhibits chymotrypsin and trypsin, and inhibitor I inhibits chymotrypsin. Transgenic grasses expressing these genes have proven to be broad-spectrum resistant to many insects. In recent years, the laboratory of Mr. Qi Zhengwu of the Shanghai Institute of Biochemistry in China has also isolated the protease inhibitor gene from the Sagittaria and Cucurbitaceae crops, and is being transformed with protein engineering to obtain a more insecticidal and broad-spectrum insect-resistant gene. .
Insect feeding trials have proven that the Su Yunjin insecticidal protein plus protease inhibitors can increase insect killing by 2-20 times. It can be expected that the future development trend will be to transfer both types of genes to plants at the same time in order to increase the resistance to insects and to solve insect resistance problems.
(d) Anti-weedy aspects:
The decrease in crop damage due to weeds in agriculture increased from 8% in the 1940s to 12%. Most of the herbicides currently widely used are non-selective herbicides. Therefore it can only be used before sowing. The development of herbicide-resistant crops through genetic engineering can not only reduce the application of chemical herbicides, reduce environmental pollution, but also give greater flexibility in the selection of crops for crop rotation or intercropping.
Glyphosate is currently the most widely used non-selective herbicide that kills 76 of the 78 species of weeds in the world, is non-toxic to humans and animals, and is easily decomposed by soil microorganisms.
Herbicide-resistant transgenic plants are expected to be one of the earliest commercially engineered plants. However, before application, it is also necessary to consider whether the viability and yield of transgenic plants will be reduced, the potential for crossing with weeds, and the possibility that the crop itself will turn into weeds.
Advances in Practical Phases of Research and Development of Transgenic Plants In order to ensure human health and environmental safety, countries in the field trials of genetically modified plants need to be approved by the relevant government departments. For example, China's State Science and Technology Commission has issued the "Genetic Safety Management Measures." The Ministry of Agriculture is also working out the "Agricultural Biological Genetic Engineering Implementation Regulations." Units and individuals engaged in research and development in this area must provide Details of safety evaluation. From the general trend, the number of field-tested transgenic plants is greatly increased. Since the field trials of each genetically modified plant in 1986, up to 1992, there have been 675 field trials approved worldwide. In 1986, there were only 5 cases. By 1992, it had rapidly increased to 244 cases. Field trials were conducted in 28 countries, including 250 in the United States, 112 in France, 76 in Canada, 54 in Belgium, 42 in the United Kingdom, 33 in the Netherlands, and 108 in the other 22 countries. Of these 675 cases, the vast majority were applied by private companies, and 60 companies accounted for 501 cases. In terms of crops, there were the largest number of potatoes (134 cases), followed by rape (122 cases), tobacco (96 cases), soybean (27 cases), loquat (23 cases), and other crops were less than 10 cases, such as rice 5 example. Among the transgenic plants, herbicide resistance was the most common (247), followed by quality improvement (116 cases), antiviral (104 cases), insect resistance (89 cases), marker gene (81 cases) and disease resistance. (20 cases) and so on. This shows that plant genetic engineering has entered the actual stage of crop improvement. It is predicted that the construction of tobacco, tomatoes, potatoes, rape, cotton, soybeans, etc., can be put on the market within 5-10 years.
Main factors affecting the commercialization of genetically modified crop varieties
1. The question of safety, that is, the effect of certain characteristics transferred on the use of the final product, especially as a food, has no adverse effects on the human body.
2. Mobility of genetically modified DNA, ie whether this DNA will be transferred to other crops or weeds, causing environmental and ecological problems.
3. The post-effects of other agricultural measures, the impact on agriculture and the export of agricultural products from developing countries, etc.
4, the public acceptance, that is, psychological factors.
In short, the research and development of transgenic plants has made great progress, but there are still many issues that deserve further discussion. In spite of this, its commercialization prospect is bright and brilliant.
The 863 Program for the evaluation of the current status of research and development of plant genetic engineering in China has played a key role in promoting the development of plant genetic engineering in China. At present, from the perspective of technological strength, equipment and equipment, it has already had the ability to track the world's advanced level.
However, as a whole, the basic research, development and application of plant biotechnology in China are still two fatal weaknesses that must be strengthened. China's plant genetic engineering
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