Genetic Transformation

Lecture 1

Transgenic plants are those that carry additional, stably integrated, and expressed foreign gene(s) usually transferred from unrelated organisms. The whole process of introduction, integration and expression of foreign gene(s) in the host is called genetic transformation. Combined use of recombinant DNA technology, gene transfer methods and tissue culture techniques has led to the efficient production of transgenics in a wide variety of crop plants. In fact, transgenesis has emerged as a novel tool for carrying out single gene breeding or transgenic breeding of crop plants. The major advantage is that the useful genes cloned from viruses, bacteria, fungi, insects, animals, human beings, unrelated plants or even the genes synthesized in the laboratory can be introduced into plants. Unlike conventional hybridization, here only the cloned gene(s) of agronomic interest are being introduced into plants without the co-transfer of other undesirable genes from the donor. The recipient genotype is least disturbed thereby setting aside the need for back crosses. Moreover, gene pyramiding can be achieved through co-transformation for different genes with similar effects for obtaining durable resistance. Transgenic varieties belonging to seven crops are now being grown in 21 countries over an area of about 110 million hectares. Some important examples are listed below:Tomato Because tomatoes are one of the world's most popular vegetables, they have benefited from a long history of genetic improvement that continues in the transgenic age. Lycopene, a naturally occurring constituent of tomato, is a nutritional factor related to Vitamin A. Tomato varieties with transgenically enhanced lycopene content are under investigation. Another trait of interest is delayed ripening. Tomatoes that ripen slower can remain on the vine longer and develop improved flavor, compared to commercial varieties that are picked at the green stage. The Flavr-Savr tomato, one of the earliest approved transgenic crop varieties, was a delayed ripening variety. Antioxidant tomatoes Flavonols are powerful antioxidants with the ability to

neutralize harmful tissue-damaging molecules circulating in the body. Some foods, such as onions and tea, are naturally rich in flavonols, and several current research projects involving GM crops aim to increase beneficial antioxidant levels in other food plants. Scientists at Unilever have inserted a petunia gene into tomatoes which 1

increases flavonol production up to 78 times over the relatively low levels normally found in the fruits. Taste is not affected, and 65% of the flavonols are retained when the tomatoes are processed into paste. Researchers have coined the term functional foods for items such as this, where conventional breeding or transgenic technology have enhanced levels of compounds in plant or animal products with health benefits beyond basic nutritional requirements. Salt-tolerant tomato Researchers at the University of California at Davis have produced a GM tomato which can grow in soil irrigated by water containing up to 200 millimolar sodium chloride, 50 times the salt content normally tolerated by unmodifed tomatoes. The tomato contains a gene from thale cress (Arabidopsis thaliana) which segregates sodium ions from intracellular water for storage in vacuoles. Fruit from the salt-tolerant tomatoes tastes normal -- the salt is stored in vacuoles in the leaves. Cavity-fighting apples Medical researchers at Guys Hospital in London and scientists at the Horticultural Research Institute in Kent, U.K., are developing apples with a bacterial gene coding for a protein which prevents decay-causing bacteria from attaching to teeth. Results to date suggest that eating the GM apples could significantly reduce cavity formation. Golden Rice Millions of people in the world suffer from Vitamin A deficiency, which leads to vision impairment and increased susceptibility to diarrhea, respiratory diseases, and measles. Rice is a staple food in many countries, particularly in Asia, but does not contain Vitamin A or its immediate precursors. By inserting two genes from daffodil and produced rice capable of synthesizing beta-carotene, the precursor of Vitamin A. This rice variety is now being crossed into adapted varieties, with field tests possible in a year or two. Iron-pumping rice Iron-deficient alkaline soils are common in many of the worlds arid regions. Cereals such as corn and barley release an iron chelator known as deoxymugineic acid (DMA) when grown in such soils. DMA binds the iron and transports it across the root membrane, enhancing uptake by the plant. Rice normally produces very small quantities of DMA, making it an ineffective scavenger of iron in alkaline soils. Japanese researchers have transformed rice plants with two barley genes coding for enzymes which synthesize DMA. The GM rice plants have

greatly enhanced DMA production and iron uptake, and yield up to four times more than non-GM rice when grown in iron-poor soils. Proteinaceous rice AmA 1 gene cloned from Amaranthus has been introduced into potato that show 200 times of recombinant protein, material is in field trials at CPRI, India and will be released in coming year. At PAU, India scientists aim to introduce AmA 1 gene into commercial rice varieties for which funding from DBT is available for three years. Canola a major oilseed crop. Transgenic research has focused on improving the nutritional quality of canola oil by enhancing the Vitamin E content or by modifying the balance of fatty acids. Plant-based vaccines Food crops engineered to produce edible vaccines against infectious diseases would make vaccination more readily available to children around the world. Because of their palatability and adaptation to tropical and subtropical environments, bananas have received considerable research attention as a vehicle for vaccine delivery. Transgenic bananas containing inactivated viruses that cause cholera, hepatitis B, and diarrhea have been produced and are currently undergoing evaluation. Sunflower A disease-resistance trait, an anti-pest trait, and a herbicide-resistance trait are all being pursued, but no commercial varieties are now on the market. White mold (Sclerotinia) is a serious problem for sunflower producers in some areas. Resistance to this disease would expand the area in which sunflowers can be grown and might improve yields in areas of current cultivation. A commercially available cultivar is not expected before 2005. Resistance to the Argentina looper, an insect that eats sunflower leaves, is also being investigated. Some research has been done on developing sunflowers that can tolerate being sprayed with the weed killer Roundup. This trait would allow a farmer to spray his field for weeds while not killing the crop. Coffee and tea Decaffeinated coffee is now made by treating coffee beans to remove the caffeine. One method uses organic solvents to extract the caffeine, which causes some consumers to be concerned that residues from the solvents will remain in the coffee they drink. Other methods are criticized for removing some of the desirable, flavor-producing components along with the undesirable caffeine. Two 3

scientists in Hawaii and Scotland have identified different genes that lead to the production of caffeine in coffee beans and tea leaves. If these genes can be "turned off" in some plants, coffee and tea trees could be developed that would produce naturally decaffeinated products with full flavor and aroma. The harvesting of coffee beans now requires many passes through the orchards because the beans ripen at different times. A scientist in Hawaii is developing a method for making all the beans ripen at the same time so that harvesters could pick all the beans during one pass through the orchard. Grapes and wine (Vitis vinifera) are susceptible to several diseases that reduce the amount and the quality of wine grapes and table grapes or even kill the vine. Genes that confer resistance to particular diseases would reduce the cost of battling diseases in the vineyard. Researchers at the University of Florida have patented a method for producing grape vines that carry a silkworm gene to provide protection from Pierce's disease, a fatal bacterial disease that affects grapes and several other plants. Tobacco Nicotine-free tobacco is now being grown for a projected introduction of nicotine-free cigarettes. Previous attempts to make low-nicotine products removed some of the flavor along with the nicotine. Genetically engineered nicotine-free tobacco doesn't synthesize nicotine in the leaf. Trees Forest trees such as poplar, aspen, and spruce have been transformed with various genes to provide resistance to insects, tolerance to herbicides, and higher levels of the commercial product. For example, reducing the lignin content of a tree can make it easier to recover wood pulp. Corn hybrids resistant to corn rootworm Corn rootworm (Diabrotica spp.) is a serious pest of corn in many U.S. growing areas. It damages roots of young corn seedlings, resulting in reduced growth and poor standability of the plant. This insect is responsible for the application of the largest amount of insecticide to U.S. corn fields. What's more, to control this pest the insecticide must be applied directly to the soil, where it may leave residues or leach into the ground water. By replacing these chemical insecticides, corn rootworm resistant hybrids may provide major benefits to environmental quality.