Transgenic Crop Plants: Volume 2: Utilization and Biosafety

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Development of transgenic crop plants, their utilization for improved agriculture, health, ecology and environment and their socio-political impacts are currently important fields in education, research and industries and also of interest to policy makers, social activists and regulatory and funding agencies. This work prepared with a class-room approach on this multidisciplinary subject will fill an existing gap and meet the requirements of such a broad section of readers.

Volume 2 with 13 chapters contributed by 41 eminent scientists from nine countries deliberates on the utilization of transgenic crops for resistance to herbicides, biotic stress and abiotic stress, manipulation of developmental traits, production of biofuel, biopharmaceuticals and algal bioproducts, amelioration of ecology and environment and fostering functional genomics as well as on regulations and steps for commercialization, patent and IPR issues, and compliance to concerns and compulsions of utilizing transgenic plants.

Help Centre. My Wishlist Sign In Join. Michler Editor , Albert G. Hence, identification and testing of crops for GM contents is important for identity and legitimacy of transgene to simplify the international trade. Normally, molecular identification is performed at three different levels, i. In this chapter, current scenario of GM crops and different molecular testing tools are described in brief.

Gene Editing - Technologies and Applications. Biotechnology is a set of scientific tools in which living organisms are used for the welfare of mankind. This technique is efficiently used to modify and improve plants, animals and other microorganisms to increase their value. Biotechnology has a very wide range of applications and almost every field of daily science get benefit from this technology. Application of biotechnology in the field of agriculture has been practiced for a long time as people have wanted to improve agriculturally important crops by selection and breeding.

In s with the advancements in molecular biology, researchers were able to modify DNA which is a chemical building block and specify the features of living organisms at molecular level. This genetic information is coded in the form of DNA or genes. Genes from any living organisms human, animal, plant and microorganism could be easily manipulated and transferred into other organisms to enhance their value. Organisms artificially modified at genome level using genetic engineering tools are termed as genetically modified organisms GMOs. Microorganisms, i. Genetic engineering also has a great role in the field of agriculture by developing the transgenic crops for various traits.

For example, a useful gene from bacteria, fungi and animals etc. After transformation, the transgenes replicate with indigenous plant genes and produce specific protein [ 4 ]. Biotechnology supports in practical exploitation of genetic material for the betterment of mankind. By using latest trends in genetic engineering one can create the new face of existing cultivars with improved and desirable characteristics.

In addition to the improvement of agronomic traits, scientists are also looking in the production and expression of commercially valuable protein in plants like spider silk protein and polymers used in surgery [ 5 ]. A huge number of human vaccines, antigens and other pharmaceutical products are very efficiently expressing in transgenic plants. GMO offer many benefits to humans, but at the same time people also worry about the possible threats of using GMOs.

These risks include the possible introduction of allergens in GM foods and transfer of selection marker genes which are normally antibiotic resistant genes to gut flora [ 6 , 7 , 8 ]. With the introduction of foreign genes, there are also some biosafety issues linked with GM crops. Such crops are often unintentionally or intentionally used for food and feed production. In some conditions, GM crops spread globally by trading, transportation and storage either intentionally or unintentionally and contaminate GM free items.

Many countries have very strict rules and regulations for the development, cultivation, commercialization and labeling of GM crops and is also a trade barrier in some situations [ 9 ]. For example, USA has an optional labeling of GM in food items, whereas European Union has very strict rules for approval, cultivation and use of GM crops, including a compulsory labeling system [ 10 ].

They require very comprehensive information about such crops like type of targeted crop and transgene, safety for humans, environment, animals and effects on other related non-modified crops [ 11 , 12 , 13 ]. The increase in GM crop production has been coupled with an intricate and asynchronous international regulatory approval system, requiring identification and testing of food and agricultural products for the presence of GM content to simplify international trade.

Molecular identification of GM crops confirms the identity and type of modified product at each stage and assures compliance with import for GM food and feed [ 14 ]. The testing of GM crops could be performed in open field or under controlled laboratory conditions that depends upon type of samples and sensitivity of test performed. Normally, molecular identification and testing of GM crops is performed at three different stages, i. Each testing level has its own importance in testing the nature and type of GM crops. Generalized GM development methodology, global status, testing methods, possible biosafety issues and other benefits etc.

The rapid acceptance of GM crops shows the significant benefits realized by large and small growers in both developed and under-developed countries growing GM crops commercially. USA is leading in the area under GM crops with In , 24 countries planted Despite the possible health risks, cultivation area of GM crops is regularly increasing and introduction of new GM crops is continued. There are 29 different crops and fruit trees in 42, which countries have been successfully modified for various traits.

The cultivation area under stacked traits, i. Soybean, maize and cotton are major crops developed with stacked traits [ 16 ]. Countries approving GM crops for food, feed and general cultivation are also increasing every year. Plants, in which one or more foreign genes are introduced artificially instead of plant getting them under natural conditions of cross-breeding or normal recombination, are known as GM plants.

The introduced gene, known as transgene, could be from identical species or from different species within the same kingdom or other kingdom [ 17 ]. The process of introducing the transgene is called as genetic transformation that has become an important tool for crop improvement. Different steps are involved in the genetic transformation work like selection and identification of gene of interest transgene , isolation from source organisms, cloning into suitable plasmid vector.

Followed by development of expression vector containing all regulatory elements, i. In addition, another gene cassette of selection is also the part of expression vector which serves as the primary selection of putative transgenic cells on artificial plant media. Normally two types of selection markers are used, antibiotic and visual selection markers, which depend upon the type of work.

Final expression cassette is multiplied in suitable bacterial media and verified using various molecular biology techniques before transformation [ 19 ]. Integration of final expression cassette into plant can normally be achieved by two methods: i direct DNA delivery system, i. Both methods have successfully been used for the introduction of transgenes in plants [ 20 ].

Following genetic transformation, the transformed tissues are initially screened for transgene integration using selective plant tissue culture media. The regenerated plantlets on selective media supposed to have the transgenes and called as putative transgenics. Because there are three possibilities that the developed plantlets may be i true transgenics ii escapees iii mutants. The overall methodology for gene isolation, cloning, transformation and selection of putative transgenics has been shown in Figure 1.

Introduction of GM crops and their products in markets required to be monitored and need to know the presence and type of GM elements. Labeling rules and trade requirements vary from country to country which necessitates for the development of reliable methods for the detection, identification and quantification of GM crop varieties and their products.

GM crops can be tested by identifying either transgenes at DNA level, at transcriptional level by mRNA of transgene or using resulting transprotein. There are many other methods like chromatography and mass spectrometry etc. An overview of test methods used for detection and identification for GM crops has been given in Figure 2. Every test method has its own significance and value towards the final conclusion of GM crops.

A brief summary of these methods has been shown in Table 2. Qualitative analysis comprises of specific detection of target DNA sequence in test samples. Qualitative results clearly validate the presence or absence of GM elements under study, comparative to suitable controls and within the detection limits of analytical technique used, and test portion analyzed [ 21 , 22 ].

This method has found very broad and wide applications in GMO detection as commonly accepted tool for regulatory purposes. PCR process is basically comprised on three main steps, i. In first step the double stranded DNA is separated into two single strands, primers then identify their homologous sequence and are annealed to each strand in second step. Third and final step involves making two identical copies of original DNA strand by adding exact nucleotides with the help of DNA polymerase at an appropriate temperature.

Amplification of target gene occur in-vitro through a reaction catalyzed by a DNA polymerase in the presence of oligonucleotide primers and deoxyribonucleoside triphosphates in a defined reaction buffer [ 23 , 24 ]. This amplified DNA can be visualized by using gel electrophoresis techniques.

The results of this method will be either positive or negative for specific GM elements. There are four testing methods which includes i Target-taxon specific ii Screening iii Construct-specific and iv Event-specific, these methods are generally used for the detection and identification of GM crops using PCR. Selection of specific and suitable primers is the most critical step in GMO detection which depends upon the testing method used. Brief detail of qualitative PCR based testing methods is given below:. Ontiveros, C. Abarca, A. Ortiz, M. Ortiz, L. Lina, F. Villalobos, G. Characterization of cry genes in a mexican Bacillus thuringiensis strain collection.

Bt transgenic crops: risks and benefits. Felsot, T. Goode, M. Hammig, D. Comparative environmental impacts of biotechnology-derived and traditional soybean, corn, and cotton crops. Council for Agricultural Science and Technology. Iowa, USA. Mandaokar, A. Shukla, D.

Pattanayak, P. Naik, R. Bacillus thuringiensis cry1Ab gene confers resistance to potato against Helicoverpa armigera Hubner. Potato Res. Sardana, H. Agrobacterium-transformed rice plants expressing synthetic cryIA b and cryIA c genes are highly toxic to striped stem borer and yellow stem borer. Surewicz, P. Carey, M. Pozsgay, T. A view of fungal ecology. Mycologia Douches, W. Li, E. Combining engineered Bt -cry3A and natural resistance mechanisms in potato for control of Colorado potato beetle. Kluwer, Dordrecht, p. Effects of Bt Bacillus thuringiensis transgenic cotton on the dynamics of pest population and their enemies.

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Potential for the environmental impact of transgenic crops. Berry, G. Biological parameters of convergent lady beetle Coleoptera: Coccinellidae feeding on aphids Homoptera: Aphididae on transgenic potato. Li, Q. Xue, M. Abo-El-Saad, D. Transgenic rice plants harboring an introduced potato proteinase inhibitor II gene are insect resistant. Gene flow and introgression from domesticated plants into their wild relatives. Entomological Society of America. Symposium Summaries. December , San Diego, California.

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    Hamilton, E. Burgess, R. Transgenic potato plants with enhanced resistance to the tomato moth, Lacanobia oleracea , growthroom trials. Genetic engineering of plants for insect resistance, p. Rechcigl eds. Biological and biotechnological control of insect pests. Lewis, Boca Raton, p. Hellmich, C. Effects of transgenic Bacillus thuringiensis corn grain on B. Picard-Nizou, E. Grallien, B. Zaccomer, L. Effects of proteinase inhibitor ingestion on survival, learning abilities and digestive proteinases of the honeybee.

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    • The evolutionary potential of crop pests. Sustainability of transgenic insecticidal cultivars: integrating pest genetics and ecology. A summary of research on the environmental impact of Bt cotton in China. Xue ed. Published by Greenpeace, 26p. Insect-resistant transgenic plants in a multi-trophic context. Plant J. Reeds, D. Burial and seed survival in Brassica napus subsp. Oleifera and Sinapis arvensis including a comparison of transgenic and non-transgenic lines of the crop.

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      Transgenic Crop Plants: Volume 2: Utilization And Biosafety by Chittaranjan Kole

      Watson, J. No detection of cry1Ac protein in soil after multiple years of transgenic Bt cotton Bollgard use. Transgenic host plant resistance and non-target effects, p. Burrows eds. Genetically engineered organisms. Assessing environmental and human health effects. Baumgartner, P. Effects of transgenic Bacillus thuringiensis corn-fed prey on mortality and development time of immature Chrysoperla carnea Neuroptera: Chrysopidae.

      Moar, M. Pusztai-Carey, A. Toxicity of Bacillus thuringiensis cry1Ab toxin to the predator Chrysoperla carnae. Prey-mediated effects of cry1Ab toxin and protoxin and cry2A protoxin on the predator Chrysoperla carnea. Powell, A. Gatehouse, J. Gatehouse, L. Gatehouse, Y. Shi, W. Hamilton, A. Merryweather, C. Newell, J. Timans, W. Peumans, E.

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      Expression of snowdrop lectin in transgenic tobacco plants results in added protection against aphids. Feldman, F. Gould, G. Kennedy, G. Naturally occurring biological controls in genetically engineered crops, p. Barbosa ed. Conservation biological control. Academic Press, London, p. Sato, M. Bruchid resistance of transgenic azuki bean expressing seed alpha-amylase inhibitor in the common bean.

      Global review of commercialized transgenic crops, Bacillus thuringiensis var. Dickburt, L. Buysse, C. Piens, B. Saey, A. Gossele, A. Transgenic corn expressing a cry9C insecticidal protein from Bacillus thuringiensis protected from European corn borer damage. Crop Sci. Billings, Q. Chen, J. Lashomb, G. Transformation of eggplant with synthetic cryIIIA gene produces a high level of resistance to the Colorado potato beetle. Scriber, J. Toxicity of Bacillus thuringiensis var. Bonade-Bottino, C. Transgenic plants for insect resistance. Plant Sci. Plant defense strategies and host-plant resistance, p.

      Kogan ed. Daniell, S. Varma, S. Garczynski, F. Overexpression of the Bacillus thuringiensis Bt cry2Aa2 protein in chloroplasts confers resistance to plants against susceptible and Bt -resistant insects.

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      USA L Beland, C. Bowman, N. Carozzi, R. Crenshaw, L. Crosslaind, J. Dawson, N. Desai, M. Hill, M. Kadwell, K. Launis, K. Lewis, D. Maddox, D. Mc Pherson, M. Meghiji, E. Merlin, R. Rhodes, G. Warren, M. Field perfomance of elite transgenic corn plants expressing insecticidal protein derived from Bacillus thuringiensis.

      BioTechnology Mandaokar, K. Sreenivasu, S. Chakrabarti, S. Bisaria, S. Sharma, S. Insect-resistant transgenic brinjal plants. Field manual of techniques in invertebrate pathology. Dordrecht, Kluwer, p. Augustin, G. Pilate, V. Delplanque, D. Toxicity to Chrysomela tremulae Coleoptera: Chrysomelidae of a transgenic poplars expressing a cysteine proteinase inhibitor.

      Use of learned odours by a parasitic wasp in accordance with host and food needs. Nature Transgenic pollen harms monarch larvae. Biodiversity and structure of ground beetle assemblages Coleoptera Carabidae in Bt corn and its effects on non target insects. Agraria Bachiocoltura Furlanis, B. Effects of Bt corn on Rhopalosiphum padi L.

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      Agraria Bachicoltura Gene flow from cultivated to wild raspberries in Scotland: developing a basis for risk assessment for testing and deployment of transgenic cultivars. Burgess, H. Gatehouse, C. Voisey, E. Effects of ingestion of a Bacillus thuringiensis toxin and a trypsin inhibitor on honeybee fligth activity and longevity. Apidologie Goyal, A. Shukla, S. Bisaria, R. Bhalla, V. Reddy, A. Chaurasia, R. Sharma, I. Crop Prot. Riazuddin, N. Loc, A. Expression of multiple insecticidal genes confers broad resistance against a range of different rice pests.

      Mele, R. Gabarra, J. Vassal, E. Influence of the developmental stage of transgenic rice plants cv. Senia expressing the cry1B gene on the level of protection against the striped stem borer Chilo suppressalis. Plant Cell Rep. Ecology of transgenic crops: Genetically engineered plants might generate weed problems and affect non-target organisms, but measuring the risk is difficult. Svab, D. Schaaf, P. Hoogan, D. Amplification of a chimeric Bacillus gene in chloroplasts leads to an extraordinary level of an insecticidal preotein in tobacco.

      McCabe, D. Russll, D. Robison, K. Stable tranformation of Populus and incorporation of pest resistance by electric discharge particle acceleration. Field assessment of the effects of a microbial pest control agent on non-target Lepidoptera. Comparison of fumonisin concentration in kernels of transgenic Bt corn hybrids and nontransgenic hybrids. Plant Dis. National Academy Press.

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      Washington, DC, p. Losey, O. Transgenic insecticidal corn: beyond insecticidal toxicity to ecological complexity. BioScience Oviposition of European corn borer Lepidoptera: Pyralidae and impact of natural enemy populations in transgenic versus isogenic corn. Engineering of insect-resistant plants with Bacillus thuringiniesis crystal protein genes. Insect control with transgenic plants expressing Bacillus thuringiensis crystal proteins, p. Coziel eds. Taylor and Francis, London, p.

      Fuchs, D. Dean, S. Modification of the coding sequence enhances plant expression of insect control protein genes.