T-DNA
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T-DNA
The transfer DNA (abbreviated T-DNA) is the transferred DNA of the tumor-inducing (Ti) plasmid of some species of bacteria such as ''Agrobacterium tumefaciens'' and ''Agrobacterium rhizogenes(actually an Ri plasmid)''. The T-DNA is transferred from bacterium into the host plant's nuclear DNA genome. The capability of this specialized tumor-inducing (Ti) plasmid is attributed to two essential regions required for DNA transfer to the host cell. The T-DNA is bordered by 25-base-pair repeats on each end. Transfer is initiated at the right border and terminated at the left border and requires the ''vir'' genes of the Ti plasmid. The bacterial T-DNA is about 24,000 base pairs long and contains plant-expressed genes that code for enzymes synthesizing opines and phytohormones. By transferring the T-DNA into the plant genome, the bacterium essentially reprograms the plant cells to grow into a tumor and produce a unique food source for the bacteria. The synthesis of the plant hormones auxin ...
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Ti Plasmid
A tumour inducing (Ti) plasmid is a plasmid found in pathogenic species of ''Agrobacterium'', including ''A. tumefaciens, ''A. rhizogenes'', ''A. rubi'' and ''A. vitis''. Evolutionarily, the Ti plasmid is part of a family of plasmids carried by many species of Alphaproteobacteria. Members of this plasmid family are defined by the presence of a conserved DNA region known as the ''repABC'' gene cassette, which mediates the replication of the plasmid, the partitioning of the plasmid into daughter cells during cell division as well as the maintenance of the plasmid at low copy numbers in a cell. The Ti plasmids themselves are sorted into different categories based on the type of molecule, or opine, they allow the bacteria to break down as an energy source. The presence of this Ti plasmid is essential for the bacteria to cause crown gall disease in plants. This is facilitated via certain crucial regions in the Ti plasmid, including the ''vir'' region, which encodes for virulence g ...
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Agrobacterium Tumefaciens
''Agrobacterium radiobacter'' (more commonly known as ''Agrobacterium tumefaciens'') is the causal agent of crown gall disease (the formation of tumours) in over 140 species of eudicots. It is a rod-shaped, Gram-negative soil bacterium. Symptoms are caused by the insertion of a small segment of DNA (known as the T-DNA, for 'transfer DNA', not to be confused with tRNA that transfers amino acids during protein synthesis), from a plasmid into the plant cell, which is incorporated at a semi-random location into the plant genome. Plant genomes can be engineered by use of ''Agrobacterium'' for the delivery of sequences hosted in T-DNA binary vectors. ''Agrobacterium tumefaciens'' is an Alphaproteobacterium of the family Rhizobiaceae, which includes the nitrogen-fixing legume symbionts. Unlike the nitrogen-fixing symbionts, tumor-producing ''Agrobacterium'' species are pathogenic and do not benefit the plant. The wide variety of plants affected by ''Agrobacterium'' makes it of gr ...
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Transfer DNA Binary System
A transfer DNA (T-DNA) binary system is a pair of plasmids consisting of a T-DNA binary vector and a ''vir'' helper plasmid. The two plasmids are used together (thus ''binary'') to produce genetically modified plants. They are artificial vectors that have been derived from the naturally occurring Ti plasmid found in bacterial species of the genus ''Agrobacterium'', such as '' A. tumefaciens''. The binary vector is a ''shuttle vector'', so-called because it is able to replicate in multiple hosts (e.g. ''Escherichia coli'' and ''Agrobacterium''). Systems in which T-DNA and ''vir'' genes are located on separate replicons are called T-DNA binary systems. T-DNA is located on the binary vector (the non-T-DNA region of this vector containing origin(s) of replication that could function both in ''E. coli'' and ''Agrobacterium'', and antibiotic resistance genes used to select for the presence of the binary vector in bacteria, became known as vector backbone sequences). The replicon containi ...
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Crown Gall Tumor
''Agrobacterium'' is a genus of Gram-negative bacteria established by H. J. Conn that uses horizontal gene transfer to cause tumors in plants. '' Agrobacterium tumefaciens'' is the most commonly studied species in this genus. ''Agrobacterium'' is well known for its ability to transfer DNA between itself and plants, and for this reason it has become an important tool for genetic engineering. Nomenclatural History Leading up to the 1990s, the genus ''Agrobacterium'' was used as a wastebasket taxon. With the advent of 16S sequencing, many ''Agrobacterium'' species (especially the marine species) were reassigned to genera such as ''Ahrensia'', ''Pseudorhodobacter'', ''Ruegeria'', and ''Stappia''. The remaining ''Agrobacterium'' species were assigned to three biovars: biovar 1 ('' Agrobacterium tumefaciens''), biovar 2 (''Agrobacterium rhizogenes''), and biovar 3 (''Agrobacterium vitis''). In the early 2000s, ''Agrobacterium'' was synonymized with the genus ''Rhizobium''. This move ...
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Agrobacterium
''Agrobacterium'' is a genus of Gram-negative bacteria established by H. J. Conn that uses horizontal gene transfer to cause tumors in plants. ''Agrobacterium tumefaciens'' is the most commonly studied species in this genus. ''Agrobacterium'' is well known for its ability to transfer DNA between itself and plants, and for this reason it has become an important tool for genetic engineering. Nomenclatural History Leading up to the 1990s, the genus ''Agrobacterium'' was used as a wastebasket taxon. With the advent of 16S sequencing, many ''Agrobacterium'' species (especially the marine species) were reassigned to genera such as ''Ahrensia'', ''Pseudorhodobacter'', ''Ruegeria'', and ''Stappia''. The remaining ''Agrobacterium'' species were assigned to three biovars: biovar 1 (''Agrobacterium tumefaciens''), biovar 2 (''Agrobacterium rhizogenes''), and biovar 3 (''Agrobacterium vitis''). In the early 2000s, ''Agrobacterium'' was synonymized with the genus ''Rhizobium''. This move pr ...
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Agrobacterium Rhizogenes
''Rhizobium rhizogenes'' (formerly ''Agrobacterium rhizogenes'') is a Gram-negative soil bacterium that produces hairy root disease in dicotyledonous plants. ''R. rhizogenes'' induces the formation of proliferative multiple-branched adventitious roots at the site of infection, so-called 'hairy roots'. In the rhizosphere, plants may suffer from wounds by soil pathogens or other sources. This leads to the secretion of phenolic compounds like acetosyringone which have chemotactic effects that attract the bacteria. Under such conditions, certain bacterial genes are turned on leading to the transfer of its T-DNA from its root-inducing plasmid (Ri plasmid) into the plant through the wound. After integration and expression, ''in vitro'' or under natural conditions, the hairy root phenotype is observed, which typically includes overdevelopment of a root system that is not completely geotropic, and altered (wrinkled) leaf morphology, if leaves are present. Bacterial genes may be retaine ...
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Transgenic Crops
Genetically modified crops (GM crops) are plants used in agriculture, the DNA of which has been modified using genetic engineering methods. Plant genomes can be engineered by physical methods or by use of ''Agrobacterium'' for the delivery of sequences hosted in T-DNA binary vectors. In most cases, the aim is to introduce a new trait to the plant which does not occur naturally in the species. Examples in food crops include resistance to certain pests, diseases, environmental conditions, reduction of spoilage, resistance to chemical treatments (e.g. resistance to a herbicide), or improving the nutrient profile of the crop. Examples in non-food crops include production of pharmaceutical agents, biofuels, and other industrially useful goods, as well as for bioremediation. Farmers have widely adopted GM technology. Acreage increased from 1.7 million hectares in 1996 to 185.1 million hectares in 2016, some 12% of global cropland. As of 2016, major crop (soybean, maize, canola a ...
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Genetic Engineering
Genetic engineering, also called genetic modification or genetic manipulation, is the modification and manipulation of an organism's genes using technology. It is a set of technologies used to change the genetic makeup of cells, including the transfer of genes within and across species boundaries to produce improved or novel organisms. New DNA is obtained by either isolating and copying the genetic material of interest using recombinant DNA methods or by artificially synthesising the DNA. A construct is usually created and used to insert this DNA into the host organism. The first recombinant DNA molecule was made by Paul Berg in 1972 by combining DNA from the monkey virus SV40 with the lambda virus. As well as inserting genes, the process can be used to remove, or "knock out", genes. The new DNA can be inserted randomly, or targeted to a specific part of the genome. An organism that is generated through genetic engineering is considered to be genetically modified (GM) an ...
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Opines
Opines are low molecular weight compounds found in plant crown gall tumors or hairy root tumors produced by pathogenic bacteria of the genus ''Agrobacterium'' and ''Rhizobium''. Opine biosynthesis is catalyzed by specific enzymes encoded by genes contained in a small segment of DNA (known as the T-DNA, for 'transfer DNA'), which is part of the Ti plasmid (in ''Agrobacterium'') or Ri plasmid (in ''Rhizobium''), inserted by the bacterium into the plant genome. The opines are used by the bacterium as an important energy, carbon and nitrogen source. Each strain of ''Agrobacterium'' and ''Rhizobium'' induces and catabolizes a specific set of opines, this set typifying the Ti plasmid and Ri plasmid. There are some 30 different opines described so far. Chemical structure Chemically, opines fall into two major structural classes: 1. The vast majority are secondary amine derivatives resulting from the reduction of the imine formed by condensation of an amino acid, either with a ke ...
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Transgenic Plant
Genetically modified plants have been engineered for scientific research, to create new colours in plants, deliver vaccines, and to create enhanced crops. Plant genomes can be engineered by physical methods or by use of ''Agrobacterium'' for the delivery of sequences hosted in T-DNA binary vectors. Many plant cells are pluripotent, meaning that a single cell from a mature plant can be harvested and then under the right conditions form a new plant. This ability can be taken advantage of by genetic engineers; by selecting for cells that have been successfully transformed in an adult plant a new plant can then be grown that contains the transgene in every cell through a process known as tissue culture. Research Much of the advances in the field genetic engineering has come from experimentation with tobacco. Major advances in tissue culture and plant cellular mechanisms for a wide range of plants has originated from systems developed in tobacco. It was the first plant to be geneti ...
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Hairy Root Culture
Hairy root culture, also called transformed root culture, is a type of plant tissue culture that is used to study plant metabolic processes or to produce valuable secondary metabolites or recombinant proteins, often with plant genetic engineering. A naturally occurring soil bacterium ''Agrobacterium rhizogenes'' that contains root-inducing plasmids (also called Ri plasmids) can infect plant roots and cause them to produce a food source for the bacterium, opines, and to grow abnormally. The abnormal roots are particularly easy to culture in artificial media because hormones are not needed in contrast to adventitious roots, and they are neoplastic, with indefinite growth. The neoplastic roots produced by ''A. rhizogenes'' infection have a high growth rate (compared to untransformed adventitious roots), as well as genetic and biochemical stability. Currently the main constraint for commercial utilization of hairy root culture is the development and up-scaling of appropriate (bioreacto ...
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Horizontal Gene Transfer
Horizontal gene transfer (HGT) or lateral gene transfer (LGT) is the movement of genetic material between Unicellular organism, unicellular and/or multicellular organisms other than by the ("vertical") transmission of DNA from parent to offspring (reproduction). HGT is an important factor in the evolution of many organisms. HGT is influencing scientific understanding of higher order evolution while more significantly shifting perspectives on bacterial evolution. Horizontal gene transfer is the primary mechanism for the spread of antibiotic resistance in bacteria, and plays an important role in the evolution of bacteria that can degrade novel compounds such as human-created Bactericide, pesticides and in the evolution, maintenance, and transmission of virulence. It often involves Temperateness (virology), temperate bacteriophages and plasmids. Genes responsible for antibiotic resistance in one species of bacteria can be transferred to another species of bacteria through various m ...
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