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Mechanisms of Infection Vir Gene

Mechanisms of Infection Vir Gene

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Published by Sathiyaraj
Agrobacteriaum mediated gene transfer a overview
Agrobacteriaum mediated gene transfer a overview

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Published by: Sathiyaraj on Nov 21, 2007
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07/16/2013

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 Agrobacterium tumefaciens
 Agrobacterium tumefaciens
is a ubiquitous soil borne pathogen responsible for Crown Gall disease (right), affecting many higher species of plant. The pathogen is a problem for agriculture all over the world. DNA transfer from
 Agrobacteriumtumefaciens
to eukaryotic cells is the only known example of inter-kingdom DNAtransfer (Dumas
et al.
, (2001). It is therefore unsuprising that
 Agrobacterium
has evolvedits own unique and specialised system to accomplish this task, which is of great interestto plant scientists. The information contained in this website will explore the history andall that we know about this remarkable micro-organism.This website aims to give an insight into the bacterium,with information about the research that has been carried out inthe past, and how the knowledge we have gained can be appliedto help alleviate problems facing food production around theworld. Other potential uses of the technology are also reviewed,such as using our knowledge of 
 Agrobacterium
to produce a biocontrol agent and producing effective transformation vectorssuch as the pGreen binary vector. This website also includes acomprehensive list of references for anyone wishing to readfurther into any aspect of the research surrounding the bacterium.The study of transgenic or genetically modified plants is currently attractingsignificant interest because of its potential applications in industry, the environment, inthe home and in medicine. Most genetic modification is perceived as being the insertionof foreign DNA into the genome of the target organism, and hence there is much debateas to whether this will have any unseen consequences in the future, and whether it ismorally acceptable. There are also many problems associated with current methods of transgenic plant production, most notably the low transformation frequency encountered,or the failure of transgenic plants to grow at all. The use of 
 Agrobacterium tumefaciens
intransgenic plant production has arisen from the need to find an effective vector system tosuccessfully integrate the gene of interest into the correct area of the plant genome.In recent years, the subject of uncontrolled cellular growth;
Cancer 
, has been thefocus of considerable attention as a result of its life threatening nature in humans.Extensive studies have unlocked information and provided a comprehensive knowledge base of undifferentaiated tissue growth in mammals, however little attention has been paid to cancer found in the plant kingdom.
 Agrobacterium tumefaciens
causes one of themost common plant tumours, commonly known as Crown Gall disease which affects awide variety of plants. Much research has taken place in an effort to understand themechanism by which the bacterium infects the plant cell, in the hope that the bacterium
 
can be used as a vector system to transfer foreign DNA into a plant genome with a higher degree of success than is currently seen with traditional methods.Crown Gall disease is a common plant pathogen, affecting over 600 types of  plants.
 Agrobacterium tumefaciens
is a ubiquitous micro-organism that can be found inmost soil samples. The disease mostly affects monocotyledonous species, such as woodyand herbaceous plants and can be identified by the appearance of tumnours or galls of varying size and shape on the lower stem and main roots of the plant. Crown Gall diseasecan affect many commercially important and valuable crops such as Grapes, Rice andSugar Beet.
Current Problems
Plant disease is currently a major problem facing the developed world. Currentlyas much as 30% of the yearly total production of food crops is lost due to plant disease.At the current time, there is enough food produced to feed the population, but only just. If the predicted population increase over the next 2-3 decades take place, it will benecessary to increase food production to meet demand. As such a large proportion of crops are lost to plant pathogens each year, there is currently much interest in developingstrategies to increase plants natural resistance to pathogenic attack.
 A. tumefaciens
canremain dormant in the soil over winter, and can live saprophytically for many years.The diagram below gives some of the applications for plant transformationtechnology.
 
Characteristics of 
 Agrobacteriumtumefaciens
 Agrobacterium tumefaciens
is a gramnegative, motile, rod shaped bacterium whichis non sporing, and is closely related to the N-fixing rhizobium bacteria which form rootnodules on leguminous plants. The bacteriumis surrounded by a small number o peritricious flagella. Virulent bacteria containone or more large plasmids, one of whichcarries the genes for tumour induction and isknown as the Ti (tumour inducing) plasmid.The Ti plasmid also contains the genes thatdetermine the host range and the symptoms,which the infection will produce. Without thisTi plasmid, the bacterium is described as beingnon virulent and will not be able to cause disease on the plant.Crown Galls first appear as small, white, soft protrusions, initially found at the base of the plant stem. As the tumours enlarge, the surface takes on a mottled dark brownappearance due to the death and decay of the peripheral cells. The tumour usually appearseither as a swelling of the plant tissue, or as a separate mass of tissue close to the plantsurface, joined only by a narrow neck of tissue. Tumours can either be soft and spongyand may crumble on touch, or can be hard and appear knobbly or knotty. Some tumourscan reach up to 30cm in diameter; though 5-10cm is more common (see cross section below). The tumours may rot away in the autumn, only to re-appear again the followingspring.When infected with the bacterium, plants may also become stunted, produce smallchlorotic leaves, and are more susceptible to extreme environmental conditions such aswinter cold and wind.
 A. tumefaciens
is most well known for its ability to integrate a small part of the Ti plasmid into the host plant genome, which causes the plant cells to become cancer cellsand produce specific compounds called Opines, which the bacterium utilise as a carbonsource. This property means that many textbooks class
 A. Tumefaciens
as a genetic parasite, since the bacterium redirects the metabolic activities of the plant to producecompounds specific to the bacterium. It is this process which gives
 A. Tumefaciens
its potential to be used as a tool for plant transformation.
Disease Control
As
 A. tumefaciens
is so ubiquitous in the soil, there is currently much interest inways to try and prevent the bacterium from infecting susceptible plants, either through

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