Editing Agrobacterium (section)

==Uses in biotechnology==
{{See also|Horizontal gene transfer}}
{{See also|Agroinfiltration}}

The ability of ''Agrobacterium'' to transfer [[gene]]s to [[plant]]s and fungi is used in [[biotechnology]], in particular, [[genetic engineering]] for [[plant improvement]]. Genomes of plants and fungi can be engineered by use of ''Agrobacterium'' for the delivery of sequences hosted in [[Transfer DNA binary system|T-DNA binary vectors]]. A modified Ti or Ri plasmid can be used. The plasmid is 'disarmed' by deletion of the tumor inducing genes; the only essential parts of the T-DNA are its two small (25 base pair) border repeats, at least one of which is needed for plant transformation.<ref name=Montagu1977/><ref name=Joos1983/> The genes to be introduced into the plant are cloned into a plant binary vector that contains the T-DNA region of the disarmed [[plasmid]], together with a selectable marker (such as [[antimicrobial resistance|antibiotic resistance]]) to enable selection for plants that have been successfully transformed. Plants are grown on media containing antibiotic following transformation, and those that do not have the T-DNA integrated into their genome will die. An alternative method is [[agroinfiltration]].<ref name=Thomson>{{cite journal |title=Genetic Engineering of Plants |volume=3 |journal=Biotechnology |author=Thomson JA |url=http://www.eolss.net/sample-chapters/c17/e6-58-03-04.pdf |access-date=17 July 2016 |archive-url=https://web.archive.org/web/20170117110839/http://www.eolss.net/sample-chapters/c17/e6-58-03-04.pdf |archive-date=17 January 2017 |url-status=live }}</ref><ref>{{cite journal | vauthors = Leuzinger K, Dent M, Hurtado J, Stahnke J, Lai H, Zhou X, Chen Q | title = Efficient agroinfiltration of plants for high-level transient expression of recombinant proteins | journal = Journal of Visualized Experiments | volume = 77 | issue = 77 | date = July 2013 | pmid = 23913006 | pmc = 3846102 | doi = 10.3791/50521 }}</ref>

[[File:Transformation with Agrobacterium.JPG|thumb|right|Plant (''[[Solanum|S. chacoense]]'') transformed using ''Agrobacterium''. Transformed cells start forming calluses on the side of the leaf pieces]]
[[Transformation (genetics)|Transformation]] with ''Agrobacterium'' can be achieved in multiple ways. [[Protoplast]]s or alternatively leaf-discs can be incubated with the ''Agrobacterium'' and whole plants regenerated using [[plant tissue culture]]. In [[agroinfiltration]] the ''Agrobacterium'' may be injected directly into the leaf tissue of a plant. This method transforms only cells in immediate contact with the bacteria, and results in transient expression of plasmid DNA.<ref>{{cite journal | vauthors = Shamloul M, Trusa J, Mett V, Yusibov V | title = Optimization and utilization of ''Agrobacterium''-mediated transient protein production in ''Nicotiana'' | journal = Journal of Visualized Experiments | issue = 86 | date = April 2014 | pmid = 24796351 | pmc = 4174718 | doi = 10.3791/51204 }}</ref>

Agroinfiltration is commonly used to transform tobacco (''[[Nicotiana]]''). A common transformation protocol for ''[[Arabidopsis thaliana|Arabidopsis]]'' is the floral dip method:<ref>{{cite journal | vauthors = Clough SJ, Bent AF | title = Floral dip: a simplified method for ''Agrobacterium''-mediated transformation of ''Arabidopsis thaliana'' | journal = The Plant Journal | volume = 16 | issue = 6 | pages = 735–43 | date = December 1998 | pmid = 10069079 | doi = 10.1046/j.1365-313x.1998.00343.x | s2cid = 410286 }}</ref> An [[inflorescence]] is dipped in a suspension of ''Agrobacterium'', and the bacterium transforms the [[germline]] cells that make the female [[gametes]]. The [[seed]]s can then be screened for antibiotic resistance (or another marker of interest). Plants that have not integrated the plasmid DNA will die when exposed to the antibiotic.<ref name="Thomson" />

''Agrobacterium'' is listed as being the vector of genetic material that was transferred to these USA GMOs:<ref>[http://www.cfsan.fda.gov/~lrd/biocon.html#list The FDA List of Completed Consultations on Bioengineered Foods] {{webarchive |url=https://web.archive.org/web/20080513162330/http://www.cfsan.fda.gov/~lrd/biocon.html#list |date=May 13, 2008 }}</ref>
* [[Soybean]]
* [[Cotton]]
* [[Maize]]
* [[Sugar Beet]]
* [[Alfalfa]]
* [[Wheat]]
* Rapeseed Oil ([[Canola]])
* [[Creeping bentgrass]] (for animal feed)
* Rice ([[Golden Rice]])

The [[Transformation (genetics)|transformation]] of fungi using ''Agrobacterium'' is used primarily for research purposes,<ref>{{cite journal | vauthors = Michielse CB, Hooykaas PJ, van den Hondel CA, Ram AF | s2cid = 23959400 | title = Agrobacterium-mediated transformation as a tool for functional genomics in fungi | journal = Current Genetics | volume = 48 | issue = 1 | pages = 1–17 | date = July 2005 | pmid = 15889258 | doi = 10.1007/s00294-005-0578-0 | hdl = 1887/3763935 | hdl-access = free }}</ref><ref>{{cite journal | vauthors = Idnurm A, Bailey AM, Cairns TC, Elliott CE, Foster GD, Ianiri G, Jeon J | title = ''Agrobacterium''-mediated transformation of fungi | journal = Fungal Biology and Biotechnology | volume = 4 | pages = 6 | date = 2017 | pmid = 28955474 | pmc = 5615635 | doi = 10.1186/s40694-017-0035-0 | doi-access = free }}</ref> and follows similar approaches as for plant transformation.  The [[Transfer DNA binary system|Ti plasmid system]] is modified to include DNA elements to select for transformed fungal strains, after co-incubation of ''Agrobacterium'' strains carrying these plasmids with fungal species.