Plant Transformation Using Agrobacterium tumefaciens
In nature, when a plant is wounded, Agrobacterium (found in soil frequently) can attach to the wound site and then proceeds to transform the cell it encounters. The wound site on the plant produces sugars and phenolic compounds, which not only signal the opportunity for pathogenesis but also aid in the induction of transcription of virulence genes. These virulence genes are located on the Tumor-inducing (Ti) plasmid. The Ti plasmid also contains transferred DNA (T-DNA). Agrobacterium can effectively transfer the T-DNA to the host plant and then cause 2 outcomes. First, they may code for a plant hormone that causes tumor growth, then they code for enzymes that produce opines, which are metabolized by agrobacterium. The result is a growth (called a ‘crown gall’) that is perfect for the agrobacterium to proliferate.
Scientists have discovered that they can use this natural ability of agrobacterium to insert genes of interest into the plant cells. The T-DNA region of the Ti plasmid that causes crown gall formation can be replaced with the genes of interest scientists want to study. Scientists have also discovered that different strains of Agrobacterium (i.e. GV3101, AGL-1, etc) work well for different types of plant species.
Plant Transformation Using Agrobacterium tumefaciens
There are several advantages for using agrobacterium mediated transformation over other transformation methods. These include(but not limited to):
- Reduction in transgene copy number
- Increased stability of intact introduced gene
Agrobacterium Mediated Plant Transformation Process Outline
Below are the basic steps in the Agrobacterium-mediated Plant Transformation Process:
- Isolate genes of interest from the source organism
- Create a functional transgenic construct that includes:
a. The gene of interest
b. Expression Promoters
c. Codon optimization that increases protein production (if needed)
d. Marker genes to track gene expression in the host plant - Insert transgene into Ti plasmid
- Insert T-DNA containing plasmid into Agrobacterium
- Mix newly transformed agro cells with plant cells to allow transfer of the T-DNA into the plant chromosome
- Regenerate the transformed cells into genetically modified plants
- Test at various stages (greenhouse, field, lab) to ensure trait performance.
Agrobacterium Competent Cells
Using Propriety methods, Intact Genomics has developed and manufactured a variety of Agrobacterium chemically and electroporation competent cells with high efficiency. Agrobacterium strains include EHA105, LBA4404, GV3101 and AGL-1. All are useful for plant transgenic operations in a variety of species.
Custom Agrobacterium cells also available!
Product Links | Efficiency | Application & Benefits |
GV3101 Chemically Competent Agrobacterium | ≥1.0 x 10^5 | Transgenic operations of Arabidopsis thaliana, tobacco, corn, potato, and other plants |
GV3101 ElectroCompetent Agrobacterium | ≥1.0 x 10^7 | |
AGL1 Chemically Competent Agrobacterium | ≥1.0 x 10^5 | Transgenic operations of Arabidopsis thaliana, maize and other monocots |
AGL1 ElectroCompetent Agrobacterium | ≥1.0 x 10^7 | |
LBA4404 Chemically Competent Agrobacterium | ≥1.0 x 10^5 | Transgenic operations of tomatoes, tobacco and other plants |
LBA4404 ElectroCompetent Agrobacterium | ≥1.0 x 10^7 | |
EHA105 Chemically Competent Agrobacterium | ≥1.0 x 10^5 | Transgenic operations of rice, tobacco and other plants |
EHA105 ElectroCompetent Agrobacterium | ≥1.0 x 10^7 | |
C58C1 Chemically Competent Agrobacterium | ≥1.0 x 10^5 | Transgenic operations that involve potatoes, tobacco and other plants |
C58C1 ElectroCompetent Agrobacterium | ≥1.0 x 10^7 | |
Chemically Competent Agrobacterium Combo Pack | Various | For applications that require multiple agrobacterium competent cell strains |
ElectroCompetent Agrobacterium Combo Pack | Various | |
GV3101 (pSoup) ElectroCompetent Agrobacterium | ≥1.0 x 10^7 | Transgenic operations of Arabidopsis thaliana, tobacco, corn, potato, and other plants |
GV3101 (pSoup-p19) ElectroCompetent Agrobacterium | ≥1.0 x 10^7 | Transgenic operations of Arabidopsis thaliana, tobacco, corn, potato, and other plants |
AGL1 (pSoup) ElectroCompetent Agrobacterium | ≥1.0 x 10^7 | Transgenic operations of Arabidopsis thaliana, maize and other monocots |
EHA105 (pSoup) ElectroCompetent Agrobacterium | ≥1.0 x 10^7 | Transgenic operations of rice, tobacco and other plants |
- Gelvin B. S. (2003) Agrobacterium-Mediated Plant Transformation: the Biology behind the “Gene-Jockeying” Tool. Microbiology and Molecular Biology Reviews 67(1): 16–37
- Jones D.H., Doherty A, and Wu H. (2005) Review of methodologies and a protocol for the Agrobacterium-mediated transformation of wheat. Plant Methods 1: 5
- Kumar K.K. , Maruthasalam S., Loganathan M., Sudhakar D. and Balasubramanian P. (2005) An Improved Agrobacterium-Mediated Transformation Protocol for Recalcitrant Elite Indica Rice Cultivars. Plant Molecular Biology Reporter 23: 67–73