Additional Information, References & Resources

Genotypes of some useful Agrobacterium tumefaciens strains

Warning!! Agrobacterium use is constrained in jurisdictions such as the USA, and it may be unwise to use it in any research that might result in a product for sale or import into the USA. You may wish to use the Transbacter system instead for freedom to operate. For information, see the  TransBacter Project.

• LBA4404 (Ach5 pTiAch5) Sm/Sp(R) in the virulence plasmid (from Tn904); all T-DNA of pTiAch5 eliminated in pAL4404 (Hoekema et al., 1983).
• EHA101, genotype C58 pTiBo542; T-region::aph, Km(R); A281 derivative harboring pEHA101, T-DNA replaced with nptII, elimination of T-DNA boundaries uncofirmed, super-virulent (Hood et al., 1986).
• EHA105 is a Km(S) derivative of EHA101 (Hood et al., 1993).
• AGL1, genotype is AGL0 (C58 pTiBo542) recA::bla, T-region deleted Mop(+) Cb(R) [AGL0 is an EHA101 with the T-region deleted, which also deletes the aph gene] (Lazo et al., 1991).
• A281, reconstructed strain, derivative of A136 (cured C58) harboring pTiBo542, super-virulent (Hood et al., 1986).

Types

• Ach5 .......... agrocinopine, octopine type
• B6S3, A6 .... octopine type
• Bo542 ........ leucinopine, succinamopine, agropine type, vir weaker than A281
• C58, T37 .... nopaline types
• A281 ......... succinamopine, leucinopine, agrocinopine

Antibiotics

• Chloramphenicol, 100 µg/mL for strain AGL1, 10 µg/mL for LBA4404, 25 µg/mL for EHA105 and for E. coli.
• Kanamycin, 50 µg/mL for both Agrobacterium and E. coli.
• For selection of transformed rice plants we use hygromycin at 50 µg/mL and 25 µg/mL for tobacco.

Selected References

• Chen L, Zhang S, Beachy RN, Fauquet CM (1998) A protocol for consistent, large-scale production of fertile transgenic rice plants. Plant Cell Reports 18:25-31
• Christou P (1991) Production of transgenic rice (Oryza sativa L.) plants from agronomically important indica and japonica varieties via electric discharge particle acceleration of exogenous DNA into immature zygotic embryos. Biotechnology 9:957-962
• Christou P (1997) Rice transformation: bombardment. Plant Mol Biol 35:197-203.
• Deblaere R, Reynaerts A, Hofte H, Hernalsteens JP, Leemans J, and Van Montagu M (1987) Vectors for cloning in plant cells. Meth Enzymol 153:277-292
• Hajdukiewicz,P, Svab, Z, Maliga, P., (1994) The small versatile pPZP family of Agrobacterium binary vectors for plant transformation. Plant Mol Biol 25:989-994
• Hiei Y, Ohta S, Komari T, Kumashiro T (1994) Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J 6:271-282
• Hoekema A, Hirsch PR, Hooykaas PJJ, Schilperoort RA (1983) Binary vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature 303:179-180
• Hood EE, Helmer GL, Fraley RT, Chilton MD (1986) The hypervirulence of Agrobacterium tumefaciens A281 is encoded in a region of pTiBo542 outside of T-DNA. J Bac 168:1291-1301
• Hood EE, Gelvin SB, Melchers S, Hoekema A (1993) New Agrobacterium helper plasmids for gene transfer to plants (EHA105). Trans Res 2:208-218
• Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: Beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901-3907
• Klapwijk PM, van Breukelen J, Korevaar K, Ooms G, Schilperoort RA (1980) T ransposition of Tn904 encoding streptomycin resistance into octopine Ti plasmid of Agrobacterium tumefaciens. J Bac 141:129-136
• Lazo GR, Stein PA, Ludwig RA (1991) A DNA transformation-competent Ara bidopsis genomic library in Agrobacterium. BioTechnology 9:963-967
• Ohta S, Mita S, Hattori T, Nakamura K (1990) Construction and expression in tobacco of a beta-glucuronidase (GUS) reporter gene containing an intron within the coding sequence. Plant Cell Physiol 31:805-814
• Ooms G, Hooykaas PJJ, Van Veen RJM, Van Beelen P, Regensburg-Tunk JG, Schilperoort RA (1982) Octopine Ti-plasmid deletion mutants of Agrobacterium tumefaciens with emphasis on the right side of the T-region. Plasmid 7 :15-29
• Peralta EG, Hellmiss R, Ream W (1986) Overdrive, a T-DNA transmission enhancer on the A. tumefaciens tumour-inducing plasmid. EMBO J 5:1137-1142
• Porath, J. (1992). Immobilized metal ion affinity chromatography. Protein Expre Purif 3:263-281
• Siemering KR, Golbik R, Sever R, Haseloff J (1996) Mutations that suppress the thermosensitivity of green fluorescent protein. Curr Biol 6:1653-1663
• Tanaka A, Mita S, Ohta S, Kyozuka J, Shimamoto K, Nakamura K (1990) Enhancement of foreign gene expression by a dicot intron in rice but not in tobacco is correlated with an increased level of mRNA and an efficient splicing of the intron. Nucl Acids Res 18:6767-6770

Other Resources

• List of TransBacter strains available (will be linked soon)
• Read Frequently Asked Questions about requesting CAMBIA materials
• Read Frequently Asked Questions about pCAMBIA Vectors
• GUSPlus information in the BioForge GUSPlus Project.

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