ISSN:
1432-203X
Keywords:
Key wordsAgrobacterium-mediated transformation
;
Datura meteloides
;
Preculture period
;
Transgene expression
Source:
Springer Online Journal Archives 1860-2000
Topics:
Biology
Notes:
Abstract A transformation system is described for Datura meteloides using the supervirulent Agrobacterium tumefaciens strain 1065, carrying both the β-glucuronidase (gusA) and neomycin phosphotransferase II (nptII) genes between the T-DNA border sequences of the binary vector. The importance of conditions such as the preculture period of the plant tissues, wounding, bacterial dilution and incubation time were evaluated in terms of transgenic plant production. A preculture period of 2–3 days, using a 1:20 or 1:10 (vol:vol) dilution of an overnight bacterial culture, resulted in optimum shoot regeneration, with 48% from a total of 576 explants regenerating transformed shoots. Expression of the gusA and nptII genes was confirmed by a GUS fluorometric assay and by NPTII ELISA. Southern analysis revealed the integration of both transgenes, which segregated as dominant Mendelian traits in seed progeny.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1007/s002990050621
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