RT article
T1 Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus
A1 Domínguez, Alfons
A1 Guerri, José
A1 Cambra, Mariano
A1 Navarro, Luis
A1 Moreno, Pedro
A1 Pena, Leandro
AB The coat protein gene of citrus tristeza virus (CTV) has been introduced into Mexican lime (Citrus aurantifolia Swing.) plants by using an improved Agrobacterium-mediated genetic transformation system. Internodal stem segments from greenhouse-grown seedlings were co-cultivated with A. tumefaciens strain EHA 105 carrying the binary plasmid pBI 121/CTV-CP in a medium rich in auxins that provided the explant cells with the proper treatment to shift them to a competent state for transformation. The transformation frequency was enhanced, and this allowed us to recover 42 transgenic plants from 1200 explants. Regenerated shoots were identified as transformants by performing beta-glucuronidase (GUS) assays and subsequently by PCR amplifications of the CTV-CP transgene. Southern analyses revealed that at least one copy of the CTV-CP gene was integrated in all PCR positive plants. Interestingly, 70% of them had linked T-DNAs arranged at one locus. Copy number of the CTV-CP gene varied from one to six among the transgenic lines. Half of them showed truncated T-DNAs in which the left border was lost. Expression of the CTV-CP transgene was demonstrated in 38 out of 42 plants by western analysis and DASI-ELISA. No correlation was found between coat protein expression and transgene copy number or integration pattern.
SN 0721-7714
YR 2000
FD 2000
LK http://hdl.handle.net/20.500.11939/5148
UL http://hdl.handle.net/20.500.11939/5148
LA en
NO Dominguez, A., Guerri, J., Cambra, M., Navarro, L., Moreno, P., Pena, L. (2000). Efficient production of transgenic citrus plants expressing the coat protein gene of citrus tristeza virus. Plant Cell Reports, 19(4), 427-433.
DS MINDS@UW
RD Aug 15, 2022