Genetic transformation of loblolly pine using mature zygotic embryo expiants byAgrobacterium tumefaciens |
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| Authors: | Tang Wei |
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| Institution: | (1) Forest Biotechnology Group, Department of Forestry, North Carolina State University, 27695-7247 Raleigh, NC, USA |
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| Abstract: | Agrobacterium tumefaciens strain LBA 4404 carrying pBI121 plasmid was used to transform mature zygotic embryos of three genotypes (E-Hb, E-Ma, and
E-Mc) of loblolly pine. The results demonstrated that the expression frequency of β-glucuronidase reporter gene (GUS) varied
among genotypes after mature zygotic embryos were infected withAgrobacterium tumefaciens cultures. The highest frequency (27.8%) of GUS expressing embryos was obtained from genotype E-Mc with mean number of 21.9
blue GUS spots per embryo. Expression of β-glucuronidase reporter gene was observed on cotyledons, hypocotyls, and radicles
of transformed mature zygotic embryos, as well as on organogenic callus and regenerated shoots derived from co-cultivated
mature zygotic embryos. Nineteen regenerated transgenic plants were obtained from GUS expression and kanamycin resistant calli.
The presence and integration of the GUS gene was confirmed by polymerase chain reaction (PCR) and Southern blot analysis.
These results suggested that an efficientAgrobacterium tumefaciens-mediated transformation protocol for stable integration of foreign genes into loblolly pine has been developed and that this transformation
system could be useful for the future studies on transferring economically important genes to loblolly pine. |
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| Keywords: | Pinus taeda L Genetic transformation Agrobacterium tumefaciens β -glucuronidase gene Polymerase chain reaction Southern blot |
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