Production of selectable marker-free transgenic tobacco plants using a non-selection approach: chimerism or escape, transgene inheritance, and efficiency View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2009-03

AUTHORS

Baochun Li, Claire Xie, Hui Qiu

ABSTRACT

Public concern and metabolic drain were the main driving forces for the development of a selectable marker-free transformation system. We demonstrated here the production of transgenic tobacco plants using a non-selection approach by Agrobacterium tumefaciens-mediated transformation. A. tumefaciens-infected leaf explants were allowed to produce shoots on a shoot induction medium (SIM) containing no selective compounds. Up to 35.1% of the A. tumefaciens-infected leaf explants produced histochemically GUS(+) shoots, 3.1% of regenerated shoots were GUS(+), and 72% of the GUS(+) shoots were stably transformed by producing GUS(+) T1 seedlings. When polymerase chain reaction (PCR) was used to screen the regenerated shoots, 4% of the shoots were found to be PCR(+) for the transgene and 65% of the PCR(+) shoots were stable transformants. Also, generation of PCR(+) escapes decreased linearly as the number of subculture increased from one to three on SIM containing the antibiotic that kills the Agrobacterium. Twenty-five to 75% of the transformants were able to transmit transgene activity to the T1 generation in a Mendelian 3:1 ratio, and a transformation efficiency of 2.2-2.8% was achieved for the most effective binary vector. These results indicated that majority of the GUS(+) or PCR(+) shoots recovered under no selection were stable transformants, and only one-third of them were chimeric or escapes. Transgenes in these transgenic plants were able to transmit the transgene into progeny in a similar fashion as those recovered under selection. More... »

PAGES

373-386

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/s00299-008-0640-8

DOI

http://dx.doi.org/10.1007/s00299-008-0640-8

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1017682965

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/19018535


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48 schema:description Public concern and metabolic drain were the main driving forces for the development of a selectable marker-free transformation system. We demonstrated here the production of transgenic tobacco plants using a non-selection approach by Agrobacterium tumefaciens-mediated transformation. A. tumefaciens-infected leaf explants were allowed to produce shoots on a shoot induction medium (SIM) containing no selective compounds. Up to 35.1% of the A. tumefaciens-infected leaf explants produced histochemically GUS(+) shoots, 3.1% of regenerated shoots were GUS(+), and 72% of the GUS(+) shoots were stably transformed by producing GUS(+) T1 seedlings. When polymerase chain reaction (PCR) was used to screen the regenerated shoots, 4% of the shoots were found to be PCR(+) for the transgene and 65% of the PCR(+) shoots were stable transformants. Also, generation of PCR(+) escapes decreased linearly as the number of subculture increased from one to three on SIM containing the antibiotic that kills the Agrobacterium. Twenty-five to 75% of the transformants were able to transmit transgene activity to the T1 generation in a Mendelian 3:1 ratio, and a transformation efficiency of 2.2-2.8% was achieved for the most effective binary vector. These results indicated that majority of the GUS(+) or PCR(+) shoots recovered under no selection were stable transformants, and only one-third of them were chimeric or escapes. Transgenes in these transgenic plants were able to transmit the transgene into progeny in a similar fashion as those recovered under selection.
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