The use of the two T-DNA binary system to derive marker-free transgenic soybeans View Full Text


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Article Info

DATE

2000-11

AUTHORS

Aiqiu Xing, Zhangyuan Zhang, Shirley Sato, Paul Staswick, Tom Clemente

ABSTRACT

A binary vector, pPTN133, was assembled that harbored two separate T-DNAs. T-DNA one contained a bar cassette, while T-DNA two carried a GUS cassette. The plasmid was mobilized into the Agrobacterium tumefaciens strain EHA101. Mature soybean cotyledonary node explants were inoculated and regenerated on medium amended with glufosinate. Transgenic soybeans were grown to maturity in the greenhouse. Fifteen primary transformants (T0) representing 10 independent events were characterized. Seven of the 10 independent T0 events co-expressed GUS. Progeny analysis was conducted by sowing the T1 seeds and monitoring the expression of the GUS gene after 21 d. Individual T1 plants were subsequently scored for herbicide tolerance by leaf painting a unifoliate leaf with a 100 mgl−1 solution of glufosinate and scoring the leaf 5 d post application. Herbicide-sensitive and GUS-positive individuals were observed in four of the 10 independent events. Southern blot analysis confirmed the absence of the bar gene in the GUS positive/herbicide-sensitive individuals. These results demonstrate that simultaneous integration of two T-DNAs followed by their independent segregation in progeny is a viable means to obtain soybeans that lack a selectable marker. More... »

PAGES

456-463

References to SciGraph publications

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  • 1993-11. Transposition Mediated Re–positioning and Subsequent Elimination of Marker Genes from Transgenic Tomato in NATURE BIOTECHNOLOGY
  • 1994-03. Transformation Systems for Generating Marker–Free Transgenic Plants in NATURE BIOTECHNOLOGY
  • 1999-11. Transformation of soybean [Glycine max (L.) Merrill] using proliferative embryogenic tissue maintained on semi-solid medium in IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY - PLANT
  • 1985-12. Frequencies of simultaneous transformation with different T-DNAs and their relevance to the Agrobacterium/plant cell interaction in MOLECULAR GENETICS AND GENOMICS
  • 1996-06. Production of transgenic soybean lines expressing the bean pod mottle virus coat protein precursor gene in PLANT CELL REPORTS
  • 1999-01. The use of glufosinate as a selective agent in Agrobacterium-mediated transformation of soybean in PLANT CELL, TISSUE AND ORGAN CULTURE (PCTOC)
  • 1999-05. Selectable marker-free transgenic plants without sexual crossing: transient expression of cre recombinase and use of a conditional lethal dominant gene in PLANT MOLECULAR BIOLOGY
  • 1999-01. Marker Gene Controversy in Transgenic Plants in JOURNAL OF PLANT BIOCHEMISTRY AND BIOTECHNOLOGY
  • 1987-11. Segregation of genes transferred to one plant cell from two separate Agrobacterium strains in PLANT MOLECULAR BIOLOGY
  • 1991-09. Two T-DNA's co-transformed intoBrassica napus by a doubleAgrobacterium tumefaciens infection are mainly integrated at the same locus in THEORETICAL AND APPLIED GENETICS
  • 1988-08. Stable Transformation of Soybean (Glycine Max) by Particle Acceleration in NATURE BIOTECHNOLOGY
  • 1998-04. Co-transformation with one Agrobacterium tumefaciens strain containing two binary plasmids as a method for producing marker-free transgenic plants in PLANT CELL REPORTS
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