Abstract
A new and simple method for wounding cotyledonary node cells of soybean [Glycine max (L) Merrill] was developed for obtaining a high frequency of transformants. Soybean seeds were germinated for 1 day, and the cotyledonary node cells of half-seeds were wounded mechanically by using a multi-needle consisting of thin 30 fibers. The wounded half-seeds were inoculated with Agrobacterium tumefaciens cells harboring a recombinant DNA that contained the bar and sgfp genes conferring phosphinothricin (PPT)-resistance and green fluorescent protein (GFP) activity, respectively. The inoculated explants were selected on medium containing 5 or 3 mg PPT/l. The transformation efficiency of soybean was up to 12%. Polymerase chain reaction and genomic Southern blot analysis confirmed stable integration of the transgenes in the genome of the PPT-resistant plants. GFP analysis revealed that the transgenes were highly expressed in the plantlets. Adult plants were resistant to 100 mg PPT/l applied on the leaves, demonstrating their herbicide-resistance.
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This work was supported by grants from Shandong Provincial Education Department and from Laiyang Agricultural College, China.
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An erratum to this article can be found at http://dx.doi.org/10.1007/s10529-006-9229-x
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Xue, RG., Xie, HF. & Zhang, B. A multi-needle-assisted transformation of soybean cotyledonary node cells. Biotechnol Lett 28, 1551–1557 (2006). https://doi.org/10.1007/s10529-006-9123-6
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DOI: https://doi.org/10.1007/s10529-006-9123-6