A multi-needle-assisted transformation of soybean cotyledonary node cells

Authors: Xue, Ren-Gao; Xie, Hong-Feng; Zhang, Biao

Source: Biotechnology Letters, Volume 28, Number 19, October 2006 , pp. 1551-1557(7)

Publisher: Springer

Buy & download fulltext article:

Price: $47.00 plus tax (Refund Policy)

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.

Keywords: Agrobacterium tumefaciens; Cotyledonary node; Gene transformation; Multi-needle; Soybean; Wounding

Document Type: Research article

DOI: http://dx.doi.org/10.1007/s10529-006-9123-6

Affiliations: 1: Email: xuerengao@163.com

Publication date: 2006-10-01