A novel Agrobacterium rhizogenes-mediated transformation method of soybean [Glycine max (L.) Merrill] using primary-node explants from seedlings

作者:Olhoft Paula M; Bernal Libby M; Grist Leslie B; Hill D Steven; Mankin S Luke; Shen Yuwei; Kalogerakis Mary; Wiley Hunt; Toren Effie; Song Hee Sook; Hillebrand Helke; Jones Todd*
来源:In Vitro Cellular and Developmental Biology - Plant, 2007, 43(6): 536-549.
DOI:10.1007/s11627-007-9050-9

摘要

A novel Agrobacterium rhizogenes-mediated transformation method using a primary-node explant from Dairyland cultivar 93061 was developed for soybean using the disarmed Agrobacterium strain SHA17. Transformed plants regenerated from explants inoculated with SHA17 were fertile and phenotypically normal. In a comparative experiment, regeneration frequencies were not significantly different between explants inoculated with A. rhizogenes strain SHA17 and Agrobacterium tumefaciens strain AGL1; however, a 3.5-fold increase in transformation efficiency [(number of Southern or TaqMan-positive independent events/total number of explants inoculated)x100] was found for explants cocultured with SHA17 compared to AGL1 (6.6 and 1.64%, respectively). Southern analysis of 48 T-0 plants suggested that 37.5, 23, and 39.6% of the T-0 plants contained 1, 2, and 3 or more T-DNA fragments integrated into the genome, respectively. Additionally, T-1 progeny analysis of 8 independent events resulted in typical Mendelian inheritance of T-DNA genes. Of seven T-0 plants that had two or more T-DNA fragments, six contained multiple loci segregating in T-1 progenies. Further analysis of four lines confirmed the presence of PAT, GUS, and/or DsRED2 proteins in transgenic plants that were encoded on the T-DNA into the T-2 generation.

  • 出版日期2007-12