Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation.
The utility of transformation for soybean improvement requires an efficient system for production of stable transgenic lines. We describe here an improved cotyledonary node method using an alternative explant for Agrobacterium tumefaciens-mediated soybean transformation. We use the term "half-seed" to refer to this alternative cotyledonary explant that is derived from mature seed of soybean following an overnight imbibition and to distinguish it from cotyledonary node derived from 5-7-day-old seedlings. Transformation efficiencies using half-seed explants ranged between 1.4 and 8.7% with an overall efficiency of 3.8% based on the number of transformed events that have been confirmed in the T1 generation by phenotypic assay using the herbicide Liberty (active ingredient glufosinate) and by Southern analysis. This efficiency is 1.5-fold higher than the cotyledonary node method used in our laboratory. Significantly, the half-seed system is simple and does not require deliberate wounding of explants, which is a critical and technically demanding step in the cotyledonary node method.
Paz
Margie M
MM
Department of Agronomy, Iowa State University, Ames, IA 50011-1010, USA.
Martinez
Juan Carlos
JC
Kalvig
Andrea B
AB
Fonger
Tina M
TM
Wang
Kan
K
D001593:Benzyl Compounds / Q000494:pharmacology; D018548:Cotyledon / Q000196:embryology* / Q000235:genetics; D005822:Genetic Vectors; D018520:Plant Shoots / Q000502:physiology; D011687:Purines / Q000494:pharmacology; D012038:Regeneration / Q000187:drug effects; D012231:Rhizobium / Q000235:genetics*; D012639:Seeds / Q000196:embryology* / Q000235:genetics; D013025:Glycine max / Q000196:embryology* / Q000235:genetics*; D046509:Tissue Culture Techniques / Q000379:methods*; D014170:Transformation, Genetic
D001593:Benzyl Compounds; D011687:Purines; C480551:benzylaminopurine