Efficient Agrobacterium tumefaciens-mediated transformation of soybeans using an embryonic tip regeneration system.
Here, we report the establishment of an efficient, in vitro, shoot organogenesis, regeneration system for soybeans [Glycine max (L.) Merr.]. Mature soybean seeds were soaked for 24 h, the embryonic tips were collected and cultured on MSB5 medium supplemented with 3.5 mg l(-1) N6-benzylaminopurine (BAP) for 24 h, and explants were transferred to MSB5 medium supplemented with 0.2 mg l(-1) BAP and 0.2 mg l(-1) indolebutyric acid. Use of embryonic tips yielded a higher regeneration frequency (87.7%) than regeneration systems using cotyledonary nodes (40.3%) and hypocotyl segments (56.4%) as starting materials. Regenerated embryonic tips were inoculated with Agrobacterium tumefaciens strain EHA105, which contains the binary vector pCAMBIA2301, and cultured for 20 h. Our results showed that the T-DNA transfer efficiency reached up to 78.2% and the transformation efficiency reached up to 15.8%. These data indicate that the embryonic tip regeneration system can be used for efficient, effective Agrobacterium-mediated transformation.
Liu
Hai-Kun
HK
Shanghai Institute of Plant Physiology & Ecology, Shanghai Institute of Biology Science, CAS, 300 Fenglin Road, 200032 Shanghai, China.
Yang
Chao
C
Wei
Zhi-Ming
ZM
D016960:Agrobacterium tumefaciens / Q000235:genetics*; D004269:DNA, Bacterial; D018506:Gene Expression Regulation, Plant; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D005838:Genotype; D030821:Plants, Genetically Modified; D012038:Regeneration; D012639:Seeds / Q000235:genetics / Q000254:growth & development; D013025:Glycine max / Q000235:genetics* / Q000254:growth & development / Q000382:microbiology; D014170:Transformation, Genetic; D019076:Transgenes
D004269:DNA, Bacterial; C036483:T-DNA