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Efficient Agrobacterium tumefaciens-mediated transformation of soybeans using an embryonic tip regeneration system

Hai-Kun Liu, Chao Yang and Zhi-Ming Wei
Planta
Vol. 219, No. 6 (October 2004), pp. 1042-1049
Published by: Springer
Stable URL: http://www.jstor.org/stable/23388620
Page Count: 8
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Efficient Agrobacterium tumefaciens-mediated transformation of soybeans using an embryonic tip regeneration system
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Abstract

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.

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