Efficient soybean regeneration and Agrobacterium‐mediated transformation using a whole cotyledonary node as an explant. (26th June 2014)

• Record Type: Journal Article
• Authors: Zhang, Fuli; Chen, Can; Ge, Honglian; Liu, Jinmei; Luo, Yunling; Liu, Kun; Chen, Long; Xu, Kedong; Zhang, Yi; Tan, Guangxuan; Li, Chengwei
• Is Part Of: Biotechnology and applied biochemistry. Volume 61:Number 5(2014)
• Page Start: 620
• Page End: 625
• Publication Date: 2014-06-26
• Subjects: Biotechnology -- Periodicals; Biochemical engineering -- Periodicals; Biochemistry -- Periodicals; Biochemistry -- Periodicals; Genetic Techniques -- Periodicals; Microbiological Techniques -- Periodicals; 660.6
• DOI: 10.1002/bab.1207
• Languages: English
• ISSNs: 0885-4513

Abstract

<abstract abstract-type="main"> <title>Abstract</title> <p>An optimized regeneration and <italic>Agrobacterium</italic>‐mediated transformation protocol based on whole cotyledonary node explants was developed in soybean (<italic>Glycine max</italic>) cultivar Zhong Huang 13. Adding 6‐benzylaminopurine (BAP) in a germinating medium could significantly increase regeneration efficiency; the optimal BAP concentration for shoot formation was 0.5 mg/L. The concentrations of plant growth regulators in a shoot induction medium were optimized by the orthogonal test [L₉(3³)]. The best combination for shoot regeneration was a medium of Murashige &amp; Skoog salts with B5 vitamins (MSB) supplemented with 3.5 mg/L BAP, 0.2 mg/L indole‐3‐butyric acid (IBA), and 0.2 mg/L kinetin (KT). Under this favorable condition, one node could regenerate 28–30 shoots. Soybean whole cotyledonary nodes were transformed by inoculation with <italic>A. tumefaciens</italic> strain EHA105 harboring a vector pBI121 containing a β‐glucuronidase gene (<italic>gus</italic>). GUS assay, polymerase chain reaction, and Southern blot analysis indicated that the <italic>gus</italic> gene was transformed into soybean plants with 23.1% transformation efficiency. Transgenic plants could be obtained within 5–6 weeks, which was about 4 weeks less than that of a traditional single cotyledonary node method.</p> </abstract>