Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette

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• 作者：Ming Liu (1)
  Jun Yang (1)
  Yun-qing Cheng (2)
  Li-jia An (1)
  
• 关键词：Soybean ; Glycine max (L.) Merrill ; In planta ovary ; drip transformation ; Optimization ; Style cutting ; Minimal gus reporter gene cassette ; Q78
• 刊名：Journal of Zhejiang University SCIENCE B
• 出版年：2009
• 出版时间：December 2009
• 年：2009
• 卷：10
• 期：12
• 页码：870-876
• 全文大小：508KB
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• 作者单位：Ming Liu (1)
  Jun Yang (1)
  Yun-qing Cheng (2)
  Li-jia An (1)
  
  1. Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian, 116024, China
  2. Institute of Eco-environmental Sciences, Jilin Normal University, Siping, 136000, China
  
• ISSN：1862-1783

文摘

Soybean transformation by ovary-drip was improved by optimizing the length of the transformation pathway by cutting the styles. These modifications facilitated soybean transformation manipulation and improved transformation reproducibility and efficiency. Using a linear minimal gus gene cassette as the foreign DNA, a maximum transformation frequency of 11% was obtained in flowers of the soybean cultivar ‘Liaodou 14-with their styles mostly removed, whereas removal of only the stigma, partial style cutting and partial ovary cutting gave transformation frequencies of 0%, 1%, and 2%, respectively. An average transformation frequency of 8.2% was obtained when 619 flowers from three soybean cultivars (‘Liaodou 14- ‘Liaodou 13- and ‘Tiefeng 29- were transformed by this optimized method. Southern blotting analysis showed that the gus reporter gene (encoding β-glucuronidase) was stably inherited with a simple pattern. Reverse transcription-polymerase chain reaction (RT-PCR) and GUS staining confirmed the expression of the gus gene in transgenic plants.