Development and optimization of agroinfiltration for soybean

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• 作者：Jessica L. King ; John J. Finer ; Leah K. McHale
• 关键词：Agrobacterium tumefaciens ; Sonication ; Agroinfiltration ; Glycine max ; Vacuum infiltration
• 刊名：Plant Cell Reports
• 出版年：2015
• 出版时间：January 2015
• 年：2015
• 卷：34
• 期：1
• 页码：133-140
• 全文大小：2,281 KB
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  19. Finer JJ, Larkin KM (2
• 作者单位：Jessica L. King (1)
  John J. Finer (2)
  Leah K. McHale (1)
  
  1. Department of Horticulture and Crop Science, The Ohio State University, Columbus, OH, 43210, USA
  2. Department of Horticulture and Crop Science, Ohio Agricultural Research and Development Center, The Ohio State University, Wooster, OH, 44691, USA
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Cell Biology
  Plant Sciences
  Biotechnology
  Plant Biochemistry
  
• 出版者：Springer Berlin / Heidelberg
• ISSN：1432-203X

文摘

Key message Agroinfiltration is an efficient method to study transgene expression in plant tissue. In this study, sonication followed by vacuum infiltration is shown to increase agroinfiltration-mediated GUS expression in soybean. Abstract Agroinfiltration, a valuable tool for rapid analysis of gene function, has been used extensively on leaf tissue of Nicotiana benthamiana and several other plant species. However, the application of this approach for gene functionality studies in soybean has been largely unsuccessful. Improvements in agroinfiltration of many plants have been achieved through a variety of approaches to allow better delivery, penetration and infection of Agrobacterium to interior leaf tissues. In this work, an agroinfiltration approach was developed for transient expression in soybean utilizing sonication followed by vacuum infiltration of intact seedlings. The optimal infiltration buffer, sonication time, and vacuum conditions for agroinfiltration of soybean were evaluated by monitoring expression of an introduced β-glucuronidase (GUS) reporter gene. The developed method included the use of an infiltration buffer (10?mM 2-(N-morpholino)ethanesulfonic acid sodium salt, 10?mM MgCl2, 100?μM acetosyringone) supplemented with the reducing agent dithiothreitol, with 30?s sonication followed by vacuum infiltration. These techniques were further applied to evaluate five different Agrobacterium strains and six different plant genetic backgrounds. Among the Agrobacterium strains examined, J2 produced the highest levels of GUS activity and ‘Peking-was the most responsive genotype.