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Sonication Effects and Transient Gene Expression Following Agrobacterium Transformation

B.-M. Lee, C. Zapata, S. H. Park, K. M. El-Zik and R. H. Smith


The efficiency of Agrobacterium-mediated transformation of cotton is low. In an attempt to enhance infection and transformation of the cotton seedling shoot apex using Agrobacterium tumefaciens LBA4404 as the vector, sonication treatment to enhance wounding and transformation efficiency was investigated. The pat gene which encodes resistance to the herbicide, glufosinate, was used as a selectable marker. Survival of the shoot apices on 2.5 mg/l of glufosinate ammonium, ppt, a level which kills all apices not expressing resistance, was used to indicate efficacy of treatment. Sonication treatment can enhance wounding of the tissue by creating fissures or channels throughout the targeted tissue. Initial sonication treatments of 0, 5, 10, 15, 30, and 60 seconds at 60 watts, 47 KHz, show an increase in shoot apex survival between 5 to 15 seconds sonication treatment over the control (o second) shoot apices.

Reprinted from Proceedings of the 1998 Beltwide Cotton Conferences pp. 488 - 489
©National Cotton Council, Memphis TN

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Document last modified Sunday, Dec 6 1998