Ames, Iowa
Iowa State University
Establishment of robust maize transformation systems for the public sector --Wang, K, Frame, B, Gelvin, S1, Wang, S-J1, Kaeppler, H2, Akula, C2, Thompson, W3, Nguyen, T3, Allen, G3

1Department of Biological Sciences, Purdue University, West Lafayette, IN 47907-1392
2Department of Agronomy, University of Wisconsin, Madison, WI 53706
3Department of Botany, North Carolina State University, Raleigh, NC 27607

Recent advancements in molecular biology have enabled the large scale, rapid identification and isolation of plant genes. The determination of the function for thousands of genes, and the application of that knowledge to crop improvement, is now one of the major challenges facing plant biologists. Plant transformation is a key technology for functional analysis of genes via complementation, over-expression, or gene silencing strategies and will be equally essential in the application of genomic technology to crop improvement.

For most public research groups, maize transformation technology remains non-straightforward because they do not have access to the physical or human resources necessary to establish an efficient transformation effort. In addition, serious limitations exist with the technology, including low efficiency and throughput, difficulty in inbred line transformation, unpredictable transgene copy numbers, gene rearrangements and undesirable levels of transgene silencing through generations.

The long-term goal of this research is to establish a robust maize transformation system to enable the maize research community for future functional genomic research as well as crop improvement. Our specific objectives are:

1) Developing a routine Agrobacterium-mediated transformation system. We will conduct experiments to systematically optimize transformation and regeneration parameters for Agrobacterium-mediated maize transformation using non-super binary vector systems.

2) Enhancing transgene integration and expression. We will transfer and express the Arabidopsis and maize rat (resistant to Agrobacterium transformation) genes/proteins in maize to evaluate their effects in enhancing transgene integration. We will also evaluate the effect of tobacco and maize MARs (Matrix Attachment Regions) on the efficiency of our transformation protocols and on the frequency of somatic silencing events in maize.

3) Investigating germline transformation protocols. We will investigate tissue culture-independent transformation protocols such as meristem transformation and female gametophyte transformation.

4) Exploring inbred line transformation. We will conduct research to improve inbred line transformation on B73, H99, Oh43 and W22.

In addition, we will facilitate transfer of improved protocols to the public sector by providing a more efficient transformation service at the Plant Transformation Facility at Iowa State University and organizing transformation workshops during the course of this program. It is also our intention that any vector systems, reliable transformation protocols and information generated from this research program will be made available for the maize community upon request.
 
 


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