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Widholm, Jack M. Luo, Guangbin 2011-05-07T14:10:50Z 2011-05-07T14:10:50Z 10000-01-01 1993 Factors which affect the transformation efficiency of Agrobacterium tumefaciens-mediated soybean transformation were studied. It was found that pre-induction of A. tumefaciens A281 using acetosyringone significantly increased the efficiency of transformation and this could be boosted further by octopine. The application of these pre-induction conditions led to the improvement in recovery of putative transformed soybean shoots although no fertile plants were recovered. The co-transformation of T-strands from a wild type Ti plasmid pTiBo542 and T-strands of a binary vector pZA-7 in a virulent A. tumefaciens strain A281 containing a binary vector pZA-7 was studied. When transformation with the wild type T-strands was selected for, 82% of the transformants also contained the T-strands from the binary vector. The frequency of co-expression of genes contained on these two different T-strands was found to be 50%. The expression of two genes in the T-region of pZA-7 was random, although both genes were driven by a 35S promotor and terminated by a polyadenylation signal of the nopaline synthase gene and closely linked in the same T-DNA. Differential DNA methylation might be the cause of this inactivation of one of the two closely linked genes. The high frequency of co-transformation led to the development of a simple system for the expression of genes of interest in soybean tissues. A simple, rapid and reliable system, the miniprep-PCR system, was developed for PCR analysis using very small amounts of plant tissue (10-50mg). The reliability of this system is ensured by the inclusion of an internal control into the system. Made available in DSpace on 2011-05-07T14:10:50Z (GMT). No. of bitstreams: 2 license.txt: 4922 bytes, checksum: 910b249b4beec47e7ab768910c8f966f (MD5) 9314908.pdf: 4950426 bytes, checksum: 9f66fc9ec8e129cd08e28b418beff838 (MD5) Previous issue date: 1993 Item marked as restricted to the 'UIUC Users [automated]' Group (id=2) by Howard Ding (hding2@illinois.edu) on 2011-05-07T15:03:50Z Item is restricted indefinitely. Restriction data tranferred 2014-07-01T11:30:27-05:00 Original Data Group with Access UIUC Users [automated] Release Date: none Reason: ETDs are only available to UIUC Users without author permission Restricted to the U of I community indefinitely during batch ingest of legacy ETDs U of I Only AAI9314908 (UMI)AAI9314908 http://hdl.handle.net/2142/23344 eng Copyright 1993 Luo, Guangbin Biology, Genetics Soybean transformation and analysis of transformed tissues using PCR text Agronomy Agronomy University of Illinois at Urbana-Champaign Dissertation Ph.D.