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Description
Title: | Soybean transformation and analysis of transformed tissues using PCR |
Author(s): | Luo, Guangbin |
Doctoral Committee Chair(s): | Widholm, Jack M. |
Department / Program: | Agronomy |
Discipline: | Agronomy |
Degree Granting Institution: | University of Illinois at Urbana-Champaign |
Degree: | Ph.D. |
Genre: | Dissertation |
Subject(s): | Biology, Genetics |
Abstract: | 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. |
Issue Date: | 1993 |
Type: | Text |
Language: | English |
URI: | http://hdl.handle.net/2142/23344 |
Rights Information: | Copyright 1993 Luo, Guangbin |
Date Available in IDEALS: | 2011-05-07 |
Identifier in Online Catalog: | AAI9314908 |
OCLC Identifier: | (UMI)AAI9314908 |
This item appears in the following Collection(s)
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Dissertations and Theses - Agronomy
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Graduate Dissertations and Theses at Illinois
Graduate Theses and Dissertations at Illinois