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Isolation of Novel Agrobacterium and Transient Expression Assays in Soybean (Glycine max) and Sunflower (Helianthus annuus)

Benzle, Kyle Arthur
http://rave.ohiolink.edu/etdc/view?acc_num=osu1405555898

Abstract Details

2014, Master of Science, Ohio State University, Horticulture and Crop Science.
Many laboratories routinely use Agrobacterium for the generation of transgenic plants; however, only a few disarmed bacterial strains are widely available and none of these strains were isolated and selected based on evaluation of transformation efficiency in target species. For this research, new Arobacterium strains were isolated from crown galls of various plants in Ohio and from rhizospheric soil throughout the Midwest US. These wild-type strains were isolated by plating gall and soil extracts on a semi-selective medium and screening the isolates for the presence of virG using PCR. The efficiency of plant transformation was evaluated by transforming hypocotyl and cotyledonary tissues of sunflower and soybean seedlings and in proliferative embryogenic tissue of soybean and then quantifying GFP expression. In sunflower, seedling cotyledonary tissue was not responsive to any of the strains tested, however hypocotyl tissues were very responsive with the highest transformation rates obtained with EHA105. With sunflower hypocotyl tissues transformed with disarmed strain EHA105, greater than 75% of the transformed cells were located in the vascular tissues. In soybean seedling tissues, tissue-specific transformation was not observed with any strain as transformed cells were evenly distributed throughout target tissue. With soybean hypocotyl, cotyledon, and embryogenic tissues, a single strain from a soybean field in North Dakota gave 5-10x higher transformation rates than EHA105, while a strain from Ohio soil gave 3-5x higher rates than EHA105.
John Finer (Advisor)
Leah McHale (Committee Member)
Finer Kim (Committee Member)
65 p.
Plant Sciences
Agrobacterium

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Citations

  • Benzle, K. A. (2014). Isolation of Novel Agrobacterium and Transient Expression Assays in Soybean (Glycine max) and Sunflower (Helianthus annuus) [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1405555898

    APA Style (7th edition)

  • Benzle, Kyle. Isolation of Novel Agrobacterium and Transient Expression Assays in Soybean (Glycine max) and Sunflower (Helianthus annuus). 2014. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1405555898.

    MLA Style (8th edition)

  • Benzle, Kyle. "Isolation of Novel Agrobacterium and Transient Expression Assays in Soybean (Glycine max) and Sunflower (Helianthus annuus)." Master's thesis, Ohio State University, 2014. http://rave.ohiolink.edu/etdc/view?acc_num=osu1405555898

    Chicago Manual of Style (17th edition)

osu1405555898
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© 2014, all rights reserved.
This open access ETD is published by The Ohio State University and OhioLINK.