Agrobacterium tumefaciens mediated transformation of orchid tissue with the sense and antisense coat protein genes from the odontoglossum ringspot virus

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dc.contributor.advisor Vann, Carolyn N. en_US
dc.contributor.author Hutchinson, Chad M. en_US
dc.date.accessioned 2011-06-03T19:35:53Z
dc.date.available 2011-06-03T19:35:53Z
dc.date.created 1992 en_US
dc.date.issued 1992
dc.identifier LD2489.Z78 1992 .H88 en_US
dc.identifier.uri http://cardinalscholar.bsu.edu/handle/handle/184299
dc.description.abstract This research was an attempt to use a dicot transformation vector to transform a monocot. The initial purpose of this thesis was to transform orchids with the sense and antisense coat protein genes from the Odontoglossum ringspot virus (ORSV) in an effort to mitigate viral symptoms in transgenic plants using the transformation vector, Agrobacterium tumefaciens. However, it soon became apparent that much time would be needed to develop a transformation protocol. The transformation vectors used included the Agrobacterium tumefaciens disarmed strain LBA4404 with the binary plasmid pB1121, the disarmed strain At699 with the binary plasmid pCNL65, and the wild-type strain Chry5. The marker gene on the binary plasmids of both disarmed strains was p-glucuronidase (GUS).Several transformation protocols were used in an effort to determine if this transformation system would work on orchids. Transformation was not achieved even though a number of experimental conditions were varied. These included using two different types of orchid tissue, callus and protocorms; using two different species of orchids, Cattleya Chocolate Drop x Cattleytonia Kieth Roth and Cymbidium maudidum; varying the time the plant tissue was exposed to the bacteria from 1 hour to 96 hours; performing experiments with and without the wound signal molecule acetosyringone; and exposing the tissue to the virulent strains of A. tumefaciens mentioned previously.This research also developed GUS assay conditions necessary to decrease the number of false positives due to bacterial contamination. These conditions included chloramphenicol in the GUS assay buffer.
dc.description.sponsorship Department of Biology
dc.format.extent ix, 95 leaves : ill. ; 28 cm. en_US
dc.source Virtual Press en_US
dc.subject.lcsh Orchids -- Diseases and pests -- Control. en_US
dc.subject.lcsh Tobacco mosaic virus -- Control. en_US
dc.subject.lcsh Orchids -- Disease and pest resistance -- Genetic aspects. en_US
dc.subject.lcsh Bacterial transformation. en_US
dc.subject.lcsh Genetic transformation. en_US
dc.subject.lcsh Orchids -- Genetic engineering. en_US
dc.title Agrobacterium tumefaciens mediated transformation of orchid tissue with the sense and antisense coat protein genes from the odontoglossum ringspot virus en_US
dc.description.degree Thesis (M.S.)
dc.identifier.cardcat-url http://liblink.bsu.edu/catkey/834608 en_US


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  • Master's Theses [5454]
    Master's theses submitted to the Graduate School by Ball State University master's degree candidates in partial fulfillment of degree requirements.

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