| Abstract Detail
Recent Topics Posters Ricigliano, Vincent [1], Howarth, Dianella [2]. Development of an Agrobacterium-mediated transformation for Valerianella locusta. An Agrobacterium-mediated transformation system is being developed for Valerianella locusta. The current work focuses on establishing a reproducible selection and regeneration system that enables the recovery of transformants. This approach can be applied to non-model species that lack transgenic systems. A high efficiency in-vitro propagation system was developed and used as the starting point for optimizing agrobacterium-mediated transformation. Agrobacterium tumefaciens strain EHA105 harboring binary vector pBI121 was used for all transformation experiments. Early transformation events were monitored using beta-glucuronidase (GUS) as a reporter gene. Neomycin phosphotransferase (nptII) was employed as a selectable marker. The effects of explant type, age,orientation, pre-culture and co-cultivation on transformation efficiency were investigated. Sensitivity to kanamycin was established by kanamycin sensitivity assay. 14-day-old cotyledons displayed the highest regeneration efficiency and were used as explants for transformation experiments. Transient beta-glucuronidase activity was detected by GUS assay in 100% of explants that were co-cultivated with agrobacterium for 72 hours on callus induction media. However,developing an agrobacterium-mediated transformation system presents many challenges. Emphasis must be placed on enhancing regeneration parameters for transformed cells while minimizing escapes. Optimizing transformation conditions requires the investigation of many variables that contribute to the final transformation efficiency, or percent of explants that eventually give rise to intact transgenic individuals. It is possible to take a systematic approach to the development of a stable transformation system that provides transgenic lines for the study of non-model species. Broader Impacts:
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1 - St Johns University, 8000 Utopia Pkwy, Queens, NY, 11439, USA
2 - St. John\'s University, Department Of Ecology & Evolutionary Biology, St. Albert Hall Rm 257, 8000 Utopia Pkwy, Jamiaca, NY, 11439, USA
Keywords:
Agrobacterium
Transformation.
Presentation Type: Recent Topics Poster
Session: P
Location: Battelle South/Convention Center
Date: Monday, July 9th, 2012
Time: 5:30 PM
Number: PRT012
Abstract ID:1305 |
