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United States Department of Agriculture

Agricultural Research Service

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Research Project: ESTABLISH AN EFFICIENT PROTOCOL FOR THE GENETIC TRANSFORMATION OF LEAFY SPURGE (EUPHORBIA ESCULA) VIA AGROBACTERIUM TUMEFACIENS

Location: Sunflower Research

Project Number: 5442-21220-026-04
Project Type: Specific Cooperative Agreement

Start Date: Sep 01, 2011
End Date: Aug 31, 2013

Objective:
Develop an Agrobacterium-mediated transformation system for leafy spurge.

Approach:
Initially, the Plant Transformation Research Center (PTCR) will use the healthy in vitro leafy spurge plants and the methodology supplied by the USDA for the propagation and the transformation experiments. The next steps will be the evaluation of explants regeneration capacity after co-cultivation with Agrobacterium on the different selection systems. Agrobacterium tumefacienens strains AGLO1, EAH105, GV3101 containing the binary plasmids pBI121, pCambia 1301, and pCambia 3300 will be used for the transformation experiments. The transformation vectors are under regulation of the CaMV 35S constitutive promoter, but the selectable marker gene in pBI121 is nptII for Kanamycin resistance, in pCambia 1301 is hpt for hygromycin resistance, and in pCambia 3300 is glufosinate that confers resistance to Basta herbicide. Confirmation of the transgenic tissue will be performed using genomic PCR with specific primers for the amplification of the corresponding selectable marker genes.

Last Modified: 7/28/2014
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