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Abstract Detail


Stress Tolerance

Zwenger, Sam R. [1], Hoffmann, Erich [2], Basu, Chhandak [2].

Transformation of Arabidopsis thaliana with heat stress-related genes from a Copaifera officinalis using a novel "binary library" approach.

Plant transformation studies often require gene characterization, cloning and ligation into a binary vector, which can be tedious and sometimes unsuccessful. Novel ways of generating thousands of genes of interest and ligating them into a binary vector might therefore be extremely useful in plant biotechnology. To address this issue, a small cDNA library of C. officinalis was generated using a plant binary vector. After sequencing reactions the 96-well plates were obtained and stored at -80°C. Individual wells from the seqeuncing plates contained E. coli harboring unique vector-insert constructs and were later selected (according to BLASTx results) and grown overnight for subsequent plasmid miniprep. Individual Agrobacterium tumefaciens (EHA105) lines harboring individual contstructs were then generated and used to transform Arabidopsis thaliana using the floral dip method. Many of the sequences identified from the library were heat stress-related, therefore a second component of this research was assessing the degree to which C.officinalis heat-stress associated genes provided thermotolerance intransgenic A. thaliana. Various stress assays, which included both in vitro and in vivo assays, were performed to help determine the degree of thermotolerance. While all A. thaliana lines expressed the transgenes, only some lines were thermotolerant. The findings have important plant biotechnological applications. Additionally, since global climate change is predicted to result in elevated temperatures over the next century, there are possible agricultural implications. Taken together, the results suggest that binary libraries facilitate high-throughput transformation of plants using choice-selected genes of interest and can provide thousands of vector-insert constructs to investigate. Future studies should consider the transcriptional state of an organism prior to binary library construction in order to obtain choice vector-insert constructs.

Broader Impacts:


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1 - University of Northern Colorado, Biological Sciences, Ross Hall, Room 2480, Campus Box 92, Greeley, CO, 80631, USA
2 -

Keywords:
binary library
plant biotechnology
heat stress
Agrobacterium-mediated transformation.

Presentation Type: Poster:Posters for Topics
Session: P
Location: Khorassan Ballroom/Chase Park Plaza
Date: Monday, July 11th, 2011
Time: 5:30 PM
Number: PST003
Abstract ID:691


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