Poster: Signaling, cell-to-cell
Abs #
438: Isolation and Characterization of Mutants Defective in Inositol Polyphosphate 5-Phosphatase (5PTase) Genes in Arabidopsis thaliana
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Presenter: |
Robinson, Jamille M., scoop804@vt.edu |
Authors | Robinson, Jamille M. (A) Ercetin, Mustafa (A) Gunesekera, Bhadra (A) Stab, Ben (A) Gillaspy, Glenda (A) | | Affiliations: |
(A): Virginia Tech
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Inositol Polyphosphate 5-Phosphatases (5PTases) are enzymes that remove a 5-phosphate from second messengers including inositol (1,4,5) trisphosphate and phosphatidylinositol (4,5) bisphosphate. The overall goal of this project is to identify and isolate several 5PTase knockout mutants in Arabidopsis thaliana and to observe the growth and signaling phenotypes of these mutants. To identify potential knock-out mutants, we queried the T-DNA mutagenized populations from SALK T-DNA Express and TMRI collections. Transgenic seed containing T-DNA insertions within exons of the At5PTase2, 3, 6, 8, 9, 10, 11 and 14 genes were obtained and grown to maturity in soil under short day conditions. When plants were 3 weeks old, leaf tissue was removed, genomic DNA was isolated and PCR reactions were performed to ascertain the presence of the wildtype locus and the T-DNA insertion. Homozygous mutants in At5PTase2, 10 and 11 were identified and grown to maturity. These plants contained no obvious growth alterations or defects. Sequencing of T-DNA insertion PCR products is underway to verify the genomic location of each insertion. RNA is being examined to ascertain whether the T-DNA insertion results in lower or absent mRNA expression. Our results suggest that expression is suppressed, although in one case the T-DNA insertion allowed for initiation of a shorter transcript. To examine whether the At5PTase 2, 10 and 11 mutants are altered in signal transduction pathways, we are focusing on stimulating knock-out plants with light and abscisic acid. These studies will provide functional, in planta data to complement ongoing biochemical approaches aimed at understanding 5PTase function.
