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1. Wu, Di. Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine.

Degree: PhD, Chemistry and Biochemistry, 2008, Georgia Tech

myo-Inositol-1,2,3,4,5,6-hexakisphosphate (InsP6), also called phytic acid, is ubiquitous in eukaryotic cells and the most abundant inositol phosphate derivative. Loblolly pine (LP, Pinus taeda) constitutes the primary commercial species in the southern forest of U.S. Somatic embryogenesis (SE) is an effective technique to maintain the desirable genetic composition of the progeny and to accomplish the efficiency of propagation. SE can also serve as a tool for study of plant development. Unlike angiosperm embryos with attached cotyledons as seed storage organs, the diploid conifer embryo is surrounded by the unattached haploid female gametophyte (FG). In LP SE, FG tissue is absent in the embryogenic tissue culture. We found that extracts from early-stage FG stimulate growth and multiplication of early-stage somatic embryos, whereas FG water extracts from late stage contain substance(s) inhibitory to early-stage somatic embryo growth (DeSilva et al., 2007). We now present the isolation and identification of the inhibitory substance as InsP6 by means of water extraction, two gel filtrations and two ion exchange FPLC chromatographies. The results represent the first complete structural characterization of InsP6 from a natural product using LC/MS, LC/MS/MS, exact MS, 1D- and 2D-NMR analyses. We also report that there is a good correlation between the amount of InsP6 purified from FG tissue (1.3 nmoles per full-term FG) and the amount of InsP6 which inhibits somatic embryo growth. This novel approach of isolating and characterizing InsP6 from plant tissue, and investigating its role on SE can allow us to improve SE technology by circumventing current bottleneck, to elucidate enigmatic functions of InsP6 in plants, and most importantly, to utilize this molecule properly. Advisors/Committee Members: Dr. Sheldon May (Committee Chair), Dr. Donald Doyle (Committee Member), Dr. Gerald Pullman (Committee Member), Dr. James Powers (Committee Member), Dr. Nicholas Hud (Committee Member).

Subjects/Keywords: NMR; LC/MS; FPLC; Female gametophyte; Loblolly pine; Somatic embryogenesis; Myo-inositol hexakisphosphate; Loblolly pine; Somatic embryogenesis; Phytic acid

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APA (6th Edition):

Wu, D. (2008). Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine. (Doctoral Dissertation). Georgia Tech. Retrieved from http://hdl.handle.net/1853/28252

Chicago Manual of Style (16th Edition):

Wu, Di. “Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine.” 2008. Doctoral Dissertation, Georgia Tech. Accessed January 18, 2021. http://hdl.handle.net/1853/28252.

MLA Handbook (7th Edition):

Wu, Di. “Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine.” 2008. Web. 18 Jan 2021.

Vancouver:

Wu D. Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine. [Internet] [Doctoral dissertation]. Georgia Tech; 2008. [cited 2021 Jan 18]. Available from: http://hdl.handle.net/1853/28252.

Council of Science Editors:

Wu D. Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine. [Doctoral Dissertation]. Georgia Tech; 2008. Available from: http://hdl.handle.net/1853/28252

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