Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine
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...
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ndltd-GATECH-oai-smartech.gatech.edu-1853-282522013-01-07T20:31:39ZDiscovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pineWu, DiNMRLC/MSFPLCFemale gametophyteLoblolly pineSomatic embryogenesisMyo-inositol hexakisphosphateLoblolly pineSomatic embryogenesisPhytic acidmyo-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.Georgia Institute of Technology2009-06-08T19:33:16Z2009-06-08T19:33:16Z2008-03-04Dissertationhttp://hdl.handle.net/1853/28252 |
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NMR LC/MS FPLC Female gametophyte Loblolly pine Somatic embryogenesis Myo-inositol hexakisphosphate Loblolly pine Somatic embryogenesis Phytic acid |
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NMR LC/MS FPLC Female gametophyte Loblolly pine Somatic embryogenesis Myo-inositol hexakisphosphate Loblolly pine Somatic embryogenesis Phytic acid Wu, Di Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine |
description |
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. |
author |
Wu, Di |
author_facet |
Wu, Di |
author_sort |
Wu, Di |
title |
Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine |
title_short |
Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine |
title_full |
Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine |
title_fullStr |
Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine |
title_full_unstemmed |
Discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine |
title_sort |
discovery and characterization of a signaling molecule regulating somatic embryogenesis in loblolly pine |
publisher |
Georgia Institute of Technology |
publishDate |
2009 |
url |
http://hdl.handle.net/1853/28252 |
work_keys_str_mv |
AT wudi discoveryandcharacterizationofasignalingmoleculeregulatingsomaticembryogenesisinloblollypine |
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1716475103306317824 |