Biosynthesis of Steroidal Glycoakaloids in Solanum chacoense Bitter

dc.contributor.authorMweetwa, Alice Mutitien
dc.contributor.committeechairTokuhisa, James G.en
dc.contributor.committeememberVeilleux, Richard E.en
dc.contributor.committeememberFlinn, Barry S.en
dc.contributor.committeememberJelesko, John G.en
dc.contributor.committeememberShang, Chaoen
dc.contributor.departmentHorticultureen
dc.date.accessioned2014-03-14T20:14:54Zen
dc.date.adate2009-09-02en
dc.date.available2014-03-14T20:14:54Zen
dc.date.issued2009-07-24en
dc.date.rdate2013-05-20en
dc.date.sdate2009-08-08en
dc.description.abstractSteroidal glycoalkaloids (SGAs) are secondary metabolites produced by approximately 350 species in the Solanaceae family. SGAs are reported to be important for pest resistance and flavor enhancement at low concentrations but are toxic to humans and other mammals at high concentrations. Studies on sterol / SGA biosynthesis have implicated squalene synthase as a key regulatory enzyme because it catalyzes an irreversible step from the mevalonic acid pathway. However, the regulatory mechanisms of squalene synthase are not yet known. A study was conducted to elucidate the distribution pattern of SGAs and to clone the squalene synthase gene in order to determine a relationship between SGAs and gene expression levels. Solanum chacoense, a wild potato species was used as a model plant from which tissues were harvested at specified developmental stages and analyzed for SGA content. The results from the SGA analysis suggest a qualitative and quantitative tissue- and age-dependent accumulation of SGAs. Regenerative tissues such as, axiliary shoots, flowers and floral buds had the highest levels of 88, 49 and 63 µmole/g DW, respectively. The roots, stems and tubers showed the lowest amounts of SGAs of 1 to 8, 5 to 15 and 7 to 15 µmole/g DW, respectively. Stolons and tubers accumulated higher amounts of α-chaconine (59 to 67%) than α-solanine (61 to 64%) at all developmental stages analyzed. On the other hand, in young expanding, fully expanded, and old senescing leaves where leptine and leptinines tend to dominate, α-solanine and α-chaconine together accounted for only 8 to 15%, 7 to 15%, and 8 to 45%, respectively. Plant organs that showed the highest biosynthetic activity for SGA production also had high levels of transcripts coding for genes of isoprenoid biosynthesis. The results from the cloning and characterization of squalene synthase suggest that the cloned cDNA fragment is a putative S. chacoense squalene synthase gene with an open reading frame / predicted protein precursor of 411 amino acids. The cloned cDNA has high similarity (68-100%) to known plant squalene synthase genes and contains six deduced peptide domains observed in other species. The 3â untranslated regions of floral buds, young leaves (early vegetative stage), and fully expanded leaves (anthesis) were different in length with, 249, 335, and 202 nucleotides, respectively. The Southern blot analysis suggests a single copy gene although the existence of a gene family cannot be ruled out.en
dc.description.degreePh. D.en
dc.identifier.otheretd-08082009-204852en
dc.identifier.sourceurlhttp://scholar.lib.vt.edu/theses/available/etd-08082009-204852/en
dc.identifier.urihttp://hdl.handle.net/10919/28572en
dc.publisherVirginia Techen
dc.relation.haspartAliceMweetwaFinal.pdfen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjectsqualene synthaseen
dc.subjectsolanineen
dc.subjectchaconineen
dc.subjectwild potato speciesen
dc.subjectsecondary metabolitiesen
dc.subjectsolanidineen
dc.titleBiosynthesis of Steroidal Glycoakaloids in Solanum chacoense Bitteren
dc.typeDissertationen
thesis.degree.disciplineHorticultureen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.leveldoctoralen
thesis.degree.namePh. D.en

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