title: Effects of down-regulating ornithine decarboxylase upon putrescine-associated metabolism and growth in Nicotiana tabacum L.
abstract: Transgenic plants of Nicotiana tabacum L. homozygous for an RNAi construct designed to silence ornithine decarboxylase (ODC) had significantly lower concentrations of nicotine and nornicotine, but significantly higher concentrations of anatabine, compared with vector-only controls. Silencing of ODC also led to significantly reduced concentrations of polyamines (putrescine, spermidine and spermine), tyramine and phenolamides (caffeoylputrescine and dicaffeoylspermidine) with concomitant increases in concentrations of amino acids ornithine, arginine, aspartate, glutamate and glutamine. Root transcript levels of S-adenosyl methionine decarboxylase, S-adenosyl methionine synthase and spermidine synthase (polyamine synthesis enzymes) were reduced compared with vector controls, whilst transcript levels of arginine decarboxylase (putrescine synthesis), putrescine methyltransferase (nicotine production) and multi-drug and toxic compound extrusion (alkaloid transport) proteins were elevated. In contrast, expression of two other key proteins required for alkaloid synthesis, quinolinic acid phosphoribosyltransferase (nicotinic acid production) and a PIP-family oxidoreductase (nicotinic acid condensation reactions), were diminished in roots of odc-RNAi plants relative to vector-only controls. Transcriptional and biochemical differences associated with polyamine and alkaloid metabolism were exacerbated in odc-RNAi plants in response to different forms of shoot damage. In general, apex removal had a greater effect than leaf wounding alone, with a combination of these injury treatments producing synergistic responses in some cases. Reduced expression of ODC appeared to have negative effects upon plant growth and vigour with some leaves of odc-RNAi lines being brittle and bleached compared with vector-only controls. Together, results of this study demonstrate that ornithine decarboxylase has important roles in facilitating both primary and secondary metabolism in Nicotiana.
authors.affiliation: School of Biological Sciences, Monash University, Melbourne, Victoria 3800, Australia Department of Molecular Ecology, Max Planck Institute for Chemical Ecology, 07745 Jena, Germany.
authors.forename: Heidi L
authors.initials: HL
authors.lastname: Dalton
authors.affiliation: School of Biological Sciences, Monash University, Melbourne, Victoria 3800, Australia.
authors.forename: Cecilia K
authors.initials: CK
authors.lastname: Blomstedt
authors.affiliation: Deakin University, Centre for Regional and Rural Futures (CeRRF), Geelong, Victoria 3216, Australia.
authors.forename: Alan D
authors.initials: AD
authors.lastname: Neale
authors.affiliation: School of Biological Sciences, Monash University, Melbourne, Victoria 3800, Australia.
authors.forename: Ros
authors.initials: R
authors.lastname: Gleadow
authors.affiliation: The UWA Institute of Agriculture, The University of Western Australia, Crawley, WA 6009, Australia.
authors.forename: Kathleen D
authors.initials: KD
authors.lastname: DeBoer
authors.affiliation: Deakin University, Centre for Regional and Rural Futures (CeRRF), Geelong, Victoria 3216, Australia john.hamill@deakin.edu.au.
authors.forename: John D
authors.initials: JD
authors.lastname: Hamill
mesh_terms: D015536:Down-Regulation; D018506:Gene Expression Regulation, Plant; D009955:Ornithine Decarboxylase / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D018517:Plant Roots / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D011700:Putrescine / Q000378:metabolism*; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D059467:Transcriptome
chemical_list: D010940:Plant Proteins; D009955:Ornithine Decarboxylase; D011700:Putrescine
keywords: Alkaloid; ODC; PMT; QPT; RNAi.; gene expression; phenolamide; polyamine; putrescine