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The A622 gene in Nicotiana glauca (tree tobacco): evidence for a functional role in pyridine alkaloid synthesis

DeBoer, Kathleen D., Lye, Jessica C., Aitkin, Campbell D., Su, Angela K.K. and Hamill, John D. 2009, The A622 gene in Nicotiana glauca (tree tobacco): evidence for a functional role in pyridine alkaloid synthesis, Plant molecular biology, vol. 69, no. 3, pp. 299-312, doi: 10.1007/s11103-008-9425-2.

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Title The A622 gene in Nicotiana glauca (tree tobacco): evidence for a functional role in pyridine alkaloid synthesis
Author(s) DeBoer, Kathleen D.
Lye, Jessica C.
Aitkin, Campbell D.
Su, Angela K.K.
Hamill, John D.
Journal name Plant molecular biology
Volume number 69
Issue number 3
Start page 299
End page 312
Total pages 14
Publisher Springer-Verlag
Place of publication Berlin, Germany
Publication date 2009-02
ISSN 1573-5028
Keyword(s) anabasine
anatabine
hairy roots
nicotine
RNAi
transgenic
Summary Nicotiana glauca (Argentinean tree tobacco) is atypical within the genus Nicotiana, accumulating predominantly anabasine rather than nicotine and/or nornicotine as the main component of its leaf pyridine alkaloid fraction. The current study examines the role of the A622 gene from N. glauca (NgA622) in alkaloid production and utilises an RNAi approach to down-regulate gene expression and diminish levels of A622 protein in transgenic tissues. Results indicate that RNAi-mediated reduction in A622 transcript levels markedly reduces the capacity of N. glauca to produce anabasine resulting in plants with scarcely any pyridine alkaloids in leaf tissues, even after damage to apical tissues. In addition, analysis of hairy roots containing the NgA622-RNAi construct shows a substantial reduction in both anabasine and nicotine levels within these tissues, even if stimulated with methyl jasmonate, indicating a role for the A622 enzyme in the synthesis of both alkaloids in roots of N. glauca. Feeding of Nicotinic Acid (NA) to hairy roots of N. glauca containing the NgA622-RNAi construct did not restore capacity for synthesis of anabasine or nicotine. Moreover, treatment of these hairy root lines with NA did not lead to an increase in anatabine levels, unlike controls. Together, these results strongly suggest that A622 is an integral component of the final enzyme complex responsible for biosynthesis of all three pyridine alkaloids in Nicotiana.
Language eng
DOI 10.1007/s11103-008-9425-2
Field of Research 059999 Environmental Sciences not elsewhere classified
Socio Economic Objective 970105 Expanding Knowledge in the Environmental Sciences
HERDC Research category C1.1 Refereed article in a scholarly journal
Persistent URL http://hdl.handle.net/10536/DRO/DU:30064358

Document type: Journal Article
Collection: Office of the Deputy Vice-Chancellor Research
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