Analysis of wound-induced gene expression in Nicotiana species with contrasting alkaloid profiles
Steven J. Sinclair A B , Richard Johnson A and John D. Hamill A CA School of Biological Sciences, Monash University, PO Box 18, Melbourne, Vic. 3800, Australia.
B Current address: Flora Ecology Research, Arthur Rylah Institute for Environmental Research (ARI), Department of Sustainability and Environment, 123 Brown St Heidelberg, Vic. 3084, Australia.
C Corresponding author; email: john.hamill@sci.monash.edu.au
Functional Plant Biology 31(7) 721-729 https://doi.org/10.1071/FP03242
Submitted: 9 December 2003 Accepted: 7 April 2004 Published: 22 July 2004
Abstract
We determined the capacity of three Nicotiana (Solanaceae) species with very different alkaloid profiles (Nicotiana sylvestris Speg & Comes, Nicotiana alata Link & Otto and Nicotiana glauca Grah.) to increase their alkaloid contents in both leaf and root tissues following foliage damage. We also investigated the transcriptional responses of genes encoding enzymes important for alkaloid biosynthesis, namely quinolinate phosphoribosyltransferase (QPT), putrescine N-methyltransferase (PMT), ornithine decarboxylase (ODC) and the putative alkaloid biosynthetic gene A622. In response to wounding of foliage in the well studied ‘model’ species N. sylvestris, a rise, approximately 2-fold, in leaf nicotine levels was observed several days after a 4–5-fold increase in the transcript levels of all genes in the roots. In contrast, leaf tissues of the ornamental tobacco N. alata showed very low levels of any pyridine alkaloid, even when analysed 1 week after wounding, correlating with a general lack of transcript abundance representing any of these genes in leaves or roots following foliage damage. However, addition of methyl jasmonate to cultured roots of N. alata did produce elevated levels of nicotine and anatabine raising the possibility that components of the leaf–root wound signalling system in N. alata are different from those in N. sylvestris. Wounding of the tree tobacco N. glauca, was followed by a 2-fold increase in anabasine levels several days later. This increase followed a large rise in transcript levels of ODC, QPT and A622, though not PMT, in wounded leaves, but not in non-wounded leaves or roots. These data support the hypothesis that N. glauca is able to produce increased anabasine levels following wounding in its foliage, setting it apart from N. sylvestris where induced alkaloid production takes place in roots. We discuss the possibility that increased transcript levels detected by ODC and A622 probes play important roles in anabasine synthesis in N. glauca.
Keywords: anabasine, nicotine, nicotine synthase, ornithine decarboxylase, putrescine N-methyltransferase, quinolinate phosphoribosyltransferase, transcript abundance.
Acknowledgments
We thank Karen Cane, Kathleen deBoer, Angela Lidgett (Monash University), Graeme Newell and Joanne Potts (ARI) for advice and assistance. This research was supported by Australian Research Council (ARC) grant A19701779. SS also acknowledges the receipt of an ARC-funded post-graduate scholarship.
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