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Plant and Cell Physiology Advance Access originally published online on May 19, 2008
Plant and Cell Physiology 2008 49(7):1003-1012; doi:10.1093/pcp/pcn077
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Jasmonate-Induced Nicotine Formation in Tobacco is Mediated by Tobacco COI1 and JAZ Genes

Tsubasa Shoji, Takayuki Ogawa and Takashi Hashimoto*

Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara, 630-0192 Japan

*Corresponding author: E-mail, hasimoto{at}bs.naist.jp; Fax, +81-743-72-5529.


  Abstract

Biosynthesis of many plant alkaloids is enhanced by endogenous accumulation and exogenous application of jasmonates, but the general and specific signaling components are not well understood. In Arabidopsis, jasmonate-induced ZIM-domain-containing (JAZ) proteins have recently been found to be critical transcriptional repressors linking CORONATINE INSENSTIVE1 (COI1)-mediated jasmonate perception and jasmonate-regulated transcriptional regulation. Insect herbivory on tobacco leaves activates the jasmonate signaling pathway, leading to up-regulation of nicotine biosynthesis genes in roots. We show here that roots of COI1-silenced tobacco plants are insensitive to growth inhibition by methyl jasmonate, and do not activate nicotine biosynthesis genes after jasmonate treatment or wounding of leaves. Tobacco JAZ proteins appeared to be rapidly degraded after jasmonate treatment, whereas a C-terminally truncated form lacking the conserved Jas motif did not. When the non-degradable JAZ forms were expressed in tobacco hairy roots, jasmonate induction of nicotine biosynthesis was strongly inhibited. Formation of tobacco alkaloids in jasmonate-elicited tobacco BY-2 cells was also effectively suppressed by the COI1 RNAi (RNA interference) construct and by the dominant-negative truncated JAZ constructs. In addition, jasmonate-mediated induction of nicotine biosynthesis genes was diminished by treatment with a proteasome inhibitor MG132. These results indicate that jasmonate-triggered, COI1-mediated degradation of JAZ repressors activates transcriptional regulation of nicotine biosynthesis genes in tobacco roots.

Keywords: COI1 - Jasmonate - JAZ - Nicotine - Tobacco

Abbreviations: COI1, CORONATINE INSENSTIVE1; EST, expressed sequence tag; GUS, β-glucuronidase; JA, jasmonate; JAZ, JASMONATE ZIM-domain; MeJA, methyl jasmonate; PI-II, proteinase inhibitor-II; PMT, putrescine N-methyltransferase; QPT, quinolinate phosphoribosyltransferase; RNAi, RNA interference; RT–PCR, reverse transcription–PCR; SCF, Skp1–Cullin–F-box; TPI, trypsin proteinase inhibitor

The nucleotide sequences reported in this paper have been submitted to DDBJ under accession numbers AB433899 (NtCOI1), AB433896 (NtJAZ1), AB433897 (NtJAZ2) and AB433898 (NtJAZ3).

(Received April 11, 2008; Accepted May 17, 2008)
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T. Shoji and T. Hashimoto
Why does Anatabine, But not Nicotine, Accumulate in Jasmonate-Elicited Cultured Tobacco BY-2 Cells?
Plant Cell Physiol., August 1, 2008; 49(8): 1209 - 1216.
[Abstract] [Full Text] [PDF]


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