Abstract

Benzoxazolin-2(3H)-one (BOA) is an allelochemical most commonly associated with monocot species, formed from the O-glucoside of 2,4-dihydroxy-2H-1,4-benzoxazin-3(4H)-one by a two-step degradation process. The capacity of Arabidopsis to detoxify exogenously supplied BOA was analyzed by quantification of the major known metabolites BOA-6-OH, BOA-6-O-glucoside, and glucoside carbamate, revealing that detoxification occurs predominantly through O-glucosylation of the intermediate BOA-6-OH, most likely requiring the sequential action of as-yet-unidentified cytochrome P450 and UDP-glucosyltransferase activities. Transcriptional profiling experiments were also performed with Arabidopsis seedlings exposed to BOA concentrations, representing I(50) and I(80) levels based on root elongation inhibition assays. One of the largest functional categories observed for BOA-responsive genes corresponded to protein families known to participate in cell rescue and defense, with the majority of these genes potentially associated with chemical detoxification pathways. Further experiments using a subset of these genes revealed that many are also transcriptionally induced by a variety of structurally diverse xenobiotic compounds, suggesting they comprise components of a coordinately regulated, broad specificity xenobiotic defense response. The data significantly expand upon previous studies examining plant transcriptional responses to allelochemicals and other environmental toxins and provide novel insights into xenobiotic detoxification mechanisms in plants.

Highlights

  • Benzoxazolin-2(3H)-one (BOA) is an allelochemical most commonly associated with monocot species, formed from the O-glucoside of 2,4-dihydroxy-2H-1,4benzoxazin-3(4H)-one by a two-step degradation process

  • Transcriptional profiling experiments were performed with Arabidopsis seedlings exposed to BOA concentrations, representing in 10-day-old seedlings by 50% (I50) and I80 levels based on root elongation inhibition assays

  • The genes encoding all of the enzymes required for the biosynthesis of the benzoxazinoid 2,4-dihydroxy-7-methoxy2H-1,4-benzoxazin-3(4H)-one have been identified in corn via transposon tagging, and they represent the first known example of a plant secondary metabolic pathway organized as a gene cluster [9]

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Summary

THE JOURNAL OF BIOLOGICAL CHEMISTRY

21867–21881, 2005 Printed in U.S.A. Detoxification and Transcriptome Response in Arabidopsis Seedlings Exposed to the Allelochemical Benzoxazolin-2(3H)-one*□S. Benzoxazolin-2(3H)-one (BOA) is an allelochemical most commonly associated with monocot species, formed from the O-glucoside of 2,4-dihydroxy-2H-1,4benzoxazin-3(4H)-one by a two-step degradation process. One recent exception is the use of DNA microarrays to characterize the transcriptome response of Arabidopsis seedlings exposed to (Ϫ)-catechin, an allelochemical identified in root exudates of Centaurea maculosa [6]. Benzoxazinoids and their benzoxazolinone derivatives represent one of the more intensively studied classes of allelochemicals [7, 8].

Benzoxazolinone Detoxification Response in Arabidopsis
EXPERIMENTAL PROCEDURES
RESULTS AND DISCUSSION
Glucoside carbamate
Primer pairsa
NAC domain protein family
Element involved in light responsiveness
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