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Plant Physiol, April 2001, Vol. 125, pp. 1688-1699
Study of the Role of Antimicrobial Glucosinolate-Derived
Isothiocyanates in Resistance of Arabidopsis to Microbial
Pathogens1
Koenraad F.M.-J.
Tierens,
Bart P.H.J.
Thomma,
Margreet
Brouwer,
Jürgen
Schmidt,
Katherine
Kistner,
Andrea
Porzel,
Brigitte
Mauch-Mani,
Bruno P.A.
Cammue,* and
Willem F.
Broekaert2
F.A. Janssens Laboratory of Genetics, Katholieke
Universiteit Leuven, Kardinaal Mercierlaan 92, B-3001 Heverlee-Leuven,
Belgium (K.F.M.-J.T., B.P.H.J.T., M.B., B.P.A.C., W.F.B.); Institut
für Pflanzenbiochemie, Weinberg 3, D-06210 Halle (Saale),
Germany (J.S., K.K., A.P.); Department of Biology/Plant Biology,
University of Fribourg, 3 Route Albert Gockel, CH-1700 Fribourg,
Switzerland (B.M.-M.); and Flanders Interuniversity Institute for
Biotechnology, Belgium (B.P.A.C.)
Crude aqueous extracts from Arabidopsis leaves were subjected to
chromatographic separations, after which the different fractions were
monitored for antimicrobial activity using the fungus Neurospora crassa as a test organism. Two major fractions were obtained
that appeared to have the same abundance in leaves from untreated
plants versus leaves from plants challenge inoculated with the fungus Alternaria brassicicola. One of both major antimicrobial
fractions was purified to homogeneity and identified by 1H
nuclear magnetic resonance, gas chromatography/electron impact mass
spectrometry, and gas chromatography/chemical ionization mass
spectrometry as 4-methylsulphinylbutyl isothiocyanate (ITC). This
compound has previously been described as a product of
myrosinase-mediated breakdown of glucoraphanin, the predominant
glucosinolate in Arabidopsis leaves. 4-Methylsulphinylbutyl ITC was
found to be inhibitory to a wide range of fungi and bacteria, producing
50% growth inhibition in vitro at concentrations of 28 µM for the most sensitive organism tested
(Pseudomonas syringae). A previously identified
glucosinolate biosynthesis mutant, gsm1-1, was found to
be largely deficient in either of the two major antimicrobial
compounds, including 4-methylsulphinylbutyl ITC. The resistance of
gsm1-1 was compared with that of wild-type plants after
challenge with the fungi A. brassicicola,
Plectosphaerella cucumerina, Botrytis
cinerea, Fusarium oxysporum, or
Peronospora parasitica, or the bacteria Erwinia carotovora or P. syringae. Of the tested
pathogens, only F. oxysporum was found to be
significantly more aggressive on gsm1-1 than on wild-type plants. Taken together, our data suggest that
glucosinolate-derived antimicrobial ITCs can play a role in the
protection of Arabidopsis against particular pathogens.
1
This work was partially supported by the
Vlaams Instituut voor Bevordering van het
Wetenschappelijk-Technologisch Onderzoek in de Industrie (grant no.
G961052). K.T.F.M.-J. is the recipient of a predoctoral fellowship of
this fund. B.P.H.J.T. is a research assistant of the fund Fonds voor
Wetenschappelijk Onderzoek-Vlaanderen.
2
Present address: Cropdesign, Technologiepark 3, B-9052
Gent, Belgium.
*
Corresponding author; e-mail bruno.cammue{at}agr.kuleuven.ac.be;fax
32-16-32-19-66.
© 2001 American Society of Plant Physiologists
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