Induced plant responses to pathogen attack. Analysis and heterologous expression of the key enzyme in the biosynthesis of phytoalexins in soybean (Glycine max L. Merr. cv. Harosoy 63).
In soybean (Glycine max L.), pathogen attack induces the formation of glyceollin-type phytoalexins. The biosynthetic key enzyme is a reductase which synthesizes 4,2', 4'-trihydroxychalcone in co-action with chalcone synthase. Screening of a soybean cDNA library from elicitor-induced RNA in lambda gt11 yielded two classes of reductase-specific clones. The deduced proteins match to 100% and 95%, respectively, with 229 amino acids sequenced in the purified plant protein. Four clones of class A were expressed in Escherichia coli, and the proteins were tested for enzyme activity in extracts supplemented with chalcone synthase. All were active in 4,2',4'-trihydroxychalcone formation, and the quantification showed that shorter lengths of the cDNAs at the 5' end correlated with progressively decreasing enzyme activities. Genomic blots with DNA from plants capable of 4,2',4'-trihydroxychalcone synthesis revealed related sequences in bean (Phaseolus vulgaris L.) and peanut (Arachis hypogaea L.), but not in pea (Pisum sativum L.). No hybridization was observed with parsley (Petroselinum crispum) and carrot (Daucus carota) which synthesize other phytoalexins. The reductase protein contains a leucine-zipper motif and reveals a marked similarity with other oxidoreductases most of which are involved in carbohydrate metabolism.
Welle
R
R
Institut für Biologie II, Biochemie der Pflanzen, Universität Freiburg, Federal Republic of Germany.
Schröder
G
G
Schiltz
E
E
Grisebach
H
H
Schröder
J
J
D000217:Acyltransferases / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D002599:Chalcone / Q000031:analogs & derivatives / Q000378:metabolism; D047188:Chalcones; D004247:DNA / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D015151:Immunoblotting; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D010936:Plant Extracts / Q000096:biosynthesis*; D012689:Sequence Homology, Nucleic Acid; D012717:Sesquiterpenes; D013025:Glycine max / Q000201:enzymology / Q000235:genetics*; D013729:Terpenes; D014162:Transfection; D000094162:Phytoalexins
D047188:Chalcones; D010936:Plant Extracts; D012717:Sesquiterpenes; D013729:Terpenes; D002599:Chalcone; D004247:DNA; C040920:isoliquiritigenin; D010088:Oxidoreductases; D000217:Acyltransferases; C020141:flavanone synthetase; D000094162:Phytoalexins