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Chalcone isomerase cDNA cloning and mRNA induction by fungal elicitor, wounding and infection
Author(s) -
Mehdy Mona C.,
Lamb Christopher J.
Publication year - 1987
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1987.tb02396.x
Subject(s) - elicitor , biology , chalcone isomerase , complementary dna , chalcone synthase , cloning (programming) , microbiology and biotechnology , gene expression , genetics , gene , computer science , programming language
The environmentally regulated synthesis of phenylpropanoid natural products was studied by examining the expression of the gene encoding chalcone isomerase (CHI). This enzyme catalyzes a step common to the synthesis of flavonoid pigments and isoflavonoid phytoalexins. A λgt11 library was constructed using mRNA from cell cultures of bean ( Phaseolus vulgaris L.) treated with fungal elicitor. Two positive clones were obtained by screening 10 5 recombinants with an antiserum to purified bean CHI. The identity of the cloned sequences was confirmed by hybrid‐select translation and the production of antigenic polypeptides from transcripts synthesized in vitro . Addition of elicitor to cell cultures resulted in the rapid accumulation of CHI mRNA, with maximum levels achieved 3–4 h after elicitation. CHI mRNA also accumulated during the natural infection of hypocotyls with the fungal pathogen Colletotrichum lindemuthianum , and in mechanically wounded hypocotyls. The kinetics of accumulation of CHI mRNA in response to these environmental signals were strikingly similar to those of mRNAs encoding two other phenylpropanoid pathway enzymes, phenylalanine ammonialyase and chalcone synthase. In contrast to the multi‐gene families encoding these two enzymes, chalcone isomerase is encoded by a single gene which is regulated by several environmental stimuli.