Induction of chalcone isomerase gene expression in plant cells in response to fungal cell wall components

By:

MEHDY, M.C

Contributor(s):

LAMB, C.J

Material type: Article

ArticleLanguage: English Description: p. 425-435Subject(s): LOC classification:

41174

In: In: Firtel, R.A.; Davidson, E.H. (eds.). Molecular approaches to developmental biologySummary: Plants respond to irradiation by producing protective anthocyanin pigments and to infection by producing phytoalexin antibiotics. Synthesis of anthocyanins and some flavonoid pigments is also developmentally regulated and tissue-specific in leaves and flowers. Chalcone isomerase (CHI) catalyzes the formation of an intermediate common to both flavonoid pigment and isoflavonoid phytoalexin biosynthetic pathways and elevated CHI enzyme activity is correlated with the accumulation of these compounds in many plants. To determine the molecular basis for enzyme induction, we constructed a lambda gt 11 cDNA library prepared from mRNA from Phaseolus vulgaris cell cultures treated with a fungal cell wall preparation (elicitor). Two positive clones were obtained by screening 10(5) recombinants with an antiserum that had been raised against purified CHI. The identity of the cloned sequence was confirmed by showing that the largest clone (CHI 1) encoded polypeptides which were precipitated by the antiserum. Blot hybridization of RNA from elicitor treated cell cultures with labeled CHI l showed substantial accumulation of a 0.95 kb RNA within 3-4 h. The kinetics of CHI mRNA accumulation are similar to those of phenylalanine ammonia-lyase and chalcone synthase mRNAs, which encode other enzymes common to both pathways. CHI l hybridized to single fragments on genomic DNA blots suggesting a single gene. The structure and expression of the gene is being investigated to elucidate the regulatory sequences conferring developmental and environmental inducibility. (AS)

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Item type	Current library	Collection	Call number	Copy number	Status	Date due	Barcode	Item holds
Book Chapters	CIAT Library Document collection CINFOS	Document Collection CINFOS	41174 (Browse shelf(Opens below))	c.1	Short Loan		100059679

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Plants respond to irradiation by producing protective anthocyanin pigments and to infection by producing phytoalexin antibiotics. Synthesis of anthocyanins and some flavonoid pigments is also developmentally regulated and tissue-specific in leaves and flowers. Chalcone isomerase (CHI) catalyzes the formation of an intermediate common to both flavonoid pigment and isoflavonoid phytoalexin biosynthetic pathways and elevated CHI enzyme activity is correlated with the accumulation of these compounds in many plants. To determine the molecular basis for enzyme induction, we constructed a lambda gt 11 cDNA library prepared from mRNA from Phaseolus vulgaris cell cultures treated with a fungal cell wall preparation (elicitor). Two positive clones were obtained by screening 10(5) recombinants with an antiserum that had been raised against purified CHI. The identity of the cloned sequence was confirmed by showing that the largest clone (CHI 1) encoded polypeptides which were precipitated by the antiserum. Blot hybridization of RNA from elicitor treated cell cultures with labeled CHI l showed substantial accumulation of a 0.95 kb RNA within 3-4 h. The kinetics of CHI mRNA accumulation are similar to those of phenylalanine ammonia-lyase and chalcone synthase mRNAs, which encode other enzymes common to both pathways. CHI l hybridized to single fragments on genomic DNA blots suggesting a single gene. The structure and expression of the gene is being investigated to elucidate the regulatory sequences conferring developmental and environmental inducibility. (AS) eng