Identification of a cytochrome P450 cDNA (CYP98A5) from Phaseolus vulgaris, inducible by 3,5-dichlorosalicylic acid and 2,6-dichloro isonicotinic acid

J Plant Physiol. 2007 Apr;164(4):421-8. doi: 10.1016/j.jplph.2006.02.006. Epub 2006 Apr 3.

Abstract

To investigate the involvement of cytochrome P450s in the metabolism of plants treated with xenobiotic agrochemicals, bean leaves were treated with 3,5-dichlorosalicylic acid (DC-SA), a priming agent of plant defense and 2,6-dichloroisonicotinic acid (DC-INA), a chemical inducer of systemic acquired resistance. Through the use of directed differential display reverse transcription polymerase chain reactions, a differentially expressed cDNA amplicon, found to be up-regulated by both DC-SA and DC-INA treatment, was identified as a cytochrome P450 cDNA, CYP98A5. The nucleotide sequence indicates extensive homology to 3'-hydroxylases of p-coumaroyl esters. Dot blot analysis of leaves treated with various SA and isonicotinic acid derivatives showed enhanced expression of CYP98A5 due to DC-SA and DC-INA. Northern blot analysis of a time-dependent induction study of CYP98A5 in treated bean leaves indicated that DC-SA induces CYP98A5 mRNA transcripts earlier than DC-INA. Both inducers resulted in high transcript levels 24-48 h after treatment. The up-regulation of CYP98A5 is supportive of the conditioning and sensitizing effects of DC-SA and DC-INA to elicit a more rapid and effective defense response.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Chlorobenzoates
  • Cloning, Molecular
  • Cytochrome P-450 Enzyme System / biosynthesis
  • Cytochrome P-450 Enzyme System / genetics*
  • DNA Primers / genetics
  • DNA, Complementary / genetics
  • DNA, Complementary / isolation & purification
  • DNA, Plant / genetics
  • DNA, Plant / isolation & purification
  • Enzyme Induction / drug effects
  • Genes, Plant / drug effects
  • Isonicotinic Acids / pharmacology*
  • Molecular Sequence Data
  • Phaseolus / drug effects
  • Phaseolus / enzymology*
  • Phaseolus / genetics*
  • Plant Leaves / drug effects
  • Plant Leaves / enzymology
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Plant / genetics
  • RNA, Plant / metabolism
  • Salicylates / pharmacology*
  • Sequence Homology, Amino Acid

Substances

  • Chlorobenzoates
  • DNA Primers
  • DNA, Complementary
  • DNA, Plant
  • Isonicotinic Acids
  • RNA, Messenger
  • RNA, Plant
  • Salicylates
  • 2,6-dichloroisonicotinic acid
  • Cytochrome P-450 Enzyme System
  • cytochrome P-450 CYP98A1 (Sorghum bicolor)
  • 3,5-dichlorosalicylic acid