Abstract

BackgroundWithin the Arabidopsis genome, there are 272 cytochrome P450 monooxygenase (P450) genes. However, the biological functions of the majority of these P450s remain unknown. The CYP709B family of P450s includes three gene members, CYP709B1, CYP709B2 and CYP709B3, which have high amino acid sequence similarity and lack reports elucidating biological functions.ResultsWe identified T-DNA insertion-based null mutants of the CYP709B subfamily of genes. No obvious morphological phenotypes were exhibited under normal growth conditions. When the responses to ABA and salt stress were studied in these mutants, only the cyp709b3 mutant showed sensitivity to ABA and salt during germination. Under moderate salt treatment (150 mM NaCl), cyp709b3 showed a higher percentage of damaged seedlings, indicating a lower tolerance to salt stress. CYP709B3 was highly expressed in all analyzed tissues and especially high in seedlings and leaves. In contrast, CYP709B1 and CYP709B2 were highly expressed in siliques, but were at very low levels in other tissues. Under salt stress condition, CYP709B3 gene expression was induced after 24 hr and remained at high expression level. Expression of the wild type CYP709B3 gene in the cyp709b3 mutant fully complemented the salt intolerant phenotype. Furthermore, metabolite profiling analysis revealed some differences between wild type and cyp709b3 mutant plants, supporting the salt intolerance phenotype of the cyp709b3 mutant.ConclusionsThese results suggest that CYP709B3 plays a role in ABA and salt stress response and provides evidence to support the functions of cytochrome P450 enzymes in plant stress response.

Highlights

  • Within the Arabidopsis genome, there are 272 cytochrome P450 monooxygenases (P450s) monooxygenase (P450) genes

  • We identified two mutant alleles of the CYP709B2 gene, one of which has an insertion in the second exon (SALK_020401, cyp709b2-1) and the other in the third intron (SALK_087806, cyp709b2-2) (Figure 1A)

  • CYP709B3 gene expression under 150 mM NaCl stress was checked, we found that CYP709B3 gene expression was not induced at the early stage of salt treatment; the expression was induced after 24 h and remained high at later time points (Figure 6A)

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Summary

Introduction

Within the Arabidopsis genome, there are 272 cytochrome P450 monooxygenase (P450) genes. The CYP709B family of P450s includes three gene members, CYP709B1, CYP709B2 and CYP709B3, which have high amino acid sequence similarity and lack reports elucidating biological functions. Cytochrome P450 monooxygenases (P450s) are universal enzymes present in most organisms from bacteria to plants and humans. They catalyze the oxidation of various substrates through activation of molecular oxygen. The A-type clade is specific to plants while the non-A-type clade is a divergent group with members showing more similarity to non-plant P450s [4,5]. Plant P450s catalyze biosynthetic steps for a wide range of plant metabolites, including pigments, defense-related compounds, UV protectants, lignin, fatty acids and phytohormones [2,3,6,7,8]

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