Molecular cloning and expression of a cucumber (Cucumis sativus L.) heme oxygenase-1 gene, CsHO1, which is involved in adventitious root formation

Gene. 2011 Oct 15;486(1-2):47-55. doi: 10.1016/j.gene.2011.07.008. Epub 2011 Jul 19.

Abstract

Our previous work showed that in cucumber (Cucumis sativus), auxin rapidly induces heme oxygenase (HO) activity and the product of HO action, carbon monoxide (CO), then triggers the signal transduction events leading to adventitious root formation. In this study, the cucumber HO-1 gene (named as CsHO1) was isolated and sequenced. It contains four exons and three introns and encodes a polypeptide of 291 amino acids. Further results show that CsHO1 shares a high homology with plant HO-1 proteins and codes a 33.3 kDa protein with a 65-amino transit peptide, predicting a mature protein of 26.1 kDa. The mature CsHO1 was expressed in Escherichia coli to produce a fusion protein, which exhibits HO activity. The CsHO1:GFP fusion protein was localized in the chloroplast. Related biochemical analyses of mature CsHO1, including Vmax, Km, Topt and pHopt, were also investigated. CsHO1 mRNA was found in germinating seeds, roots, stem, and especially in leaf tissues. Several well-known adventitious root inducers, including auxin, ABA, hemin, nitric oxide donor sodium nitroprusside (SNP), CaCl(2), and sodium hydrosulfide (NaHS), differentially up-regulate CsHO1 transcripts and corresponding protein levels. These results suggest that CsHO1 may be involved in cucumber adventitious rooting.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Cloning, Molecular
  • Cucumis sativus / enzymology*
  • Cucumis sativus / genetics*
  • Cucumis sativus / growth & development
  • DNA Primers / genetics
  • DNA, Plant / genetics
  • Exons
  • Gene Expression Regulation, Developmental / drug effects
  • Gene Expression Regulation, Enzymologic / drug effects
  • Gene Expression Regulation, Plant / drug effects
  • Genes, Plant*
  • Heme Oxygenase-1 / chemistry
  • Heme Oxygenase-1 / genetics*
  • Heme Oxygenase-1 / metabolism
  • Introns
  • Kinetics
  • Molecular Sequence Data
  • Molecular Weight
  • Phylogeny
  • Plant Growth Regulators / pharmacology
  • Plant Roots / drug effects
  • Plant Roots / enzymology
  • Plant Roots / growth & development
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • RNA, Plant / genetics
  • RNA, Plant / metabolism
  • Sequence Homology, Amino Acid

Substances

  • DNA Primers
  • DNA, Plant
  • Plant Growth Regulators
  • RNA, Messenger
  • RNA, Plant
  • Heme Oxygenase-1