Isolation, molecular cloning and antimicrobial activity of novel defensins from common chickweed (Stellaria media L.) seeds

Biochimie. 2011 Mar;93(3):450-6. doi: 10.1016/j.biochi.2010.10.019. Epub 2010 Nov 4.

Abstract

Two novel highly homologous defensins, Sm-AMP-D1 and Sm-AMP-D2, were isolated from seeds of common chickweed Stellaria media L. (family Cariophyllaceae). They show sequence homology to defensins of the Brassicaceae plants and display strong inhibitory activity against phytopathogenic fungi and oomycetes in the micromolar range (IC(50)≤1μM). The cDNA sequences coding for Sm-AMP-D1 and Sm-AMP-D2 were obtained. They code for highly homologous precursor proteins, consisting of a signal peptide of 32 amino acid residues and the mature peptide domain of 50 amino acid residues. The Sm-AMP-D1 and Sm-AMP-D2 precursors differ by two amino acids: one in the signal peptide region, and the other, in the mature peptide domain. Two Sm-D1-encoding genes were identified in S. media genome by PCR amplification from the genomic DNA using Sm-D1-specific primers. They contain a single 599-bp intron in the signal peptide domain and differ from each other by nucleotide substitutions in the intron and 3'-untranslated regions, while the coding sequences are well conserved. One of the genes matched perfectly the sm-D1 cDNA sequence. The sm-D genes show promise for engineering pathogen resistance in crops and expand our knowledge on weed genomics.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antifungal Agents / chemistry
  • Antifungal Agents / isolation & purification*
  • Antifungal Agents / pharmacology
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Defensins / chemistry
  • Defensins / genetics*
  • Defensins / isolation & purification
  • Defensins / pharmacology*
  • Fungi / drug effects
  • Genome, Plant / genetics
  • Genomics
  • Molecular Sequence Data
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Plant Proteins / isolation & purification*
  • Plant Proteins / pharmacology
  • Seeds / chemistry*
  • Seeds / genetics
  • Sequence Analysis, DNA
  • Stellaria / chemistry*
  • Stellaria / genetics

Substances

  • Antifungal Agents
  • DNA, Complementary
  • Defensins
  • Plant Proteins