Isolation and characterization of Stemphylium sedicola SBU-16 as a new endophytic taxol-producing fungus from Taxus baccata grown in Iran

FEMS Microbiol Lett. 2012 Mar;328(2):122-9. doi: 10.1111/j.1574-6968.2011.02488.x. Epub 2012 Jan 10.

Abstract

In this study, a total of 25 endophytic fungi were successfully isolated from the inner bark of Taxus baccata grown in Iran by the aseptic technique. Genomic DNA was extracted from isolated endophytic fungi and subjected to polymerase chain reaction (PCR) analysis for the presence of the Taxus taxadiene synthase (ts) gene, which encodes the enzyme catalyzing the first committed step of taxol biosynthesis. Four of 25 isolated endophytic fungi isolates showed PCR positive for the ts gene. Subsequently, taxol and 10-deacetylbaccatin III (10-DAB III) were extracted from culture filtrates and mycelia of the PCR positive isolates and analyzed by high-performance liquid chromatography and mass spectrometry. The analysis showed that one isolate (SBU-16) produced taxol (6.9 ± 0.2 μg L(-1) ) and its intermediate compound, 10-DAB III (2.2 ± 0.1 μg L(-1) ). The isolate SBU-16 was identified as Stemphylium sedicola SBU-16, according to its morphological characteristics as well as the internal transcribed spacer nuclear rDNA gene sequence analysis. Interestingly, this is the first report of the genus Stemphylium as a taxol-producing taxon.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ascomycota / classification
  • Ascomycota / isolation & purification*
  • Ascomycota / metabolism
  • Chromatography, High Pressure Liquid
  • Culture Media / chemistry
  • DNA, Fungal / genetics
  • DNA, Ribosomal Spacer / genetics
  • Endophytes / classification
  • Endophytes / isolation & purification*
  • Endophytes / metabolism
  • Genes, Fungal
  • Iran
  • Isomerases / genetics
  • Mass Spectrometry
  • Mycelium / metabolism
  • Paclitaxel / biosynthesis*
  • Phylogeny
  • Polymerase Chain Reaction
  • Taxoids / chemistry
  • Taxoids / isolation & purification*
  • Taxus / growth & development
  • Taxus / microbiology*

Substances

  • Culture Media
  • DNA, Fungal
  • DNA, Ribosomal Spacer
  • Taxoids
  • 10-deacetylbaccatine III
  • Isomerases
  • taxa-4(5),11(12)-diene synthase
  • Paclitaxel