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A mixed community of actinomycetes produce multiple antibiotics for the fungus farming ant Acromyrmex octospinosus

Jörg Barke1, Ryan F Seipke1, Sabine Grüschow2, Darren Heavens3, Nizar Drou3, Mervyn J Bibb4, Rebecca JM Goss2, Douglas W Yu15 and Matthew I Hutchings16*

Author Affiliations

1 School of Biological Sciences, University of East Anglia, Norwich, Norwich Research Park, NR4 7TJ, UK

2 School of Chemistry, University of East Anglia, Norwich, Norwich Research Park, NR4 7TJ, UK

3 The Genome Analysis Centre, Norwich, Norwich Research Park, NR4 7UH, UK

4 Department of Molecular Microbiology, John Innes Centre, Norwich, Norwich Research Park, NR4 7UH, UK

5 State Key Laboratory of Genetic Resources, and Evolution, Ecology, Conservation and Environment Center (ECEC), Kunming Institute of Zoology, Chinese Academy of Sciences, Kunming, Yunnan 650223, China

6 School of Medicine, Health Policy and Practice, University of East Anglia, Norwich, Norwich Research Park, NR4 7TJ, UK

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BMC Biology 2010, 8:109  doi:10.1186/1741-7007-8-109

Published: 26 August 2010

Additional files

Additional file 1:

Detecting candicidin biosynthesis genes using polymerase chain reaction (PCR). PCR analysis of antifungal producers using primers against candicin biosynthesis genes fscM and fscP. Sequence identities to Haeder et al. [12]: fscM gene, S4 = 100%, S5 = 99%; fscP gene: S4 = 98% and S5 = 98%

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Additional file 2:

Streptomyces and Pseudonocardia strains identified in this study. The Pseudonocardia and Streptomyces strains isolated in this study are listed with the Acromyrmex octospinosus colony they were isolated from (1,2 or 3), the accession numbers for their 16 S ribosomal DNA (rDNA) sequences, the top National Center for Biotechnology Information Blast hits for each of their 16 S rDNA sequences and the percentage identity to these BLAST hits. Also noted are the results from polymerase chain reaction testing for the candicidin biosynthetic genes fscM and fscP using primers from a previous study [12] and the nystatin-like Pseudonocardia polyene biosynthetic gene nppDIII using the primer set RFS84 (CAGATCCGCTTCTACCAGG) and RFS85 (CGCACCGAGTGCATCTG).

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Additional file 3:

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) identification of candicidin in S4 extracts. Analysis of S4-derived extracts. Left panel (A), ultraviolet spectrum extracted at RT 8.3 min (see panel B) from the S4 extract. The absorption maxima match those previously reported for candicidin D [12]. Right panel (B), LC-MS analysis of S4 extract. Ion chromatograms extracted for the molecular ion of candicidin D (m/z 1109.6) are shown. (C), MS2 analysis of the extracted ion m/z 1109.6. The fragmentation pattern of the antifungal compound from Streptomyces S4 perfectly matched the fragmentation of candicidin as reported previously [12]. The ions highlighted in the Haeder et al. study [12] are labelled in a larger font.

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Additional file 4:

genome sequencing data for Pseudonocardia P1. Summary of the Pseuodonocardia sp. P1 draft genome sequence output obtained by 454 pyrosequencing

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Additional file 5:

Identification of the nystatin P1 biosynthetic gene cluster. Tiling of Pseudonocardia sp. P1 contigs (GenBank accession ADUJ00000000) to the NPP biosynthetic gene cluster from P. autotrophica (GenBank accession EU108007). *The negative value for PP100949 denotes that the contig extends 4517 bp beyond the nystatin-like Pseudonocardia polyene biosynthetic gene cluster. **Negative values indicate that adjacent contigs overlap.

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Additional file 6:

MS3 data for nystatin P1. The spectrum shows the fragmentation data of the m/z 1088 → 326 ion. The most frequently observed fragmentation corresponds to loss of water: m/z 308 (-1 H20), m/z 290 (-2 H20), m/z 272 (-1 H20). The m/z 146 product ion is consistent with a mycosamine sugar after loss of the hexose (mass difference 180).

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