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Applied and Environmental Microbiology, December 2008, p. 7243-7251, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.01243-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Widespread Distribution and Identification of Eight Novel Microcystins in Antarctic Cyanobacterial Mats{triangledown}

Susanna A. Wood,1,2 Doug Mountfort,1* Andrew I. Selwood,1 Patrick T. Holland,1 Jonathan Puddick,2 and S. Craig Cary2,3

Cawthron Institute, Private Bag 2, Nelson 7042, New Zealand,1 Department of Biological Sciences, University of Waikato, Private Bag 3105, Hamilton, New Zealand,2 College of Marine and Earth Studies, University of Delaware, Lewes, Delaware 199583

Received 4 June 2008/ Accepted 1 October 2008

The microcystin (MC) content and cyanobacterial community structure of Antarctic microbial mat samples collected from 40 ponds, lakes, and hydroterrestrial environments were investigated. Samples were collected from Bratina Island and four of the Dry Valleys, Wright, Victoria, Miers, and Marshall. Enzyme-linked immunosorbent assays (ELISAs), liquid chromatography-mass spectrometry (LC-MS), and protein phosphatase 2A (PP-2A) inhibition assays resulted in the identification of low levels (1 to 16 mg/kg [dry weight]) of MCs in all samples. A plot of indicative potencies of MCs (PP-2A inhibition assay/ELISA ratio) versus total MCs (ELISA) showed a general decrease in potency, as total MC levels increased, and a clustering of values from discrete geographic locations. LC-tandem MS analysis on selected samples identified eight novel MC congeners. The low-energy collisional activation spectra were consistent with variants of [D-Asp3] MC-RR and [D-Asp3] MC-LR containing glycine [Gly1] rather than alanine and combinations of homoarginine [hAr2] or acetyldemethyl 3-amino-9-methoxy-2,6,8-trimethyl-10-phenyl-4,6-decadienoic acid (acetyldemethyl ADDA) [ADMAdda5] substitutions. Nostoc sp. was identified as a MC producer using PCR amplification of a region of the 16S rRNA gene and the aminotransferase domain of the mcyE gene. Automated ribosomal intergenic spacer analysis (ARISA) was undertaken to enable a comparison of cyanobacterial mat community structure from distant geographical locations. Two-dimensional multidimensional scaling ordination analysis of the ARISA data showed that in general, samples from the same geographic location tended to cluster together. ARISA also enabled the putative identification of the MC-producing Nostoc sp. from multiple samples.

* Corresponding author. Mailing address: Cawthron Institute, Coastal and Freshwater Resources, 98 Halifax Street East, Private Bag 2, Nelson 7042, New Zealand. Phone: 64-03-5482319. Fax: 64-03-5469464. E-mail: doug.mountfort{at}cawthron.org.nz

{triangledown} Published ahead of print on 10 October 2008.

Applied and Environmental Microbiology, December 2008, p. 7243-7251, Vol. 74, No. 23
0099-2240/08/$08.00+0     doi:10.1128/AEM.01243-08
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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