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Diversity of mcyD and mcyA genes in Cyanobacteria of Lake Menomin
Kang Vang, Juliana Lucchesi, Dr. Stephen Nold, University of Wisconsin-Stout
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Figure 1. Cyanobacterial blooms
Overview
•Lake Menomin is heavily polluted
with phosphorous and experiences
frequent (and smelly!) cyanobacterial
blooms (Fig. 1)
•Lake Menomin is part of the Red
Cedar watershed in Menomonie, WI
(Fig. 2)
Figure 2. Lake Menomin
Figure 3. a, Raw sequence data; b, Sequences edited and aligned with closest
relatives from GenBank
Clone Designation
22 of our mcyD sequences
080428jn3_07A_SP6-mcyD-II-14
080428jn3_07F_SP6-mcyD-II-6
080428jn3_07H_SP6-mcyD-III-1
080428jn3_09A_SP6-mcyD-IV-10
080428jn5_07A_T7-mcyD-II-14
080428jn5_07H_T7-mcyD-III-1
080428jn5_09A_T7-mcyD-IV-10
Taxon
Microcystis aeruginosa NIES-843 DNA, complete genome
Microcystis wesenbergii AICB702 polyketide synthase (mcyD) gene, partial cds
Microcystis aeruginosa AICB682 polyketide synthase (mcyD) gene, partial cds
Microcystis aeruginosa AICB682 polyketide synthase (mcyD) gene, partial cds
Microcystis aeruginosa PCC 7806 genome sequencing data, contig C322
Microcystis aeruginosa AICB682 polyketide synthase (mcyD) gene, partial cds
Microcystis aeruginosa PCC 7806 genome sequencing data, contig C322
Microcystis aeruginosa PCC 7806 genome sequencing data, contig C322
Max.
Identity (%)
≥95%
97%
91%
93%
96%
97%
97%
96%
Figure 4. mcyD closest GenBank relatives
Figure 7. mcyA nucleic acid neighbor-joining phylogenetic tree
•Cyanobacterial blooms release
toxins called microcystins that are
toxic to plants, animals, and humans
Conclusions
•Sequences fell within two lines of
descent: Planktothrix and Microcystis
aeruginosa
Methods
•DNA was extracted from Lake Menomin
water that contained cyanobacteria.
•Microcystin genes mcyA and mcyD
were amplified using the polymerase
chain reaction (PCR).
Figure 5. mcyD nucleic acid neighbor-joining phylogenetic tree
•This suggests low diversity in the
cyanobacterial community
•Controlling cyanobacterial populations
may be easier than in a highly diverse
community
•PCR product was ligated into plasmids
•After purifying plasmid DNA, genes
were sequenced (Fig. 3a), compiled (Fig.
3b), and compared to closest relatives in
the GenBank database (Fig. 4).
•Neighbor Joining phylogenetic trees
were constructed from nucleic acid
(Figs. 5, 7) and amino acid (Fig. 6) data.
Accession No.
AP009552.1
HM132461.1
HM132458.1
HM132458.1
AM778952.1
HM132458.1
AM778952.1
AM778952.1
Reference
Hotto, A.M. et. al. Molecular characterization of
potential microcystin-producing cyanobacteria in Lake
Ontario embayments and nearshore waters. Applied
and Environmental Microbiology 73 (2007): 4570-4578.
Figure 6. mcyD amino acid neighbor-joining phylogenetic tree