Open Access

Genomics insights into production of 2-methylisoborneol and a putative cyanobactin by Planktothricoides sp. SR001

  • Shu Harn Te1,
  • Boon Fei Tan2,
  • Chek Yin Boo1,
  • Janelle Renee Thompson2, 3 and
  • Karina Yew-Hoong Gin1, 4Email author
Standards in Genomic Sciences201712:35

https://doi.org/10.1186/s40793-017-0247-1

Received: 22 December 2016

Accepted: 26 May 2017

Published: 5 June 2017

Abstract

Planktothricoides is a free-living filamentous cyanobacterium belonging to the order Oscillatoriales and the family Phormidiaceae, capable of forming bloom in fresh and brackish waters. A unicyanobacterial non-axenic culture dominated by Planktothricoides sp. SR001 was obtained from a freshwater reservoir in Singapore. The draft genome presented here is the first tropical freshwater Planktothricoides sp. ever sequenced. The genome of 7.0Mbp contains 5,776 genes predicted using the JGI IMG pipeline. The whole genome sequence allows identification of genes encoding for nitrogen-fixation, accessory photosynthetic pigments and biosynthesis of an off-flavor compound, 2-methylisoborneol, which has been experimentally verified here based on metabolite detection. In addition, strain SR001 genome contains an operon putatively involved in the production of a linear tripeptide cyanobactin related to viridisamide A and aeruginosamide, with the later known to possess anti-microbial or cytotoxic effect.

Keywords

Phormidiaceae Planktothricoides 2-methylisoborneol Viridisamide A Cyanobactin

Introduction

Managing cyanobacterial blooms is a growing concern worldwide due to increasing anthropogenic pollution and climate change that lead to eutrophication of marine, estuarine and fresh waters [1, 2]. Secondary metabolites produced by cyanobacteria are one of the emerging pollutants causing environmental degradation, economic losses and negative impacts on drinking and recreational waters [35]. Amongst the metabolites, odiferous terpenes are commonly detected in many cyanobacterial species, for example geosmin and 2-MIB which give earthy and muddy smells, are responsible for most of the taste and odor issues for water resources and subverting consumers’ confidence on the safety of treated water [6, 7]. Cyanobacterial toxins such as microcystins, cylindrospermopsins and saxitoxins, produced via the nonribosomal peptide synthetase or polyketide synthase, have been shown to cause intoxication cases in livestock and human [8]. Other than non-ribosomal peptides and polyketides, some cyanobacteria also produce bioactive compounds such as cyanobactins via post-ribosomal peptide synthesis.

Many cyanobacterial genera, such as Aphanizomenon , Oscillatoria , Phormidium , Lyngbya , Pseudanabaena , Planktothrix and Planktothricoides identified as the common off-flavor producers [6, 9], are also commonly found in freshwater bodies in Singapore. Planktothricoides is a bloom-forming planktonic-filamentous cyanobacterium which occurs naturally in freshwater and estuarine aquatic systems [10]. The genus was originally classified as Planktothrix under the family of Phormidiaceae due to their high morphological similarity; but it was later designated as a new genus because they are phylogenetically distinct cyanobacteria based on the 16S rRNA gene analysis [11]. Planktothricoides spp. have been occasionally detected in cyanobacterial blooms, either as dominating or co-occurring taxa [1214]. They can produce taste-and-odor compounds (e.g. 2-MIB) and substances toxic to aquatic biota [15]. We report here the first draft genome of Planktothricoides sp. (SR001) which was isolated from a Singapore freshwater reservoir to facilitate molecular and physiological characterizations for a better understanding of their ecological roles in aquatic ecosystems for future study.

Organism Information

Classification and features

Planktothricoides sp. SR001 examined in this study was isolated from a reservoir located at the north-east part of Singapore. The reservoir receives water from its catchment with a mixed land use comprised of residential, industry and reserved lands. The water body was under eutrophic or hypereutrophic state (Carlson trophic index 63-75) characterized with high levels of chl-a and total phosphorus [16]. Two off-flavor compounds, 2-MIB and geosmin, exhibited concentration range from undetectable to 53.1 ng/L in reservoir water, frequently exceeded the olfactory thresholds of 4 – 10 ng/L for drinking water [17]. The phytoplankton community was dominated by cyanobacteria including genera capable of odor synthesis such as Pseudanabaena , Planktothrix and Planktothricoides [18, 19]. Isolation attempts were carried out to capture species responsible for off-flavor production. To obtain unicyanobacterial culture, grab water samples collected from the reservoir were examined under an inverted microscope (Leica DFC450 C) to identify target cyanobacteria. Filaments of Planktothricoides were picked using a sterile pipette and washed with sterile water before transferring into nutrient-enriched MLA medium [20]. After multiple transfers, a unicyanobacterial culture containing Planktothricoides as the dominant species was obtained for morphological and genomic characterization.

Morphological identification of Planktothricoides sp. SR001 was determined based on the common morphology characteristics for Phormidiaceae family (Table 1), i.e. filaments are solitary, straight, free-floating and unbranched; cells in the filament have cylindrical shape; are shorter than wide and similar in shape [21]. This was followed by intergeneric identification based on phenotypic features of which the end of the trichome (filament of plankton) is attenuated and without calyptra (Fig. 1), differentiating Planktothricoides from the genus Planktothrix [10]. The average filament length was 282.4 (±93.9) μm; width of individual cell was 8.13 (±0.92) μm and length was 5.48 (±2.01) μm; and cell width to length ratio was 1.68 (± 0.65) μm. Gas vesicles were spread along the filament near the edge of the cell. The single copy 16S rRNA gene of 1497 bp (locus tag:AM228_RS28415) identified for strain SR001 is >99% identical to those in different strains of Planktothricoides raciborskii (e.g., strain NIES-207, NR_040858.1), and form a congruent monophyletic clade with other Planktothricoides strains but is distinctive from clades containing Planktothrix spp., Arthrospira spp. and Oscillatoria spp. (Fig. 2).
Table 1

Classification and general features of Planktothricoides strain SR001 [38]

MIGS ID

Property

Term

Evidence codea

 

Classification

Domain bacteria

TAS [39]

  

Phylum Cyanobacteria

TAS [40]

  

Class Cyanophyceae

TAS [41]

  

Order Oscillatoriales

TAS [42]

  

FamilyPhormidiaceae

TAS [21]

  

Genus Planktothricoides

TAS [11]

  

Species Unknown

 
  

Strain SR001

 
 

Gram stain

Negative

TAS [43]

 

Cell shape

Filamentous / thallous

IDA

 

Motility

Motile / free-floating

IDA

 

Sporulation

Not reported

 
 

Temperature range

Not reported

 
 

Optimum temperature

Not reported

 
 

pH range; Optimum

Not reported

 
 

Carbon source

Phototrophic

IDA

MIGS-6

Habitat

Freshwater

IDA

MIGS-6.3

Salinity

0.015% NaCl (w/v)

 

MIGS-22

Oxygen requirement

Aerobic

IDA

MIGS-15

Biotic relationship

Free-living

IDA

MIGS-14

Pathogenicity

Non-pathogen

IDA

MIGS-4

Geographic location

North-eastern region, Singapore

IDA

MIGS-5

Sample collection

March 2014

IDA

MIGS-4.1

Latitude

1.401577

IDA

MIGS-4.2

Longitude

103.927010

IDA

MIGS-4.4

Altitude

Not Applicable

 

aEvidence codes - IDA: Inferred from Direct Assay; TAS: Traceable Author Statement (i.e., a direct report exists in the literature); NAS: Non-traceable Author Statement (i.e., not directly observed for the living, isolated sample, but based on a generally accepted property for the species, or anecdotal evidence). These evidence codes are from the Gene Ontology project [44]

Fig. 1

Photomicrograph of a trichome of Planktothricoides sp. SR001 under bright field and 40× magnification. Cell attenuation was observed at the end of trichome

Fig. 2

Neighbor joining tree of the 16S rRNA gene of strain SR001 and selected 16S rRNA sequences of the Oscillatoriales. All 16S rRNA sequences were aligned using MUSCLE [45], manually curated, following which neighbor joining tree was constructed using the Tamura-Nei Model. Bootstrap values are labelled in each branch node and the 16S rRNA of Hapalosiphon welwitschii UH IC-52-3 was used as the outgroup

Extended feature descriptions

Strain SR001 was sub-cultured through multiple transfers in MLA medium containing 6.2 mg phosphorus and 28 mg nitrogen per liter of medium. Incubation was conducted in a plant growth chamber (Percival) at 25 °C with light intensity 24.5 ± 2.0 μmol photons m-2s-1 and a dark/light cycle of 12 hours. The growth rate of strain SR001 was monitored spectrophotometrically with optical density at 680 nm, and also with biomass inferred from chl-a concentration. Both measurements demonstrated similar growth rates of 0.12 day-1 as illustrated in Fig. 3. The presence of accessory photosynthetic pigments commonly found in cyanobacteria including PC, APC and PE were assessed using methods described previously [22]. All three phycobilin pigments including PE, which are not found in earlier study of Planktothricoides [11], were detected in the late-exponential-phase culture with a PC:APC:PE:chl-a ratio of 1.4:2.8:0.7:1.0. Biochemical analysis of two cyanobacterial toxins, microcystins and cylindrospermopsin, were tested negative for strain SR001 using commercial ELISA kits (Abraxis, LLC). However, metabolite profiling using a GC-MS/MS triple quadrupole system (Agilent 7000 GC QQQ) with automated SPME extraction [7] detected 2-MIB but not geosmin during culture growth. Laboratory experiments were conducted to investigate the effects of environmental variables on strain SR001, as studies have shown that light intensity and temperature could alter the off-flavor production rates of cyanobacteria [23, 24]. Triplicate cultures cultivated under different light and temperature conditions were sampled three times during exponential phase, and 2-MIB concentration and cell biovolume were measured. It is worth noting that the culture of strain SR001 was able to tolerate a wide range of light intensity and temperature differences – from 10 to 100 μmol photons m-2 s-1 and from 18 to 38 °C. Significant reduction in 2-MIB was observed when light intensity increased from 10 to 50 μmol photons m-2 s-1 as illustrated in Fig. 3B. A 2-fold decrease in 2-MIB per biovolume was detected when light intensity doubled (independent T-test, P < 0.05) but no further decrease was found thereafter. In contrast, no significant difference (independent T-test, P > 0.05) in 2-MIB production was observed for the temperature range of 25 – 38 °C (Fig. 3c), indicating that the effect of temperature on the 2-MIB content per biovolume of strain SR001 was negligible.
Fig. 3

a Growth curves of Planktothricoides sp. SR001 fitted with dose-response model. Estimated growth rates are 0.00524 h-1 and 0.00513 h-1 based on optical density and chl-a, respectively; b Changes of 2-MIB content for Planktothricoides sp. SR001 under different light intensity and, c temperature conditions. 2-MIB contents are normalized to cell biovolume

Genome sequencing information

Genome project history

The project information and the associated MIGS 2.0 compliance [25] are provided in Table 2. Strain SR001 was selected for sequencing because it is capable of producing 2-MIB, an off-flavor which is known to reduce water palatability. Furthermore, the genome of Planktothricoides is currently underrepresented in public database. This work provides a standard draft genome, of which the assembled contigs have been deposited in NCBI database under the accession LIUQ00000000.
Table 2

Project information

MIGS ID

Property

Term

MIGS 31

Finishing quality

Draft

MIGS-28

Libraries used

Illumina Truseq Nano DNA Library Prep Kit

MIGS 29

Sequencing platforms

HiSeq Rapid V2 sequencing Run

MIGS 31.2

Fold coverage

150

MIGS 30

Assemblers

CLC Genomics Workbench 8.0

MIGS 32

Gene calling method

Prodigal

 

Locus Tag

AM228

 

Genbank ID

LIUQ00000000

 

GenBank Date of Release

August 31, 2015

 

GOLD ID

Ga0099329

 

BIOPROJECT

PRJNA29364

MIGS13

Project relevance

Cyanobacterial ecology, Environmental

Growth conditions and genomic DNA preparation

Our laboratory observation demonstrated that Planktothricoides sp. SR001 was able to grow in nitrogen-free MLA media, consistent with the genus of Oscillatoria in the same Order (Oscillatoriales) [26]. However, this might not be a generic physiological feature for all Planktothricoides as studies have shown that nitrogen fixing capacity is strain-dependent for Planktothrix , another member of Oscillatoriales [27, 28]. Individual trichomes were grown in nitrogen-free MLA media to select against non-nitrogen-fixing species. Strong association of strain SR001 with co-occurring heterotrophic bacteria resulted in a non-axenic unicyanobacterial culture which was maintained in MLA media incubated at 25 °C with a light intensity of 20 ± 5 μmol photons m-2s-1. Total DNA was isolated from the culture fluid using MO BIO PowerWater DNA Isolation Kit (MO BIO), following which the DNA quality and concentration were determined using Qubit 3.0 (Invitrogen).

Genome sequencing and assembly

The total isolated DNA was used in the construction of a paired-end library using a Illumina TruSeq Nano DNA Library Prep Kit with an insert size of 550 bp, and subsequently sequenced with Illumina HiSeq 2000 applying the 250 bp paired-end sequencing protocol at Singapore Centre for Environmental Life Sciences Engineering. Adaptors and reads with quality score <0.01 and length <150 bp were removed using CLC Genomics Workbench V.8 (CLC-Bio, USA), yielded 9,839,009 paired-reads with average read length of 251 bp. The reads were then subjected to de novo assembly with CLC Genomics Workbench V8.0 using default kmer size. The mini-metagenome was assembled into 5,572 scaffolds (764 - 1,110,006 bp) with mean lengths (N50) of 86,064 bp and average length of 8,294 bp. The genome of strain SR001 was extracted from this mini-metagenome using MetaBAT [29], after which the extracted genome was confirmed for completeness and purity using CheckM [30]; thus revealing that the genome has 100% coverage of single copy genes and no evidence for sequence contamination or intra-strain genomic heterogeneity.

Genome annotation

Gene prediction was performed using Prodigal [31] as part of the Joint Genomic Institute IMG automated genome annotation pipeline [32] and the NCBI Prokaryotic Genome Annotation Pipeline [33]. Additionally, gene clusters encoding secondary metabolite biosynthesis were predicted using AntiSMASH 3.0 [34].

Genome Properties

The draft genome of 43.5% GC is 7.0 Mbp contained in 165 scaffolds (1017 – 297,434 bp; Table 3). The N50 and L50 of the 165 scaffolds are 108,940 and 22, respectively. Annotation using the NCBI Prokaryotic Genome Annotation Pipeline [33] predicted 5,776 total genes (Table 3). Complete genome statistics and COG annotation of protein coding genes are presented in Tables 3 and 4, respectively.
Table 3

Genome statistics

Attribute

Value

% of total

Genome size (bp)

7,066,705

100.0

DNA coding (bp)

5,499,857

77.8

DNA G + C (bp)

3,066,802

43.4

DNA scaffolds

165

-

Total genes

5,776

100.0

Protein coding genes

5,049

87.4

RNA genesa

53

0.9

Pseudo genesa

676

11.7

Genes with function predictionb

3,816

64.1

Genes assigned to COGsb

2,714

45.6

Genes with Pfam domainsb

4,079

68.6

Genes with signal peptidesb

159

2.7

Genes with transmembrane helicesb

1,204

20.2

CRISPR repeatsa

12

-

aGenome statistics obtained using the NCBI Prokaryotic Genome Annotation Pipeline [33]

bGenome statistics obtained using the JGI IMG pipeline [32]

Table 4

Number of genes associated with general COG functional categories

Code

Value

%age

Description

J

189

6.33

Translation, ribosomal structure and biogenesis

A

1

0.03

RNA processing and modification

K

94

3.15

Transcription

L

52

1.71

Replication, recombination and repair

B

2

0.07

Chromatin structure and dynamics

D

32

1.07

Cell cycle control, Cell division, chromosome partitioning

V

120

4.02

Defense mechanisms

T

241

8.07

Signal transduction mechanisms

M

212

7.1

Cell wall/membrane biogenesis

N

40

1.34

Cell motility

U

32

1.07

Intracellular trafficking and secretion

O

151

5.05

Posttranslational modification, protein turnover, chaperones

C

137

4.59

Energy production and conversion

G

116

3.88

Carbohydrate transport and metabolism

E

197

6.59

Amino acid transport and metabolism

F

72

2.41

Nucleotide transport and metabolism

H

185

6.19

Coenzyme transport and metabolism

I

75

2.51

Lipid transport and metabolism

P

157

5.25

Inorganic ion transport and metabolism

Q

58

1.94

Secondary metabolites biosynthesis, transport and catabolism

R

479

16.03

General function prediction only

S

222

7.43

Function unknown

-

3236

54.39

Not in COGs

The total is based on the total number of protein coding genes in the genome COG was obtained from the JGI IMG pipeline [32]

Insights from the genome sequence

Planktothricoides is an important cyanobacterial species as several members of this genus are known to produce taste-and-odor compounds, as well as toxins that are harmful to aquatic biota [15]. Using antiSMASH 3.0.5 [34], complete gene clusters encoding biosynthesis of 2-MIB was detected in the genome of strain SR001 (Fig. 4A). In addition, a putative cyanobactin gene cluster with identical gene organization to the reference of viridisamide A was also found in the genome (Fig. 4a). No other toxins/off-flavors – i.e. microcystin, geosmin and cylindrospermopsin genes were detected using antiSMASH or tBLASTn using reference genes. The 2-MIB biosynthesis gene cluster contains homologous cnbA, mtf, mic, and cnbB genes [AM228_RS20060 to AM228_RS20075] with amino acid similarity of 85-93% compared to those detected in Planktothricoides raciborskii CHAB3331 (HQ830028). This finding is consistent with the detection of 2-MIB metabolite in strain SR001 culture fluids (Fig. 3b and c).
Fig. 4

a Gene cluster in the genome of strain SR001 encoding biosynthesis of a putative cyanobactin related to viridisamide A. The patABCDEFG labelled for viridisamide A was named according to Fig. 4 a [35], and their predicted functions are listed in Table 5; b Alignment of cyanobactin precursor peptide according to [35]. Conserved motif LAELSEE is underlined, whereas conserved variable regions that are cleaved to form final cyanobactin are boxed

The putative viridisamide A gene cluster detected in strain SR001 contains eight genes with >60% amino acid similarity to those in the viridisamide A gene cluster first described for Oscillatoria nigro-viridis PCC 7112 [35]. Viridisamide A is a linear tripeptide (cyanobactin) and the organization of the gene cluster encoding for this cyanobactin is highly identical to that encoding for aeruginosamide identified in Microcystis PCC 9432 [35]. The leader sequences of the precursor peptides of viridisamide A, aeruginosamide and several other cyanobactins are highly conserved, although each sequence uniquely contains more than one variable core region that are modified and cleaved to form the final structural variants [35]. Evidently, the leader sequence of the precursor peptide in strain SR001 (AM228_RS10425) is also highly conserved compared to other cyanobactins and contains the highly-conserved motif LAELSEE in the leader sequence (Fig. 4b and Table 5). The core variable regions of the precursor peptide of strain SR001, however, are distinctive from those of viridisamide A and aeruginosamide (Fig. 4b); thus, suggesting that the cyanobactin produced is likely to be structurally different from the two linear cyanobactins. The final structures of this cyanobactin of strain SR001 is currently unknown. Like the gene clusters encoding viridisamide A and aeruginosamide [35], the putative cyanobactin gene cluster contains genes predicted to encode for thiazoline oxidase adjacent to a predicted c-terminal protease gene, thus suggesting that the cyanobactins of strain SR001 may contain a c-terminal bound to a thiazole. The functions of both viridisamide A and aeruginosamide have not been established.
Table 5

Gene cluster encoding cyanobactins detected in strain SR001 genome

Gene namea

Locus tag

Predicted function

Amino acid identity (%)

Oscillatoria nigro-viridis PCC 7112

Microcystis aeruginosa PCC9432

PatA

AM228_RS

N-terminal protease

90

73

PatB

AM228_RS

Hypothetical

82

92

PatC

AM228_RS

Hypothetical

78

44

PatD

AM228_RS

Heterocyclase

83

88

PatE

AM228_RS

Precursor

84

84

PatF1

AM228_RS

Methyltransferase

76

95

PatF2

AM228_RS

Putative prenyl transferase

73

83

PatG

AM228_RS

C-terminal protease/Thiazoline oxidase

80

75

aNaming according to [35]

Strain SR001 was isolated from a surface water sample and is likely a free-living planktonic cyanobacterial species. The genome carries multiple genes essential for movement within the water column including genes predicted to encode for gas vesicles important for buoyancy regulation [36], and pilus and twitching motility important for photo- and chemotaxis [37]. Energy is primarily derived through photosynthesis, with a predicted capability to harvest a broad spectrum of sunlight with different wavelengths, based on annotation of genes encoding alpha- and beta-subunits of phycocyanin (e.g., AM228_RS09220 and AM228_RS09225) and allo-phycocyanin (e.g., AM228_RS19895 and AM228_RS19900). The presence of different pigmentation likely confers ecological advantage for competitive growth in environments with fluctuating sunlight. Nitrogen is likely derived through N2 fixation, under some circumstances evidenced by annotation of multiple nitrogenase genes in the genome (e.g., AM228_RS02340, AM228_RS22395), and growth in nitrogen-free media. Genes for utilization of additional nitrogen sources are predicted in the genome including ammonium [ammomium transporters, e.g., AM_SR11610], urea [urease (e.g., ureABCDEF, AM228_RS18855 to RS18880); urea transporter (RS18565 to RS1885650)] and nitrate (nitrate transporters, e.g., AM228_RS00860), indicating the strain is versatile in utilizing different nitrogen sources.

Conclusions

This first draft genome sequence of Planktothricoides sp. will facilitate genetic insights into the genus of Planktothricoides which is currently under-described. The bioinformatic analysis revealed gene clusters encoding for nitrogenases, 2-MIB, PC, APC, which are in agreement with experimental data or physiological observations. In addition, a putative cyanobactin, likely related to viridisamide, was detected in the genome. Presence of genes encoding for nitrate, ammonia and urea transporters together with nitrogenases indicate that strain SR001 has evolved a variety of strategies that allow them to grow with different nitrogen sources. The genome presented here enables sequence analysis and comparative genomics to drive further research on the ecology and physiology of cyanobacterial strains that may impact water quality.

Abbreviations

2-MIB: 

2-methylisoborneol

APC: 

Allo-phycocyanin

chl-a: 

Chlorophyll-a

L50: 

Number of scaffolds whose summed length is N50

N50: 

Mean of contig/scaffold lengths

PC: 

Phycocyanin

PE: 

Phycoerythrin

Declarations

Acknowledgements

We thank Maxine Allayne Darlene Mowe from National University of Singapore for isolating the strain, PUB for their support in logistic arrangement and NUS Environmental Research Institute (NERI) for their administrative support.

Funding

This research grant is supported by the Singapore National Research Foundation under its Environmental & Water Technologies Strategic Research Programme and administered by PUB, the Singapore’s National Water Agency (Grant number: 1102-IRIS-14-02). BFT and JRT were supported by the National Research Foundation Singapore through the Singapore MIT Alliance for Research and Technology’s (SMART) Center for Environmental Sensing and Modeling (CENSAM) research program.

Authors’ contributions

SHT participated in the design of study, culture characterization and drafted the manuscript. BFT participated in the design of study, genome assembly, sequence data analysis and drafted the manuscript. CYB participated in culture characterization and conducting control experiment. KYG and JRT participated in the design of the study, edited the manuscript and supervised the work. All authors read and approved the final manuscript.

Competing interests

The authors declare that they have no competing interests.

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Authors’ Affiliations

(1)
NUS Environmental Research Institute, National University of Singapore
(2)
Singapore Centre for Environmental Sensing and Modelling, Singapore-MIT Alliance for Research and Technology Centre
(3)
Department of Civil and Environmental Engineering, Massachusetts Institute of Technology
(4)
Department of Civil and Environmental Engineering, National University of Singapore

References

  1. O’Neil JM, Davis TW, Burford MA, Gobler CJ. The rise of harmful cyanobacteria blooms: The potential roles of eutrophication and climate change. Harmful Algae. 2012;14:313–34.View ArticleGoogle Scholar
  2. Paerl HW, Paul VJ. Climate change: Links to global expansion of harmful cyanobacteria. Water Res. 2012;46:1349–63.View ArticlePubMedGoogle Scholar
  3. Chorus I, Bartram J. Toxic cyanobacteria in water : a guide to their public health consequences, monitoring, and management. 1999. E & FN Spon.View ArticleGoogle Scholar
  4. Chorus I, Falconer IR, Salas HJ, Bartram J. Health risks caused by freshwater Cyanobacteria in recreational waters. J. Toxicol. Environ. Heal. Part B [Internet]. 2000 [cited 2016 Sep 13];34:1093–7404. Available from: http://dx.doi.org/10.1080/109374000436364
  5. Dodds WK, Bouska WW, Eitzmann JL, Pilger TJ, Pitts KL, Riley AJ, et al. Eutrophication of U.S. Freshwaters: Analysis of Potential Economic Damages. Environ. Sci. Technol. [Internet]. American Chemical Society; 2009 [cited 2016 Sep 13];43:12–9. Available from: http://pubs.acs.org/doi/abs/10.1021/es801217q
  6. Wang Z, Xu Y, Shao J, Wang J, Li R. Genes associated with 2-methylisoborneol biosynthesis in cyanobacteria: isolation, characterization, and expression in response to light. PLoS One [Internet]. Public Library of Science; 2011 [cited 2016 Sep 12];6:e18665. Available from: http://www.ncbi.nlm.nih.gov/pubmed/21490938
  7. Tan BF, Te SH, Boo CY, Gin KY-H, Thompson JR, Te S, et al. Insights from the draft genome of the subsection V (Stigonematales) cyanobacterium Hapalosiphon sp. Strain MRB220 associated with 2-MIB production. Stand. Genomic Sci. [Internet]. BioMed Central; 2016 [cited 2016 Sep 12];11:58. Available from: http://standardsingenomics.biomedcentral.com/articles/10.1186/s40793-016-0175-5
  8. Carmichael WW. Human and Ecological Risk Assessment: An International Journal Health Effects of Toxin-Producing Cyanobacteria: &quot; The CyanoHABs &quot; Health Effects of Toxin-Producing Cyanobacteria: &quot; The CyanoHABs &quot; Hum. Ecol. Risk Assess. An Int. J. Hum. Ecol. Risk Assess. [Internet]. 2001 [cited 2016 Sep 13];7:1393–407. Available from: http://dx.doi.org/10.1080/20018091095087.
  9. Watson SB. Cyanobacterial and eukaryotic algal odor compounds: signals or by-products? A review of their biological activity. Phycologia [Internet]. The International Phycological Society Phycologia Business Office, Allen Press, 810 East 10th Street, P.O. Box 1897, Lawrence, KS 66044-8897 ; 2003 [cited 2016 Sep 13];42:332–50. Available from: http://www.phycologia.org/doi/abs/10.2216/i0031-8884-42-4-332.1
  10. Komárek J, Komárková J. Taxonomic review of the cyanoprokaryotic genera Planktothrix and Planktothricoides Taxonomický přehled rodů Planktothrix a Planktothricoides (Cyanoprokaryota). Czech Phycol Olomouc. 2004;4:1–18.Google Scholar
  11. Suda S, Watanabe MM, Otsuka S, Mahakahant A, Yongmanitchai W, Nopartnaraporn N, et al. Taxonomic revision of water-bloom-forming species of oscillatorioid cyanobacteria. Int J Syst Evol Microbiol. 2002;52:1577–95.PubMedGoogle Scholar
  12. Wu X, Jiang J, Wan Y, Giesy JP, Hu J. Cyanobacteria blooms produce teratogenic retinoic acids. Proc. Natl. Acad. Sci. [Internet]. National Academy of Sciences; 2012 [cited 2016 Sep 13];109:9477–82. Available from: http://www.pnas.org/cgi/doi/10.1073/pnas.1200062109
  13. Zhong-Xing W, Bo-Shi Y, Xin P, Gong-Liang Y, Jia-Li Q, (reli@ihb.ac.cn) LR-H. Planktothricoides, a newly recorded genus of water bloom forming cyanophyta in China. Plant Sci J. 2008;26:461-5.Google Scholar
  14. Yamamoto Y, Shiah F-K, Chen Y-L. Importance of large colony formation in bloom-forming cyanobacteria to dominate in eutrophic ponds. Int. J. Limnol. [Internet]. EDP Sciences; 2011 [cited 2016 Sep 13];47:167–73. Available from: http://www.limnology-journal.org/10.1051/limn/2011013
  15. Guidi-Rontani C, Jean MRN, Gonzalez-Rizzo S, Bolte-Kluge S, Gros O. Description of new filamentous toxic Cyanobacteria (Oscillatoriales) colonizing the sulfidic periphyton mat in marine mangroves. FEMS Microbiol. Lett. [Internet]. 2014 [cited 2016 Sep 13];359:173–81. Available from: http://www.ncbi.nlm.nih.gov/pubmed/25088450
  16. Carlson RE. A trophic state index for lakes1. Limnol. Oceanogr. [Internet]. 1977 [cited 2016 Sep 6];22:361–9. Available from: http://doi.wiley.com/10.4319/lo.1977.22.2.0361
  17. Xie Y, He J, Huang J, Zhang J, Yu Z. Determination of 2-Methylisoborneol and Geosmin Produced by Streptomyces sp. and Anabaena PCC7120.Google Scholar
  18. Wang Z, Xu Y, Shao J, Wang J, Li R. Genes associated with 2-methylisoborneol biosynthesis in cyanobacteria: isolation, characterization, and expression in response to light. PLoS One [Internet]. 2011 [cited 2016 Aug 26]; Available from: http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0018665
  19. Su M, Yu J, Zhang J, Chen H, An W, Vogt RD, et al. MIB-producing cyanobacteria (Planktothrix sp.) in a drinking water reservoir: Distribution and odor producing potential. Water Res. 2015;68:444–53.View ArticlePubMedGoogle Scholar
  20. Bolch CJS, Blackburn SI. Isolation and purification of Australian isolates of the toxic cyanobacterium Microcystis aeruginosa Kütz. J. Appl. Phycol. [Internet]. 1996 [cited 2017 Feb 23];8:5–13. Available from: http://link.springer.com/10.1007/BF02186215
  21. Anagnostidis K, Komárek J. Modern approach to the classification system of cyanophytes. 3-Oscillatoriales. Arch Hydrobiol. 1988;80:327–472.Google Scholar
  22. Sobiechowska-Sasim M, Stoń-Egiert J, Kosakowska A. Quantitative analysis of extracted phycobilin pigments in cyanobacteria-an assessment of spectrophotometric and spectrofluorometric methods. J. Appl. Phycol. [Internet]. Springer; 2014 [cited 2016 Sep 13];26:2065–74. Available from: http://www.ncbi.nlm.nih.gov/pubmed/25346572
  23. Zhang T, Li L, Song L, Chen W. Effects of temperature and light on the growth and geosmin production of Lyngbya kuetzingii (Cyanophyta). J. Appl. Phycol. [Internet]. 2009 [cited 2016 Sep 12];21:279–85. Available from: http://link.springer.com/10.1007/s10811-008-9363-z
  24. Li Z, Hobson P, An W, Burch MD, House J, Yang M. Earthy odor compounds production and loss in three cyanobacterial cultures. Water Res. [Internet]. 2012 [cited 2016 Sep 12];46:5165–73. Available from: http://www.ncbi.nlm.nih.gov/pubmed/22818951
  25. Field D, Garrity G, Gray T, Morrison N, Selengut J, Sterk P, et al. The minimum information about a genome sequence (MIGS) specification. Nat Biotechnol. 2008;26:541–7.View ArticlePubMedPubMed CentralGoogle Scholar
  26. Carpenter EJ, Price CC. Marine oscillatoria (Trichodesmium): explanation for aerobic nitrogen fixation without heterocysts. Science (80-.). [Internet]. 1976 [cited 2017 Feb 23];191:1278–80. Available from: http://www.ncbi.nlm.nih.gov/pubmed/1257749
  27. Dolman AM, Rücker J, Pick FR, Fastner J, Rohrlack T, Mischke U, et al. Cyanobacteria and Cyanotoxins: The Influence of Nitrogen versus Phosphorus. Bertilsson S, editor. PLoS One [Internet]. Public Library of Science; 2012 [cited 2017 Feb 25];7:e38757. Available from: http://dx.plos.org/10.1371/journal.pone.0038757
  28. Pancrace C, Barny M-A, Ueoka R, Calteau A, Scalvenzi T, Pédron J, et al. Insights into the Planktothrix genus: Genomic and metabolic comparison of benthic and planktic strains. Sci. Rep. [Internet]. Nature Publishing Group; 2017 [cited 2017 Feb 25];7:41181. Available from: http://www.ncbi.nlm.nih.gov/pubmed/28117406.
  29. Kang DD, Froula J, Egan R, Wang Z, PeerJ I. MetaBAT, an efficient tool for accurately reconstructing single genomes from complex microbial communities. Peer J Inc. 2015;3:e1165.View ArticleGoogle Scholar
  30. Parks DH, Imelfort M, Skennerton CT, Hugenholtz P, Tyson GW. CheckM: assessing the quality of microbial genomes recovered from isolates, single cells, and metagenomes. Genome Res. 2015;25:1043-55. doi:10.1101/gr.186072.114.
  31. Hyatt D, Chen G-L, Locascio PF, Land ML, Larimer FW, Hauser LJ. Prodigal: prokaryotic gene recognition and translation initiation site identification. BMC Bioinfo. 2010;11:119.View ArticleGoogle Scholar
  32. Markowitz VM, Chen I-MA, Palaniappan K, Chu K, Szeto E, Pillay M, et al. IMG 4 version of the integrated microbial genomes comparative analysis system. Nucleic Acids Res. 2014;42:D560–7.View ArticlePubMedGoogle Scholar
  33. Tatusova T, DiCuccio M, Badretdin A, Chetvernin V, Ciufo S, Li W. Prokaryotic Genome Annotation Pipeline. National Center for Biotechnology Information (US); 2013.Google Scholar
  34. Weber T, Blin K, Duddela S, Krug D, Kim HU, Bruccoleri R, et al. antiSMASH 3.0--a comprehensive resource for the genome mining of biosynthetic gene clusters. Nucleic Acids Res. 2015;43:W237–43.View ArticlePubMedPubMed CentralGoogle Scholar
  35. Leikoski N, Liu L, Jokela J, Wahlsten M, Gugger M, Calteau A, et al. Genome Mining Expands the Chemical Diversity of the Cyanobactin Family to Include Highly Modified Linear Peptides. Chem Biol. 2013;20:1033–43.View ArticlePubMedGoogle Scholar
  36. Walsby AE. Gas vesicles. Microbiol Rev Am Soc Microbiol. 1994;58:94–144.Google Scholar
  37. Schuergers N, Nürnberg DJ, Wallner T, Mullineaux CW, Wilde A. PilB localization correlates with the direction of twitching motility in the cyanobacterium Synechocystis sp. PCC 6803. Microbiology. 2015;161:960–6.View ArticlePubMedGoogle Scholar
  38. Field D, Amaral-Zettler L, Cochrane G, Cole JR, Dawyndt P, Garrity GM, et al. The Genomic Standards Consortium. PLoS Biol. [Internet]. Public Library of Science; 2011 [cited 2016 Nov 23];9:e1001088. Available from: https://doi.org/10.1371/journal.pbio.1001088
  39. Woese CR, Kandlert O, Wheelis ML. Towards a natural system of organisms: Proposal for the domains Archaea, Bacteria, and Eucarya. Evolution. 1990;87:4576–9.Google Scholar
  40. STANIER RY, SISTROM WR, HANSEN TA, WHITTON BA, CASTENHOLZ RW, PFENNIG N, et al. Proposal to Place the Nomenclature of the Cyanobacteria (Blue-Green Algae) Under the Rules of the International Code of Nomenclature of Bacteria. Int. J. Syst. Evol. Microbiol. [Internet]. Microbiology Society; 1978 [cited 2017 Feb 27];28:335–6. Available from: http://www.microbiologyresearch.org/content/journal/ijsem/10.1099/00207713-28-2-335
  41. Schaffner JH. The Classification of Plants, IV. Ohio J Sci Acad Sci. 1909;9:446–55. Available from: https://kb.osu.edu/dspace/bitstream/handle/1811/1629/V09N04_446.pdf;jsessionid=6C85E8297FBD3CC2BDED7CA2616DE9B4?sequence=1.Google Scholar
  42. Cavalier-Smith T. The neomuran origin of archaebacteria, the negibacterial root of the universal tree and bacterial megaclassification. Int J Syst Evol Microbiol. 2002;52:7–76.View ArticlePubMedGoogle Scholar
  43. Hoiczyk E, Hansel A. Cyanobacterial Cell Walls: News from an Unusual Prokaryotic Envelope. J. Bacteriol. [Internet]. American Society for Microbiology; 2000 [cited 2016 Sep 12];182:1191–9. Available from: http://jb.asm.org/cgi/doi/10.1128/JB.182.5.1191-1199.2000
  44. Ashburner M, Ball CA, Blake JA, Botstein D, Butler H, Cherry JM, et al. Gene ontology: tool for the unification of biology. The Gene Ontology Consortium. Nat. Genet. [Internet]. NIH Public Access; 2000 [cited 2016 Nov 23];25:25–9. Available from: http://www.ncbi.nlm.nih.gov/pubmed/10802651
  45. Edgar RC. MUSCLE: multiple sequence alignment with high accuracy and high throughput. Nucleic Acids Res. 2004;32:1792–7.View ArticlePubMedPubMed CentralGoogle Scholar

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