biochemical screening and lipid profiling from cyanobacterial...
TRANSCRIPT
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Biochemical screening and lipid profiling from
cyanobacterial diversity of rice fields of Manipur
4.1. Introduction
Cyanobacteria are photosynthetic prokaryotes possessing the ability to synthesize
chlorophyll-a besides their ability to form the phycobilins. Phycobilins are high concentration of
pigment occurring under some conditions which leads to the bluish colour of the organisms and
hence known, cyanobacteria or blue-green algae. They are grouped under the gram-negative
bacteria and their morphology varies from unicellular to multicellular. Some species have unique
cells called heterocysts, which are capable of fixing atmospheric nitrogen. Cyanobacteria are
known to be one of the promising supplements to nitrogenous fertilizer, but the process biological
nitrogen fixation, mediated through the enzyme nitrogenase may be inhibited in presence of
readily available nitrogen sources.
Chlorophyll provides a chelating agent activity which can be used in ointment, treatment
for pharmaceutical benefits especially liver recovery and ulcer treatment. It also repair cells,
increases haemoglobin in blood and faster the cell growth (Puotinen, 2009). Carotenoids are
widely used as natural colourant for food, drug and cosmetic products. They served as a second
line density protein. Cyanobacteria, in general possess all the known phycobiliproteins
(phycocyanin, phycoerythrin, phycoerythrocyanin and allophycocyanin). Among them,
phycocyanin and phycoerthrin are commercially valuable. Phycocyanin is used in water-
insoluble, dairy products and soft drinks (Cohen, 1986). Cyanobacteria are considered as a system
of photo-production of ammonia by their diazoptrophic potential. Mutant strains of Anabaena
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variabilis fixed N2 and liberated NH4+ into the media (Kerby et al., 1986). The high content
protein indicates relatively good amino acid profile and low metal content enabled the use of algal
biomass as feed supplement. The use of microalgae in industry encourage the development of
better cultivation system in order to optimize the production of algae rich in active substances
such as vitamins, protein, amino acids, fatty acids and trace elements.
Fatty acids, in general are of commercial value and many are pharmaceutical agents
(Tanticharoen et al., 1994). The cyanobacterium Spirulina has commercially exploited in several
countries (Benemann, 1988; Venkataraman and Becker, 1985). It has been used as food for many
centuries in central America. For the past two decades, Spirulina platensis has been a focus of
interesting among researchers in various fields because of its commercial importance as a source
of proteins, vitamins, essential amino acids and fatty acids (Ciferri and Tiboni, 1985; Vonshak
and Richmond, 1988; Vonshak, 1990; Tanticharoen et al., 1994) and more recently, for its
potential in therapeutic effects (Amha Belay et al., 1993).
The merits of an organism for commercial exploitation are maximum yield and utility of
cellular constituents (Borowitzka and Borowitzka, 1988). Higher growth rate and nutrient profile
of cyanobacteria make them a potentially valuable source of nutrients (Cannell, 1989). Growth of
a living organism is defined as an increase in mass or size accompanied by synthesis of
macromolecules, leading to the production of newly organized structure. Microbiologists use a
variety of techniques to quantify microbial growth other than determining direct cell count. These
include the measurement of macromolecules in the cell (Healy and Henzdel, 1976), the cell quota
of specific elements (Rhee and Gotham, 1980) or the kinetic parameters for nutrient uptake
(Zevenboom et al., 1982). Although relative incorporation, into protein was constant, the absolute
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rate of protein incorporation increased at higher growth rate because the photosynthetic rate
increased. Chlorophyll-a is another component of biomass, which can be estimated as a measure
of growth (Kobayasi, 1961).
Venkataraman and Mahadevaswamy (1992) pointed out that good culture management
with suitable strain is one of the basic needs to get promising yields with quality material on
commercial scale. Therefore, cultivation techniques are to be improved with the main objective of
obtaining higher algal biomass that exhibits specific qualities (Lobban and Herrison, 1994). Mass
cultivation of cyanobacteria is essentially a complex process involving a large number of
variables for successful growth of essential requirements of the organism as possible. The
limitations imposed in the cultivation process be due to physical factors like light, nutrients,
temperature, pH and physiological (organism-environmental-interrelationship) and economic
constraints.
The environmental factors may be either physiological such as acidity and pH or
chemical which provide all the raw materials used for structural cells and protoplasmic synthesis
of cyanobacterial cells (Becker, 1994). Physical and chemical factors such as temperature, acidity
and light (Lobban and Herrison, 1994), aeration (Chen and Johns, 1991) or nutrient concentration
(Bjornsater and Wheeler, 1990) influence the biochemical composition, physiological status and
ultrastructure of the cyanobacteria. Culture medium has been found to play a significant role in
the growth kinetic of algae, since it has to stimulate the natural conditions as closely as possible.
Cyanobacteria are known to exhibit a wide adaptability to pH, but for mass cultivation of
cyanobacteria, it is essential to determine their optimal conditions.
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4.2. Materials and methods
Biochemical composition: Screening for chlorophyll-a, total soluble proteins, total
carbohydrates, phycobiliproteins, carotenoids, ammonia excretion and ARA activity were
investigated in 15th
days and 30th
days old cultures of all heterocystous cyanobacteria. Non
heterocystous cyanobacteria were executed for lipids profiling and fatty acid composition
accordingly.
4.2.1. Preparation of inoculums: A loopful cyanobacterial biomass were inoculated in 250 ml
conical flasks and incubated in culture room at 28±2⁰C of 4KLux light intensity provided by
white fluorescent tubes 14/10 light dark phase. 15 days old biomass homogenized uniformly.
5 ml well homogenized cyanobacterial inoculum of each strain was inoculated into 250 ml
conical flasks with 100 ml culture medium in duplicate for 15th
day and 30th
day investigation.
Every day agitation was done to avoid clumping of the cells and for faster growth.
4.2.2. Biochemical estimation of chlorophyll-a: Estimation of chlorophyll-a was determined by
adapting the method described by Mckinney (1941). Cyanobacteria possess chlorophyll-a and it
is important for evaluation of growth and photosynthetic rate and participate directly in the light
requiring reactions of photosynthesis. 10 ml of homogenized algal suspension was taken in
centrifuge tube and done centrifugation at 7000 rpm for 10 mins and then discarded the
supernatant and transferred the algal pellet to a test tube and added 10 ml 90% methanol. Shaked
the contents and placed the tubes covered with aluminium foil in a water bath at 60˚C for 30 mins.
The absorbance from supernatant was measured at 665 nm against methanol blank. Calculation
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was done by using the following equation, chl-a (µg/ml) = (O.D.665 x 13.42) where, 13.42 is the
extinction co-efficient at 665 nm.
4.2.3. Biochemical estimation of total soluble proteins: Estimation of total soluble proteins was
determined by the method described by Herbert et al. (1971). Proteins make up large fraction of
actively growing cyanobacteria and have vast applicability for using as value added products and
other pharmaceutical applications. 0.5 ml homogenized algal suspension was taken and added 0.5
ml of solution NaOH (40 gm/1000 ml=1N) then incubated in water bath for 15 mins at 100°C.
Cooled in running tap water and added 2.5 ml of 5% Na2CO3=5 gm/100 ml and 0.5%
CuSO4.5H2O in 1% sodium potassium tartarate. Finally added 0.5 ml of 1:1 Folin Ciocalteu’s
reagent and shaked the content well in a vortex shaker and incubated at room temperature for 15
mins.
Standard curve preparation: Dissolved 10 mg
BSA in a standard flask and make up upto 100
ml (conc.100 µg/ml). 5 different protein
concentrations were taken along with a blank
solution. The blue absorbance was measured at
650 nm against blank. A standard curve having
X-axis as concentration of protein in µg/ml and
Y-axis as absorbance (O.D.) was drawn and found out the concentration of unknown sample from
the optical density obtained.
Fig.-4: Standard graph for total soluble proteins
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4.2.4. Biochemical estimation of total carbohydrates: Total sugar was estimated following the
method described by Spiro (1966). 0.2 ml of the homogenized algal cell suspension was taken and
added 0.8 ml of distil water and 4 ml anthrone reagent (dissolved 0.1 g of anthrone and 1.0 g of
thiourea in freshly prepared 100 ml of 75% sulphuric acid) and shaken gently. Tubes were kept in
a boiling water bath for 15 min by covering the mouth of tube with aluminium foil to prevent
evaporation. After cooling the tubes the absorbance was measured at 620 nm against blank and
total sugar content was calculated from the reference of standard graph.
Standard graph of total carbohydrates: Standard
graph was prepared with different concentration of
glucose i.e. 10-100 µg/ml following the method of
Spiro (1966). The optical density and concentration of
glucose were plotted on Y-axis and X-axis respectively
to get the standard graph and the corresponding
concentration of carbohydrates content of unknown
algal culture was calculated from the graph.
4.2.5. Biochemical estimation of phycobiliproteins: Phycobiliproteins are a family of highly
soluble and reasonably stable fluorescent proteins. These proteins contain covalently linked
tetrapyrrole groups that play a biological role in collecting light and through fluorescence
resonance energy transfer, conveying it to a special pair of chlorophyll molecules located in the
photosynthetic reaction center and also serve for nitrogen storage of cyanobacteria. Estimation of
phycobiliproteins was determined by the method described by Bennett and Bogorad (1973). 10 ml
algal suspension was centrifuged at 7000 rpm for 10 mins. The pellets were suspended in 5 ml
Fig.- 5: Standard graph for total carbohydrates
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phosphate buffer. The contents were repeatedly freezed in 4ºC and thawed at room temperature.
The supernatants were pooled and the absorbance was measured at 562 nm, 615 nm and 652 nm
respectively.
Calculation was done using the following equation:
C-Phycocyanin= (A615-0.474 x A652)/ 5.34; C-Allophycocyanin= (A652-0.208 x A615)/ 5.09
C-Phycoerythrin= (A562-(2.41(PC))-(0.849(APC)/ 9.62
Where 5.34, 5.09 and 9.62 are the extinction co-efficient
4.2.6. Biochemical estimation of total carotenoids: Carotenoids belong to the category of
tetraterpenoids. Structurally they are in the form of a polyene chain. Carotenoids have important
functions in photosynthesis, nutrition and protection against photooxidative damage and hence
used as food colourants and vitamin-A precursor possessing anti-cancer properties. In present
study, estimation of carotenoid was determined by the method described by Jensen (1978). 10 ml
homogenized algal suspension was taken and centrifuged at 6500 rpm for 10 min. Discarded the
supernatant and added 3 ml 85% acetone and subjected to repeat freezing and thawing until the
pellet becomes colourless. Measured the volume of the extract and make up the final volume upto
10 ml with 85% acetone and read the O.D. at 450 nm using 85% acetone as blank and calculated
the total amount of carotenoid in µg/ml.
Calculation: C= (D x V x f) x 10 /2500; Where, D= O. D. at 450nm; V= volume of the extract;
f = dilution factor; average extinction co-efficient of pigment is 2500
4.2.7. Biochemical estimation of ammonia excretion: Ammonia excretion by cyanobacteria is
important to study the nitrogen contribution to the soil i.e. the amount of nitrogen trapped from
the atmosphere. The more ammonia excreted, the more free nitrogen fixed. In present
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investigation, ammonia excretion was determined by the method described by Solorzano, 1969.
05 ml culture filtrate obtained by filtration of homogenized algal suspension filtered through
Whatmann’s filter paper was taken. Added 0.2 ml mixed phenol (2 g of reagent grade phenol
dissolved 100 ml of 95% ethyl alcohol) thereafter added 0.2 ml reagent-A (0.15 g of sodium
nitroprusside dissolved in 30 ml of distilled water and stored in amber colour bottle). Finally 0.5
ml reagent-B (10 g of trisodium citrate and 0.5 g of NaOH in 50 ml of distilled water with 20 ml
of 1.5 N sodium hypochlorite soln), mixed thoroughly with the aid of vortex shaker and kept for
1 hour for development of blue colour in dark place. The absorbance was measured at 640
against blank. Calculation was done in µg/ml = O.D.640 x 300 where 300 is the extinction co-
efficient.
4.2.8. Lipid profiling: In present investigation for lipid profiling from non heterocystous
filamentous cyanobacteria, the method followed as per described by Bligh and Dyer 1959. 15 ml
of 2% H2SO4 in methanol solution was added to total biomass (300 mg) in round bottom (RB)
flask. Refluxed the RB flask containing the biomass in heating mantle at 10°C for 4 hours.
Transferred the FAME solution to a separating funnel then added ethyl acetate and distil water to
the FAME solution contained inside the separating funnel. Two aqueous phase layer is formed.
Then separated out the lower phase and the upper phase was retained and washed with distil water
till it gives a pH 7.0 (checking through pH strip). Separated the extract into a conical flask and put
excess amount of sodium sulphate (Na2SO4) into it and kept for 20 min. Extract was transferred to
50 ml RB flask and rotaevaporated at 65°C. Rinsed the RB flask containing the FAME by putting
few drops of dichloro-methane and transferred the solution to a vial. Took 1µl sample from vial
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using micro syringe and injected in GC. From the standard (SUPELCOTM
37 components) and its
retention time, the fatty acid content was identified.
4.2.9. Determination of acetylene reduction activity: Cyanobacteria have the ability to fix
atmospheric nitrogen by the help of nitrogenase enzyme mandatory present in heterocysts and
sometimes and in specific conditions active in vegetative cells too. Acetylene reduction activity
was determined by the method described by Hardy et al., 1973. A known volume of algal biomass
was taken into 13 ml capacity test tubes. Stopper the tubes and remove the gas phase equivalent to
10% of the remaining volume of the tubes and injected equivalent volume of acetylene (C2H2).
Vials were incubated for 120 min under light conditions (4 Klux) at 28±2°C interval shake was
done. A gas sample of known volume (0.1 to 1.0 ml) was withdrawn with gas tight syringe and
injected into injection port of the gas chromatograph.
Calculations:
(i) amount of ethylene (C2H4) produced=Cmole
(ii) calculation volume of vial=dml
(iii) substract volume of algal sample ‘e’ ml from ‘d’ to get volume of gas phase in vial
volume of gas phase in vial (d-e)=fml
(iv) nmole C2H4/ vial =fxc for 1½ h incubate.
acetylene reduction activity=fxcx60/90xX nmole C2H4/µg chl-a/hr
Statistical analysis: Result were analyzed for standard deviation by using Microsoft office excel
sheet
Reference and equipments used for biochemical characterization and lipid profiling:
SN Materials methods and reference Equipments with brand name
1 Chlorophyll a: Mckinney, (1941) Ø Homogenizer Remi RQ127A
Ø Vortex shaker –Vortexer GeNei
Ø Water bath NSW-125
Ø Spectrophotometer Shimadzu UV-
2 Total soluble protein: Herbert et al.,
(1971) 3 Total sugar content: Spiro, (1966)
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4 Phycobiliproteins: Bennett and
Bogorad, (1973)
1800
Ø Refrigerated centrifuge Eppendorf
5430R
Ø Weighing balance-Mettler Toledo
New Classic MS
Ø GC-FID Thermo scientific Chemito
Ceres 800 plus
Ø Reflux Heating menthel LABCD
India
Ø Rotary evaporator-Hahnvapor HS-
2005V
5 Carotenoids: Jensen, (1978)
6 Ammonia excretion: Solorzano,
(1969) 7 Nitrogenase activity: Hardy et al.,
(1973) 8 Lipid profiling: Bligh and Dyer,
(1959)
4.3. Results
One hundred twelve (112) cyanobacterial strains belong to 16 genera including non
heterocystous (07 genera 40 strains) and heterocystous (09 genera 72 strains) namely;
Myxosarcina (01), Limnothrix (03), Phormidium (16), Oscillatoria (01), Spirulina (01), Lyngbya
(08) Plectonema (10) Cylindrospermum (04), Anabaena (32), Nostoc (13), Aulosira (02),
Scytonema (03), Calothrix (09), Microchaete (06), Dichothrix (02) and Westiellopsis (01) were
isolated from rice fields of Manipur and incorporated in present study. These unialgal
cyanobacterial strains were investigated for chl-a, ammonia excretion, total carotenoids, total
soluble proteins, total carbohydrates and phycobiliproteins of 15th
and 30th
days old growth
(table 7-9). Seventy two (72) heterocystous cyanobacteria were investigated for acetylene
reduction activity of 15th
and 30th
days old growth (table 10); however forty (40) non-
heterocystous strains were deeply characterized for fatty acid composition and lipid profiling
(table-11).
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Table-7: Screening for chl-a and total soluble proteins from cyanobacterial
strains of rice fields of Manipur
Strains name with code no. chlorophyll-a (µg/ml) total soluble protein (µg/ml)
15th
day 30th
day 15th
day 30th
day
Microchaete uberrima BTA001 7.84±1.12 12.3±1.10 27.6±1.50 126.6±10.4
Phormidium mucosum BTA002 5.64±0.24 8.12±0.92 22.6±4.50 130.0±0.01
Anabaena variabilis BTA003 2.32±0.15 4.23±0.31 23.6±1.02 59.6±2.51
Anabaena doliolum BTA004 4.19±0.19 4.80±0.02 89.3±1.92 93.0±1.90
Limnothrix vacuolifera BTA005 3.95±0.75 5.92±0.32 35.6±1.15 98.0±2.30
Anabaena variabilis BTA006 6.19±2.12 8.34±0.12 27.0±1.88 78.0±1.73
Microchaete grisea BTA007 7.02±0.23 10.8±2.04 64.0±1.08 116.6±5.77
Anabaena circinalis BTA008 2.54±0.38 4.35±0.00 3.00±0.00 8.00±0.00
Plectonema radiosum BTA009 3.51±0.28 5.62±0.32 61.0±6.02 119.0±9.64
Plectonema boryanum BTA010 4.96±0.32 5.90±0.43 51.6±1.04 15.6±1.62
90
Nostoc hatei BTA012 5.82±0.31 6.00±2.12 42.3±0.57 76.0±3.40
Phormidium tenue BTA013 5.93±1.54 5.91±2.04 93.3±2.30 120.3±1.52
Anabaena oryzae BTA014 2.45±0.27 4.00±1.90 10.1±3.29 42.3±1.52
Calothrix wembaerensis BTA015 2.49±0.22 2.67±1.09 54.6±1.25 63.0±1.09
Plectonema boryanum BTA016 3.81±0.62 4.29±1.23 4.30±0.57 123.3±5.77
Anabaena oryzae BTA017 2.09±0.36 3.09±0.09 17.6±3.78 24.3±1.27
Anabaena oryzae BTA018 5.15±0.90 6.09±0.00 33.6±1.15 124.6±1.15
Anabaena spiroides BTA019 4.33±0.43 7.90±0.08 33.2±2.20 72.3±1.41
Phormidium faveolarum BTA020 0.36±0.06 2.09±0.90 15.0±0.07 22.2±2.61
Anabaena ambigua BTA021 3.23±0.08 6.09±0.03 45.0±1.54 65.3±1.65
Anabaena anomala BTA023 8.58±0.75 9.00±0.01 47.8±3.44 95.0±2.64
Calothrix javanica BTA024 5.89±1.07 14.4±1.17 67.9±2.56 103.0±2.33
Calothrix geitonos BTA025 12.1±1.19 15.0±1.12 48.3±3.42 83.6±2.08
Calothrix marchica BTA026 13.2±1.33 16.0±1.09 82.3±2.50 117.0±1.00
Nostoc muscorum BTA027 8.97±1.05 9.00±1.09 75.2±1.32 120.0±0.00
Dichothrix orsiniana BTA028 4.84±0.58 6.09±0.00 17.5±2.56 33.3±1.52
Nostoc parmelioides BTA029 10.6±2.16 12.0±1.10 52.6±2.30 74.6±1.10
Anabaena fertilissima BTA030 1.70±0.86 2.10±0.06 4.32±2.35 6.00±4.35
Anabaena oryzae BTA031 9.21±1.04 14.9±1.04 64.3±1.33 89.6±4.61
Plectonema radiosum BTA032 1.39±0.55 2.04±0.07 5.66±4.04 14.6±5.50
Plectonema litorale BTA033 1.94±0.20 3.80±1.09 12.0±0.75 17.2±1.32
Anabaena ambigua BTA034 3.09±0.03 17.3±1.20 106.0±1.64 90.3±8.08
Anabaena fertilissima BTA035 11.6±3.59 4.48±0.48 85.3±4.16 92.3±1.32
91
Anabaena variabilis BTA036 18.2±1.96 8.21±1.52 76.6±1.27 56.0±2.86
Nostoc hatei BTA037 13.3±2.21 5.51±0.60 51.3±4.04 115.0±5.00
Nostoc carneum BTA038 19.1±0.29 19.1±0.29 83.6±1.52 77.0±3.60
Myxosarcina burmensis BTA040 6.71±1.95 5.64±0.62 58.3±4.50 53.3±7.63
Anabaena iyengarii BTA041 19.7±0.42 0.51±0.08 34.6±1.62 74.0±10.4
Phormidium arthurensis BTA042 0.74±0.13 12.5±0.69 61.6±1.15 59.3±1.36
Anabaena variabilis BTA043 3.24±0.14 5.24±0.00 19.3±1.50 27.5±2.52
Microchaete loktakensis BTA044 9.29±1.54 10.5±0.54 58.3±5.68 94.0±6.55
Microchaete uberrima BTA045 13.0±0.75 14.2±0.01 42.6±4.22 51.6±13.6
Calothrix marchica BTA046 7.02±0.32 9.05±0.10 78.3±2.22 103.6±7.76
Plectonema nostocorum BTA047 3.44±1.11 4.39±1.15 129.0±3.46 90.2±2.36
Phormidium fragile BTA048 5.26±0.17 11.1±0.55 129.0±2.64 95.6±3.22
Microchaete tenera BTA049 18.4±1.76 14.2±1.36 103.3±2.54 89.6±10.0
Anabaena oryzae BTA050 11.5±1.60 12.4±1.30 110.3±13.4 103.6±15.7
Phormidium purpurascens BTA081 3.08±0.10 5.71±0.13 4.20±1.20 1.00±1.00
Limnothrix redekei BTA082 4.08±0.04 10.1±0.26 128.0±1.73 112.0±3.62
Phormidium kuetzingianum BTA083 1.01±0.00 3.84±0.02 40.3±5.50 55.7±6.60
Anabaena spiroides BTA084 10.4±3.55 31.1±3.55 108.2±1.85 83.0±2.06
Lyngbya laxespiralis BTA085 4.57±0.22 8.23±0.12 92.6±14.4 98.3±2.88
Phormidium tenue BTA086 6.40±0.10 10.3±0.00 59.6±4.50 78.0±2.94
Nostoc muscorum BTA087 20.8±1.66 17.2±0.00 67.0±0.00 90.3±4.50
Plectonema notatum BTA088 2.78±0.28 4.10±0.27 96.0±0.92 87.6±1.32
Scytonema bohneri BTA106 1.06±0.07 3.27±1.02 99.0±1.73 93.3±2.46
92
Plectonema notatum BTA108 1.79±0.33 0.86±0.15 65.6±6.02 56.7±3.21
Phormidium incrustatum BTA118 2.15±0.19 4.21±0.22 80.0±14.7 81.3±1.54
Limnothrix redekei BTA123 4.06±1.26 5.32±0.12 22.0±1.00 25.0±1.20
Phormidium valderianum BTA162 3.53±0.43 4.32±1.22 33.2±1.20 49.6±3.21
Scytonema guyanense BTA167 0.67±0.68 2.32±0.02 27.6±3.21 37.6±2.22
Nostoc commune BTA168 3.15±0.03 6.23±1.02 42.7±2.50 39.7±4.60
Oscillatoria agardhii BTA170 4.15±0.36 7.12±0.12 21.7±1.20 28.6±3.21
Spirulina platensis BTA174 10.8±0.02 10.0±0.12 115.6±2.45 130.3±1.89
Calothrix castellii BTA177 3.45±1.37 4.01±0.03 61.3±4.21 56.3±5.20
Lyngbya connectens BTA178 8.45±1.36 11.2±1.13 26.7±3.15 37.6±4.50
Lyngbya nordgardhii BTA184 2.55±0.07 3.69±0.47 104.0±1.78 66.3±1.20
Scytonema schmidtii BTA186 3.03±0.14 4.01±0.16 20.7±1.36 23.6±1.35
Aulosira aenigmatica BTA188 0.88±1.33 14.1±0.00 31.6±1.15 90.0±1.73
Phormidium tenue BTA189 2.47±0.14 4.54±0.01 129.0±7.93 93.3±2.06
Aulosira bombayensis BTA190 0.38±0.00 11.8±0.01 2.66±0.57 122.0±7.21
Plectonema notatum BTA194 5.13±0.25 20.7±0.07 72.0±2.20 116.3±1.15
Calothrix marchica BTA195 4.45±0.97 6.25±0.82 113.0±7.76 80.6±2.36
Limnothrix mirabilis BTA199 2.68±1.06 4.22±1.00 39.6±2.36 49.6±1.32
Nostoc calcicola BTA204 0.89±0.04 13.8±0.03 50.0±1.80 87.6±1.15
Anabaena circinalis BTA205 1.11±0.36 6.24±0.03 24.6±2.51 117.3±4.50
Calothrix marchica BTA206 2.57±1.08 6.23±0.10 112.0±7.00 98.4±3.50
Calothrix clavata BTA218 3.92±0.31 9.66±0.03 97.3±1.53 112.6±6.65
Phormidium tenue BTA222 0.37±0.11 10.5±0.01 82.7±2.10 92.6±1.01
93
Anabaena circinalis BTA246 1.75±0.16 3.31±0.22 39.0±4.58 54.0±0.00
Lyngbya martensiana BTA436 3.29±0.02 2.49±0.40 114.0±1.38 83.3±5.77
Lyngbya digueti BTA475 2.46±1.82 5.65±0.91 120.6±2.30 78.3±1.96
Phormidium fragile BTA521 2.97±0.07 12.1±0.01 90.6±2.65 87.6±7.02
Anabaena circinalis BTA561 1.64±0.74 3.04±0.02 82.6±5.20 73.2±2.36
Anabaena sp. BTA564 1.22±0.12 10.5±2.64 20.4±6.92 36.1±4.26
Plectonema notatum BTA565 6.04±0.86 9.36±1.15 126.6±2.88 85.6±5.50
Phormidium autumnale BTA587 1.33±0.19 3.04±0.30 40.6±6.02 58.0±3.46
Lyngbya aestuarii BTA597 0.87±0.13 4.26±0.87 48.3±1.89 75.0±0.00
Anabaena variabilis BTA600 0.20±0.20 0.67±0.20 58.6±2.26 66.3±1.32
Lyngbya allorgei BTA606 10.7±0.14 11.6±2.64 15.6±1.32 18.3±1.42
Lyngbya martensiana BTA640 2.17±1.37 4.21±0.24 17.8±2.30 21.6±2.45
Westiellopsis prolifica BTA645 1.15±0.01 2.51±0.26 26.7±1.20 35.3±1.22
Anabaena orientalis BTA653 1.10±0.19 2.10±0.10 32.6±1.87 42.6±3.24
Phormidium stagnina BTA855 0.85±0.00 2.98±0.32 19.6±1.32 24.7±3.63
Anabaena variabilis BTA880 2.45±1.16 6.23±1.01 21.6±2.34 31.6±4.50
Anabaena oryzae BTA881 1.24±0.30 3.63±0.19 96.6±11.5 84.0±8.18
Anabaena fertilissima BTA883 1.08±0.06 0.91±0.17 29.3±3.44 81.0±1.54
Nostoc ellipsosporum BTA902 0.28±0.07 0.80±0.05 46.3±1.42 44.6±6.11
Anabaena constricta BTA903 2.11±0.41 0.75±0.42 66.7±3.03 98.3±2.02
Cylindrospermum muscicola
BTA904
0.65±0.02 0.89±0.05 10.5±2.30 11.3±7.50
Cylindrospermum doryphorum
BTA905
1.07±0.50 3.50±0.11 39.6±2.74 54.0±173
Cylindrospermum indentatum
BTA906
3.23±0.19 4.94±0.05 55.3±1.40 66.6±2.90
94
Anabaena oryzae BTA919 2.90±0.78 1.72±0.40 42.3±3.41 65.6±6.65
Nostoc ellipsosporum BTA923 2.48±0.45 1.72±0.53 55.7±7.21 87.6±12.7
Microchaete grisea BTA926 9.96±0.76 7.38±0.90 29.4±9.50 54.0±13.5
Anabaena anomala BTA927 3.85±0.56 0.69±0.50 57.3±0.10 87.0±0.00
Nostoc verrucosum BTA939 4.83±0.60 4.28±0.68 34.3±2.08 28.7±3.20
Nostoc calcicola BTA942 1.79±0.27 3.53±0.44 24.7±4.81 19.6±5.21
Nostoc piscinale BTA947 4.22±0.95 25.2±0.05 69.6±1.12 57.0±5.19
Cylindrospermum indicum BTA960 0.58±0.04 4.21±0.01 18.7±1.23 67.3±5.00
Anabaena variabilis BTA990 2.64±0.04 2.64±0.40 27.6±2.00 37.6±2.10
Dichothrix baueriana BTA1059 2.71±0.05 6.68±1.05 39.0±4.58 90.6±9.01
Table-8: Screening for ammonia excretion and total carbohydrates from
cyanobacterial strains of rice fields of Manipur
Strains name with code no. Ammonia excretion
(µg/ml)
Total carbohydrates
content (µg/ml)
15th
day 30th
day 15th
day 30th
day
Microchaete uberrima BTA001 1.00±0.34 4.10±0.62 91.6±2.46 46.3±3.19
Phormidium mucosum BTA002 5.20±0.62 4.11±0.01 96.6±1.76 52.4±3.00
Anabaena variabilis BTA003 20.1±2.12 9.30±1.82 13.0±1.73 36.3±11.6
Anabaena doliolum BTA004 21.1±180 17.7±1.37 41.0±3.46 50.2±4.36
Limnothrix vacuolifera BTA005 1.90±1.53 0.39±1.00 38.0±7.21 42.6±2.46
Anabaena variabilis BTA006 7.20±2.07 18.3±2.34 73.6±6.50 48.0±10.8
Microchaete grisea BTA007 14.8±1.21 14.7±1.08 16.0±3.46 43.6±1.49
Anabaena circinalis BTA008 13.8±2.61 27.9±1.42 48.6±14.2 31.0±9.64
95
Plectonema radiosum BTA009 1.20±0.79 2.00±1.00 59.2±4.52 66.2±4.36
Plectonema boryanum BTA010 2.03±1.01 1.10±0.80 71.2±5.36 82.3±3.00
Nostoc hatei BTA012 12.4±1.21 56.5±6.57 11.0±1.73 41.6±1.65
Phormidium tenue BTA013 45.1±4.90 75.5±1.28 53.6±2.01 43.0±3.46
Anabaena oryzae BTA014 17.4±3.60 24.5±1.46 38.3±1.70 49.6±4.04
Calothrix wembaerensis BTA015 11.7±2.60 14.8±1.82 23.0±3.60 37.6±3.21
Plectonema boryanum BTA016 0.90±0.00 3.10±1.22 61.2±4.33 72.0±4.00
Anabaena oryzae BTA017 16.6±0.46 10.7±0.34 40.0±5.19 39.0±1.35
Anabaena oryzae BTA018 0.60±0.00 23.4±5.47 30.2±1.36 35.3±2.30
Anabaena spiroides BTA019 8.80±1.40 13.5±0.79 15.3±0.57 54.3±0.57
Phormidium faveolarum BTA020 0.40±0.17 5.50±0.11 47.3±1.04 49.0±1.22
Anabaena ambigua BTA021 4.10±2.70 3.13±1.80 43.3±6.35 45.2±3.65
Anabaena anomala BTA023 11.7±0.30 5.40±0.00 27.3±8.08 22.6±3.05
Calothrix javanica BTA024 10.8±0.30 26.8±2.44 25.7±1.52 29.6±8.96
Calothrix geitonos BTA025 10.1±3.45 6.00±0.30 27.0±0.00 41.0±5.29
Calothrix marchica BTA026 7.80±1.03 18.5±1.65 21.0±1.73 26.0±7.54
Nostoc muscorum BTA027 130.8±1.58 18.9±3.13 36.6±0.57 49.3±10.6
Dichothrix orsiniana BTA028 4.95±0.45 41.7±2.59 9.60±4.72 22.6±2.88
Nostoc parmelioides BTA029 17.1±4.58 11.1±2.07 24.3±2.51 49.0±1.90
Anabaena fertilissima BTA030 12.1±4.84 8.09±1.20 25.0±2.60 49.0±12.0
Anabaena oryzae BTA031 41.8±1.21 35.7±12.7 14.6±1.52 35.6±1.52
Plectonema radiosum BTA032 0.60±0.79 9.00±1.96 38.2±3.52 43.5±2.50
Plectonema litorale BTA033 0.80±0.18 5.00±0.01 25.3±1.30 30.3±1.67
96
Anabaena ambigua BTA034 33.6±0.90 38.1±4.35 13.0±2.64 24.0±1.00
Anabaena fertilissima BTA035 57.3±1.08 29.1±5.50 20.0±0.01 45.0±6.55
Anabaena variabilis BTA036 112.0±1.25 18.2±3.23 19.6±3.21 35.6±1.09
Nostoc hatei BTA037 169.3±1.03 19.2±3.11 15.0±1.00 46.0±8.00
Nostoc carneum BTA038 92.7±1.09 17.9±1.27 24.6±6.42 30.6±1.32
Myxosarcina burmensis BTA040 36.1±3.00 33.5±9.45 22.0±1.00 28.3±5.13
Anabaena iyengarii BTA041 44.4±1.73 14.6±4.25 10.6±2.08 2.00±1.73
Phormidium arthurensis BTA042 52.4±1.14 44.5±2.66 20.6±8.14 15.0±0.00
Anabaena variabilis BTA043 47.4±1.00 39.0±1.20 27.6±1.22 25.6±1.00
Microchaete loktakensis BTA044 126.9±1.34 24.9±6.41 18.6±2.51 40.0±4.35
Microchaete uberrima BTA045 56.5±1.40 41.5±5.75 47.0±8.80 36.2±4.20
Calothrix marchica BTA046 30.3±1.12 33.0±1.58 24.6±1.20 16.2±2.20
Plectonema nostocorum BTA047 43.9±0.20 39.7±0.45 54.3±4.93 33.6±12.3
Phormidium fragile BTA048 34.0±1.63 27.6±1.56 95.0±5.00 28.6±6.65
Microchaete tenera BTA049 12.6±2.40 8.20±6.47 31.0±7.21 29.0±4.36
Anabaena oryzae BTA050 4.20±1.20 0.70±0.69 22.6±3.20 27.3±4.30
Phormidium puspurascens BTA081 33.4±0.17 11.4±3.36 16.0±7.21 21.3±1.36
Limnothrix redekei BTA082 42.3±1.08 39.1±1.09 13.3±2.88 33.0±1.40
Phormidium kuetzinggianum
BTA083
38.5±1.19 27.9±7.86 31.3±4.04 21.3±11.8
Anabaena spiroides BTA084 34.0±1.01 48.7±1.05 34.6±5.85 37.3±4.22
Lyngbya laxespiralis BTA085 28.1±5.41 22.6±1.21 49.0±6.92 45.2±2.63
Phormidium tenue BTA086 59.3±2.19 33.4±2.29 29.6±1.15 31.6±2.16
97
Nostoc muscorum BTA087 92.3±1.86 13.3±0.86 38.3±2.63 44.6±11.5
Plectonema notatum BTA088 63.5±4.54 40.2±2.50 42.6±4.12 46.3±6.02
Scytonema bohneri BTA106 58.2±1.01 49.5±1.20 18.3±3.78 30.6±1.61
Plectonema notatum BTA108 47.5±1.14 31.8±1.19 66.0±3.46 20.6±2.51
Phormidium incrustatum BTA118 25.1±5.53 27.1±0.17 36.2±3.36 39.4±2.26
Limnothrix redekei BTA123 16.2±1.15 12.5±1.00 28.6±1.02 19.2±1.10
Phormidium valderianum BTA162 55.6±1.25 30.0±1.30 22.3±1.11 25.2±3.56
Scytonema guyanense BTA167 41.2±6.17 23.5±1.67 19.3±2.75 15.7±3.70
Nostoc commune BTA168 35.3±1.60 32.5±1.20 14.6±3.76 17.2±4.62
Oscillatoria agardhii BTA170 93.8±2.52 72.0±1.39 18.2±3.05 21.5±4.00
Spirulina platensis BTA174 84.4±0.09 58.8±0.01 80.0±0.01 90.5±0.03
Calothrix castellii BTA177 41.0±7.88 36.0±1.80 32.6±2.10 44.6±1.36
Lyngbya connectens BTA178 111.3±1.34 90.3±1.20 14.6±1.40 21.7±1.00
Lyngbya nordgardhii BTA184 62.0±0.00 46.7±1.23 32.3±0.57 31.0±7.93
Scytonema schmidtii BTA186 15.1±3.55 12.2±1.35 23.6±1.35 27.6±0.25
Aulosira aenigmatica BTA188 37.5±3.24 15.0±1.80 13.3±3.05 16.0±1.30
Phormidium tenue BTA189 134.0±2.97 116.0±2.59 34.3±9.71 18.3±5.00
Aulosira bombayensis BTA190 24.7±4.70 18.8±3.45 12.6±3.21 9.00±1.73
Plectonema notatum BTA194 39.7±2.25 79.1±4.65 9.33±0.57 34.3±8.96
Calothrix marchica BTA195 32.7±5.60 35.2±1.30 27.6±6.42 29.9±3.26
Limnothrix mirabilis BTA199 108.4±8.30 60.4±2.30 31.5±3.00 26.5±4.25
Nostoc calcicola BTA204 13.6±5.28 11.1±2.10 15.6±4.50 2.33±1.52
Anabaena circinalis BTA205 28.0±8.16 22.5±1.03 8.00±6.35 11.6±2.08
98
Calothrix marchica BTA206 22.3±9.00 19.0±2.01 56.6±3.21 98.7±4.01
Calothrix clavata BTA218 31.3±1.32 18.8±4.67 19.0±1.73 20.0±9.53
Phormidium tenue BTA222 43.0±3.01 68.9±1.08 39.0±5.19 38.6±7.09
Anabaena circinalis BTA246 27.4±1.08 11.0±4.02 8.33±3.05 10.6±5.68
Lyngbya martensiana BTA436 24.7±7.18 71.9±1.38 36.0±6.24 32.3±1.52
Lyngbya digueti BTA475 13.4±3.04 19.0±1.41 24.3±5.77 10.3±4.50
Phormidium fragile BTA521 43.3±8.70 63.7±1.22 27.0±3.60 30.0±9.53
Anabaena circinalis BTA561 44.1±9.80 35.2±5.20 42.6±3.63 36.3±1.36
Anabaena sp. BTA564 4.50±1.87 13.6±6.92 47.6±3.02 51.6±2.02
Plectonema notatum BTA565 63.2±9.86 84.5±2.29 29.3±5.03 58.6±0.35
Phormidium autumnale BTA587 15.7±0.79 24.6±2.10 21.3±4.50 19.6±3.51
Lyngbya aestuarii BTA597 28.1±1.49 36.0±2.40 39.5±2.56 32.3±1.52
Anabaena variabilis BTA600 13.6±2.85 12.4±3.20 19.2±4.60 22.3±1.36
Lyngbya allorgei BTA606 12.0±2.07 21.5±2.55 35.6±1.22 37.7±3.62
Lyngbya martensiana BTA640 38.0±5.76 47.0±9.15 37.4±3.44 39.5±1.22
Westiellopsis prolifica BTA645 4.10±0.17 6.90±1.82 42.6±3.20 45.7±3.10
Anabaena orientalis BTA653 10.5±5.49 7.23±1.37 12.3±1.22 6 .22±1.30
Phormidium stagnina BTA855 39.3±6.28 24.6±3.22 29.3±2.10 32.6±0.12
Anabaena variabilis BTA880 51.4±6.86 41.7±1.36 29.3±2.10 32.6±0.12
Anabaena oryzae BTA881 32.2±1.35 28.5±8.25 21.3±2.51 13.6±4.16
Anabaena fertilissima BTA883 24.9±4.95 17.7±7.28 12.6±1.63 15.5±2.00
Nostoc ellipsosporum BTA902 10.6±1.99 36.5±3.62 9.33±0.57 13.5±0.66
Anabaena constricta BTA903 10.1±4.37 12.8±5.85 16.3±2.51 18.4±3.41
99
Cylindrospermum muscicola
BTA904
25.5±5.90 23.2±9.27 5.66±2.88 9.03±4.22
Cylindrospermum doryphorum BTA905 6.79±0.45 3.25±0.11 27.6±5.50 18.3±1.20
Cylindrospermum indentatum BTA906 11.6±6.06 23.4±4.25 14.5±5.02 09.2±1.36
Anabaena oryzae BTA919 16.3±5.11 26.1±3.38 19.3±2.51 34.2±1.36
Nostoc ellipsosporum BTA923 15.2±4.59 24.3±3.45 18.6±2.08 27.6±1.36
Microchaete grisea BTA926 10.8±1.37 12.8±6.32 32.3±4.72 41.1±5.22
Anabaena anomala BTA927 5.60±5.30 46.2±1.20 42.0±7.00 52.0±3.44
Nostoc verrucosum BTA939 15.0±2.10 19.2±2.86 14.6±0.57 44.0±2.45
Nostoc calcicola BTA942 65.4±1.22 66.8±3.91 24.0±6.92 34.0±2.00
Nostoc piscinale BTA947 53.0±5.30 5.90±1.75 19.0±2.64 48.6±2.88
Cylindrospermum indicum BTA960 44.1±1.06 49.2±1.00 45.6±4.20 56.0±3.22
Anabaena variabilis BTA990 8.80±2.17 12.3±4.93 47.5±3.11 61.2±1.30
Dichothrix baueriana BTA1059 17.7±2.74 46.2±7.20 31.5±0.10 26.0±0.00
100
Str
ain
s n
am
e a
nd
co
de
no.
P
hyco
bil
ipro
tein
s (µ
g/m
l)
Caro
ten
oid
s (µ
g/m
l)
15
th d
ay
30
th d
ay
15
th
3
0th
PE
P
C
AP
C
PE
P
C
AP
C
Mic
roch
aet
e u
ber
rim
a B
TA
00
1
12
.1±
0.8
2
7.3
4±
0.5
8
3.2
9±
0.4
4
15
.9±
4.1
9
15
.2±
3.8
1
6.7
8±
0.7
4
18
.6±
5.6
5
28
.6±
5.0
0
Ph
orm
idiu
m m
uco
sum
BT
A0
02
1
.09
±1.0
5
11
.2±
1.4
1
3.6
0±
1.5
8
9.3
6±
0.1
0
33
.1±
2.7
5
14
.6±
0.7
3
27
.8±
2.1
6
24
.2±
2.0
0
An
ab
aen
a v
ari
abil
is B
TA
00
3
5.2
3±
1.9
5
29
.5±
1.9
2
34
.2±
1.7
5
11
.2±
1.5
7
32
.6±
1.8
6
38
.9±
1.2
3
5.3
7±
1.4
0
19
.8±
2.4
1
An
ab
aen
a d
oli
olu
m B
TA
00
4
4.1
1±
2.7
8
38
.9±
1.7
4
12
.5±
1.5
9
13
.3±
0.6
2
31
.3±
1.9
2
36
.1±
1.6
1
6.1
5±
0.6
8
26
.5±
1.9
1
Lim
no
thri
x va
cuo
life
ra B
TA
00
5
3.2
1±
0.5
2
14
.9±
6.2
6
5.5
3±
0.5
8
7.0
3±
1.0
4
22
.2±
1.3
2
14
.1±
3.2
5
7.8
2±
1.1
9
26
.5±
1.9
1
An
ab
aen
a v
ari
abil
is B
TA
00
6
15
.5±
3.5
8
30
.3±
1.7
0
42
.5±
2.6
1
12
.3±
1.3
2
22
.4±
1.3
2
24
.7±
1.6
9
9.8
4±
0.8
9
39
.3±
1.7
6
Mic
roch
aet
e g
rise
a B
TA
00
7
34
.5±
3.1
4
35
.7±
2.5
2
19
.5±
1.6
0
60
.7±
1.7
8
70
.1±
1.7
6
47
.3±
1.8
9
34
.5±
3.1
9
42
.1±
1.7
5
An
ab
aen
a c
irci
na
lis
BT
A00
8
17
.3±
1.5
8
44
.3±
1.2
0
24
.3±
1.4
0
23
.4±
2.0
0
55
.3±
1.0
0
18
.2±
0.0
1
43
.5±
5.2
0
53
.7±
1.3
0
Ple
cto
nem
a r
adio
sum
BT
A0
09
0
.87
±0.4
1
3.7
7±
1.2
9
1.3
6±
0.6
5
4.2
0±
0.7
8
6.4
3±
1.0
8
5.5
6±
1.2
0
16
.3±
0.5
2
16
.3±
0.5
2
Ple
cto
nem
a b
ory
an
um
BT
A0
10
2
.55
±0.3
3
11
.2±
1.8
1
6.8
6±
3.1
5
4.5
6±
0.3
1
1
4.2
±1
.20
7.3
2±
1.0
0
17
.4±
2.8
7
17
.2±
2.0
0
Nost
oc
ha
tei
BT
A0
12
2.5
2±
0.7
7
77
.5±
1.7
4
25
.7±
2.6
0
11
.3±
1.0
8
64
.2±
3.8
3
20
.7±
1.8
5
19
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4.8
2
28
.5±
5.2
2
Ph
orm
idiu
m t
enu
e B
TA
01
3
14
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1.4
5
1
02
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1.9
2
5.3
±2.2
5
18
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3.4
5
11
2.6
±1
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32
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2.0
0
25
.0±
3.4
0
32
.7±
2.6
2
An
ab
aen
a o
ryza
e B
TA
014
6
.69
±1.4
6
61
.9±
1.8
1
14
.3±
3.3
3
7.7
8±
1.0
9
48
.2±
1.7
3
15
.3±
1.7
6
7.3
7±
4.5
6
6.2
2±
2.8
4
Ca
loth
rix
wem
ba
eren
sis
BT
A0
15
1.6
7±
1.3
6
36
.1±
0.7
4
8.4
1±
1.0
2
6.5
4±
0.9
1
32
.9±
1.4
8
13
.9±
1.0
6
12
.8±
0.3
4
20
.0±
0.7
2
Ple
cto
nem
a b
ory
an
um
BT
A0
16
0
.85
±0.3
6
5.0
1±
2.5
5
1.9
7±
0.5
1
3.7
3±
0.3
0
28
.6±
0.2
5
11
.9±
0.9
8
14
.3±
0.5
3
14
.3±
0.5
3
T
ab
le-9
: S
cree
nin
g f
or
ph
yco
bil
ipro
tein
s an
d c
aro
ten
oid
s fr
om
cya
nob
act
eria
l st
rain
s of
rice
fie
lds
of
Ma
nip
ur
101
An
ab
aen
a o
ryza
e B
TA
01
7
12
.6±
1.6
4
43
.1±
1.9
6
10
.1±
0.8
5
3.0
2±
0.6
7
7.8
1±
2.1
9
6.5
0±
0.9
6
7.7
0±
0.1
3
8.8
7±
1.2
1
An
ab
aen
a o
ryza
e B
TA
01
8
6.9
5±
1.3
3
10
.4±
2.0
3
1.0
2±
0.3
6
17
.6±
0.9
1
29
.8±
1.7
3
9.5
9±
0.7
7
18
.4±
3.6
7
18
.6±
3.4
6
An
ab
aen
a s
pir
oid
es B
TA
01
9
6.7
4±
0.9
8
28
.4±
1.5
5
12
.3±
1.0
5
3.4
0±
0.7
8
19
.3±
3.5
3
8.4
7±
2.6
5
4.5
3±
0.9
6
2.8
3±
0.3
3
Ph
orm
idiu
m f
ave
ola
rum
BT
A0
20
1.0
5±
0.2
1
2.3
7±
0.2
2
1.7
1±
0.3
5
2.7
4±
0.1
6
3.1
2±
0.1
6
4.0
4±
1.8
9
1.5
5±
0.1
0
2.8
1±
0.2
8
An
ab
aen
a a
mb
igua
BT
A0
21
5
.79
±0.9
0
16
.7±
1.3
9
17
.7±
2.3
0
9.3
2±
0.2
0
21
.0±
1.0
3
19
.2±
1.4
2
13
.1±
1.6
0
2.8
1±
0.2
8
An
ab
aen
a a
no
ma
la B
TA
02
3
6.5
2±
0.7
4
16
.3±
1.3
9
15
.2±
1.0
0
4.1
4±
0.3
6
55
.8±
4.0
9
24
.1±
2.9
7
59
.7±
6.4
1
24
.2±
0.7
6
Ca
loth
rix
ja
van
ica B
TA
02
4
10
.7±
1.7
8
19
.9±
1.7
5
9.8
0±
1.2
7
80
.9±
0.3
7
27
.9±
3.3
8
20
.0±
2.0
5
50
.5±
4.6
5
26
.3±
1.8
2
Ca
loth
rix
gei
ton
os
BT
A0
25
1
1.4
±1.4
0
22
.5±
1.4
5
9.4
8±
1.3
0
12
.3±
2.5
1
19
.9±
3.2
6
13
.9±
2.1
3
33
.5±
6.3
6
23
.9±
0.7
8
Ca
loth
rix
ma
rchic
a B
TA
02
6
25
.3±
1.5
5
49
.1±
1.9
3
28
.2±
1.9
7
25
.3±
1.5
4
49
.1±
1.9
8
28
.2±
1.9
7
27
.1±
6.1
3
13
.1±
0.6
8
Nost
oc
musc
oru
m B
TA
027
2
.52
±0.2
0
79
.2±
1.9
3
20
.1±
1.5
4
4.8
5±
0.9
3
14
8.3
±1
.24
32
.7±
1.5
4
38
.2±
1.5
7
19
.4±
2.2
5
Dic
hoth
rix
ors
inia
na
BT
A0
28
1
.27
±0.4
3
21
.2±
0.7
6
9.5
5±
0.5
4
2.4
2±
0.4
6
17
.3±
1.1
7
8.1
0±
0.9
7
24
.2±
3.3
5
8.4
7±
0.0
6
Nost
oc
pa
rmel
ioid
es B
TA
02
9
15
.5±
0.3
0
14
.7±
0.4
8
4.6
4±
0.4
4
15
.3±
1.7
3
14
.5±
1.2
9
7.7
0±
2.8
9
26
.5±
0.4
5
14
.3±
1.5
2
An
ab
aen
a f
erti
liss
ima
BT
A0
30
1
8.2
±3.0
2
25
.7±
1.0
2
16
.2±
1.0
0
22
.0±
1.4
0
30
.3±
1.0
7
18
.7±
1.8
0
14
.6±
3.2
1
19
.5±
1.3
0
An
ab
aen
a o
ryza
e B
TA
03
1
14
.2±
0.3
4
33
.5±
1.9
8
4.4
9±
1.0
5
24
.2±
7.7
8
76
.8±
2.4
1
10
.2±
4.2
1
9.3
1±
2.6
0
29
.2±
2.0
0
Ple
cto
nem
a r
adio
sum
BT
A0
32
0
.32
±0.0
0
0.6
5±
0.3
1
0.8
0±
0.5
1
4.8
8±
2.1
3
12
.0±
4.7
1
10
.1±
5.3
9
0.8
8±
0.4
4
7.2
0±
0.6
5
Ple
cto
nem
a l
itora
le B
TA
03
3
0.9
4±
0.3
6
1.8
2±
0.8
9
2.5
5±
1.5
8
4.9
9±
1.0
5
6.2
8±
1.0
9
8.0
1±
2.5
1
0.3
4±
0.0
8
8.0
2±
1.5
5
An
ab
aen
a a
mb
igua
BT
A0
34
1
5.7
±1.9
5
50
.8±
0.9
7
40
.2±
4.0
8
10
.6±
6.2
0
11
2.7
±1
.46
20
.6±
7.8
4
13
.8±
1.5
5
38
.4±
2.3
0
An
ab
aen
a f
erti
liss
ima B
TA
03
5
7.9
7±
3.8
6
62
.5±
4.5
3
54
.4±
7.6
2
21
.1±
5.4
0
15
0.4
±1
.12
13
0.7
±1
.46
29
.6±
1.9
9
31
.7±
1.0
0
102
An
ab
aen
a v
ari
abil
is B
TA
03
6
1.4
3±
0.3
4
33
.5±
1.9
9
4.4
9±
1.0
5
4.2
9±
0.7
6
63
.7±
7.7
5
3.8
5±
1.9
4
18
.2±
1.5
6
31
.3±
2.6
0
Nost
oc
ha
tei
BT
A0
37
6.9
3±
5.8
1
30
.7±
7.9
6
19
.4±
1.6
4
4.3
4±
2.1
0
16
.7±
5.3
8
10
.3±
5.4
7
10
.4±
2.5
5
21
.5±
2.5
0
Nost
oc
carn
eum
BT
A038
20.0
±4.0
9
82.7
±8.3
5
58.7
±9.3
9
11.2
±1.8
5
98.8
±8.4
4
39.9
±6.3
2
30.9
±3.9
5
58.9
±1.8
0
Myx
osa
rcin
a b
urm
ensi
s B
TA
04
0
8.8
6±
0.8
5
19
.0±
1.3
9
23
.8±
1.5
3
5.9
5±
1.9
0
12
.3±
3.3
0
16
.4±
3.7
7
7.3
2±
1.4
0
19
.2±
0.9
2
An
ab
aen
a i
yeng
ari
i B
TA
04
1
8.0
7±
2.9
8
51
.2±
1.0
2
50
.5±
1.5
1
10
.2±
1.3
8
66
.3±
1.0
3
45
.0±
1.0
0
25
.6±
2.3
3
27
.4±
1.2
2
Ph
orm
idiu
m a
rth
ure
nsi
s B
TA
04
2
3.2
7±
0.7
5
4.7
5±
0.3
0
6.2
8±
0.4
6
69
.2±
7.4
0
28
6.1
±1
.00
46
.8±
4.3
7
5.1
2±
0.3
5
30
.4±
6.5
0
An
ab
aen
a v
ari
abil
is B
TA
04
3
7.3
8±
1.3
6
47
.3±
0.1
1
23
.0±
1.3
6
13
.2±
1.0
9
57
.2±
1.0
3
36
.8±
0.0
9
16
.3±
1.3
2
21
.3±
1.4
2
Mic
roch
aet
e lo
kta
kensi
s B
TA
04
4
5.3
6±
0.7
0
10
.5±
1.0
8
18
.2±
1.1
0
9.3
1±
1.0
0
18
.8±
5.5
9
18
.5±
8.2
0
20
.6±
1.4
4
50
.2±
1.2
0
Mic
roch
aet
e u
ber
rim
a B
TA
04
5
5.5
6±
0.5
0
10
.8±
0.3
4
7.5
6±
0.4
1
11
.0±
0.1
0
15
.2±
0.4
1
9.0
8±
0.7
1
38
.4±
1.6
5
36
.3±
4.2
0
Ca
loth
rix
ma
rchic
a B
TA
04
6
40
.9±
4.8
0
70
.1±
5.1
7
50
.5±
2.1
1
51
.2±
1.2
0
63
.2±
1.0
9
46
.2±
0.0
1
44
.9±
2.2
0
51
.3±
1.3
0
Ple
cto
nem
a n
ost
oco
rum
BT
A0
47
7.6
3±
0.9
5
11
.95
±1
.8
16
.6±
2.8
9
3.2
6±
2.0
0
7.9
5±
4.3
0
6.0
5±
5.4
9
27
.0±
5.7
8
47
.2±
7.1
3
Ph
orm
idiu
m f
ragil
e B
TA
04
8
19
.5±
1.4
0
46
.4±
3.4
0
38
.8±
2.8
1
6.5
4±
0.9
8
17
.2±
1.9
2
14
.5±
4.2
7
27
.0±
3.5
3
55
.2±
2.0
7
Mic
roch
aet
e te
ner
a B
TA
04
9
4.9
4±
2.2
0
7.7
2±
2.5
7
4.5
7±
1.5
8
25
.1±
3.1
0
47
.7±
5.7
6
43
.8±
1.8
2
45
.2±
5.4
3
37
.6±
5.2
0
An
ab
aen
a o
ryza
e B
TA
05
0
42.7
±1.4
0
36
.8±
1.9
3
9.3
1±
3.0
1
53
.4±
1.5
0
72
.7±
1.2
5
39
.6±
3.6
4
47
.7±
2.5
0
32
.3±
1.3
0
Ph
orm
idiu
m p
urp
ura
scen
s B
TA
08
1
7.2
0±
1.8
1
15
.6±
1.0
2
19
.2±
1.0
3
11
.2±
1.0
8
16
.2±
1.1
1
25
.6±
0.0
1
56
.3±
1.3
0
66
.5±
1.5
6
Lim
no
thri
x re
dek
ei B
TA
082
15.5
±2.3
0
15.9
±2.7
4
13.2
±2.5
0
23.8
±2.5
0
40.5
±1.0
8
22.8
±8.0
1
19.6
±3.4
9
30.7
±1.7
3
Ph
orm
idiu
m k
uet
zingia
num
BT
A0
83
4.3
2±
0.5
5
5.5
0±
0.5
5
9.8
0±
2.4
3
1.6
5±
0.3
4
0.3
7±
1.6
7
4.4
5±
0.8
9
5.0
7±
0.6
6
18.1
±0.8
3
103
An
ab
aen
a s
pir
oid
es B
TA
08
4
23
.4±
0.2
0
35
.0±
3.4
2
0.1
4±
0.5
9
73
.9±
2.7
0
61
.3±
2.1
1
1.3
8±
2.1
4
12
.2±
1.2
0
40
.3±
2.3
0
Lyn
gbya
laxe
spir
ali
s B
TA
08
5
6.9
9±
0.9
0
43
.4±
4.6
5
24
.2±
3.1
5
26
.2±
1.5
0
97
.2±
3.4
2
75
.1±
2.9
2
17
.7±
1.6
6
22
.4±
1.3
6
Ph
orm
idiu
m t
enu
e B
TA
086
0
.54
±0.4
0
26
.7±
5.1
7
2.2
0±
1.3
2
3.5
3±
0.4
3
32
.6±
1.1
3
6.3
0±
1.0
2
12
.1±
1.2
0
17
.3±
2.3
0
Nost
oc m
usc
oru
m B
TA
087
1
4.7
±5.5
0
12
3.6
±1
.8
65
.4±
1.5
2
43
.9±
1.3
0
27
3.1
±5
.68
15
9.7
±3
.29
60
.8±
4.2
0
80
.3±
1.4
0
Ple
cto
nem
a n
ota
tum
BT
A0
88
7
.66
±1.2
5
9.8
8±
1.5
5
12
.2±
1.8
7
2.4
6±
0.2
4
18
.9±
1.9
0
0.6
9±
1.9
7
7.4
0±
0.8
6
22
.2±
1.5
0
Scy
ton
ema b
oh
ner
i B
TA
10
6
1.8
1±
0.2
3
1.9
8±
0.9
6
2.4
6±
0.8
5
2.8
2±
1.0
3
6.2
8±
1.0
9
4.2
0±
1.3
0
6.1
7±
0.9
3
11
.2±
0.7
2
Ple
cto
nem
a n
ota
tum
BT
A1
08
7
.09
±3.0
9
9.9
9±
5.0
7
12
.9±
1.0
5
3.7
3±
1.9
2
11
.9±
2.8
5
5.7
7±
2.9
3
12
.6±
4.7
1
13
.4±
2.1
3
Ph
orm
idiu
m i
ncr
ust
atu
m B
TA
11
8
0.2
2±
0.1
0
1.3
8±
0.3
9
0.0
5±
0.2
7
1.9
8±
0.7
0
21
.0±
2.4
9
7.5
1±
0.9
0
28
.4±
1.9
7
24
.6±
0.8
2
Lim
no
thri
x re
dek
ei B
TA
123
1
3.2
±0.7
0
8.9
0±
0.5
7
2.9
5±
0.9
1
16
.2±
0.8
0
6.2
0±
0.3
2
3.0
3±
1.0
9
30
.2±
1.3
3
28
.4±
0.8
1
Ph
orm
idiu
m v
ald
eria
nu
m B
TA
16
2
2.2
7±
0.9
0
39
.9±
0.3
2
8.9
9±
0.1
4
5.3
2±
0.1
0
35
.0±
0.1
7
10
.6±
1.0
2
22
.9±
1.2
0
34
.6±
1.4
0
Scy
ton
ema g
uya
nen
se B
TA
16
7
1.8
2±
1.0
0
3.1
9±
1.0
3
4.3
7±
1.4
6
2.5
0±
1.0
1
6.1
7±
1.0
9
3.4
2±
1.5
0
10
.4±
3.9
6
15
.6±
4.9
4
Nost
oc
com
mu
ne
BT
A1
68
0.9
9±
1.0
2
3.1
8±
1.0
0
1.0
6±
1.0
2
1.2
0±
0.0
1
6.0
2±
1.0
0
3.4
2±
1.5
0
32
.4±
2.4
6
25
.6±
3.4
2
Osc
illa
tori
a a
ga
rdhii
BT
A1
70
2
3.6
±2.1
0
25
0.1
±1
.2
68
.4±
3.6
6
28
.7±
3.1
0
48
.7±
1.2
7
72
.4±
1.6
3
10
.8±
1.9
1
46
.5±
4.6
7
Sp
iruli
na
pla
tensi
s B
TA
174
2
7.5
±0.0
5
47
.1±
0.0
0
45
.5±
0.0
2
8.7
2±
0.0
0
19
6.1
±0
.01
56
.8±
0.0
2
20
.0±
0.0
0
55
.1±
0.0
0
Ca
loth
rix
cast
elli
i B
TA
177
0
.44
±0.4
0
1.5
8±
0.6
1
1.7
2±
0.5
8
1.4
7±
0.5
1
2.4
2±
0.0
0
3.7
0±
1.0
2
16
.1±
1.5
0
26
.2±
0.1
2
Lyn
gbya
co
nnec
tens
BT
A17
8
2.1
5±
0.8
0
4.1
5±
2.0
8
3.5
2±
0.7
2
6.1
2±
0.1
2
12
.0±
2.1
1
0.0
5±
0.1
3
22
.7±
1.3
9
18
.6±
1.1
1
Lyn
gbya
ag
ard
hii
BT
A1
84
3.1
3±
0.7
8
3.8
4±
0.8
2
5.0
6±
1.3
0
6.0
8±
1.0
6
8.0
0±
1.9
0
8.9
3±
1.6
9
10
.9±
0.8
7
30
.3±
1.9
8
Scy
ton
ema
sch
mid
tii
BT
A1
86
1
.37
±0.0
8
2.0
0±
0.0
1
2.5
6±
1.0
1
2.7
3±
0.0
2
3.7
5±
0.0
5
4.3
2±
1.1
1
19
.6±
2.8
3
21
.4±
2.5
0
104
Au
losi
ra a
enig
ma
tica
BT
A1
88
0
.46
±0.2
4
2.6
9±
2.6
3
3.1
3±
2.7
9
3.3
1±
4.1
8
0.9
2±
0.2
4
1.4
5±
0.3
9
10
.8±
2.4
2
15
.7±
3.4
3
Ph
orm
idiu
m t
enu
e B
TA
189
1
4.8
±1.2
5
25
.1±
6.6
9
20
.6±
3.7
3
2.3
5±
0.4
9
20
.9±
3.0
7
3.8
0±
0.7
4
35
.8±
1.2
8
40
.3±
2.4
4
Au
losi
ra b
om
ba
yensi
s B
TA
19
0
0.6
1±
0.1
6
0.9
7±
0.2
9
1.3
2±
0.3
0
6.2
0±
0.8
9
8.0
6±
1.5
9
1.8
4±
0.2
2
12
.7±
3.2
3
13
.8±
2.6
6
Ple
cto
nem
a n
ota
tum
BT
A1
94
3
.43
±0.3
2
29
.3±
3.9
8
6.0
4±
0.4
6
2.0
6±
0.8
3
2.5
5±
1.0
4
2.6
8±
0.4
8
30
.9±
2.6
6
45
.9±
2.3
1
Ca
loth
rix
ma
rchic
a B
TA
19
5
22
.9±
5.4
5
23
.6±
1.4
0
16
.1±
0.8
0
25
.6±
1.3
6
22
.3±
1.2
0
21
.0±
0.9
0
27
.6±
1.6
0
66
.4±
1.3
6
Lim
no
thri
x m
ira
bil
is B
TA
19
9
1.2
2±
0.1
0
77
.7±
1.0
5
1.2
3±
0.4
9
2.2
2±
0.1
2
82
.3±
1.0
7
2.3
2±
1.2
2
18
.9±
3.2
2
31
.0±
1.3
0
Nost
oc
calc
ico
la B
TA
20
4
3.9
9±
3.4
1
16
.8±
5.7
2
5.3
5±
0.4
8
0.5
0±
0.1
7
11
.5±
1.5
8
4.4
1±
0.0
6
20
.1±
2.2
4
44
.6±
3.1
2
An
ab
aen
a c
irci
na
lis
BT
A20
5
5.2
3±
0.4
2
8.2
9±
0.8
0
2.3
2±
0.4
0
4.2
8±
0.4
5
11
.6±
1.0
8
4.4
7±
0.0
6
16
.0±
0.6
1
17
.5±
1.4
2
Ca
loth
rix
ma
rchic
a B
TA
20
6
27
.2±
3.1
3
27
.1±
3.3
6
18
.3±
2.5
3
25
.2±
1.1
3
29
.2±
1.6
9
21
.0±
0.4
3
26
.3±
3.1
0
38
.1±
4.1
3
Ca
loth
rix
cla
vata
BT
A2
18
20
.5±
1.0
1
41
.4±
1.0
6
21
.7±
1.3
5
5.0
5±
0.3
8
7.0
0±
1.2
3
2.5
4±
0.4
3
22
.1±
3.3
0
24
.6±
4.5
0
Ph
orm
idiu
m t
enu
e B
TA
22
2
6.3
7±
5.3
7
40
..6
±6
.71
9.3
0±
1.5
2
1.1
0±
1.4
6
1.9
6±
1.0
1
1.8
7±
0.7
6
60
.9±
2.0
9
66
.1±
1.9
9
An
ab
aen
a ci
rcin
ali
s B
TA
24
6
7.3
6±
1.0
0
39
.3±
1.2
0
18
.0±
1.0
4
11
.6±
4.4
8
42
.3±
1.5
0
24
.2±
1.0
6
6.6
9±
0.8
6
10
.5±
0.8
4
Lyn
gbya
mart
ensi
an
a B
TA
43
6
5.1
0±
1.0
2
49
.3±
2.0
8
8.0
3±
1.0
4
8.1
3±
1.2
7
55
.9±
4.0
7
14
.1±
0.5
9
17
.2±
1.7
9
31
.8±
0.4
1
Lyn
gbya
d
igu
eti
BT
A4
75
8.3
2±
5.0
0
27
.3±
1.0
4
26
.6±
1.3
4
10
.1±
3.0
1
55
.9±
4.0
7
14
.1±
0.5
9
4.2
9±
1.1
6
7.7
4±
0.6
4
Ph
orm
idiu
m fr
agil
e B
TA
52
1
10
.8±
4.1
1
90
.2±
1.1
0
30
.2±
1.2
0
13
.2±
0.8
8
11
1.2
±1
.17
38
.8±
4.5
1
20
.3±
1.1
1
45
.2±
4.1
1
An
ab
aen
a ci
rcin
ali
s B
TA
56
1
6.1
6±
1.6
0
16
.4±
3.4
3
4.8
5±
1.3
6
5.1
2±
1.2
2
18
.3±
1.4
2
6.2
2±
1.3
0
22
.7±
2.3
0
29
.6±
1.4
0
An
ab
aen
a s
p.
BT
A5
64
16
.6±
7.8
0
48
.5±
1.3
8
44
.5±
3.2
2
12
.4±
2.3
0
24
.4±
3.9
1
8.3
2±
1.9
3
19
.0±
0.5
8
12
.9±
0.9
9
Ple
cto
nem
a n
ota
tum
BT
A5
65
5
.32
±3.4
3
12
.7±
3.8
9
10
.5±
1.3
6
7.9
5±
1.4
3
17
.7±
2.7
2
15
.3±
2.6
3
16
.7±
3.8
4
22
.0±
0.3
3
105
Ph
orm
idiu
m
autu
mn
ale
BT
A5
87
8.0
2±
3.5
2
15
.2±
1.0
9
11
.0±
3.6
6
10
.2±
1.5
4
24
.5±
6.3
1
18
.1±
3.0
5
4.2
6±
0.5
3
6.6
0±
0.4
0
Lyn
gbya
aes
tua
rii
BT
A597
4
.22
±1.3
6
7.3
2±
3.4
4
5.3
3±
1.1
1
5.9
1±
0.6
9
9.1
5±
1.7
4
8.6
2±
1.0
2
1.9
9±
0.4
8
5.7
9±
0.1
3
An
ab
aen
a va
ria
bil
is B
TA
60
0
3.9
4±
1.6
0
5.4
7±
0.8
6
5.9
4±
0.7
8
7.3
2±
3.5
4
7.2
2±
1.3
6
8.3
2±
3.4
0
2.5
0±
0.1
0
5.6
0±
1.4
0
Lyn
gbya
a
llo
rgei
BT
A6
06
7.8
0±
4.0
0
82
.8±
1.2
0
38
.9±
1.4
0
3.7
0±
3.6
0
71
.1±
3.6
5
30
.2±
1.2
8
16
.2±
3.5
4
30
.7±
12
.7
L
yng
bya
ma
rten
sia
na
B
TA
64
0
3.6
0±
2.9
0
0.8
7±
0.1
1
4.5
3±
0.8
6
5.8
7±
4.3
0
12
.8±
5.0
0
15
.2±
1.6
9
5.0
3±
1.1
3
16
.4±
2.5
0
Wes
tiel
lop
sis
pro
lifi
ca
BT
A6
45
0
.48
±0.0
0
4.2
2±
0.0
8
0.6
1±
0.3
2
2.4
2±
1.3
2
3.3
3±
1.0
0
2.5
0±
0.0
1
2.3
0±
0.9
0
12
.1±
0.5
5
An
ab
aen
a o
rien
tali
s B
TA
65
3
18
.7±
9.0
0
15
.0±
5.3
5
3.4
9±
2.3
9
32
.7±
4.5
0
22
.6±
5.9
0
10
.2±
6.2
0
11
.7±
2.6
5
16
.8±
4.6
6
Ph
orm
idiu
m s
tag
nin
a B
TA
85
5
3.9
2±
1.9
0
24
.5±
8.7
3
32
.4±
1.4
3
6.9
1±
0.1
0
25
.3±
2.1
1
15
.3±
0.1
0
30
.5±
6.0
9
21
.1±
1.3
0
An
ab
aen
a v
ari
abil
is B
TA
88
0
8.4
0±
5.3
0
24
.6±
8.7
5
32
.4±
1.8
5
4.6
7±
8.1
0
21
.2±
1.2
9
26
.6±
2.4
6
12
.8±
3.6
0
18
.7±
9.9
0
An
ab
aen
a o
ryza
e B
TA
88
1
6.2
0±
5.2
1
12
.6±
4.2
2
9.6
0±
3.6
2
7.0
5±
6.3
1
18
.8±
1.6
2
16
.8±
1.4
7
6.8
2±
11
.5
13
.1±
0.8
6
A
na
ba
ena f
erti
liss
ima
BT
A8
83
2
.53
±0.3
0
33
.5±
5.6
8
9.0
4±
1.3
5
4.3
6±
1.8
0
19
.5±
6.9
6
14
.5±
4.6
7
6.0
1±
0.1
8
1.0
6±
0.0
4
N
ost
oc
elli
pso
spo
rum
BT
A9
02
3
.23
±1.3
0
19
.3±
4.2
0
11
.5±
3.2
0
6.0
0±
1.3
2
28
.5±
5.3
6
19
.6±
4.1
1
1.3
0±
0.7
0
0.4
2±
0.0
6
A
na
ba
ena co
nst
rict
a B
TA
90
3
4.2
7±
2.0
0
17
.8±
5.3
2
9.5
6±
1.1
8
7.5
1±
0.2
0
16
.5±
3.3
4
21
.2±
6.3
0
4.5
2±
0.4
3
0.5
7±
0.1
6
Cyl
ind
rosp
erm
um
m
usc
ico
la
B
TA
90
4
5.2
3±
0.9
0
10
.3±
0.7
9
18
.0±
2.2
2
7.5
1±
0.2
0
16
.5±
3.3
4
21
.2±
6.3
0
1.0
5±
0.0
0
2.6
6±
0.1
9
Cyl
ind
rosp
erm
um
dory
phoru
m
BT
A9
05
3.0
5±
1.2
0
9.6
1±
2.2
0
6.9
9±
2.4
4
6.5
0±
1.4
0
24
.6±
3.2
2
11
.9±
3.2
0
1.5
2±
0.3
0
1.0
2±
0.2
2
Cyl
ind
rosp
erm
um
in
den
tum
BT
A9
06
2.1
3±
0.9
0
7.6
7±
1.1
7
4.9
3±
1.0
1
16
.4±
5.6
0
43
.4±
6.3
4
53
.7±
1.1
6
20
.1±
2.6
8
4.2
7±
0.4
6
106
An
ab
aen
a o
ryza
e B
TA
91
9
9.2
5±
1.8
0
27
.3±
0.6
1
7.8
6±
0.2
3
12
.2±
1.6
0
20
.7±
1.2
7
7.7
5±
3.4
6
24
.9±
3.6
3
2.6
6±
0.1
9
Nost
oc
ell
iposp
oru
m B
TA
92
3
6.5
2±
2.1
0
18
.2±
1.9
0
4.0
5±
0.9
9
5.1
4±
0.3
0
9.7
4±
0.5
9
12
.9±
1.1
1
8.3
3±
5.8
2
2.0
4±
0.4
3
Mic
roch
aet
e g
rise
a B
TA
92
6
19
.8±
5.8
0
20
.3±
3.6
8
14
.5±
2.1
7
30
.4±
6.1
0
46
.1±
6.1
6
36
.6±
4.6
5
43
.7±
2.3
2
11
.4±
1.5
7
An
ab
aen
a a
no
mala
BT
A92
7
20
.9±
3.6
0
31
.8±
4.5
7
15
.6±
1.6
8
28
.3±
5.7
0
52
.9±
5.9
0
22
.1±
3.2
0
43
.7±
2.3
2
11
.4±
1.5
7
Nost
oc
ver
ruco
sum
BT
A93
9
7.4
6±
0.8
8
21
.4±
2.4
4
4.8
8±
0.7
3
6.9
9±
1.0
7
13
.6±
1.4
9
16
.0±
1.4
4
14
.8±
2.1
9
16
.6±
1.8
3
Nost
oc
calc
ico
la B
TA
94
2
10
.1±
1.4
6
18
.1±
1.8
5
18
.0±
8.2
2
7.9
7±
0.8
2
16
.3±
0.4
4
12
.8±
1.3
3
4.2
8±
2.1
5
7.3
2±
1.8
3
Nost
oc
pis
cin
ale
BT
A94
7
1.7
2±
0.5
0
49
.1±
4.0
1
19
.0±
1.4
3
15
.6±
2.4
0
1
28
.7±
1.8
9
72
.9±
2.4
9
25
.5±
2.5
0
32
.6±
1.3
0
Cyl
ind
rosp
erm
um
in
dic
um
BT
A9
60
2.2
4±
0.7
0
37
.4±
6.0
8
21
.2±
4.7
0
10
.3±
0.5
0
67
.3±
0.0
4
79
.2±
1.4
9
4.6
7±
0.9
6
5.6
7±
1.0
0
An
ab
aen
a va
ria
bil
is B
TA
99
0
7.5
5±
0.8
0
23
.3±
1.7
4
19
.2±
1.2
6
10
.0±
7.6
0
36
.8±
1.9
3
41
.9±
1.8
3
4.4
9±
0.9
0
7.7
9±
1.6
0
Dic
hoth
rix
ba
uer
ian
a B
TA
10
59
5
.63
±4.2
0
7.3
6±
5.2
0
6.2
0±
3.2
0
9.5
2±
1.7
1
10
.3±
0.9
8
8.0
1±
3.7
1
9.3
1±
1.8
1
21
.3±
0.7
4
107
Strains name and code no. Biomass production and ARA activity
15th
day observation 30th
day observation
Chl-a
(µg/ml)
nmole
C2H4/µg of Chl-
a/hr H4/µg
Chl-a
(µg/ml)
nmole C2 of
Chl-a/hr
H4/µg
Microchaete uberrima BTA001 4.21 2.34±0.01 7.84 0.06±0.01
Anabaena variabilis BTA003 5.15 1.09±0.02 8.72 0.80±0.01
Anabaena doliolum BTA004 2.99 2.20±0.00 4.15 0.24±0.03
Anabaena variabilis BTA006 2.64 3.48±0.03 2.99 0.40±0.09
Microchaete grisea BTA007 5.78 1.22±0.01 10.5 0.15±0.04
Anabaena circinalis BTA008 4.94 1.18±0.04 5.42 0.15±0.06
Nostoc hatei BTA012 1.52 1.46±0.01 3.51 0.37±0.00
Anabaena oryzae BTA014 8.27 2.05±0.05 12.1 0.08±0.01
Calothrix wembaerensis BTA015 1.65 1.97±0.02 4.15 0.60±0.02
Anabaena oryzae BTA017 0.60 1.16±0.03 1.72 0.75±0.05
Anabaena oryzae BTA018 3.41 1.14±0.08 5.15 0.31±0.02
Anabaena spiroides BTA019 0.15 4.00±0.02 0.32 4.48±0.01
Anabaena ambigua BTA021 0.45 1.33±0.01 1.87 0.13±0.09
Anabaena anomala BTA023 0.58 0.06±0.03 1.90 0.22±0.01
Calothrix javanica BTA024 1.77 1.90±0.04 2.12 0.94±0.10
Calothrix geitonos BTA025 1.59 1.75±0.02 3.11 0.26±0.01
Calothrix marchica BTA026 1.59 1.50±0.03 2.65 0.72±0.04
Nostoc muscorum BTA027 4.43 0.07±0.02 7.23 0.21±0.04
Dichothrix orsiniana BTA028 0.92 1.74±0.02 0.78 3.33±0.06
Nostoc parmeliodes BTA029 1.21 1.57±0.01 1.54 1.95±0.12
Anabaena fertilissima BTA030 0.15 10.0±0.05 0.68 3.08±0.01
Anabaena oryzae BTA031 1.56 0.13±0.01 1.93 0.57±0.02
Table-10: Biomass production and nitrogenase activity of heterocystous
cyanobacteria of rice fields of Manipur
108
Anabaena ambigua BTA034 1.50 1.33±0.12 1.64 2.07±0.01
Anabaena fertilissima BTA035 1.76 1.53±0.06 1.88 1.43±0.03
Anabaena variabilis BTA036 1.23 7.48±0.07 1.47 7.01±0.02
Nostoc hatei BTA037 0.56 19.9±0.02 0.68 22.9±0.09
Nostoc carneum BTA038 0.08 11.3±0.07 1.14 8.68±0.10
Anabaena iyengarii BTA041 0.46 6.10±0.08 0.94 10.0±0.02
Anabaena varibilis BTA043 1.49 2.21±0.08 1.80 6.77±0.08
Microchaete loktakensis BTA044 0.44 2.50±0.09 1.14 4.70±0.05
Microchaete uberrima BTA045 0.18 5.55±0.06 0.36 12.8±0.04
Calothrix marchica BTA046 0.55 1.80±0.04 2.72 0.88±0.03
Microchaete tenera BTA049 3.96 2.17±0.02 2.90 0.38±0.05
Anabaena oryzae BTA050 0.15 3.8±0.09 0.12 5.31±0.06
Anabaena spiroides BTA084 0.13 3.48±0.05 3.60 0.25±0.05
Nostoc muscorum BTA087 3.52 0.11±0.01 2.21 9.00±0.06
Scytonema bohneri BTA106 3.45 2.61±0.00 2.29 5.85±0.10
Scytonema guyanense BTA167 1.23 9.59±0.02 0.64 3.20±0.12
Nostoc commune BTA168 0.28 8.80±0.08 0.50 7.20±0.11
Calothrix castellii BTA177 1.57 8.78±0.03 1.79 7.90±0.03
Scytonema schmidtii BTA186 0.19 5.30±0.05 0.36 3.30±0.03
Aulosira aenigmatica BTA188 1.64 5.49±0.02 3.00 3.70±0.04
Aulosira bombayensis BTA190 1.05 8.09±0.09 1.55 7.03±0.07
Calothrix marchica BTA195 3.76 7.45±0.03 4.89 7.01±0.00
Nostoc calcicola BTA204 2.22 5.18±0.01 2.37 6.16±0.02
Anabaena circinalis BTA205 1.71 5.38±0.00 2.14 5.00±0.07
Calothrix marchica BTA206 3.51 5.87±0.04 4.62 8.03±0.09
109
T
Calothrix clavata BTA218 6.79 3.14±0.01 8.74 4.30±0.05
Anabaena circinalis BTA246 1.37 0.58±0.02 1.54 1.62±0.12
Anabaena circinalis BTA561 2.49 0.44±0.08 1.02 2.65±0.03
Anabaena sp. BTA564 8.38 0.20±0.06 0.40 8.68±0.01
Anabaena variabilis BTA600 1.10 1.09±0.02 1.12 3.66±0.06
Westiellopsis prolifica BTA645 0.89 2.58±0.08 2.06 7.57±0.02
Anabaena orientalis BTA653 0.65 5.07±0.06 0.57 1.30±0.07
Anabaena variabilis BTA880 2.27 2.33±0.07 2.56 2.72±0.02
Anabaena oryzae BTA881 1.90 6.68±0.04 2.38 8.52±0.05
Anabaena fertilissima BTA883 6.31 1.46±0.03 1.60 6.86±0.08
Nostoc ellipsosporum BTA902 1.63 6.62±0.07 1.88 11.7±0.01
Anabaena constricta BTA903 1.69 7.81±0.04 3.07 11.2±0.02
Cylindrospermum muscicola
BTA904
2.38 4.96±0.01 1.60 12.1±0.05
Cylindrospermum doryphorum
BTA905
5.28 2.29±1.20 6.32 5.08±0.05
Cylindrospermum indentum
BTA906
1.22 10.2±0.02 3.26 10.7±0.08
Anabaena oryzae BTA919 1.55 13.7±1.23 3.44 13.4±0.02
Nostoc ellipsosporum BTA923 0.89 10.0±1.90 3.26 4.38±0.09
Microchaete grisea BTA926 2.43 0.29±0.02 0.18 4.79±0.01
Anabaena anomala BTA927 0.53 12.1±0.10 0.83 13.7±0.06
Nostoc verrucosum BTA939 4.69 2.92±0.11 5.21 4.61±0.00
Nostoc calcicola BTA942 2.06 0.73±0.09 0.92 2.27±0.12
Nostoc piscinale BTA947 3.77 0.26±0.05 0.41 4.61±0.13
Cylindrospermum indicum
BTA960
0.88 2.50±0.03 0.50 3.97±0.02
Anabaena variabilis BTA990 0.63 3.17±0.12 0.40 6.50±0.18
Dichothrix baueriana BTA1059 1.45 13.2±0.01 0.64 19.5±0.16
110
Table-11: Screening for total lipid from non heterocystous cyanobacterial
strains of rice fields of Manipur
Strains name and code no. Total lipid (%)
Phormidium mucosum BTA002 1.00
Limmothrix vacuolifera BTA005 3.00
Plectonema radiosum BTA009 1.70
Plectonema boryanum BTA010 2.00
Phormidium tenue BTA013 1.00
Plectonema boryanum BTA016 3.00
Phormidium faveolarum BTA020 1.00
Plectonema radiosum BTA032 1.00
Plectonema litorale BTA033 2.00
Myxosarcina burmensis BTA040 2.00
Phormidium arthurensis BTA042 2.00
Plectonema nostocorum BTA047 3.00
Phormidium fragile BTA048
1.00
Phormidium purpurascens BTA081 1.00
Limnothrix redekei BTA082 2.00
Phormidium kuetzingianum BTA083 1.00
Lyngbya laxespiralis BTA085 3.00
Phormidium tenue BTA086 1.00
Plectonema notatum BTA088 2.00
Plectonema notatum BTA108 2.00
Phormidium incrustatum BTA118 1.00
111
Limnothrix redekei BTA123 2.00
Phormidium valderianum BTA162 1.00
Oscillatoria agardhii BTA170 1.00
Spirulina platensis BTA174 2.00
Lyngbya connectens BTA178 2.50
Lyngbya nordgardhii BTA184 3.00
Phormidium tenue BTA189 2.00
Plectonema notatum BTA194 1.00
Limnothrix mirabilis BTA199 2.00
Phormidium tenue BTA222 2.00
Lyngbya martensiana BTA436 2.00
Phormidium fragile BTA521 1.00
Plectonema notatum BTA565 2.00
Phormidium autumnale BTA587 1.00
Lyngbya aestuarii BTA597 1.00
Lyngbya allorgei BTA606 1.00
Lyngbya martensiana BTA640 2.00
Phormidium stagnina BTA855 1.00
112
Table-12: Major lipid content from non heterocystous cyanobacterial
strains of rice fields of Manipur Strains name code no. Major fatty acids content in %
Phormidium mucosum
BTA002
Capric acid methyl ester (13.9), Lauric acid methyl ester
(7.23), Pentadecanoic acid methyl ester (39.9), Myristoleic
acid methyl ester (4.09), Tricosanoic Acid Methyl Ester (3.56)
Limnothrix vacuolifera
BTA005
Lauric acid methyl ester (8.59), Myristoleic acid methyl ester
(5.04), Pentadecanoic acid methyl ester (48.2), Heptadecanoic
acid methyl ester (5.45)
Plectonema radiosum
BTA009 Capric acid methyl ester (18.0), Lauric acid methyl ester
(11.0), Myristoleic acid methyl ester (3.12), Pentadecanoic
acid methyl ester (56.4), Cis-13, 16-Docosadienoic Acid
Methyl Ester (4.29)
Plectonema boryanum
BTA010
Butyric acid methyl ester (6.99), Capric acid methyl ester
(13.2) Myristoleic acid methyl ester (8.52), Pentadecanoic acid
methyl ester (13.8), Elaidic acid methyl ester (22.3)
Phormidium tenue BTA013 Caprylic acid methyl ester (5.47), Tridecanoic acid methyl
ester (4.76), Pentadecanoic acid methyl ester (65.2), Cis-10-
Heptadecanoic acid methyl ester (6.19), Oleic acid methyl
ester (11.6)
Plectonema boryanum
BTA016
Capric acid methyl ester (6.08), Lauric acid methyl ester
(7.49),Pentadecanoic acid methyl ester (54.5), Gamma-
Linolenic Acid Methyl Ester (8.82), Cis-13, 16-Docosadienoic
Acid Methyl Ester (2.99)
Phormidium faveolarum
BTA020
Caprylic acid methyl ester (4.48), Tridecanoic acid methyl
ester (4.89), Pentadecanoic acid methyl ester (56.0), Oleic acid
methyl ester (11.8), Linoleic Acid Methyl Ester (11.8)
Plectonema radiosum
BTA032
Capric acid methyl ester (16.6), Lauric acid methyl ester
(12.0), Pentadecanoic acid methyl ester (52.0), Gamma-
Linolenic Acid Methyl Ester (7.89), Cis-13, 16-Docosadienoic
Acid Methyl Ester (4.29)
Plectonema litorale
BTA033
Capric acid methyl ester (10.1), Pentadecanoic acid methyl
ester (72.7), Cis-10- Pentadecanoic acid methyl ester (6.32),
Cis-10- Heptadecanoic acid methyl ester (3.79), Oleic acid
methyl ester (0.11)
Myxosarcina burmensis
BTA040
Capric acid methyl ester (5.92), Pentadecanoic acid methyl
ester (49.3), Cis-10- Pentadecanoic acid methyl ester (13.9),
Cis-10- Heptadecanoic acid methyl ester (11.6), Oleic acid
methyl ester (11.6)
113
Phormidium arthurensis
BTA042
Pentadecanoic acid methyl ester (55.3), Cis-11-Eicosenoic acid
methyl ester (12.2), Caprylic acid methyl ester (4.93), Cis-10-
Heptadecanoic acid methyl ester (5.66), Oleic acid methyl
ester (4.93)
Plectonema nostocorum
BTA047
Capric acid methyl ester (13.0), Myristoleic acid methyl ester
(9.11), Pentadecanoic ester (25.8), Heptadecanoic acid methyl
ester (16.1), Caproic acid methyl ester (6.72)
Phormidium fragile
BTA048
Pentadecanoic acid methyl ester (65.7), Oleic acid methyl ester
(11.2), Cis-10- Heptadecanoic acid methyl ester (6.18),
Tridecanoic acid methyl ester (4.80), Caprylic acid methyl
ester (4.78)
Phormidium fragile
BTA081
Pentadecanoic acid methyl ester (21.8), Capric acid methyl
ester (13.3) ,Cis-10- Pentadecanoic acid methyl ester (7.36),
Tricosanoic Acid Methyl Ester (40.9), Elaidic acid methyl
ester (5.17)
Limnothrix redekei
BTA082
Pentadecanoic acid methyl ester (49.3), Capric acid methyl
ester (13.5), Lauric acid methyl ester (9.32), Tridecanoic acid
methyl ester (5.75), Myristoleic acid methyl ester (5.10)
Phormidium kuetzingianum
BTA083
Pentadecanoic acid methyl ester (40.9), Capric acid methyl
ester (13.8), Lauric acid methyl ester (7.30), Tridecanoic acid
methyl ester (4.56), Myristoleic acid methyl ester (4.16)
Lyngbya laxespiralis
BTA085
Pentadecanoic acid methyl ester (68.4), Oleic acid methyl ester
(8.91), Cis-10- Pentadecanoic acid methyl ester (6.88), Cis-10-
Heptadecanoic acid methyl ester (6.07), Capric acid methyl
ester (4.85)
Phormidium tenue BTA086 Capric acid methyl ester (14.9), Pentadecanoic acid methyl
ester (25.9), Cis-10- Pentadecanoic acid methyl ester (8.32),
Stearic acid methyl ester (6.67), Tricosanoic Acid Methyl
Ester (42.5)
Plectonema notatum
BTA088
Palmitic acid methyl ester (27.9), Tridecanoic acid methyl
ester (6.68), Capric acid methyl ester (12.0), Cis-10-
Pentadecanoic acid methyl ester (9.56), Butyric acid methyl
ester (2.78)
Plectonema notatum
BTA108
Pentadecanoic acid methyl ester (21.5), Palmitic acid methyl
ester (7.19), Cis-10- Heptadecanoic acid methyl ester (8.34),
Linoleic Acid Methyl Ester (9.49), Linolenic Acid Methyl
Ester (8.47)
Phormidium incrustatum
BTA118
Lauric acid methyl ester (8.77), Pentadecanoic acid methyl
ester (47.3), Gamma- Linolenic Acid Methyl Ester (7.41),
Heptadecanoic acid methyl ester (4.36), Tricosanoic Acid
Methyl Ester (4.14)
114
Limnothrix redekei
BTA123
Capric acid methyl ester (21.9), Lauric acid methyl ester
(12.0), Pentadecanoic acid methyl ester (15.9), Caprylic acid
methyl ester (2.56), Tridecanoic acid methyl ester (4.91)
Phormidium valderianum
BTA162
Capric acid methyl ester (17.7), Lauric acid methyl ester
(13.4), Pentadecanoic acid methyl ester (5.98), Tricosanoic
Acid Methyl Ester (5.16), Myristoleic acid methyl ester (4.67)
Oscillatoria agardhii
BTA170
Capric acid methyl ester (19.5), Lauric acid methyl ester
(10.5), Pentadecanoic acid methyl ester (6.10), cis-11, 14,17-
Eicosatrienoic Acid Methyl Ester (9.10), Erucic acid methyl
ester (4.78)
Spirulina platensis
BTA174
Caprylic acid methyl ester (6.50), Pentadecanoic acid methyl
ester (60.0), Palmitic acid methyl ester (6.89), Stearic acid
methyl ester (6.10), Behenic Acid Methyl Ester (11.6)
Lyngbya connectens
BTA178
Capric acid methyl ester (7.56), Lauric acid methyl ester
(8.02),Pentadecanoic acid methyl ester (69.4), Heptadecanoic
acid methyl ester (2.49), Tricosanoic Acid Methyl Ester (1.19)
Lyngbya nordgardhii
BTA184
Pentadecanoic acid methyl ester (66.7), Cis-10- Pentadecanoic
acid methyl ester (7.96), Oleic acid methyl ester (9.91),
Linoleic Acid Methyl Ester (8.09), cis-11, 14-Eicosadienoic
Acid Methyl Ester (1.23)
Phormidium tenue
BTA189
Lauric acid methyl ester (9.89), Pentadecanoic acid methyl
ester (47.8), Capric acid methyl ester (12.4), Heptadecanoic
acid methyl ester (5.47), Elaidic acid methyl ester (1.34)
Plectonema notatum
BTA194
Pentadecanoic acid methyl ester (18.3), Cis-10- Heptadecanoic
acid methyl ester (9.00), Linoleic Acid Methyl Ester (9.67),
Linolenic Acid Methyl Ester (9.87), Cis-11-Eicosenoic acid
methyl ester (9.16)
Limnothrix mirabilis
BTA199
Capric acid methyl ester (11.9), Lauric acid methyl ester
(9.18), Pentadecanoic acid methyl ester (46.9), Palmitoleic
acid methyl ester (2.46), Heptadecanoic acid methyl ester
(4.56)
Phormidium tenue
BTA222
Capric acid methyl ester (24.4), Lauric acid methyl ester
(17.0), Pentadecanoic acid methyl ester (16.2), Cis-10-
Pentadecanoic acid methyl ester (6.76), Stearic acid methyl
ester (10.9)
Lyngbya martensiana
BTA436
Capric acid methyl ester (7.64), Lauric acid methyl ester
(5.29), Pentadecanoic acid methyl ester (47.4), Tricosanoic
Acid Methyl Ester ( 25.7), Elaidic acid methyl ester (2.94)
Lyngbya digueti
BTA475
Caprylic acid methyl ester (4.39), Capric acid methyl ester
(4.02), Lauric acid methyl ester (3.66), Tridecanoic acid
methyl ester (3.29), Pentadecanoic acid methyl ester (84.6)
115
Phormidium fragile
BTA521
Capric acid methyl ester (17.5), Lauric acid methyl ester
(11.6), Myristoleic acid methyl ester (5.68), Pentadecanoic
acid methyl ester (6.97), Tricosanoic Acid Methyl Ester (44.9)
Plectonema notatum
BTA565
Capric acid methyl ester (9.43), Pentadecanoic acid methyl
ester (22.8), Heptadecanoic acid methyl ester (12.8), Gamma-
Linolenic Acid Methyl Ester (8.41), Linolenic Acid Methyl
Ester (10.7)
Phormidium autumnale
BTA587
Capric acid methyl ester (13.8), Lauric acid methyl ester
(9.67), Pentadecanoic acid methyl ester (46.9), Palmitoleic
acid methyl ester (3.45), Tricosanoic Acid Methyl Ester (4.17)
Lyngbya aestuarii
BTA597
Caprylic acid methyl ester (8.48), Pentadecanoic acid methyl
ester (47.7), Cis-10- Heptadecanoic acid methyl ester (4.91),
Oleic acid methyl ester (34.3), Linoleic Acid Methyl Ester
(2.68)
Lyngbya allorgei
BTA606
Pentadecanoic acid methyl ester (61.8), Capric acid methyl
ester (12.3), Lauric acid methyl ester (7.89), Tridecanoic acid
methyl ester (8.99), Palmitoleic acid methyl ester (1.31)
Lyngbya martensiana
BTA640
Capric acid methyl ester (51.2), Pentadecanoic acid methyl
ester (45.1), Heptadecanoic acid methyl ester (14.2), Stearic
acid methyl ester (8.90), Oleic acid methyl ester (56.2)
Phormidium stagnina
BTA855
Capric acid methyl ester (11.3), Pentadecanoic acid methyl
ester (21.6), Cis-10- Pentadecanoic acid methyl ester (7.32),
Stearic acid methyl ester (5.49), Tricosanoic Acid Methyl
Ester (39.9)
116
Table-13: Selected and potent strains with geographical information
SN Name & code of selected strains GPS information
1 Phormidium mucosum BTA002 N24°49'26.4" E093°57'52.0" Altitude: 775 m
2 Plectonema boryanum BTA010 N24°49'26.4" E093°57'52.0" Altitude: 775 m
3 Calothrix javanica BTA024 N24°49'36.0" E093°53'25.5" Altitude: 780 m
4 Anabaena fertilissima BTA035 N24°49'26.4" E093°57'52.0" Altitude: 775 m
5 Nostoc hatei BTA037 N24°42'26.1" E093°49’09.6" Altitude: 762 m
6 Phormidium arthurensis BTA042 N24°30'12.3" E093°46'46.4" Altitude: 761m
7 Microchaete loktakensis BTA044 N24°32'21.4" E093°45'27.6" Altitude: 764 m
8 Anabaena spiroides BTA084 N24°29'25.1" E094°00'43.7" Altitude: 769 m
9 Nostoc muscorum BTA087 N24°48'14.3" E093°54'18.3" Altitude: 782 m
10 Phormidium incrustatum BTA118 N24°30'12.3" E093°46'46.6" Altitude: 761 m
11 Spirulina platensis BTA174 N24°32'21.9" E093°45'27.6" Altitude: 764 m
12 Calothrix marchica BTA206 N24°20'36.7" E093°41'50.3" Altitude: 835 m
13 Phormidium tenue BTA222 N24°30'12.3" E093°46'46.6" Altitude: 761 m
14 Nostoc piscinale BTA947 N24°29'28.7" E094°00'24.1" Altitude: 805 m
15 Dichothrix baueriana BTA1059 N24°42'09.6" E093°48'22.3" Altitude: 773 m
117
Morpho-taxo and biochemical characterization of
selected cyanobacterial strains
1. Phormidium mucosum BTA002
Division: cyanophyta; class: cyanophyceae; order: nostocales; family: oscillatoriaceae
Thallus mucilaginous, lamellated, blue green
colour, sheath diffluent, bottom dweller,
entangled, distinctly constricted and
attenuated at the ends
Chlorophyll-a: 0.50 and 5.64 µg/ml
Total soluble proteins: 22.6 and 130.0 µg/ml
Ammonia excretion: 5.20 and 4.11 µg/ml
Total carbohydrates: 96.6 and 52.4 µg/ml
Carotenoids: 27.8 and 24.2 µg/ml
Phycoerythrin: 1.09 and 9.36 µg/ml
Phycocyanin: 11.1 and 33.1 µg/ml
Allo-phycocyanin: 3.60 and 14.6 µg/ml
Nitrogenase activity: 0.01 and 0.04 nmole C2H4/µg chl-a/hr
2. Plectonema boryanum BTA010
Division: cyanophyta; class: cyanophyceae; order: nostocales; family:
scytonemataceae
Thallus blue green color, colonial forms,
initially, floating later attached to the
bottom, constriction at septa present single
and branches present
Chlorophyll-a: 1.22 and 4.96 µg/ml
Total soluble proteins: 51.6 and 15.6 µg/ml
Ammonia excretion: 2.03 and 1.10 µg/ml
Total carbohydrates: 71.2 and 82.3 µg/ml
Carotenoids: 17.4 and 17.2 µg/ml
Phycoerythrin: 2.55 and 4.56 µg/ml
Phycocyanin: 11.0 and 14.2 µg/ml
Allo-phycocyanin: 6.86 and 7.32 µg/ml
Nitrogenase activity: 0.06 and 0.02 nmole C2H4/µg chl-a/hr
Total lipid composition: 28
118
3. Calothrix javanica BTA024
Division: cyanophyta, class: cyanophyceae; order: nostocales; family: rivulariaceae
Light green colour, filaments single,
unlamellated sheath, pointed apex, basal
heterocyst
Chlorophyll-a: 10.93 and 14.3 µg/ml
Total soluble proteins: 67.9 and 103.0 µg/ml
Ammonia excretion: 10.8 and 26.8 µg/ml
Total carbohydrates: 25.7 and 29.6 µg/ml
Carotenoids: 50.5 and 26.3 µg/ml
Phycoerythrin: 10.6 and 80.9 µg/ml
Phycocyanin: 19.9 and 27.9 µg/ml
Allo-phycocyanin: 9.80 and 20.0 µg/ml
Nitrogenase activity: 1.03 and 0.09 nmole C2H4/µg chl-a/hr
4. Anabaena fertilissima BTA035
Division: cyanophyta; class: cyanophyceae; order: nostocales; family: nostocaceae
Trichomes single, bent with almost rounded
ends cells, heterocyst spherical and adjoining,
spores in long chain,
Chlorophyll-a: 11.6 and 4.48 µg/ml
Total soluble proteins: 85.3 and 92.3 µg/ml
Ammonia excretion: 57.3 and 29.1 µg/ml
Total carbohydrates: 20.0 and 45.0 µg/ml
Carotenoids: 29.6 and 31.7 µg/ml
Phycoerythrin: 7.97 and 21.1 µg/ml
Phycocyanin: 62.5 and 150.4 µg/ml
Allo-phycocyanin: 54.4 and 130.7 µg/ml
Nitrogenase activity: 5.23 and 2.43 nmole C2H4/µg chl-a/hr
119
5. Nostoc hatei BTA037
Division: cyanophyta; class: cyanophyceae; order: nostocales; family: nostocaceae
Thallus spherical, trichome broad,
irregularly curved and densely entangled
cells spherical and heterocyst single
Chlorophyll-a: 13.3 and 5.51 µg/ml
Total soluble proteins: 51.3 and 115.0 µg/ml
Ammonia excretion: 169.3 and 19.2 µg/ml
Total carbohydrates: 15.0 and 46.0 µg/ml
Carotenoids: 10.4 and 21.5 µg/ml
Phycoerythrin: 6.93 and 4.34 µg/ml
Phycocyanin: 30.7 and 10.7 µg/ml
Allo-phycocyanin: 19.4 and 10.3 µg/ml
Nitrogenase activity: 19.9 and 22.9 nmole C2H4/µg chl-a/hr
6. Phormidium arthurensis BTA042
Division: cyanophyta; class: cyanophyceae; order: nostocales; family:
oscillatoriaceae
Thallus leathery, very thin, blue green,
trichomes bent, entangled, constricted at the
cross walls, sheath colourless
Chlorophyll-a: 0.74 and 12.5 µg/ml
Total soluble proteins: 61.6 and 59.3 µg/ml
Ammonia excretion: 52.4 and 44.5 µg/ml
Total carbohydrates: 20.6 and 15.0 µg/ml
Carotenoids: 5.12 and 30.4µg/ml
Phycoerythrin: 3.27 and 69.2 µg/ml
Phycocyanin: 4.75 and 286.1µg/ml
Allo-phycocyanin: 6.28 and 46.8µg/ml
Nitrogenase activity: 0.87 and 2.88 nmole C2H4/µg chl-a/hr
120
7. Microchaete loktakensis BTA044
Division: cyanophyta; class: cyanophyceae; order: nostocales; family:
microchaetaceae
Filaments aggregated, long uniformly
cylindrical, broad at base, sheath lamellated,
heterocyst basal, cell granulated
Chlorophyll-a: 9.29 and 10.5 µg/ml
Total soluble proteins: 58.3 and 94.0µg/ml
Ammonia excretion: 126.9 and 24.9µg/ml
Total carbohydrates: 18.6 and 40.0 µg/ml
Carotenoids: 20.6 and 50.2µg/ml
Phycoerythrin: 5.36 and 9.31µg/ml
Phycocyanin: 10.5 and 18.8µg/ml
Allo-phycocyanin: 18.2 and 18.5µg/ml
Nitrogenase activity: 3.22 and 1.23 nmole C2H4/µg chl-a/hr
8. Anabaena spiroides BTA084
Division: cyanophyta; class: cyanophyceae; order: nostocales; family: nostocaceae
Trichome single, regularly spirally coiled
with thick mucilaginous sheath, cells
spherical, heterocyst subpherical
Chlorophyll-a: 10.4 and 31.1µg/ml
Total soluble proteins:108.2 and 83.0 µg/ml
Ammonia excretion: 34.0 and 48.7µg/ml
Total carbohydrates: 34.6 and 37.3 µg/ml
Carotenoids: 12.2 and 40.2µg/ml
Phycoerythrin: 23.4 and 73.9µg/ml
Phycocyanin: 35.0 and 61.3µg/ml
Allo-phycocyanin: 0.14 and 1.38 µg/ml
Nitrogenase activity: 3.48 and 1.44 nmole C2H4/µg chl-a/hr
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9. Nostoc muscorum BTA087
Division: cyanophyta; class: cyanophyceae; order: nostocales; family: nostocaceae
Thallus membranous, irregularly expanded,
filaments densely. Entangled, sheath
distinct, heterocyst spherical, spores oblong
Chlorophyll-a: 20.2 and 17.2 µg/ml
Total soluble proteins: 59.6 and 78.0µg/ml
Ammonia excretion: 92.3 and 13.3µg/ml
Total carbohydrates: 38.3 and 44.6µg/ml
Carotenoids: 55.8 and 80.3µg/ml
Phycoerythrin: 14.7 and 43.9µg/ml
Phycocyanin: 123.6 and 273.1µg/ml
Allo-phycocyanin: 65.4 and 159.7µg/ml
Nitrogenase activity: 3.26 and 1.12 nmole C2H4/µg chl-a/hr
10. Phormidium incrustatum BTA118
Division: cyanophyta; class: cyanophyceae; order: nostocales; family:
oscillatoriaceae
Thallus mucilaginous lamellated, blue green
colour, sheath diffluent, end cell-acute-
conical, calyptra
Chlorophyll-a: 2.15 and 4.21µg/ml
Total soluble proteins: 80.0 and 81.3µg/ml
Ammonia excretion: 25.1 and 27.1µg/ml
Total carbohydrates: 36.2 and 39.4µg/ml
Carotenoids: 28.4 and 24.6µg/ml
Phycoerythrin: 1.98µg/ml
Phycocyanin: 21.0µg/ml
Allo-phycocyanin: 7.51µg/ml
Total lipid compositions: 23
Nitrogenase activity: 0.05 and 0.00 nmole C2H4/µg chl-a/hr
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11. Spirulina platensis BTA174
Division: cyanophyta; class: cyanophyceae; order: nostocales; family:
oscillatoriaceae
Blue green, trichomes slightly constricted
at the cross, walls regularly spirally coil,
end cells broadly rounded
Chlorophyll-a: 10.8 and 10.0µg/ml
Total soluble proteins: 65.6 and 130.3µg/ml
Ammonia excretion: 84.4 and 58.8µg/ml
Total carbohydrates: 80.0 and 90.0 µg/ml
Carotenoids: 20.0 and 74.1µg/ml
Phycoerythrin: 27.5 and 8.72µg/ml
Phycocyanin: 47.1 and 196.1µg/ml
Allo-phycocyanin: 45.5 and 56.8 µg/ml
Nitrogenase activity: 0.04 and 0.01 nmole C2H4/µg chl-a/hr
12. Calothrix marchica BTA206
Division: cyanophyta; class: cyanophyceae; order: nostocales; family: rivulariaceae
Filaments single, with close coloured sheath,
trichome blue green distinctly constricted,
basal heterocyst
Chlorophyll-a: 2.57 and 6.23µg/ml
Total soluble proteins: 112.0 and 98.4µg/ml
Ammonia excretion: 22.3 and 19.0µg/ml
Total carbohydrates: 56.6 and 98.7µg/ml
Carotenoids: 26.3 and 38.1µg/ml
Phycoerythrin: 27.2 and 29.2µg/ml
Phycocyanin: 27.1 and 21.0µg/ml
Allo-phycocyanin: 18.3 and 21.0µg/ml
Nitrogenase activity: 0.06 and 0.02 nmole C2H4/µg chl-a/hr
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13. Phormidium tenue BTA222
Division: cyanophyta; class: cyanophyceae; order: nostocales; family:
oscillatoriaceae
Thallus pale blue green, thin membranous,
expanded, trichome straight highly
entangled, slightly constricted at the cross
walls, attenuated at the ends, sheath thin, end
cell acute conical
Chlorophyll-a: 0.37 and 10.5µg/ml
Total soluble proteins: 82.7 and 92.6µg/ml
Ammonia excretion: 43.0 and 68.9µg/ml
Total carbohydrates: 39.0 and 38.6µg/ml
Carotenoids: 60.9 and 66.1µg/ml
Phycoerythrin: 6.37 and 1.10µg/ml
Phycocyanin: 40.6 and 1.96µg/ml
Allo-phycocyanin: 9.30 and 1.87µg/ml
Nitrogenase activity: 0.06 and 0.00 nmole C2H4/µg chl-a/hr
14. Nostoc piscinale BTA947
Division: cyanophyta; class: cyanophyceae; order: nostocales; family: nostocaceae
Thallus membranous, blue green colour,
coiled, sheath distinct, heterocyst spherical
and intercalary, spores oblong
Chlorophyll-a: 4.22 and 25.2µg/ml
Total soluble proteins: 69.6 and 57.0 µg/ml
Ammonia excretion: 53.0and 5.90µg/ml
Total carbohydrates: 19.0 and 48.6µg/ml
Carotenoids: 25.5 and 32.6µg/ml
Phycoerythrin: 1.72 and 15.6 µg/ml
Phycocyanin: 49.1 and 128.7µg/ml
Allo-phycocyanin: 19.0 and72.9 µg/ml
Nitrogenase activity: 0.66 and 0.13 nmole C2H4/µg chl-a/hr
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15. Dichothrix baueriana BTA1059
Division: cyanophyta; class: cyanophyceae; order: nostocales; family: rivulariaceae
Thallus caespitose, penicillate, green in colour,
ultimate branches, sheath close to trichome,
constricted at the cross walls, heterocyst basal
Chlorophyll-a: 2.71 and 6.68µg/ml
Total soluble proteins: 39.0 and 90.6µg/ml
Ammonia excretion: 17.7 and 46.2µg/ml
Total carbohydrates: 31.5 and 26.0 µg/ml
Carotenoids: 9.31 and 21.3µg/ml
Phycoerythrin: 5.63 and 9.52µg/ml
Phycocyanin: 7.36 and 10.3µg/ml
Allo-phycocyanin: 6.20 and 8.01µg/ml
Nitrogenase activity: 13.2 and 19.5 nmole C2H4/µg chl-a/hr
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4.4. Discussion
Out of one hundred twelve (112) unialgal cyanobacterial strains screened and
characterized in present study, 15 strains namely; Phormidium mucosum BTA002,
Plectonema boryanum BTA 010, Calothrix javanica BTA024, Anabaena fertilissima
BTA035, Nostoc hatei BTA037, Phormidium arthurensis BTA042, Microchaete
loktakensis BTA044, Anabaena spiroides BTA084, Nostoc muscorum BTA087,
Phormidium incrustatum BTA118, Spirulina platensis BTA174, Calothrix marchica
BTA206, Phormidium tenue BTA222, Nostoc piscinale BTA947 and Dichothrix
baueriana BTA1059 were found to be potent candidates for commercialization.
Anabaena spiroides and Nostoc piscinale were produced highest chl-a in 30th
days old biomass ie., 31.1 µg/ml and 25.2 µg/ml respectively. Nostoc hatei and
Microchaete loktakensis were identified for potential candidates for excretion of
ammonia in culture medium during exponential growth phase as excreted 169.3 µg/ml
and 126.9 µg/ml respectively. Highest carbohydrate was produced by Calothrix
marchica ( 98.7 µg/ml) followed by Plectonema boryanum (82.3 µg/ml) in 30th days
old biomass. Spirulina platensis and Phormidium mucosum were produced almost
similar quantity of total soluble proteins ie., 130.3 µg/ml and 130.0 µg/ml respectively
after the end of growth phase. Nostoc muscorum and Phormidium tenue produced
highest arotenoids ie., 80.3 µg/ml and 66.1 µg/ml respectively in 30th
days old
biomass. Calothrix javanica and Anabaena spiroides were released highest
phycobiliprotein ie., 80.9 µg/ml and 73.9 µg/ml respectively in 30th
days old biomass.
Phormidium arthurensis and Nostoc muscorum were released highest
phycocyanin in 30th
days old biomass ie., 286.1 µg/ml and 273.1 µg/ml respectively.
Anabaena fertilissima and Nostoc muscorum were produced highest amount of allo-
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phycocyanin in 30th
days old biomass ie., 130.7 µg/ml and 159.7 µg/ml respectively.
Nostoc hatei BTA037 and Dichothrix baueriana BTA1059 were showed highest
acetylene reduction activity on 15th
day growth ie., 22.9 nmole C2H4/µg of Chl-a/hr
and 19.5 nmole C2H4/µg of Chl-a/hr respectively.
Forty (40) non heterocystous unialgal cyanobacterial strains were subjected
for lipid profiling and fatty acid composition. Plectonema boryanum BTA010 and
Phormidium incrustatum BTA118 were identified most potent strain as produced total
28 and 23 known fatty acid compositions respectively and few of them are very useful
for pharmaceutical and nutraceutical industry. The following five fatty acid
compositions were recorded in present investigation are to be comparable with
already commercialized strains. These are; lauric acid methyl ester (8.77%),
pentadecanoic acid methyl ester (47.3%), gamma-linolenic acid methyl ester (7.41%),
heptadecanoic acid methyl ester (4.36%), tricosanoic acid methyl ester (4.14%).
The cyanobacterium Spirulina has already been commercially exploited
because of its merits on yield and utility of cellular constituents (Borowitzka and
Borowitzka, 1988). Therefore, it would be appropriate if the growth characteristics
and biochemical composition of the presently investigated species could be compared
with those of Spirulina strain already commercialized worldwide. Spirulina
established at CFTRI, Mysore, India was discovered as potent candidate and
commercialized long back due to presence of protein within the range of 40-67 %
along with total lipid between 2-7 % (Tasneem Fatma et al., 1999). In the present
investigation, Plectonema boryanum BTA010 and Phormidium incrustatum BTA118
showed high lipid content including linoliec acid, gamma linolenic acid which are
very important for nutraceutical and pharmaceutical industry. The gamma linolenic
acid (GLA) were reported as a promising therapeutic agent for numerous health
127
disorders acting precursor for prostaglandin E1 an important compound necessary for
reducing inflammation and in treatment of heart disease (Biagi et al., 1991; Ghazala
et al., 2005; Tran et al., 2009; Wainwright et al., 1996).
In present investigation, Spirulina platensis BTA174 and Phormidium tenue
BTA222 were identified as potent strains as a source of single cell protein which were
contained total soluble proteins ie., 130.3 µg/ml, 130.0 µg/ml respectively and are
comparable with commercialized Spirulina strains world wide. Phycocyanin is the
major and important phycobiliproteins which exhibited anti-cancer activity,
stimulation of immune system and ability to treat ulcers and haemmorrhoidal
bleeding. Studies revealed that Phormidium arthurensis BTA042 and Nostoc
muscorum BTA087 were identified as a highest amount of phycocyanin produced
cyanobacteria ie., 286.1 µg/ml and 273.1 µg/ml respectively and comparable with
already commercialized strains for said purpose (Cohen 1986; Dainippon Patent 1980;
Dainippon Patent 1981; Borowitzka 1994).Vargas et al., 1998 reported that when
nitrogen fixing cyanobacteria were grown under diazotrophic conditions, protein,
carbohydrate and lipid content varied organism to organism.
In the present investigation, Anabaena spiroides BTA084 (25.2 µg/ml) and
Nostoc piscinale BTA947 (31.1 µg/ml) were identified as highest amount of chl-a
producing cyanobacteria which is the clear cut indication that the both cultures were
capable to produce good amount of biomass in culture conditions. Since the culture
conditions are known to be changed the biochemical composition of the algae
(Ciferri, 1983), the constituents can be improved upon further by manipulating culture
conditions. Deitary intake of carotenoids is positively correlated with
chemoprevention of cancer and other degenerative diseases. In the present study, it
was observed that Nostoc muscorum BTA087 and Phormidium tenue BTA222 were
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produced highest carotenoids as 66.1 µg/ml and 80.3 µg/ml respectively under culture
conditions which was comparable to previous workers reported the carotenoid
between 39-113 µg/ml in Phormidium laminosum (Fresnedo et al., 1991)
Synechococcus sp. (Gombos and Vigh 1986; Linden et al., 1990) and Nostoc
commune (Olie and Potts 1986; McPherson. 1986; Clement et al., 1967; Becker,
1986). The rate of acetylene reduction activity by cyanobacteria generally ranges
from 1-10 nmol C2H4/µg chl-a/hr (Fogg et al., 1973). In the present study, Nostoc
hatei BTA037 and Dichothrix baueriana BTA1059 were identified as a potent
diazotrophic cyanobacteria as these were expressed ARA as 82.9 nmole C2 of Chl-
a/hr H4/µg and 90.5 nmole C2 of Chl-a/hr H4/µg respectively in culture conditions.
Selected strains may be good candidates for their utilization as cyanobacterial
biofertilizer program.
The present investigation may be concludes that the Phormidium mucosum
BTA002, Plectonema boryanum BTA010, Calothrix javanica BTA024, Anabaena
fertilissima BTA035, Nostoc hatei BTA037, Phormidium arthurensis BTA042,
Microchaete loktakensis BTA044, Anabaena spiroides BTA084, Nostoc muscorum
BTA087, Phormidium incrustatum BTA118, Spirulina platensis BTA174, Calothrix
marchica BTA206, Phormidium tenue BTA222, Nostoc piscinale BTA947 and
Dichothrix baueriana BTA1059 are to be potent candidates for commercial
exploitation for one or other reason (table 13 followed by detailed characterization
including morpho-taxonomy). While considering as a potent cyanobacterial strains for
pharmaceutical and nutraceutical exploitation, other parameters to be worked out
thoroughly before proceeding commercialization. Therefore, detailed
characterizations are to be required for further understanding the biology and
biochemistry of individual strains accordingly.