Review of literature - Information and Library Network …shodhganga.inflibnet.ac.in/bitstream/10603/116004/8/08...Review of Literature 4 Cyanobacteria evolved on this planet ca 3.5

Review of literature

Review of Literature 4

Cyanobacteria evolved on this planet ca 3.5 billion years ago and the taxonomy

of this group is still a subject of controversy. As far as cyanobacterial taxonomy and

phylogeny are concerned, two schools are currently identifiable. One school considers

cyanobacteria as blue-green algae and the taxonomic system is purely morphology

based, while the other considers them as bacteria and follows a system similar to

bacterial taxonomy. Geitler (1932) mainly contributed to the taxonomy of cyanobacteria

based on their morphological details and he described about 1300 species of 144

genera from Germany, Austria and Switzerland. Desikachary (1959) has described

about 700 species of 85 genera from India and its sub-continents. Starmach (1966)

described about 1000 species of 91 genera. Other accumulated descriptions of

cyanobacterial genera and species such as those of Fremy (1930; 1934), Komarek (1958),

Kondrateva (1968; 1975), Bourrelly (1969; i970a; i970b), Anagnostidis and Komarek, (was,

1988), etc., have all followed the same classification system as that of Geitler (1932).

This morphology-based taxonomy is therefore referred to as the "Geitlerian" system

(Castenholz and Waterbury, 1989).

All the above systems of classification are based on mere differences in

morphological characters such as the presence of a sheath around individuals or

around a colony, pigmentation, trichome width, cell division planes, cell shape, cell

dimension, cell numbers in a colony, presence and position of heterocyst, type of

branching, presence of nannocytes, akinetes, hormogones, gas vacuoles, absence of

thylakoids, arrangement of the thylakoids, type of trichome breakage, presence or

absence of necridia, number of septa per cell, motility of spores, zones of intensive

growth ("meristems"), cells with special reproductive functions (baeocytes), etc..

However, some of these characters were found to vary with changes in environmental

condi t ions as wel l as in cul ture (Pearson and Kingsbury, 1966; Chang, 1977; Hoffmann, 1985;

Hoffmann and Demoulin, 1985; Anand, 1988; Wilmotte, 1988).

Morphological variations due to culture and environmental conditions made

Drouet (1968; 1981) to revise the cyanobacterial taxonomy drastically, where he reduced

the number of species to only 62 of 9 genera by selecting some morphological features

considered to be stable irrespective of the environmental conditions (Drouet, 1968; I98i).

This, however, was another morphology based classification system called, the

" D r o u e f sys tem (Drouet and Daily, 1956; Drouet, 1968; 1973; 1978; 1981) that d id not folloW the

"Geitlerian" system. Drouet's system of classification was based on morphological

characteristics of herbarium species and unpublished descriptions of cultures of

cyanobacteria. Drouet's basic concept was that each cyanobacterium has one


genotype and many phenotypes or ecophenes (ecological growth forms). According to

Anagnostidis and Komarek (1985), the species sensu Drouet represents polymorphic

clusters of "ecophenes" in a few, broadly conceived genera.

According to bacterial system of taxonomy, each isolate should be a type culture

and thus, Rippka et al. (1979) have provided a system of classification based on

cyanobacteria in culture, mostly following the traditional system, but, modified wherever

discriminatory characters were not determinable. This system, however, could only be

useful up to the generic level identification, where they have divided cyanobacteria into

five sections. The cyanobacteria studied in pure culture are currently placed in five

"orders" with gradual refinements (Castenholz, 1989a; 1989b; 1989c; 20OI; Castenholz and

Waterbury, 1989; Waterbury, 1989a; Waterbury and Rippka, 1989) that correspond closely to the

five groups (Sections) used by Stanier and his collaborators (Rippka, 1988; Rippka et ai.,

1979; 1981).

The position of the genus Plectonema (filament with single trichome, false branch

and no heterocyst) remains controversial throughout the history of cyanobacterial

taxonomy. Gomont (1892) has placed Plectonema along with Symploca, Lyngbya,

Phormidium, Trichodesmium, Borzia, Oscillatoria, Arthrospira and Spirulina under

Lyngbyeae of Oscillatoriaceae. Geitler (1932), Desikachary (1959) and Starmach (1966)

have placed it variously under Scytonemataceae of Hormogonales, Nostocales and

Hormogoniophyceae respectively. Several other authors like Tilden (1910), Fritsch

(1942; 1945), Bourrelly (I970a; 1970b), Hirose and Hirano (1977) and Christensen (1980) also

considered Plectonema under Scytonemataceae. However, Kondrateva (1968) has

placed it under a separate family Plectonemataceae along with three more families of

Oscillatoriales Elenk. Anagnostidis and Komarek (1988) placed it under the sub-family

Plectonematoideae of family Oscillatoriaceae, of the order Oscillatoriales. Rippka et al.

(1979) placed it under "LPP group B" along with Lyngbya and Phormidium. Castenholz

(2001) has recently assigned over 50 cultured cyanobacterial strains under the form-

genus Leptolyngbya including Plectonema, Lyngbya, Phormidium and Oscillatoria.

Wilmotte and Herdman (200i) questioned whether the genus Plectonema should be

retained or merged with Leptolyngbya.

Komarek and Anagnostidis (1989) commented that the assignments of more than

50% of the taxa deposited in culture collections are not proper. Wilmotte (1994) pointed

out that incorrect identification of strains leads to wrong comparison and interpretation

of not only taxonomic results but also studies on physiology, biochemistry or molecular


biology. Therefore, Wilmotte (1994) suggested to re-characterize all the cyanobacterial

strains maintained in the laboratories and culture collections throughout the world.

Stanier et al. (1971) concluded that the members of Chroococcales cannot be

adequately characterized or classified solely based on morphology and the taxonomy

of Chroococcales should be based on isolation, investigation and comparison of

biochemical and physiological parameters of pure strains. Lewin (1974) also believed

that the combination of morphology, biochemical and physiological features could

provide the most useful classification system.

Lyra et al. (1997) have emphasized the need to improve the taxonomical criteria of

cyanobacteria, which are mostly based on morphology. Several authors have shown

interest In utilization of biochemical characters for cyanobacterial taxonomy and

phytogeny (Lewin, 1974; Smith and Hoare, 1977; Stanier and Cohen-Bazire, 1977; Ragan and

Chapman, 1978; Hoiton, 1981). Isozymes have been investigated as possible tools for the

study of taxonomy and phylogeny in a variety of organisms (Markert, 1975). However,

relatively only few, studies have been made on algae in general or cyanobacteria in

particular (Helton, 1973). Hoiton (I98i) strongly believed that isozyme patterns of

cyanobacteria might aid in taxonomic studies at the genus and species level.

The available information related to the parameters considered for taxonomic and

phylogenetic characterization of cyanobacteria in the present study is reviewed below.

CHLOROPHYLL a

Chlorophylls are greenish pigments of several kinds and the most important one

is chlorophyll a. Although, all plants, algae and cyanobacteria that photosynthesize

contain chlorophyll a, in cyanobacteria alone chlorophyll a is the only type of

chlorophyll present. This is characteristic to cyanobacteria and is the sole reaction

center pigment. Chlorophyll molecule consists of two parts, a porphyrin head and a

long hydrocarbon, or phytol tail. A porphyrin is a cyclic tetrapyrrole, made up of four-

nitrogen containing pyrrole rings arranged in a cyclic fashion. Chlorophyll a content in

cyanobacteria reported in literature range from 0.052-2.99 % of dry weight (Table 1).

The lowest amount observed in Oscillatoria willei BDU 130511 (Saha et. ai., 2003b) and

the highest amount was reported from Aphanizomenon sp. (de Nobel et ai., 1998).

Environmental factors also cause variations in chlorophyll a content (Oiaizoia and Duerr,

1990; de Nobel et al., 1998; Gordlllo et ai, 1999; Kallb, 2002; Saha et al, 2003b).


Phytol is a mono-unsaturated diterpenoid primary alcohol and is the alcohol

moiety of chlorophyll, is a suitable precursor for the synthesis of vitamin A, p-carotene,

vitamin E, vitamin K, and vitamin K2. Phytol can be prepared readily by hydrolysis of

chlorophyll with mild acid, thus deserves commercial importance (Borowitzka, 1988).

CAROTENOIDS

Carotenoids are yellow, orange or red pigments, largely distributed in nature

namely, cyanobacteria, algae, green plants, bacteria, yeast and fungi (Goodwin, 1980). In

photosynthetic organisms, these pigments are involved in trapping light energy as

accessory pigments. A more general role applicable to both photosynthetic and non-

photosynthetic cell is protection from photodynamic action (Goodwin, 1980). There are

two basic categories of carotenoids: i) carotenes that consist of only hydrocarbon and

ii) xanthophylls that are oxygen-containing derivatives. In addition to very few bacterial

carotenoids with 30, 45 or 50 carbon atoms, C4o-carotenoids represent the majority of

more than 600 known structures. Especially bacterial carotenoids are most diverse

(Sandman, 2001). Genes controlling the synthesis of these pigments have been

studied in several organisms including Arabidopsis (Cunningham et ai., 1996; Bartiey et ai.,

1999), Brevibacterium linens (Kmbasik and Sandmann, 2000), Erwinia sp. (Misawa et a!., 1990;

Perry et at., 1986; Schnurr et al., 1991), Mycobacterium aurum A"" (Houssaini-kaqui et ai, 1992;

Viveiros et a!., 2000) and Xanthophyllomyces dendrortious (Verdoes et at.. 1999). Carotenes

and xanthophylls contribute to the yellowish colour of cyanobacteria grown at high light

intensities (Van Uere and Waisby, 1982). In cyanobacteria, major carotenoid components

are |3-carotene (80%) followed by myxoxanthophyll (Goodwin, 1980). Qualitative and

quantitative carotenoids composition was used to characterize cyanobacterial strains

(Komarek et al., 1999).

Total carotenoids content varies from species to species as reported in literature

range from 0.02-2% of dry weight (Table 2). In Spirulina platensis, Gordillo et al. (1999)

found 2.02% carotenoids, Olaizola and Duerr (1990) reported almost near 1.8%

whereas, Aakerman et al. (1992) reported a very low content (0.13% of dry weight) of

carotenoids for the same species (Table 2). However, the variation in carotenoids

content is correlated with the environmental factors especially quality and quantity of

l ight (Vesk and Jeffery, 1977; Guillard et al., 1985; Bidigare et al., 1989; Boussiba et al., 1999; Mallick

and Rai, 1999; Lakatos et ai, 2001; Kalib, 2002; Saha et al., 2003b).

Carotenoids are considered as photoprotective and also antioxidative agents

(Krinsky, 1979; Paiozza and Krinsky, 1992). A photoprotecive role of canthaxanthin has been


demonstrated by genetically engineered Synechococcus sp. (Aibrecht et ai., 2001). The

carotenolds act by quenching singlet oxygen produced during photosynthesis, owing to

their multiple conjugated double-bond system (Terao, 1989). The ketocarotenoids

echinenone is unique in normal photosynthetic cells of cyanobacteria, whilst

myxoxanthophyll Is a carotenoid glycoside, which is present in all cyanobacteria so far

examined except Phormidium spp. (Henzberg etai., 1971). Myxoxanthophyll was found to

be the major pigment constituting nearly 32% of the total carotenoids in SpiruHna. This

was followed by p-carotene (29%) and zeaxanthin (14%) in abundance (Priya Sethu,

1996). The cyanobacterial extracts containing carotenes and various other carotenoids

are frequently used as natural colouring materials. Carotenoids also have important

metabolic functions in animals and man, including conversion to vitamin A,

enhancement of the immune response and protection against diseases such as cancer

by way of scaveng ing f ree radicals (Goodwin, 1986; Bendich and Olson, 1989).

Thus, carotenoids obtained from cyanophytes have high market value, as the

commercial applications of carotenoids are many, p-carotene is used as a food

colourant and the annual market value for the pigment is about US$ 10 million

(Borowitzka, 1994). As a feed additive, P-carotene enhances the colour of the salmonoid

fish flesh (Schiedt et ai., 1985). It also enhances colour of the egg yolks (Schiedt et ai., 1985)

and improve health and fertility of cattle (Jackson et ai., I98i).

PHYCOBIUNS

In cyanobacteria and red algae, the outer surface of the thylakoids or

photosynthetic membranes is covered by rows of closely spaced granular structures.

They harvest light energy for photosynthesis in the reaction centers embedded in the

thylakoid membranes. These granular structures are composed of phycobilins with

covalently linked bile pigment chromophores, which may comprise more than 60% of

the total soluble protein, or about 20% of the total dry weight (Bennett, 1972; Bennett and

Bogorad, 1971). Phycobilins in phycobilisomes are accessory photosynthetic pigments,

consisting mainly of allophycocyanin (APC) cores surrounded by phycocyanin (PC) and

phycoerythrin (PE) on the periphery with distinct structure and absorption maxima. The

colours of the phycobilins arise from the presence of covalently attached prosthetic

groups - bilins, which are linear tetrapyrroles derived biosynthetically from heme via

biliverdin. Although PC is the major phycobilin in most of the cyanobacteria, PE and

APC may also occur in considerable quantities in some species. For example, PC and

APC were up to 17% and 11 % dry weight of Anabaena sp. and Nostoc sp. respectively

(Moreno et ai., 1995). The phycobilin composition (especially PC:PE ratio) was used by


several authors for characterization of various cyanobacterial taxa (Waterbury and Rippka,

1989; Waterbury et ai., 1986; Komarek et ai., 1999). From the characterization Study with three

simplest cyanobacteria (Cyanobium, Cyanobacterium and Synechococcus), Komarek

et al. (1999) felt that the PC/PE ratio can be considered as strain- and species-specific.

However, Schenk and Kuhfittig (1983) who studied the electrophoretic patterns of

phycobilins of 21 cyanobacterial species and could not establish any classification

schemes. Even the presence or absence of phycoerythrocyanin synthesis could not

provide any solid taxonomic information except for exclusionary character at the genus

level for filamentous, heterocystous strains (Bryant, 1982). Neilan et al. (1995) have used

intergenic spacer (IGS) between the two bilin-subunit genes of PC operon, designated

P (cpcB) and a {cpcA) as highly variable region of DNA sequence for the establishment

of genetic diversity and phylogeny of toxic cyanobacteria.

Phycocyanin content in various cyanobacteria reported in literature range from

2.1-22.4 % of dry weight (Table 3). Cohen et al (1993) found 10.4-22.4 % phycocyanin

for Spirulina platensis, while Gordillo et al. (1999) reported only 4.16% for the same

species. Malliga (1993) reported distinct variations in phycocyanin content within two

isolates of Anabaena azollae (Table 3). Allophycocyanin content of cyanobacteria

determined so far have also shown a range of variation within the genus, species and

strains. The amount reported in the literature range from 1-6.3 % of dry weight (Table

4). Similarly, phycoerythrin content of cyanobacteria, range from 0.26-8.8 % of dry

weight (Table 5).

Light intensity plays a major role on abundance and size of phycobilisomes that

accumulate and enlarge at low light intensities, while light quality can cause a dramatic

change in PBS composition. This modulation of the accessory pigment composition

occurs by synthesis or decrease of the phycobilins phycoerythrin or phycocyanin (See

Postius et al., 2001). The amount of PE increased under green light and that of

phycocyanin under red light conditions and the process is called 'complementary

chromatic adaptation' (Tandeau de Marsac and Houmard, 1993). Cyanobacteria are assigned

to three physiological groups based on their response to light quality: group 1 - PE and

PC synthesis is not changed due to light quality, group 2 - PE synthesis is regulated by

light quality and not PC, group 3 - preferential synthesis of light-harvesting pigments

with absorption spectra complementary to the quality of incident light (Tandeau de Marsac,

1983).

Phycobiliproteins particularly phycocyanin, phycoerythrin and allophycocyanin,

were introduced as a novel class of fluorescent tags in flow cytometry, cell sorting.


histochemistry and immunoassay (Giazer, 1994). Phycocyanin is sold in Japan as a blue

colourant with an annual market of 1 -2 tonnes (Borowitzka, 1994). A potential source of

phycocyanin is Spirulina platensis and the pigment content may reach 22.4% of dry

weight (Cohen et ai., 1993). The annual market for phycocyanin is US$ 5-10 million. The

value of phycocyanin used in diagnostics is > US$ 10,000/kg and that used as food

colour is > US$ 100/kg.

ABSORPTION SPECTRA

The photopigments characteristic of cyanobacteria are chlorophyll a,

phycocyanin, allophycocyanin, and in some strains phycoerythrin, and a variety of

carotenoids. Absorption maximas of whole cell methanolic extract are attributable to

chlorophyll a, C-phycocyanin, carotenoids, scytonemin, mycosporine like amino acids

(MAAs) and other UV absorbing compounds (Garcia-Pichel and Castenholz, 1991; 1993;

Garcia-Pichel et ai, 1993; Karsten and Garcia-Pichel, 1996; Adhikary and Sahu, 1998; 2000; Sinha et al.,

2000; Rath and Adhikary, 2002; Ryan et ai, 2002). The position of chlorophyll a peak in vivO is

essentially constant, at 680 to 683 nm, whereas, the position of C-phycocyanin peak

shows a greater variation, extending from 627 to 638 nm. The reason for the same in

literature is reported as a reflection of strain variations with respect to the intracellular

state of molecular aggregation of C-phycocyanin (Stanier et ai, I97i). Absorption

maximum of the isolated pigment (C-phycocyanin) from several different strains lies

within 622 and 625 nm. The absorption peaks associated with pigments in vivo axe

shifted relative to their position in solvents (Giteison et ai, 1999). Peaks detectable in the

whole cell methanolic extract of cyanobacteria and red algae could be: 665 nm

(chlorophyll a), 618 nm (phycocyanin), 475 nm (carotenoids), 436 nm (chlorophyll a),

420 nm (unknown), 362 nm (biopterin glucoside), 334 nm (MAAs), 384 nm

(scytonemin), 265 nm (unknown), 260 nm (scytonemin) and 256 nm (biopterin

glucoside) (Matsunaga et ai, 1993; Adhikary and Sahu, 1998; Sinha et ai, 2000). Scytonemin, the

cyanobacterial sheath pigment, having an in vivo absorption maximum at 370 nm, is

thought to act as a photoprotectant against UV-A radiation (Garcia-Pichel et ai, 1992).

Amount of scytonemin content is more in cyanobacteria that live in habitats exposed to

direct sunlight. Most of cyanobacteria screened so far contained one or more water

soluble, UV -absorbing, mycosporine like amino acids, however, the contents of MAAs

varies within and among strains (Garcia-Pichel and Castenholz. 1993). Both MAAs and

scytonemin content were found increasing/induced with UV illumination compared to

normal culture (Garcia-Pichel and Castenholz, 1991; 1993; Garcia-Pichel et ai, 1992; Ehling-Schuiz et

ai, 1997). Ten unicellular marine cyanobacteria were grouped into two groups based on

their relative absorption maxima (UV region with visible region) of whole cell methanolic


extract (Saha et ai, unpublished). The ratio of the peak heights of C-phycocyanin and

chlorophyll a was used to characterize cyanobacterial strains and found a correlation

with their GC content (of DNA) (Stanier et ai., 1971). Peak height represents the amount

of a particular compound and their relative proportion was found variable within the

species (Saha et ai, unpublished).

LIPIDS AND FATTY ACIDS

Lipids and fatty acids (FAs) are the principal components of cyanobacterial

membrane and cell wall, a property that they share with bacteria. However, their

oxygenic photosynthetic ability resembles that of higher algae and vascular plants (See

review by Sinha and Hader, 1996). Bacteria possess a variety of lipids, in which polar lipids

are the major ingredients of the lipid bilayer of bacterial membranes. They also

possess sphingophospholipids, but restricted to some taxa only. Whereas, micro-algal

lipids are mostly esters of glycerol and fatty acids, mainly straight-chain molecules with

an even number of carbon atoms (C14-C22). Lipids are classified into several classes

based on polarity, viz., the neutral (non-polar) lipids includes triglycerol (TG), free fatty

acids (FFA), hydrocarbons (HC) and wax esters, while polar lipids Include

phospholipids (PL), glycolipids (GL) and sulpholipids (SL). The major cellular lipids of

cyanobacteria include three glycolipids characteristic of the chloroplast: mono-

galactosyldiglyceride (MGDG), di-galactosyldiglyceride (DGDG), and

sulfoquinovosyldiglyceride (SQDG) (Nichols, 1970). Mono-galactosyldiglycerides, have

also been reported from green bacteria (Constantopoulos and Bloch, 1967; Cruden and stanier,

1970). Kenyon (1972) also has found mono- and di-galactosyldiglyceride apart from

neutral, phospholipids, free fatty acids and diglycerides within various unicellular

cyanobacterial strains. Cyanobacteria, unlike eukaryotes, usually lack sterols (Quinn and

Williams, 1983). Merritt et al. (1991) have reported a new lipid, i.e., phosphatidylglycerol

(PG) in addition to mono-galactosyldiglyceride (MGDG), di-galactosyldiglyceride

(DGDG), and sulfoquinovosyldiglyceride (SQDG) from Synechocystis PCC 6308 and

Synechococcus WH 7803. Sallal et al. (1990) have found considerable difference

between marine and freshwater species of cyanobacteria. Not only that, irrespective of

their habitat, there is variations in lipid content within the genus and species (Table 6).

The lipid content in cyanobacteria reported in literature range from 1.2-21.8% of dry

weight biomass. Hill et al. (1997) have reported 3-5% lipids in Nostoc commune while,

for the same species Vargas et al. (1998) found 8.4% lipid of dry weight. Similarly, in

Spirulina maxima (2-11%) and Spirulina platensis (3-21.8%) also wide variations in lipid

content was noticed by various authors (Table 6). Vargas et al. (1998), have found


variations in terms of lipid content within four different nitrogen-fixing cyanobacteria

(8.4-12.6% of dry weight). The relative proportion of fatty acids (Kenyon, 1972; Fisher and

Schwarzenbach, 1978; Materassi et al., 1980; Tedesco and Duerr, 1989; Floreto et al., 1996,

Subramanian, 2001; Kalib, 2002; Nkang et al., 2003; Saha et al., 2003b) and the total lipid content

(McCarthy and Patterson, 1974; Dubinsky et al., 1978; Shifrin and Chisholm, 1980; 1981; Tedesco and

Duerr, 1989; Cohen et al., 1987; Floreto et al., 1996; Subramanian, 2001; Dembitsky and Srebnik, 2002;

Kalib, 2002; Nkang et al., 2003; Saha et al., 2003b) might have varied due to change in

environmental factors.

Whole cell fatty acid profiles correspond to genetic information, and are not

influenced by plasmid information or by minor mutations; therefore, fatty acid profiles

are well correlated with current taxonomic conventions especially for bacteria (Miller and

Berger, 1985). Several thousands of bacteria were compared based on their fatty acid

profiles and an identification system based on gas chromatography is available (Miller

and Berger, 1985; Bottger, 1996). Fatty acid compositions of red, brown and green algae as

well as cyanobacteria, belonging to different taxonomic classes, orders or families and

genera, also have distinguishing features of taxonomic value (Miraiies et ai., 1990; Aknin et

al., 1992; Khotimchenko, 1993; 1995; 1998; Khotimchenko et al., 2002; Subramanian, 2001; Li et al.,

2002). in general, both the occurrence and the relative amounts of fatty acids are

constant for a particular bacterial species grown under controlled conditions, helps to

differentiate even very closely related organisms. However, the amount of fatty acid

may vary within the different growth stages as was observed in Synechocystis PCC

6308 and Synechocystis PCC 6803, but the same fatty acids were present in each

organism all through the growth period (Merritt et al., i99i). The changes in amount of

fatty acid were not statistically significant (Merritt et ai., I99i). More than 300 different

chemical structures of fatty acids have been identified so far. The variability in chain

length, double-bond position, and substituent groups has proven to be very useful for

the characterization of bacterial taxa (Suzuki et ai., 1993). The analysis by Holton and

Blecker (1970) have shown that cyanobacteria are uniquely diverse with respect to fatty

acid composition: some have a fatty acid composition similar to bacteria, some similar

to chloroplast of algae, while, some others neither to that of bacteria or of algal

chloroplast type. It was reported that bacterial type of fatty acid composition is

relatively common among unicellular cyanobacteria and filamentous cyanobacteria

possess large quantities of polyenoic fatty acids, characteristic of eukaryotic algae and

vascular plants (Kenyon and stanier, 1970). Caudales and Wells (1992) differentiated

between free-living Anabaena and Nostoc effectively based on their fatty acid

composition. De Philippis et al. (1999) have clustered Cyanothece strains of saline

environments into sub-groups based on their whole cell fatty acid profiles and


suggested that fatty acid analysis should be utilized in conjunction with other features

in order to obtain a sound classification. Merritt et al. (1991) have found similar fatty

acid composition in both isolated membranes and whole cells of Synechocystis PCC

6803. Sato et al. (1979) also made similar observations. Three marine, picoplanktonic

Synechococcus strains had a similar fatty acid composition to the freshwater strain

Synechocystis PCC 6308 (Merritt et al., 1991). Based on the fatty acid composition of 32

filamentous cyanobacteria, Kenyon et al. (1972) grouped the organisms into various

taxa with characteristics of fatty acids. As for example, Oscillatoria contain

diunsaturated fatty acids as the major fatty acid of highest degree of unsaturation.

Placement of Tolypothrix tenuis (strain 7101) with Calothrix yNas mainly based on the

presence of significant quantities of y-'inolenic and octadecatetraenoic acids (Kenyan et

al., 1972). Geitler (1932) have placed Plectonema within Scytonemataceae, based on the

presence of false branching, but, Kenyon et al. (1972), have found fatty acid profile

similar to that of Lyngbya. Likewise, Anabaena and Nostoc were characterized by

triunsaturated fatty acids (Pari<er et al., 1967; Nichols and Wood, 1968; Holton et al., 1968; Kenyon

et al., 1972). Results obtained by Kruger et al., (1995) demonstrated that fatty acid

composition is an effective taxonomic tool for solving the taxonomic problems of

Microcystis isolates. They have analysed the fatty acid composition of 14 different

Microcystis isolates and seven other species of Chroococcales, which was proved to

be consistent within isolates. Fatty acid profile was able to separate toxic forms of

Microcystis from non-toxic within the cluster (Kruger et al., 1995). Holton et al. (1968)

showed that a direct correlation exists between the morphological complexity and fatty

acid profile of cyanobacteria. Kruger et al., (1995) strongly suggested that the fatty acid

composition is a valuable taxonomic tool for unicellular cyanobacteria as presence or

absence of the colonial habit or gas vacuoles are not stable characters in culture.

Subramanian (200i) studied lipids and fatty acids profile of 38 marine cyanobacteria

wherein, he indicated the taxonomic significance of phospholipids in distinguishing

cyanobacteria up to generic level. He has drawn a phylogenetic tree based on fatty

acid profile and developed an identification system for marine cyanobacteria. Gugger

et al. (2002) have analysed the cellular fatty acid content of 22 cyanobacterial strains

representing seven genera (Anabaena, Aphanizomenon, Calothrix, Cylindrospermum,

Microcystis, Nostoc and Planktottirix) and could obtain three distinct groups formed by

the Microcystis strains, the NostodPlanktottirix strains and the

Anabaena/Aptianizomenon/Cylindrospermum strains by correspondence analysis,

however, Calothrix strain did not cluster with any of the other heterocystous

cyanobacteria. They have concluded by saying that the fatty acid composition could be

used to complement other approaches to establish a polyphasic classification of


cyanobacteria. The results obtained for group B (NostodPlanktothrix) are incongruent

with the morphologic and genetic data, but the results obtained for group C

(Anabaena/Aphanizomenon/Cylindrospermum) are in agreement with molecular

phytogeny, in contrast to morphological classification, ftezanka et al. (2003) have

studied the fatty acid composition of six freshwater cyanobacterial species

{Chroococcus minutus, Lyngbya ceylanica, Merismopedia glauca, Nodularia

sphaerocarpa, Nostoc linckia and Synechococcus aeruginosas). They have

discovered several new acids (methyl-branched-saturated and dioic acids) in

cyanobacteria and a highest number of 51 fatty acids were found in some

cyanobacterial species.

Several studies have shown that the polyunsaturated fatty acid y -linolenic acid

(GLA, 18:3(o6) is of potential pharmaceutical value for lowering low-density lipoprotein

in hypocholesteromic patients (ishikawa et ai., 1989), for alleviation of symptoms of the

pre-menstrual syndrome (Horrobin, 1983) and for treatment of atopic eczema (Biagi et ai.,

1988). The cyanobacteria Spirulina was shown to be an alternative source of GLA

(Nichols and Wood, 1968). Cohen et al. (1993) have found increase production of fatty acids

including GLA by increasing the cell concentration in Spirulina platensis. In

cyanobacteria, 016 and C 18 are the major fatty acids (Christie, 1987) and the branch

chains are minor components (Caudaies et ai., 1992). GLA was also reported from

Calothrix, Microchaete, Microcystis, and Synectiococcus strains (Kenyon, 1972; Kenyan et

ai, 1972). Cyanobacteria are heterogeneous with reference to their cellular fatty acid

composition (See stanier et al., 1971). The comparative analysis of fatty acids composition

of several cyanobacteria showed that the majority of filamentous cyanobacteria contain

large amounts of polyunsaturated fatty acids and majority of unicellular forms do not

(See Stanier et ai., 1971).

PROTEIN CONTENT AND SDS-PAGE PROFILE

Proteins are long chains of amino acids linked together by peptide bonds with

positively charged nitrogen containing amino-group at one end and a negatively

charged carboxyl group at the other end. Cyanobacteria have been reported to contain

a maximum of 71 % and a minimum of 3.2 % protein of its dry weight (Table 7).

Variation in protein content exists even within same species reported by various

authors (Table 7)- This variation sometimes is due to change in environmental

condi t ions (de Nobel et al., 1998; Gordillo et al., 1999; Kalib, 2002; Saha et al., 2003b).


Seed protein electrophoresis of Larrea amoeghinoi and L nitida showed

differences in banding pattern that was used to confirm the hybrid nature of several

specimens (Hunziker et ai, 1977, 1978; Lia et ai., 1999). Lapina and Kelner (1990) have

studied single-seed storage proteins of four fiber flax cultivars by SDS-PAGE. The

resulting 62-71 bands analysis revealed intravarietal variation. Kuniyal et al. (200i)

have studied seed polypeptide patterns to characterize populations of Aconitum atrox,

although, several polypeptide bands were common within four populations, some

polypeptides appeared to be population specific. The comparison of whole-cell protein

profile obtained by SDS-PAGE has proven to be extremely reliable for comparing and

grouping of c losely re lated bacter ial strains (Kilpper-Balzetal., 1982; Potetai, 1994; Vautermef

al., 1991; 1993). Several studies have found a correlation between high similarity in

whole-cell protein content and DNA-DNA hybridization (Castas, 1992). Whole-cell protein

patterns of Campylobacters are stable regardless of the time and methods of

preservation, growth conditions and age of the cells (Fiores et al., 1989; Hanna et ai, 1983;

Vandamme et al., 1991). However, in general, provided highly standardized culture

conditions and electrophoresis, computer-assisted numerical comparisons of protein

patterns are feasible and databases can be created for Identification purposes (Kersters

et al., 1994; Pot and Janssens, 1993; Vandamme et al., 1992). Somet imes , numerical analysis

may be hindered by the presence of distorted protein profiles or hypervariable dense

protein bands. In such cases, visual analysis is essential to interpret the similarity or

dissimilarity of protein patterns (Vandamme etai., 1996). SDS-PAGE of cell-free extracts

(protein fingerprinting) has been proved to be an efficient identification method for lactic

acid bacteria (LAB) since it generated complex and stable patterns, analysis of which,

differentiated eight species and subspecies of LAB (Perez et ai., 2000). A comparison of

two identification methods revealed that 23% of Tenerife cheese LAB isolates was

misclassified by the classical technique (Perez et ai., 2000). Klein et al. (1973) have used

electrophoretic pattern of biliproteins and soluble proteins to characterize

cyanobacteria, where they have found no taxonomic value of biliproteins, however, the

soluble protein-profiles, although somewhat variable from one experiment to another,

indeed showed taxonomic value. Schenk and Kuhfittig (i983) have studied

phycobiliprotein patterns by polyacrylamide disc-gel electrophoresis and found a

prominent heterogeneity even between strains belonging to the same species of

cyanobacteria. Hence, it was concluded that phycobiliprotein patterns could be useful

only for the identification of similar strains but not for the establishment of classification

schemes. Electrophoresis of total soluble proteins was able to distinguish one

cyanobiont and the soil Nostoc (Zimmerman and Rosen, 1992). Lyra et al. (1997) have found

a mutual relationship at the genus level within cyanobacterial species based on total


protein pattern analysis. They could find approximately 40 bands, however, most of

the variation was observed in areas with molecular weight of 50-60 kDa and 17-20

kDa. SDS-PAGE of whole-cell proteins differentiated non-heterocystous, green- and

red-pigmented Oscillatoria strains into two distinct clusters that differed from the cluster

of heterocystous strains (Lyra etai., 1997).

ISOENZYME FINGER PRINTING

Esterase (E.G. 3.1.1.x)

Esterases represent a diverse group of hydrolases catalyzing the cleavage and

formation of ester bonds. They are widely distributed in animals, plants and

microorganisms. They are stable and are even active in organic solvents (See review,

Bomscheuer, 2002). Esterase isoenzyme characterization is useful to examine the degree

of genetic diversity and relationships within as well as among the species (Bousquet et ai.

1986). Among the enzyme systems more frequently studied are the esterases, which

are species-specific, and the patterns obtained were sufficiently distinct to differentiate

strains of the s a m e species (Williams and Shah, 1980; Goullet, 1981; Goullet and Picard, 1984,

1986; Picard and Goullet, 1985; Picard et al., 1994; Pons etai, 1993;).

In insects, esterase genes have shown high rates of intraspecific and interspecific

variation (Brady and Richmond, 1992). On the basis of their affinity for the a-naphthyl and

P-naphthyl acetates used as substrates, Nascimento and de Campos Bicudo (2002)

were able to find a phylogenetic relationships of Drosophila species in the saltans

subgroup (saltans group). In addition, they have characterized and used the types of

esterase and their electrophoretic mobility for the above study. Monteiro et al. (2002)

have studied on allozyme relationships among ten species of Rhodini

(haematophagous bugs), where they were unable to detect any diagnostic locus

between R. proxilus/R. robustus pair, however, they could find only one loci (a-Est) out

of 22 loci examined, which distinguished R nasufus from R. neglectus.

Lia et al. (1999) have found twelve monomorphic loci including two loci of esterase

(Est-1 and Est-3) for the same allelic variation in seeds of Larrea ameghinoi and L.

nitida. Pasquet (1999) has used esterase (EST) and fluorescent esterase (FLE) system

in obtaining genetic relationships among subspecies of Vigna unguiculata (L.) Walp.

Kuniyal et al. (2001) have found that some of the isoenzymes of the esterase are

specific to a particular population and others are common in all populations of

Aconitum atrox, a threatened medicinal herb of Himalaya. In a study, the highest


polymorphism was obtained with esterase and acid phosphatase, where they have

identified flax and linseed cultivars by isozymes markers (Kmiickova et a/., 2002). Higher

level of diversity in esterase isoenzyme profiles was observed in Saccharum

spontaneum and its cultivated clones compared to Erianthus spp. (SBIC, 2003).

In cyanobacteria, as chemotaxonomic marker, Klein et al. (1973) have found a-

esterase based zymograms supportive to morphological criteria in recognizing four

groups of conspecific taxa: 1) Oscillatoria tenuis, O. amoena and O. animalis 2) O.

chalybea and O. formosa 3) Phiormidium foveolarum and P. luridum var. olivacea 4) P.

persjcjnum and P. ectocarpii. Baker et al (1993) although, have screened

approximately 80 enzymes for electrophorectic activity in cyanobacteria for taxonomic

purpose, they failed to detect esterase enzyme activity under extraction/assay

conditions used. However, Klein et al. (1973) could detect a maximum of 16 and a

minimum of 2 bands stained for a-esterase in cyanobacteria.

Although, the real metabolic function of this class of hydrolyzing enzymes is

unknown, they are sometimes used for biocatalysis, in food technology, antibiotics

bioconversion or in production of value added products (Venturi, 2000). In aqueous

solution, esterases catalyse the hydrolytic cleavage of esters to form the constituent

acid and alcohol whereas, in organic solutions, the transesterification reaction is taking

place (Niehaus et al., 1999).

Peroxidase (E.G. 1.11.1.7)

Peroxidases are hemoproteins that scavenge H2O2, a product of dismutation of

superoxide (O2) by superoxide dismutase. These are ascorbate peroxidase,

glutathione peroxidase and thiol-specific peroxidase based on their substrate specificity

(Asada, 1994; Yokota et al., 1988; Tichy and Vermaas, 1999). All the above types of peroxidase

activities were detected in vitro in cyanobacteria (Tichy and Venvaas, 1999; Tripathi and

Srivastava, 2001) except glutathione peroxidase, which was also detected in Oscillatoria

willei BDU 130511 (Vinoth, 2003). Though two glutathione peroxidase genes were

identified in Synechocystis sp. strain PCC 6803 (See Tichy and Venvaas, 1999), the actual

detection of the enzyme in the gel could not be made till recently. In plants, peroxidase

activity has been documented for various reactions such as pathogen defense

(Hammerschmidt et al., 1982), wound healing (Espelie et al., 1986), auxin degradation (Chlbbar

and Van Huystee, 1984; Zheng and Van Huystee, 1992), and cell differentiation (Van Huystee and

Cairns, 1982). However, in cyanobacteria, peroxidase activity was found associated with

desiccation tolerance in Gloeocapsa crepidinum BDU FW15 and nitrogen stress


alleviation in Oscillatoria willei BDU 130511 (Thpathi and Srivastava, 2001; Kalib, 2002; Saha et

al., 2003b).

Krulickova et al. (2002), although have found lower polymorphism in some

phases of plant material used for peroxidase activity, were able to differentiate 20

cultivars of flax and linseed (71% of screened cultivars) based on polymorphic

isozymes including peroxidase. Sulman et al. (1999) have observed differences in

peroxidase isoenzyme activity within black point resistant and susceptible barley

varieties during screening for black point resistant barley. Higher level of diversity were

observed among Saccharum spontaneum and cultivated clones compared to Erianthus

spp. in terms of isoenzyme profile including peroxidase (SBic, 2003). No attempt has

been made to use banding patterns of peroxidase isoenzyme for cyanobacterial

taxonomy and phylogeny.

Superoxide dismutase (E.G. 1.15.1.1)

Superoxide dismutase (SOD) is the most efficient antioxidant enzyme

distributed widely among oxygen consuming organisms, aerotolerant anaerobes, and

some obligate anaerobes (Fridovich, 1986). It was first isolated from bovine blood as a

green copper protein (Mann and KeiHn, 1938) whose, biological function was believed to be

copper storage and has been variably referred to as erythrocuprein, indophenol

oxidase and tetrazolium oxidase. Later on, it was found that SOD functions by

catalyzing the dismutation of O2" to H2O2 and O2. Three distinct types of SODs (17 to

85 kDa), based on the metal ion in their active sites, have been observed from a wide

range of organisms examined (e.g., Cu/Zn-SOD, Mn-SOD and Fe-SOD). Recently, a

new class of SOD has been described in Streptomyces griseus and S. coelicolor, the

nickel SOD (Ni-SOD) (Kim et al., 1996; 1998; Youn et al., 1996). Another two hybrid SODs

namely, Fe/Zn-SOD from Streptomyces coelicolor (Kim et ai., 1996) and Fe/Mn-SOD from

Lyngbya arboricola (Tnpathi and Snvastava, 2001) have been reported.

With a few exceptions, Cu/Zn-SODs are generally found in the cytosol of

eukaryotic cells and chloroplasts (Duke and Saiin, 1985). Sequence homologies suggest

that Mn-SOD and Fe-SOD evolved from a common ancestral form in early prokaryotes,

whereas the Cu/Zn-SOD appears to have evolved independently in the green plant line

(Bowler et a!., 1992). Generally, prokaryotes, protozoa and most algae lack Cu/Zn-SOD,

but contain Fe-SOD and/or Mn-SOD (Asada et al., 1977; Asada and Kanematsu, 1978). The

Fe-SOD enzyme has been detected in algae, cyanobacteria, and other prokaryotes as

well as in the stroma of chloroplasts (Okada et ai., 1979; Tsang et al., 1991). Mn-SOD has


also been detected in some cyanobacteria and is widely distributed among prokaryotic

and eukaryotic organisms (Okada et ai., 1979). In cyanobacteria {Anacystis nidulans,

Anabaena variabilis and Plectonema boryanum), Fe-SOD is localized in the cytosol,

whereas Mn-SOD is found primarily in the membrane fraction (Okada et ai., 1979).

Lumsden and Hall (1974, 1975) reported cyanide-insensitive SOD activity in

Spirulina platensis. They have made an electrophoretic survey of the occurrence of

SOD in photosynthetic organisms, both prokaryotic (bacteria and cyanobacteria) and

eukaryotic (red and green algae) and have confirmed the presence of cyanide-

insensitive SOD activity in all the organisms. Based on the above an evolutionary

relationship within photosynthetic organisms was hypothesized. The cyanobacteria

such as Anabaena variabilis, Anacystis nidulans, Gloeocapsa crepidinum FW 15,

Oscillatoria willei BDU 130511 and Plectonema boryanum have found to contain both

Fe-SOD and M n - S O D (Okada et ai, 1979; Kalib, 2002; Saha et ai. 2003b). Fe -SOD has also

been reported in Spirulina sp. There is a general agreement that Fe-SOD is a

constitutive enzyme in prokaryotes (Nettieton et ai. 1984). Three constitutive forms of

superoxide dismutase viz., Fe-SOD, Cu/Zn-SOD and an unidentified SOD have been

demonstrated in the cyanobacterium Synechococcus sp. WH 7803 (Chadd et ai, 1996).

However, although, the expression of Fe-SOD is thought to be constitutive, the levels

of Mn-SOD vary In response to O2 - and upon changes in growth phase (Dempie. 1991).

In cyanobacterium, Nostoc commune CHEN/1986, an active Fe-SOD was detected In

desiccated cells even after 13 years of storage. TrIpathI and Srivastava (2001) identified

five Isoforms of SOD representing one Mn-SOD, one Fe/Mn-SOD and three Fe-SOD,

In a desiccation-tolerant cyanobacterium, Lyngbya arboricola. Kalib (2002) identified the

role of Fe-SOD in protection during desiccation In Gloeocapsa crepidinum FW15, while

in another study with Oscillatoria willei BDU 130511, expression of Fe-SOD was found

related with nitrogen stress alleviation (Saha etai, 2003b).

Monteiro et al. (2002) have studied SOD along with eight other enzymes in order

to assess the degree of genetic differentiation within Rhodnius spp (haematophagous

bugs).

Lia et al. (1999) have used SOD profile for studying genetic diversity and

relationship between woody plants such as Larrea ameghinoi and Larrea nitida and

have found twelve monomorphic loci including four loci of SOD for the same allelic

variant in Larrea ameghinoi and L. nitida. Pasquet (1999) have used SOD and another

20-enzyme system to find the genetic relationships among the subspecies of Vigna

unguiculata (L.) Walp. In 23 Lathyrus species, a total of 11 different SOD bands were


detected, which gave both species specific as well as species similarity information

(Schifino-Wittmann, 2001). Krulickova et al. (2002) have observed very high SOD activity in

all analyzed phases, but with variations in isoenzyme profile of various organs at

various phases of flax and linseed cultivars. Based on the above they were able to

determine two genotypes with unique spectra Amazon and Jitka.

Even though the enzyme is ubiquitous among the taxa, very few studies only

have considered this enzyme to find the genetic diversity and their phylogenetic

relationships. As far as cyanobacterial taxonomy and phylogeny are concerned, no

study with respect to this enzyme has been undertaken. Although, Baker et al. (1993)

made an attempt to use SOD for studying the taxonomy of cyanobacterial blooms, they

were unsuccessful as they failed in enzyme extraction and staining.

Malate dehydrogenase (E.G. 1.1.1.37)

Hatam et al. (1999) have used MDH and another five-enzyme system to

characterize and differentiate Leishmania major from L tropica (protozoa). Malate

dehydrogenase banding patterns were also used to detect cell cross-contamination

and to verify the origin of the ceils of an artificial organ (Chang et ai., 2001). They have

used MDH and two more enzyme system to evaluate the reliability of human and

porcine cells. Monteiro et al. (2002) have studied MDH along with several other enzyme

systems to investigate the monophyly and paraphyly of Rhodniini spp

(haematophagous bugs).

Lia et al. (1999) have found twelve monomorphic loci including three loci of MDH

for the same allelic variant in woody plants such as Larrea ameghinoi and L. nitida.

Pasquet (1999) has studied allozyme variation including malate dehydrogenase to find

out the genetic relationships among subspecies of Vigna unguiculata. Schifino-

Wittmann (2001) has observed a total of 7 different bands for MDH among five Lathyrus

species, some of them common to all or most of the taxa while others were species

specific. Krulickova et al. (2002) have studied polymorphism of five enzyme systems in

6 flax cultivars/lines and one wild sample, where only two-enzyme system including

MDH exhibited exploitable polymorphism.

Forty four isolates of Fusobacterium nucleatum (pathogenic bacteria of oral

cavities) were examined at 21 enzyme loci including malate dehydrogenase by using

electrophoretic technique to establish an accurate genetic framework for taxonomic

purposes, where all the enzymes were polymorphic for the isolates tested (Morris et ai.,

1997).


Schenk et al. (1973) studied malate dehydrogenase isoenzyme profile in eight

cyanobacterial species with polyacrylamide disc gel electrophoresis and obtained a

maximum of 8 and a minimum of 5 bands with different relative mobility (Rm). They

have concluded that the malate dehydrogenase zymograms of cyanobacteria can be

used as "fingerprints" and used successfully in some cases as in Anabaena and

Nostoc showing shared bands, and not so successful in determining the position of

Anacystis nidulans within the order Chroococcales.

MULTIPLEX RAPD-PCR

Randomly amplified polymorphic DNA (RAPD) as a technique to reveal

nucleotide variations using random primers was developed by Williams et al. (1990) and

Welsh and McClelland (1990). RAPD markers are based on the amplification of

unknown sequences including non-coding regions of the genome, using single, short

(10-12 mer), random oligonucleotide primers. It has shown wide use such as in linkage

map construction (Grattapagiia and Sedroff, 1994), insect resistance gene localization

(Dweikat et al., 1997), identification of hybrid origin (Friesen et al., 1997), utilization in breeding

(Durham and Korban, 1994; Baril etal., 1997) and populat ion dif ferent iat ion (Hwang et ai, 2001).

Zapparoli et al. (2000) have used this method for the determination of genetic

diversity of 60 Oenococcus oeni strains from different wines. Chtourou-Ghorbel et al.

(2001) have concluded that RAPDs are equivalent to RFLPs in the estimation of genetic

diversity within the genus Lathyrus; and, because of their relative simplicity and lower

cost, RAPDs are considered more practical than RFLPs for studies on germplasm

organization and characterization. Tettelin et al. (1999) have employed multiplex RAPD-

PCR in Streptococcus pneumoniae whole genome shotgun project that closed a large

number of gaps with excellent results. Cocolin et al. (2000) have developed a multiplex-

PCR method for the amplification of sit and eaeA genes of enteropathogenic (EPEC)

and enterohemorrhagic (EHEC) Escherichia coli. Wang et al. (2002) have used

multiplex PCR for the simultaneous detection of genes coding for various toxins from

enterotoxigenic Staphylococcuc aureus. Likewise, multiplex PCR has got several other

applications viz., detection of cry9 genes in Bacillus thuringiensis strains (Ben-Dov et ai.,

1999), identification and differentiation of Campylobacterspp. (Wangetal., 2002), detection

of Shiga toxin-producing bacteria (Beianger et ai., 2002), detection of methicillln-resistant

Sfap/7y/ococc/from blood culture bottles (Louie etal., 2002), detection of Herpesvirus DNA

in clinical samples (Druce et ai., 2002), identification of pathogenic fungi (Luo and Mitchell,

2002), etc.


West and Adams (1997) have used RAPD-PCR for phenotypic and genotypic

comparison of symbiotic and free-living cyanobacteria. Neilan (1995) has combined the

concept of RAPD-PCR with multiplex-PCR, where, he used multiple primers for

randomly amplified polymorphic DNA for the identification and phylogenetic analysis of

toxigenic cyanobacteria. The results of this study showed the genetic relatedness

among the genera of bloom forming cyanobacteria similar to that obtained from 16S

rRNA gene sequences. Neilan (1995) has got strain-specific randomly amplified

polymorphic DNA profiles that discriminated all toxigenic cyanobacteria into three

taxonomic levels of genus, species and strain. Analysis of multiplex RAPD-PCR

products clearly distinguished between the genera Anabaena and Microcystis (Neilan,

1995).

Table 1. Chlorophyll a content of selected cyanobacteria reported in literature.

Cyanobacteria Chlorophyll a

(mgg "̂ dry weight)

Spirulina platensis 18.2


Spirulina platensis 9.2-14.4

Spirulina sp 17.0

Spirulina sp. 10.5

Spirulina sp. 5.4

Spirulina sp. 6.9

Oscillatoria willei BDU 130511 0.52-0.77

Phormidium valderianum BDU 30501 3.25

Aplianizomenon sp. 20.4-29.9

Nostoc calcicola BDU 40302 2.12

Nostoc commune var commune 1.8

Nostoc commune var flagelleforme 2.7

Anabaena azollae ML1 11.3-11.7


Anabaena sp. 22.4-25.7

Reference

Olaizola and Duerr, 1990

Gordillo et al., 1999

Cohen etal., 1993

Richmond, 1988

Wuefa/., 1993

Wuefa/., 1992

Durand-Chastel, 1980

Sahaefa/.,2003b

Prabaharan, 1988

deNobelefa/., 1998

Sujatha and Kaushik, 1998

Scherer and Zhong, 1991


Malliga, 1993

Malliga, 1993

deNobelefa/., 1998

Table 2. Carotenoids content of selected cyanobacteria reported in literature.

Cyanobacteria Carotenoids

(mgg ^ dry weight)

Anacystis sp. 7.67

Microcystis sp. 3.14

Synechococcus sp. 3.59

Merismopedia sp. 3.64

Spirulina maxima 6.48


Spirulina platensis 18


Spirulina sp. 1.7

Spirulina sp. 3.5

Spirulina sp. 4.5

Spirulina sp. 5

Spirulina subsalsa 0.30

Oscillatoria limnetica 3.60

Oscillatoria sp. 2.32

Oscillatoria willei BDU 130511 0.23-0.28

Phormidium sp. 2.95

Phormidium sp. 3.21-6.03

Phormidium valderianum BDU 30501 0.21

Aphanizomenon flos-aquae 4.17

Nostoc calcicola BDU 40302 7.59

Nostoc commune var commune 0.37

Nostoc commune var flagelleforme 0.53


Anabaena azollae ML2 3.3

Anabaena sp. 2.82

Anabaena variabilis 5.84

Tolypothrix tenuis 6.38

Reference

Paerl etal., 1984

Paerl etal., 1984

Paerl etal., 1984

Paerl etal., 1984

MikiefaA, 1986

Aakerman etal., 1992

Olaizola and Duerr, 1990

Gordilloefa/., 1999

Wuefa/., 1992


Wuefa/., 1993

Tornabene etal., 1985



Paerl etal., 1984

Sahaefa/.,2003b

Paerl etal., 1984

Vesk and Jeffrey, 1977

Prabaharan, 1988

Fikshdahl eta/., 1983

Sujatha and Kaushik, 1998


Scherer and Zheng, 1991

Malliga, 1993

Malliga, 1993

Paerl etal., 1984

Goodwin, 1980

Fikshdahl ef a/., 1983

Table 3. Phycocyanin content of selected cyanobacteria reported in literature.

Cyanobacteria Phycocyanin

(mg g'̂ dry weight)

Reference

Synechococcus sp. NKBG042902

Spirulina platensis

Spirulina platensis

Spirulina sp.

Spirulina sp.

Oscillatoria willei BD\J 130511

Phonvidium valderianum BDU 30501

Nostoc calcicola BDU 40302

Anabaena azollae ML1


150.0 Tai<ano et al., 1995

104.0-224.0 Cohen et al., 1993

41.6 Gordillo etal., 1999

41.5 Wuefa/., 1992

69.8 \Nuetal., 1993

79.6-99.0 Sahaefa/.,2003b

79.1 Prabaharan, 1988

21.31 Sujatha and Kaushil<, 1998

53-57 Malliga, 1993

21-36 Malliga, 1993

Table 4. Aliophycocyanin content of selected cyanobacteria reported in literature.

Cyanobacteria Aliophycocyanin Reference

(mg g "̂ dry weight)

Spirulina sp. 15.7 Wuefa/., 1992


Oscillatoria willei BDU 130511 19.1-19.4 Sahaefa/., 2003b

Phormidium valderianum BDU 30501 63.1 Prabaharan, 1988

Nostoc calcicola BDU 40302 11.67 Sujatha and Kaushilc, 1998

Anabaena azollae M L1 16-19 !\/laiiiga, 1993

Anabaena azollae ML2 10-14 Malliga, 1993

Table 5. Phycoerythrin content of selected cyanobacteria reported in literature.

Cyanobacteria Phycoerythrin

(mg g "̂ dry weight)

Reference


Anabaena azollae IVILI


67.0 Sujatha and Kaushik, 1998

2.6-87.5 Malliga, 1993

6.4-88.0 Malliga, 1993

Table 6. Lipid contents of seiected cyanobacteria reported in literature.

Cyanobacteria Lipid

(% dry weight)

Reference

Anacystis nidulans 14.3-14.8

Chroococcus minutus BDU 40401 3.4

Gloeocapsa sp. BDU 60102 4.7

Aphanocapsa sp. BDU 50261 3.9

Synechococcus elongatus BDU 30312 4.9



Myxosarcina sp. BDU 40881 11.4

Hyella caespitosa BDU 40911 14.7




Spirulina maxima 6.0-7.0

Spirulina maxima 6.0-7.0

Spirulina platensis UTEX 1928 3.2-11.8






Spirulina sp. 3.8-4.6

Spirulina sp. 6.5

Spirulina sp. 8.8

Spirulina subsalsa BDU 40303 17.3

Oscillatoria boryana BDU 92181 10.8

Oscillatoria chalybea BDU 91051 13.3

Oscillatoria curviceps BDU 92131 14.2

Oscillatoria formosa BDU 40261 16.8

Oscillatoria laete-virens BDU 40101 17.4

Oscillatoria minnesotensis BDU 70491 12.1

Oscillatoria salina BDU 10142 12.7

Oscillatoria subtilissima BDU 92183 13.2

Oscillatoria tenuis BDU 60751 11.3

Piorreckefa/., 1984

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Becker, 1994

Trubachev etal., 1976

Subramanian, 2001

Subramanian, 2001

Hudson and Karis, 1974

Miller, 1968

Switzer, 1980

Becker, 1994


Tedesco and Duerr, 1989

Piorreckefa/., 1984

Tornabene etal., 1985

Becker and Venkataraman, 1984

Becker, 1994

Tipins and Pratt, 1960

He, 1988


Wuefa/., 1993

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Subramanian, 2001

Cyanobacteria Lipid Reference

(% dry weight)

14.0 Subramanian, 2001

6.5-8.8 Sahaefa/.,2003b












11.4 Vargas etal., 1998


3.0-5.0 Hillefa/., 1997



12.8-13.7 Malliga, 1993

6.7-8.8 Malliga, 1993

4.0-7.0 Becl<er, 1994




9.0-10.0 Pushparaj etal., 1994







1.2 Kaeliler and Kennisii, 1996


Oscillatoria terebriformis BDU 81331

Oscillatoria willei BDU 130511

Phormidium tenue BDU 20391









Lyngbya confervoides BDU 40321

Microcoleus chthonoplastes BDU 91212

Anabaenopsis sp.


Nostoc commune

Nostoc commune

Nostoc paludosum



Anabaena cylindrica

Anabaena sp. BDU 41811

Anabaena variabilis

Pseudanabaena schmidlei BDU 20761

Nodularia harveyana

Nodularis sp.

Plectonema terebrans BDU 30342

Scytonema crustaceum BDU 40142

Calothrix scopulorum BDU 60101

Dichothrix baueriana BDU 40481

Mastigocoleus testarum BDU 60674

Kyrtuthrix maculans

Hapalosiphon welwitschii BD\J 91601

Table 7. Protein content of selected cyanobacteria reported in literature.

Cyanobacteria Protein

(% dry weight)

Reference

Synechococcus sp. 63.0 Becker, 1994

Synechococcus sp. 63.0 Trubachevefa/., 1976

Spirulina maxima 60.0-71.0 Becker, 1994

Spirulina maxima 65.0 Miller, 1968

Spirulina platensis 46.0-50.0 Tipins and Pratt, 1960

Spirulina platensis 46.0-63.0 Becker, 1994

Spirulina platensis 62.5 Becker and Venkataraman, 1984

Spirulina sp. 53.6-58.2 He,1988

Spirulina sp. 65.5 Durand-Chastel, 1980



Oscillatoria willei BDU 130511 22.6-37 Sahaefa/., 2003b

Anabaenopsis sp. 52.2 Vargas eta!., 1998

Nostoc calcicola BDU 40302 3.2 Sujatha and Kaushik, 1998

Nostoc commune var commune 8.1 Scherer and Zhong, 1991

Nostoc commune var flagelleforme 11.8 Scherer and Zhong, 1991

Nostoc commune 39.9 Vargas etal., 1998

Nostoc commune 50.2 Santra, 1992

Nostoc paludosum 40.4 Vargas etal., 1998

Anabaena azollae ML1 12.1-12.3 Malliga, 1993

Anabaena azollae ML2 11.3-11.6 Malliga, 1993

Anabaena cylindrica 43.0-56.0 Becker, 1994

Anabaena variabilis 42.5 Santra, 1992

Anabaena variabilis 47.2 Vargas etal., 1998

Nodularia harveyana 61-63 Pushparaj etal., 1994

Nodularia sp. 42.9 Vargas etal., 1998

Scytonema sp. 45.5 Santra, 1992

Kyrtuthrix maculans 7.3 Kaehler and Kennish, 1996

Westiellopsis prolifica 45.0 Santra, 1992

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Review of literature - Information and Library Network …shodhganga.inflibnet.ac.in/bitstream/10603/116004/8/08...Review of Literature 4 Cyanobacteria evolved on this planet ca 3.5