Int.J.Curr.Microbiol.App.Sci (2017) 6(3): 1195-1204

International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 3 (2017) pp. 1195-1204 Journal homepage: http://www.ijcmas.com

Original Research Article https://doi.org/10.20546/ijcmas.2017.603.139

Exploration of Algal Varieties from Panikhaiti Area of Guwahati using Winogradsky Column

Sushma Gurumayum* and Sushree Sangita Senapati

Department of Microbiology, College of Allied Health Sciences, Assam down town University, Panikhaiti, Guwahati – 781026, Assam, India *Corresponding author

ABSTRACT

An attempt has been made to explore different types of from Panikhaiti area of Guwahati, Assam using Winogradsky column. In order to prepare Winogradsy column, K e yw or ds

soil and water samples were collected from different locations. A transparent, clear plastic Winogradsky bottle was taken and filled with 500g of soil and over layered with 500ml of water sample. colu mn; Micro The columns were enriched with different carbon and nitrogen supplements. They were algae; Soil; covered with plastic sheets and few holes were punctured on them. These were incubated Panikhaiti . at room temperature (±28°C) in presence of sunlight. One column was kept covered with

Article Info dark paper and kept in dark as control. Observations were made weekly, for development of algal growth and microbial communities over a period of 12 months. A gradual change

Accepted: in colour of column and also on the water layer was observed over the course of incubation 20 February 2017 period. The columns started showing stratified micro ecosystems with an oxic top layer Available Online: 10 March 2017 and anoxic sub-surface layers. Algal growth prominently appeared in all the columns. The algal specimens belonged to 18 genera which included members of class Cyanophyceae, Zygnematophyceae, Ulvophyceae, , Bacillariophyceae and Euglenoidea.

Introduction

Algae are found in varied habitats ranging headquarter of Panikhaiti village. The total from wetlands, water bodies and moist soil of geographical area of village is 304.74 forest areas. Algae play important role in hectares. Panikhaiti has a total population of ecosystem as primary producers and a lot of 3,817. Guwahati is nearest town to Panikhaiti industrially significant products have been which is approximately 15km away. The developed using them. Nowadays, algae are present study covers some of the village area also desirable entities for study of carbon as well as some of the surrounding forest area. sequestration. Thus, there is a requirement to In this study, an attempt has been made to study and document the varieties of algae gather information about the algal varieties in occurring in various ecological niches and these locations. This will add to the pool of specific geographical areas. Panikhaiti village existing information about algae diversity is located in Chandrapur Tehsil of Kamrup present in Assam. Till now there is no work Metropolitan district in Assam, India. It is on detailed investigation of algal diversity in situated 15km away from district headquarter Panikhaiti area of Guwahati, Assam. Chandrapur. Chandrapur is the sub-district

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Materials and Methods Morpho-taxonomic descriptions of the different varieties of algae studied from these The study was conducted for a year from Winogradsky columns are given as follows. August, 2015 to Aug, 2016. Water and soil samples were collected from different areas of Chroococcus Panikhaiti region, Guwahati. Using these soil samples, Winogradsky columns were Kingdom: Bacteria prepared. In each of the columns, the soil was Phylum: Cyanobacteria supplemented with different carbon and Class: Cyanophyceae nitrogen sources. 500gm of soil mixed with Order: Chroococcales these supplements were overlayered with Family: Chroococcaceae water sample. These columns were covered Genus: Chroococcus with transparent plastic sheets with holes punched in them and incubated at room Chroococcus is a kind of Cyanobacteria temperature in presence of sunlight. found in freshwater. It is unicellular blue - Observations were recorded after every seven green algae. Microscopic colonies are found days. Samples of algal growth from: a) upper within an external sheath. They are usually water layer, b) biofilm on walls of the column found in colonies inside a transparent and c) surface of soil in the column were protective covering sheath which contains collected. Wet mounts of algal samples were photosynthetic pigments. It inhabits prepared and observed under bright field light underwater environments like sludge of lake microscope. and river bottoms. For photosynthesis, the algae use an extensive quantity of Results and Discussion atmospheric carbon creating free oxygen in the atmosphere. Changes were observed in the Winogradsky column around 7th day of incubation. Initially, Oscillatoria the growth of algae was seen in the upper water layer as well as on the sides of the Kingdom: Bacteria column as a biofilm. As the number of Phylum: Cyanobacteria incubation days increased, layer of growth Class: Cyanophyceae was also seen on surface of soil in the Order: Nostocales columns. Figure 1 shows the different Family: Oscillatoriaceae columns with algal growth in them. Along Genus: Oscillatoria with the algal growth, the underlying layers of soil also developed different hues of colours Oscillatoria is a filamentous blue-green or showing growth of different groups of brown-green cyanobacterium. It is named bacteria. after the oscillation in its movement. It reproduces by fragmentation. It can form long The algal specimens observed in the columns filaments of cells which can break into belonged to 18 genera which included fragments called hormogonia. The members of class Cyanophyceae, hormogonia can grow into a new, longer Zygnematophyceae, Ulvophyceaea, filament. Each filament consists of trichome Chlorophyceae, Bacillariophyceae and which is made up of rows of cells and the tip Euglenoidea (Table 1). of the trichome oscillates like a pendulum.

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This gives the filaments easier mobility in all Anabaena directions. It is also able to fix nitrogen. Kingdom: Bacteria Phormidium Phylum: Cyanobacteria Class: Hormogoneae Kingdom: Bacteria Order: Nostocales Phylum: Cyanobacteria Family: Nostocaceae Class: Cyanophyceae Genus: Anabaena Order: Oscillatoriales Family: Phormidiaceae Anabaena is a filamentous cyanobacteria and Genus: Phormidium exists as plankton. Cells are attached to form unbranched filaments which may appear It is unbranched and occurs usually in fine, randomly twisted and coiled or entangled smooth and layered filaments. Filaments are with numerous other filaments. Vegetative short cylindrical cells in a fine sheath and cells may be spherical to oblong with granular glide within the sheath. Sheaths are tube-like, contents and conspicuous, refractive firm, colourless, joined to the trichomes, pseudovacuoles. They fix nitrogen. They opened at the ends, containing always only show symbiotic relationships with certain one trichome. Trichomes are isopolar, plants. They form heterocysts which straight, coiled, usually 2-12 μm wide, convert nitrogen into ammonia. Certain uniserial, unbranched, composed of species of Anabaena have been used as cylindrical up to slightly barrel-shaped cells, effective natural fertilizer and found to be screw-like twisted towards the ends, motile productive on rice paddy fields. within and out of sheaths. End cells are tapered, with a calyptra on the terminal cell. Microcoleus Tapering may extend several cells from the end. Organelles absent from the cells, but Kingdom: Eubacteria vesicles are common. They form mats which Phylum: Cyanobacteria may be dark brown, green, purple or blue- Class: Cyanophyceae green. They can survive in streams as well as Order: Oscillatoriales rivers. Family: Microcoleaceae Genus: Microcoleus Lyngbya The organism is a terrestrial species which Kingdom: Bacteria has filaments and are often in an interwoven Phylum: Cyanobacteria mat of densely entangled trichomes in one Order: Oscillatoriales sheath. Trichomes are found arranged Genus: Lyngbya parallelly or rope-like. Sometimes only one trichome in terminal branches is present The organisms are filamentous. They share which is known as pseudo-branching, sheath the entire range of cellular types with is colorless, diffluent, varying in thickness, oscillatoria. They also produce a distinct and not lamellated, with rough with uneven persistent sheath. The sheath is thin and surface. Trichomes attenuate at the ends. End extends beyond the terminal cell of the cells are capitate, mainly with calyptras, trichome. The trichome diameters range from bluntly-rounded, conical, hemispherical or 1µm to about 80 µm. Trichomes are usually obtuse-conical. Trichomes are pale blue- nonmotile within the sheath.

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Int.J.Curr.Microbiol.App.Sci (2017) 6(3): 1195-1204 green, dirty green or olive-green, long, not conjugation. Zygote within the conjugation constricted at cross-walls. tube extends into both gametangia. Mature zygospores spherical to quadrate, or Pseudanabaena polygonal, with smooth or papillate outer surface and smooth mesospore wall layer. Kingdom: Eubacteria Mesotaenium resides mostly often as small Phylum: Cyanobacteria gelatinous masses among mosses, or on moist Class: Cyanophyceae soil or rocks; occasioanlly in acidic, Order: Synechococcales oligotrophic, aquatic habitats or in acid bogs. Family: Pseudanabaenaceae Each cell has a single axial, plate-like Genus: Pseudanabaena chloroplast with one or several pyrenoids. The cells are solitary or aggregated within Pseudanabaena species are filamentous common mucilage to form irregular colonies. bloom-forming cyanobacteria. The organisms are unicellular, colonial, or filamentous Spirogyra bacteria that conduct photosynthesis using chlorophyll a. Pseudanabaena species are Kingdom: Plantae nonheterocystous cyanobacteria. They are Phylum: Charophyta characterized by simple trichomes with a Class: Zygnematophyceae width less than 4 μm. The cells are longer Order: Zygnematales than wide, possess parietal thylakoids, contain Family: Zygnemataceae polar gas vesicles and the cross walls are Genus: Spirogyra conspicuously constricted. Most cultured Spirogyra is also known as water strains reveal gliding motility and some are silk, mermaid's tresses, and blanket weed. capable of anaerobic N2 fixation. They are filamentous green algae with Pseudanabaena spp. is a dominant group of helical or spiral arrangement of organisms found in dense water blooms. the chloroplast. It is commonly found in fresh

Mesotaenium water. In spring, Spirogyra grows under water, but when there is enough sunlight and Kingdom: Plantae warmth they produce large amounts of Phylum: Charophyta oxygen, adhering as bubbles between the Class: Zygnematophyceae tangled filaments. Spirogyra measures Order: Zygnematales approximately 10μm to 100μm in width and Family: Mesotaeniaceae may grow to several centimeters in length. It Genus: Mesotaenium develops slimy, filamentous green masses. The filamentous masses form slimy green Mesotaenium is a unicellular, short, straight, mats on the surface. Spirogyra can reproduce cylindrical, solitary alga within common both sexually and asexually. In vegetative gelatinous matrix with simple or layered reproduction, fragmentation takes place, structure. Cells size vary from10-123 µm and Spirogyra simply undergoes the long, straight or slightly curved, long or short intercalary mitosis to form new filaments. cylindric, with broadly rounded ends. Cell has one or two chloroplasts with 1 or 2 pyrenoids. Ulothrix Cell wall is two-layered. Asexual reproduction is usually by transverse cell Domain: Eukaryota division and sexual reproduction by Kingdom: Protista

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Phylum: Division: Chlorophyta Class: Ulvophyceae Class: Trebouxiophyceae Order: Ulotrichales Order: Chlorellales Family: Ulotrichaceae Family: Chlorellaceae Genus: Ulothrix Genus: Chlorella

Ulothrix is eukaryotic, unicellular filamentous Chlorella is single-cell green algae. It is green algae. They inhabit in spherical and 2 to 10 μm in diameter, without fresh and marine water. They can also survive flagella. It contains the green photosynthetic in the low temperatures of spring. The cells pigments chlorophyll-a and chlorophyll-b in are normally as broad as they are long. They its chloroplast. It multiplies rapidly through become attached to surfaces by a modified photosynthesis. It requires carbon dioxide, holdfast cell. Reproduction is vegetative. It water, sunlight, and a small amount of consists of unbranched, uniseriate filaments minerals for its reproduction. arranged end to end. They are barrel-shaped or cylindrical. The apical cell is rounded at its terminal end whereas the basal cell is elongated. It is also called the basal holdfast, Domain: Eukaryota which attaches the filament to the substratum. Kingdom: The cell wall is composed of propectin and Division: Chlorophyta cellulose. Each cell has a single girdle-like Class: Chlorophyceae and parietal chloroplast and pyrenoids are Order: present in each chloroplast. Family: Genus: Haematococcus Chlamydomonas Haematococcus are solitary, ovoid to Domain: Eukaryota ellipsoidal, with broadly rounded ends. Its Kingdom: Plantae size ranges from 5-25 μm. They can grow in Division: Chlorophyta 4-9% of moisture. They have 2 equal flagella. Class: Chlorophyceae Chloroplasts are more or less cup-shaped, Order: Chlamydomonadales sometimes reticulate. It is green in colour but Family: Chlamydomonadaceae cells often appear as orange or reddish due to Genus: Chlamydomonas extraplastidial carotenoid. Nucleus is present in the central area. Species of this genus are found in a wide variety of habitats. When observed under Oedogonium microscope, they appear as small, elliptical- oval cells. They are biflagellate, with two Kingdom: Plantae whip-like flagella. A tiny red eyespot at the Division: Chlorophyta base of the flagella is a characteristic feature Class: Chlorophyceae of this alga. There is presence of single large Order: Oedogoniales cup-shaped chloroplast. Genus: Oedogonium

Chlorella Oedogonium is found in fresh water like pools, ponds, lakes etc. The thalloid plant Domain: Eukaryota body is green, multicellular and filamentous. Kingdom: Viridiplantae 1199

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The filaments are unbranched and cells of Gonium each filament are attached end to end and form uniseriate row. The filament is Domain: Eukaryota differentiated into three types of cells which Kingdom: Plantae include basal cell, apical cell and middle cells. Division: Chlorophyta Basal Cell is the lowermost cell of the fila- Class: Chlorophyceae ment. The cell is generally colourless, which Order: Volvocales performs the function of fixation to the Family: Goniaceae substratum and called holdfast. The topmost Genus: Gonium cell of the filament is apical cell. The cell is usually rounded towards apical side and green Cells are ovoid with a single cup-shaped in colour. All the cells in between basal and chloroplast containing one pyrenoid. Each apical cells are similar. Cells have reticulate cell has two cilia, contractile vacuoles at the chloroplasts. Asexual reproduction takes base of the cilia, and an eyespot. They are place by multi- flagellate zoospore. Sexual mostly found in freshwater like pools and reproduction is advanced oogamous type. The ponds. Asexual reproduction is carried out by female gamete is known as ovum and the a formation of 2-4 zoospores in each cell and male gamete is known as antherozoids. The sexual reproduction is isogamous, by a fusion zygote undergoes meiotic division and of biciliate zoogametes. The type of nutrition produces four zoospores. is phototrophic or chemo or photo- organotrophic.

Table.1 List of algae species under different groups in the present study

Sl.No. Group Genus 1 Cyanophyceae a) Chroococcus, b) Oscillatoria, c) Phormidium, d) Lyngbya, e) Anabaena f) Microcoleus g) Pseudanabaena 2 Zygnematophyceae a) Mesotaenium, b) Sprigyra 3 Ulvophyceaea a) Ulothrix 4 Chlorophyceae a) Chlamydomonas, b) Chlorella, c) Haematococcus d) Oedogonium e) Gonium 5 Bacillariophyceae a) Pinnularia, b) Navicula 6 Euglenoidea a) Euglena

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Fig.1 (1a) Chroococcus, (1b) Oscillatoria, (1c) Phormidium, (1d) Lyngbya, (1e) Anabaena, (1f) Microcoleus, (1g) Pseuanabaena (2a) Mesotaenium, (2b) Sprigyra, (3a) Ulothrix, (4a) Chlamydomonas, (4b) Chlorella, (4c) Haematococcus, (4d) Oedogonium, (4e) Gonium, (5a) Pinnularia, (5b) Navicula, (7a) Euglena

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Fig.2 Winogradsky columns prepared with soil samples from different areas

Cells usually connected by extracellular cells, and a fragile gelatinous capsule zone matrix comprising of fibrillar sheath, a surrounding entire colony. Tripartite tripartite boundary surrounding individual boundaries of adjoining cells attached or

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Int.J.Curr.Microbiol.App.Sci (2017) 6(3): 1195-1204 fused with one another; cells more or less diadinoxanthin). Oil and volutin spherical to slightly ovoid or pyriform; two chrysolaminarin are in the form of stored food equal flagella; cup-shaped chloroplast, with products. It shows gliding movement. anterior eyespot and 1or 2 pyrenoids; 2 Reproduction occurs by cell division and contractile vacuoles. auxospore formation. Motile stages possess a single, anterior pantonematic flagellum. Pinnularia Euglena Domain: Eukaryota Phylum: Heterokontophyta Division: Eukaryota Class: Bacillariophyceae Class: Euglenoidea Order: Naviculales Order: Euglenales Family: Pinnulariaceae Family: Euglenaceae Genus: Pinnularia Genus: Euglena

It is unicellular, elongated cell. Cell wall is They are mostly found in freshwater habitats. composed of pectic substances. It is covered Under the microscope they appear as motile by mucilaginous layer. Cytoplasm is typically spindle-shaped and unicellular. They arranged in parietal layer. A large central have disc-like chloroplasts which may be vacuole with a single nucleus is present. The pigmented structures within the cell. most prominent features are the two chloroplasts present along the two sides of the In conclusion, taxonomic investigations on cells. They contain chlorophyll a, c, beta- algal flora of Assam are generally carried out carotene and diatomin or fucoxanthin in ponds, rice fields and ditches. The study pigment. Fucoxanthin gives the alga with Winogradsky column offers convenient characteristic color. One or two pyrenoids are sample development from desired locations present in the chloroplast. Their storage with help of soil and water (Fig. 2). The compound is oil or chrysolaminarin. above study resulted in identification of several algae from Panikhaiti area. Further Navicula studies on the industrial potential and environmental applications of these algal Kingdom: Chromista strains can be carried out. Phylum: Ochrophyta Class: Bacillariophyceae Acknowledgement Order: Naviculales Family: Naviculaceae Authors would like to acknowledge the Genus: Navicula support rendered by Assam down town University for conducting the above work. It is commonly known as diatoms and is commonly found in fresh water, water, in air References or on soil. Thallus is unicellular, uninucleate and diploid with radial or bilateral symmetry. Baruah, P.P., Kakati, B., and Kakati, I.A. Cell wall is silicified. They are also known as 2009. Some Fresh Water Algae of Oil golden brown algae because of their Refinery Effluent Drains of Assam, characteristic pigments like carotenoids, India. Our Nature, 7: 139-145. fucoxanthin, diatomin (diatoxanthin,

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Deka, M., and Bordoloi, R.P.M. 1991. Studies Kathajodi River after receiving sewage on the Blue-Green Algae from Rice of Cuttack and its ecological Field of Assam. Phykos, 30: 175-180. implications. Indian Hydrobiol., 8(1): Harshil, H.B., Sharma, B.M., and Upasani, 67-74. V.N. 2016. Studies on Microbial Sinha, J.P., and Mukherjee, D. 1975. On blue- Diversity of a Soda Lake in India by green algae from the paddy fields of Winogradsky Column Technique. Int. J. Bankura District of West Bengal. Curr. Microbiol. Appl. Sci., 5(4): 608- Phykos, 14: 117-118. 614. Syiem, M., Nongbri, B.B., Pinokiyo, A., Hazarika, D., and Barukial, J. 2012. Bhattacharjee, A., Nongrum, N.A., and Occurrence and Distribution of Blue Hynniewta, L. 2010. Significance of Green Algae in paddy fields of cyanobacterial diversity in different Golaghat district, Assam India. Adv. ecological conditions of Meghalaya, Plant Sci., 25(1): 341-345. India. J. Appl. Natural Sci., 2(1): 134- Jena, M., Ratha, S.K., and Adhikary, S.P. 139. 2005. Algal diversity changes in

How to cite this article:

Sushma Gurumayum and Sushree Sangita Senapati. 2017. Exploration of Algal Varieties from Panikhaiti Area of Guwahati using Winogradsky Column. Int.J.Curr.Microbiol.App.Sci. 6(3): 1195-1204. doi: https://doi.org/10.20546/ijcmas.2017.603.139

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