Freshwater diatom diversity in Deepor Beel - A Ramsar site

Keywords:
Freshwater diatoms, diversity, Frustule, Deepor Beel, Ramsar site
182-191 | JRPS | 2013 | Vol 2 | No 2

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Authors:
Lisha Gurung
1
, Alak
Kumar Buragohain
2
,
Sailendra Prasad Borah
1

and Bhaben Tanti
1*
.

Institution:
1. Department of Botany,
Gauhati University,
Guwahati-781014, Assam,
India.

2. Department of Molecular
Biology and Biotechnology,
Tezpur University, Tezpur-
784028, Assam, India

Corresponding author:
Bhaben Tanti.















Email:
bt53@rediffmail.com



Web Address:
http://www.plantsciences.info
documents/PS0055.pdf.
Dates:
Received: 12 Mar 2013 Accepted: 20 Mar 2013 Published: 29 May 2013
Article Citation:
Lisha Gurung, Alak Kumar Buragohain, Sailendra Prasad Borah and Bhaben Tanti.
Freshwater diatom diversity in Deepor Beel - A Ramsar site.
Journal of Research in Plant Sciences (2013) 2(2): 182-191
An International Scientific Research Journal
Original Research
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Journal of Research in Plant Sciences
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Journal of Research in
Plant Sciences
An International Scientific
Research Journal
ABSTRACT:

This study was conducted for an investigation of freshwater diatom diversity
in Deepor beel, a Ramsar site of Assam, India. The water samples were collected from
six different sites on the basis of habitat stratification. The study was carried out from
April 2010 to March 2011. Laboratory pure cultures in diatom specific media followed
by microscopic investigations supplemented with taxonomic keys, aided in
identification of the diatoms were undertaken. The results showed that there were 65
species of diatoms, belonging to 26 genera of which 53 were pinnate and 12 were
centric types. The dominant species were found to be Nitzschia, Pinnularia and
Navicula respectively. Abundance of diatom species provides evidence for their
important role as primary producers in this wetland ecosystem of Deepor beel of
Assam.


INTRODUCTION
Diatoms are unicellular, eukaryotic, microscopic
algae belonging to the class Bacillariophyta. They are
photosynthetic and usually appear golden-brown in
color, abundant in both freshwater as well as marine
environment (Beakes et al., 1988). The characteristic
feature of the diatom is its cell wall known as frustules,
which is heavily impregnated with amorphous silica. The
structure and arrangement of the diatom frustules could
be used as a taxonomic criterion for identification up to
their species level. They are the only organisms known to
process genetic ability to mineralize amorphous silica
into complex structures. Diatoms have recently evolved
tremendous interest for their silica nanostructures, which
have unending possibilities in the field of engineering,
electronics and medical science (Pareek et al., 2011;
Koen et al., 2003; Taylor et al., 2005).
Diatoms play very important ecological roles.
They play a key role in the global carbon cycle, fixing
25% of the carbon each year (Jones, 2007). Therefore,
this group of organism contributes a major amount in the
primary productivity of our ecosystem. Diatoms also
involve in the silica cycling pathways on the earth.
Benthic diatoms are very diverse and can live
either free floating as phytoplanktons or in the sediments,
or attached to the rocks (epilithon), sand grains
(epipsammon), and aquatic plants (epiphyton). Diatoms
living on soft sediments (epipelon) are often motile
species that are able to migrate up and down in the
sediment matrix. Attached communities can be closely
attached to the substrate; e.g., Cocconeis or attached by
mucilaginous stalks e.g., Gomphonema. Often diatoms
can group themselves into 3-dimensional communities
forming complex microscopic forests. Diatoms can also
form part of the diverse 3-dimensional microbial
communities of filamentous algal mats which form in
streams and lakes, in some cases forming perennial
layers up to 30 cm thick (Rai, 2006).

Deepor Beel, located in the vicinity of Greater
Guwahati, Assam (Latitude 267'26'', longitude
2638'49'') is an important wetland of North East India.
This wetland has enormous ecological and economic
significance (Gogoi 2007). Considering the necessity of
protection and conservation, Deepor Beel has been
included amongst the few Ramsar sites. The diatom
species are known to play critical roles in wetland
ecosystems. The diatom diversity has not been assessed
at this important Ramsar site of Assam, India.
Therefore, the present study was emphasised to explore
the diatom diversity at Deepor Beel.

MATERIALS AND METHODS
Collection of samples and culture conditions
Water and semi aquatic soil samples were
collected from six different sites of Deepor Beel
ecosystem on the basis of habitat stratification from
April 2010 to March 2011 (Figure 1). Freshly collected
samples were immediately transferred in the DM
(Diatom Medium) proposed by Beakes et al., (1988).
The medium was standardized with slight modification
and the composition was as follows: Ca(NO
3
)
2
. 4H
2
O -
0.02 g, KH
2
PO
4

- 0.0124 g, MgSO
4
.7H
2
O - 0.025 g,
NaHCO
3
- 0.0159 g, EDTAFeNa - 0.225 mg, EDTANa
2
- 0.225 mg, H
3
BO
3
- 0.248 mg, MnCl
2
.4H
2
O - 1.39 mg,
(NH
4
)6Mo
7
O
24
.4H
2
O - 1.0 mg, Cyanocobalamin -
0.04 mg, Thiamine Hcl - 0.04 mg, Biotin - 0.04 mg and
NaSiO
3
.9H
2
O - 0.114 g in 1 L distilled water, adjusted to
pH 6.8. Cultures were allowed to grow at 3K light and
18-20C for 15-20 days. Repeated subcultures were done
on solid medium to obtain pure cultures of diatom
species.
Diatom cleaning for microscopic analysis
Diatoms obtained in pure cultures were cleaned
for microscopic analysis following the protocol of
Battarbee et al., (2001) with slight modification. Equal
volume of 10% HCl solution was added to the diatom
cultures. After proper mixing, 6 ml of a 30% H
2
O
2

Gurung et al.,2013
183 Journal of Research in Plant Sciences (2013) 2(2): 182-191
solution was added, heated in a water bath at 40C for 20
min and then cooled. A drop of NH
3
was added and
centrifuged for 5 min at 4000 rpm in a 20C centrifuge
and decanted the supernatant. Cleaned diatom samples
were used to prepare permanent slides with Naphrax
(mounting medium with refractive index 1.73.
The slides were analyzed under a Light
microscope and examined for the presence of specific
genera of diatoms (Nikon, Eclipse-E200, with photo
micrographic attachment).
Identification
Taxonomic identifications and nomenclatures
were made by consulting various literatures and
monographs (Gandhi, 1955; Husted, 1959; Hendey,
1964; Patrick and Reimer, 1966; Prescott, 1975;
Desikachary, 1989; Round et al., 1990; Nautiyal et al.,
1996 and Anand, 1998). In this study, the diatoms
obtained in pure culture were identified up to genus level
only.

RESULTS AND DISCUSSION
A total of 65 species of freshwater diatoms were
recorded in the Deepor beel of Assam, India. Pure
cultures of diatoms in terms of frequency and abundance
belonged to 27 genera of which 53 are pennate and 12
are centric type. The major genera were found as
Nitzschia (10), Pinnularia (9), Navicula (7), Eunotia (5),
Cyclotella (3), Stauroneis (3), Melosira (3),
Gomphonema (2), Achanthes (2), Achmanthas (3),
Mastogloia (2), Pleurosira (2). The single species of
Gurung et al.,2013
Journal of Research in Plant Sciences (2013) 2(2): 182-191 184
Figure 1: Map showing six different sample collecting sites at Deepor Beel, Assam, India


following diatoms: Synedra, Rhopalodia, Cocconeis,
Cymbella, Luticola, Meridion, Denticula, Diploneis,
Eucocconeis, Placoneis, Stephanodiscus, Thalassiosira,
Coscinodiscus, Aulacoseira (Figure 2-7) were found on
the examined site. In this study, the diatoms obtained in
pure culture were identified up to their genus level,
because for species level identification correctly, detailed
study will be required for characterizing their
ultrastructure of frustules through scanning electron
microscopy. Further, no diatom culture collection facility
is available in India due to which identification up to
species level is very much difficult.
Out of 65 isolates obtained in pure cultures, 10
diatoms were found to be different species of Nitzschia,
representing 15% of the total diatom flora. Further, there
were 9 different species of Pinnularia, representing 14%,
followed by Navicula (11%), Eunotia (8%), Stauroneis
and Melosira representing 5% each, Cyclotella (4%),
and 3% each of Gomphonema, Achanthes, Achnanthes,
Mastogloia and Pleurosira respectively. The rest 14
Gurung et al.,2013
185 Journal of Research in Plant Sciences (2013) 2(2): 182-191
Figure 2(A-I): Pinnularia
Figure 3 (A-J): Nitzschia
diatoms were represented by only one species showing
21% out of the total diatoms obtained in pure cultures
(Figure 8).
Taxonomic descriptions of few major diatom
species found in Deepor beel during study period are as
follows:

Gurung et al.,2013
Journal of Research in Plant Sciences (2013) 2(2): 182-191 186
Figure 4: A F, Navicula, G & H - Gomphonema, I & J - Achanthes, K-M, Achmanthas
Figure 5: A E, Eunotia, F- Navicula, G Mastogloia, H- Synedra, Rhopalodia, J- Cocconeis, K- Cymbella


Class: Bacillariophyceae
Order: Bacillariales
Sub-order: Coscinodiscineae
Family: Coscinodiscaceae
Genus: Cyclotella, Kuetz., 1834
Cyclotella sp. (Fig. 7 A-C)
Valves circular; central area smooth or rarely
finely punctuate; marginal striae coarse, wedge-shaped,
diameter 12.5 m and Striae 9 in 10 m.
Class: Bacillariophyceae
Order: Eunotiales
Sub-order: Eunotiineae
Family: Eunotiaceae
Genus: Eunotia, Ehrenberg 1837
Eunotia sp. (Fig. 5 A-E)
Valves crescentric to arcuate, more or less
parallel sides and slightly narrower rounded apices; short
raphe at the ventral margin near the poles; polar nodules
small; striae parallel, 45-46 m length, 3.5-5.2 m width,
Striae 13-14 in 10 m.
Class: Bacillariophyceae
Order: Bacillariales
Sub-order: Naviculineae
Family: Naviculaceae
Genus: Navicula, Bory 1822, emend. Cleve 1894
Navicula sp. (Fig. 4 A-F)
Valves broadly elliptic; apices short rostrate and
broadly rounded; axial area narrow, parallel; central area
widened and irregular; raphe straight, median with
distinct central nodules; striae radiate through, striae
around the central area alternately long and short, 32 m
length, 15 m width, Striae 8-10 in 10 m.
Class: Bacillariophyceae
Order: Naviculales
Family: Pinnulariaceae
Genus: Pinnularia, Ehrenberg 1840
Pinnularia sp. (Fig. 2 A-I)
Valves linear or elliptic-linear with parallel or
slightly convex sides; apices slightly pointed or broadly
rounded; axial area wide, linear, large central area
somewhat elliptic and slightly wider on one side; raphe
broad and undulate with approximated central nodules
and hooked terminal fissures; striae very coarse,
punctuate, striae radiate becoming parallel convergent
towards the poles, 94 m length, 15-16 m width, Striae
9-10 in 10 m.
Gurung et al.,2013
187 Journal of Research in Plant Sciences (2013) 2(2): 182-191
Figure 6: A Luticola, B D, Stauroneis, E- Meridion, F Denticula, G- Mastogloia, H- Diploneis,
I- Eucocconeis, J- Placoneis, K- Achmanthas
Class: Bacillariophyceae
Order: Cymbellales
Family: Gomphonemaceae
Genus: Gomphonema, C.A. Agardh 1824
Gomphonema sp. (Fig. 4 G & H)
Valves broad, clavate with an apiculate head pole
and a capitate foot pole; axial area linear, widening into a
round or elliptical central area, which is wider on one
side and has an isolated pore on the primary side of the
central nodule; raphe straight with hooked terminal
fissures; striae coarsely punctate, slightly radiate, striae
shorted and wider around the central area, 48.5 m
length, 10 m width, Str 10-12 in 10 m.
Class: Bacillariophyceae
Order: Cymbellales
Family: Cymbellaceae
Genus: Cymbella, C.A. Agardh 1830
Cymbella sp. (Fig. 5 K)
Valves asymmetric, semi-lanceolate, dorsal side
convex and ventral side almost straight and slightly
expanded at the centre of the valve; apices slightly
rostrate and broadly rounded; axial area broad and
widening into an elliptic central area; raphe located
slightly closer to the ventral valve margin, very slightly
curved; striae radiate, 130 m length, 29 m width,
Striae 8-10 in 10 m.
Gurung et al.,2013
Journal of Research in Plant Sciences (2013) 2(2): 182-191 188
Figure 7: A C: Cyclotella, D & E- Pleurosira, F- Stephanodiscus, G- Thalassiosira, H- Coscinodiscus,
I and K Melosira, J- Aulacoseira


Class: Bacillariophyceae
Order: Fragilariales
Family: Fragilariaceae
Genus: Synedra Ehrenberg 1832: 87
Synedra sp. (Fig. 5 H)
Valves linear to linear lanceolate and tapering at
the poles, sometimes narrow with capitate ends; ends
rounded pseudoraphe linear, central area absent, striae
coarse. Length 95 to 156 m, breadth 7 to 9 m, striae 9
to 11 in 10 .
Class: Bacillariophyceae
Order: Bacillariales
Family: Bacillariaceae
Genus: Nitzschia, Hassall, 1845: 435
Nitzschia sp. (Fig. 3 A-J)
Frustule showed wide variation among the
species, generally narrowly linear. Some species
revealed small frustules, valves linear to linear
lanceolate, with slightly constricted, shortly wedge
shaped, acute rounded or capitates ends. Striae fine and
almost indistinct. Length 15 to 26 m, breadth 2.5 to 4.5
m, striae 32 to 35 in 10 m and punctae 10 to 13 in
10 m.
Class: Bacillariophyceae
Order: Acnanthales
Family: Achnanthaceae
Genus: Achnanthes, C.A. Agardh (1824)
Achnanthes sp. (Fig. 4 K-M)
Valves linear, lanceolate in outline, gibbous in
the middle; ends obtusely rounded. Valve with raphe
were straight with distinct central pores, axial area
narrow and linear. Central area broad reaching the side
walls. Length - 40 to 46 m, breadth 8.5 to 10 m.
Class: Bacillariophyceae
Order: Melosirales
Family: Melosiraceae
Genus: Melosira, C.A. Agardh (1824)
Melosira sp. (Fig. 7 I-K)
Frustules cylindrical, united in chains, mantle
line straight. Surface of the Mantle punctate with
puncta in spiral rows. Diameter of the cell 7 to 12 m.
Height of half cell 8 to 14 m.
Class: Bacillariophyceae
Order: Thalassiosirales
Family: Thalassiosiraceae
Genus: Thalassiosira, Hasle & Heimdal, 1970
Thalassiosira sp. (Fig 7G)
In valve view, the frustules is circular with a
diameter ranging from 3.8 to 6.0 m, averaging 4.5 m,
regular occurring bright spots, areas of greater density,
observed around the margin ca. 0.9 m apart.
Class: Bacillariophyceae
Order: Aulacoseirales
Family: Aulacoseiraceae
Genus: Aulacoseira, (Ehrenberg) Simonsen 1979
Aulacoseira sp. (Fig. 7J)
Circular valve ranging from 12 to 15 m in
diameter and the length of the mantle from 15 to 18 m,
the valve face covered with areolae, areolar rows are
usually spiral and in some population straight, spines
large and sharply pointed.
Class: Bacillariophyceae
Order: Coscinodiscales
Family: Coscinodiscaceae
Genus: Coscinodiscus, Gran and Angst 1931
Coscinodiscus sp. (Fig. 7H)
Gurung et al.,2013
189 Journal of Research in Plant Sciences (2013) 2(2): 182-191
Figure 8: Diversity of diatom flora in Deepor beel.
Cylindrical, pervalvar axis varying from about
to the whole length of the diameter. The transition from
valve face to valve mantle abrupt, forming nearly 90
degree angle, the valve centre irregular hyaline due to
variable termination of the radial rows of areolae, valve
mantle large, 30-40 m in the pervalvar direction, with
slightly larger areolae, 4-6 in 10 m.
Class: Bacillariophyceae
Order: Thalassiosirales
Family: Stephanodiscaceae
Genus: Stephanodiscus, Ehrenberg, 1845
Stephanodiscus sp. (Fig. 7F)
Cells discoid, ring of spines were present around
the valve face, areolae simple round pores, areolae of the
valve mantle slightly closer together and often occur in
diagonal rows without intervening costae, mantle edge
often vertically grooved areolae 12-18 m in 10 m.
Majority of the forms were solitary and few
colonial. Distinct numbers of dominant genera of
pennate diatoms like Nitzschia, Navicula, Gomphonema,
Cymbella, Synedra etc. were dominant throughout the
year. Further, Nitzschia and Pinnularia were found to be
common in all the selected locations. Centric diatoms
like Coscinodiscus, Stephanodiscus, Melosira and
Aulacoseira were most dominant during the spring.
Taxonomic investigations on the diatom flora of
Assam are still much limited and most studies were
performed randomly along with other algae. Northeast
India in general and Assam in particular is one of the
biodiversity hotspot which along with different
demographic and geographical patterns create diverse
types of ecological niche, supporting different types of
diatoms, thereby requires a more extensive exploration
and documentation of its diatom flora. The establishment
of a diatom database would be an essential contribution
to the conservation and exploitation of its aquatic
biodiversity (Round et al. 1990).
All diatoms were solitary and only Melosira and
Aulacoseira were found in colonial form. Particularly,
diatoms like Nitzschia, Navicula, Eunotia, Cyclotella,
Melosira, Pleurosira and, Eucocconeis were most
dominant during the monsoon whereas distinct numbers
of dominant genera of diatoms like Nitzschia, Navicula,
Cyclotella, Pleurosira, Synedra and Eucocconeis were
dominant during winter season. The Nitzschia, Navicula,
Eunotia, Cyclotella, Gomphonema, Pleurosira, Synedra,
Diploneis and Eucocconeis were found to be most
abundant in all the seasons. In this study, the diatom
diversity clearly depicts highest number of diatoms in
winter, followed by that in post monsoon, summer and
least was recorded in monsoon season. This finding is in
good agreement with some previous reports (Singh et al.
2011).

CONCLUSION
In the present investigation, 65 diatom taxa under
26 genera were obtained in pure cultures from Deepor
Beel, Assam, India, which is an evident of rich diatom
taxa in this important Ramsar site of Assam. As diatoms
play very important roles from ecological point of view,
therefore, the present basic information of the diatom
database, their distribution and diversity would form a
useful tool for further ecological assessment and
monitoring of the Deepor beel.

ACKNOWLEDGEMENT
I would like to thank Dr. G.C. Sarma,
Department of Botany, Gauhati University, Assam for
helping in identification of the specimens. F i n a n c i a l
support received from the North East Center for
Research & Development (NECRD), IGNOU, Govt. of
India, grant no. IG/NECRD/09-10/RES/PROJ/
BT/52/721 is highly acknowledged.

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