ANALYSIS OF CHOLOPHYLL CONTENT IN ADIANTUM CAPILLUS-VENERIS L. GROWING IN DIFFERENT HABITATS IN DOON VALLEY AND NEARBY AREAS

Abstract

The present study focused on the analysis of photosynthetic pigments (Chl-a,Chl-b, chlorophylla/b ratio) of Adiantum capillus-veneris L. collected from five different locations of Doon valley and nearby areas. The areas include Maldevta, Shasthradhara, Majra, Patel Nagar, Biharigarh. Out of the five sites, plants collected from two sites (Maldevta, Shasthradhara) were more of natural habitats while other three (Majra, Patel Nagar, Biharigarh) were more near human habitat. The analysis of photosynthetic pigment was carried out by using chlorophyll extraction method as described by Arnon, 1949. Conclusion The highest concentration of photosynthetic pigment was found in plants which grow in moist and shady places i.e., Maldevta and Shasthradhara near water source, while the lowest value was obtained from the plants which belongs to dry and exposed areas i.e., Patel Nagar and Biharigarh. Highest Chl a content (0.16 mg/gfw) was recorded from Maldevta while lowest 0.083 mg/gfw have been reported from Majra near human habitat. Highest Chl b value (0.13 mg/gfw) was obtained from Biharigarh region whereas lowest value (0.008 mg/gfw) was found in Shasthradhara region. Chlorophyll a/b ratio is lesser in plants which grow in shady places as seen in the study that plants in Maldevta showed least value (3.7) while highest in exposed areas i.e., Shasthradhara (20). In our investigation it has been observed that chl a/b ratio is high in plants which are found in shady places in comparison to the plants which are grown in exposed areas so these changes can be attributed to disturbances in environmental factors i.e., increase in anthropogenic activities which leads to environmental stress. Thus it can be concluded that chlorophyll loss may be associated to environmental stress and the variation in total chlorophyll can be considered as a significant indicator of pollution.

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RESEARCH ARTICLE
ANALYSIS OF CHOLOPHYLL CONTENT IN ADIANTUM CAPILLUS-VENERIS L. GROWING IN
DIFFERENT HABITATS IN DOON VALLEY AND NEARBY AREAS
1Chhaya Singh, 1,*Dr. Neha Chauhan, 1Shagun Gupta, 2Anju Rani and 1Sumita Tomar
1Division of Life Sciences, Shri Guru Ram Rai Institute of Technology and Sciences, Dehradun, India
2K.V. Subharti College of Science, Swami Vivekanand Subharti University, Meerut (UP), India
ARTICLE INFO ABSTRACT
The present study focused on the analysis of photosynthetic pigments (Chl-a,Chl-b, chlorophylla/b
ratio) of Adiantum capillus-veneris L. collected from five different locations of Doon valley and nearby
areas. The areas include Maldevta, Shasthradhara, Majra, Patel Nagar, Biharigarh. Out of the five sites,
plants collected from two sites (Maldevta, Shasthradhara) were more of natural habitats while other
three (Majra, Patel Nagar, Biharigarh) were more near human habitat. The analysis of photosynthetic
pigment was carried out by using chlorophyll extraction method as described by Arnon, 1949.
Conclusion The highest concentration of photosynthetic pigment was found in plants which grow in
moist and shady places i.e., Maldevta and Shasthradhara near water source, while the lowest value was
obtained from the plants which belongs to dry and exposed areas i.e., Patel Nagar and Biharigarh.
Highest Chl a content (0.16 mg/gfw) was recorded from Maldevta while lowest 0.083 mg/gfw have
been reported from Majra near human habitat. Highest Chl b value (0.13 mg/gfw) was obtained from
Biharigarh region whereas lowest value (0.008 mg/gfw) was found in Shasthradhara region.
Chlorophyll a/b ratio is lesser in plants which grow in shady places as seen in the study that plants in
Maldevta showed least value (3.7) while highest in exposed areas i.e., Shasthradhara (20). In our
investigation it has been observed that chl a/b ratio is high in plants which are found in shady places in
comparison to the plants which are grown in exposed areas so these changes can be attributed to
disturbances in environmental factors i.e., increase in anthropogenic activities which leads to
environmental stress. Thus it can be concluded that chlorophyll loss may be associated to
environmental stress and the variation in total chlorophyll can be considered as a significant indicator of
pollution.
Copyright © 2018, Chhaya Singh. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use,
distribution, and reproduction in any medium, provided the original work is properly cited.
INTRODUCTION
The major light absorbing pigments in green plants are
chlorophyll. They are found within the chloroplasts. They play
a major role in photosynthesis, a complex biochemical
pathway. The most common type of chlorophyll is
Chlorophyll–a. It is dark green in colour. It is found in the core
complexes of photosystems. The main leaf pigment includes
chlorophyll-a, chlorophyll-b and carotenoids that are essential
for photosynthesis process. Internal factors and environmental
conditions may be responsible for the variation in leaf
pigments (chlorophylls and carotenoids). Shaikh and Dongare
(2008) reported that chlorophyll and carotenoids content
changes with microclimatic conditions in Adiantum species.
The ratio of chlorophyll-a and chlorophyll-b in terrestrial
plants has been used as an indicator of response to light shade
conditions. The small value of chlorophyll a/b can be
considered as sensitive biomarker of pollution and
environmental stress. The qualitative and quantitative analysis
of pigments is facilitated by the absorbance properties of them.
*Corresponding author: Dr. Neha Chauhan,
Division of Life Sciences, Shri Guru Ram Rai Institute of Technology
and Sciences, Dehradun, India.
Adiantum capilus- veneris is a common fern mostly found
growing in shady and moist as well as in open and dry habitats
in Doon valley and nearby areas.
MATERIAL AND METHODS
Collection of plants: The plants were collected from five
different locations of Doon valley and nearby areas i.e.,
Maldevta, Shasthradhara, Majra, Patel Nagar and Biharigarh.
The species were collected from natural habitat as well from
human habitat.
Extraction of chlorophyll (Arnon, 1949): One gram of finely
cut fresh leaves were taken and ground with 20 – 40ml of 80%
acetone. It was then centrifuged at 5000 –10000rpm for 5mins.
The supernatant was transferred and the procedure was
repeated till the residue becomes colourless. The absorbance of
the solution was red at 645nm and 663nm against the solvent
(acetone) blank.
ISSN: 0976-3376
Asian Journal of Science and Technology
Vol. 09, Issue, 08, pp.8497-8501, August, 2018
Available Online at http://www.journalajst.com
ASIAN JOURNAL OF
SCIENCE AND TECHNOLOGY
Article History:
Received 19th May, 2018
Received in revised form
07th June, 2018
Accepted 25th July, 2018
Published online 30
th
August, 2018
Key words:
Photosynthetic pigments,
Adiantm capillus-veneris L.,
Chlorophyll,
Environmental stress.

Estimation of Chlorophyll content: The concentrations of
chlorophyll a, chlorophyll b and total chlorophyll were
calculated using the following equation:
mg chlorophyll a /gm Fr.wt [12.7 (A 663) – 2.69(A645)] X
V/1000 X W
mg chlorophyll b /gm Fr.wt [22.9(A645)- 4.68 (A663)]X
V/1000 X W
Total chlorophyll (mg/gfw)=20.2(A645)+8.03(A663) X
V/1000 X W
Where,
A = Absorbance of chlorophyll extract at the specific indicated
wavelength,
V = Final volume of the solution & W= gm of tissue extracted
RESULT AND DISCUSSION
Majority of the ferns are found growing in shady, moist
conditions, damp areas. Doon valley which is situated up to
elevation of 1500 m with tropical and subtropical vegetation
dominated by Sal forest (Chhaya et.al. 2017). Adiantum
species is mostly found in exposed areas of forest with scanty
vegetation especially Adiantum capillus-veneris. (Chhaya
et.al. 2013). Fig 1 shows the distribution of Adiantum species
at various places: Estimation of Chlorophyll was done using
fresh green fronds extracted with 80% acetone solvent. The
absorbance values of chlorophyll extracts were measured at
two different wavelengths 645nm and 663nm respectively.
The calculations of the amount of chlorophyll a, chlorophyll b,
chlorophyll a/b and total chlorophyll were based on the
absorbance values obtained, using Arnon’s (1949) equation
and were tabulated in table 1. Extraction of Chlorophyll can be
observed in Fig 1. As Blank (i), after 24 hour (ii), after
48hours (iii) and after 72hours (iv). It was observed from the
present investigation that highest Chlorophyll a concentration
(0.16mg/gfw) was reported in Maldevta and Sahastradhara
locations and least (0.083mg/gfw) in Majra (Fig: 3). Highest
chlorophyll b values (0.13mg/gfw) were showed by plants
belonging to Biharigarh location and least were (0.008mg/gfw)
observed in the case of sahatradhara. The chlorophyll contents
were greater in the shade ferns when compared to the sun ferns
(Boyce & Mohamed 1987). The greater values of total
chlorophyll were recorded from the region of Maldevta (0.204
mg/gfw) and Sahatradhara (0.205 mg/gfw) whereas lower in
Patel nagar and Biharigarh (0.058 mg/gfw). Shaikh &
Dongare(2008) showed that variation in chlorophyll and
carotenoids contents varied with microclimatic conditions in
Adiantum philippense L. Because of numerous factors
concerned with nutrition and light which can influence
chlorophyll content, and because the plants sampled were
growing under a variety of conditions (Wolf 1958). The
variation in total chlorophyll and its loss is believed to be
associated with environment stress and good indicator of stress
which is indicated by observed ratio of chlorophylla/b.
According to Lahani et.al. (2003) chlorophyll a/b ratio is lesser
in the plants that grow under the shade than in plants that were
exposed to full sunlight. But in our study deviation has been
observed as the highest chlorophyll a/b ratio (20 mg/gfw) in
case of Shasthradhara (Shady place). The reason for this
variation may be the environmental stress due to increase in
anthropogenic activities as it a tourist spot.
i ii
iii iv
Fig. 1. Shows the extraction of Chlorophyll after 24hours to 72hours of treatment with 80% acetone
8498 Asian Journal of Science and Technology, Vol. 09, Issue, 08, pp.8497-8501, August,
2018

Table 1. Estimation of Chlorophyll in Adiantum capillus-veneris of different locations in Doon Valley
S. no Name of the plant Habitat Chlorophyll a
(mg/gfw)
Chlorophyll b
(mg/gfw)
Total Chlorophyll
(mg/gfw
Chlorophyll a/b
(mg/gfw)
1. Adiantum capillus-veneris Maldevta (natural habitat/shade) 0.16 0.043 0.204 3.7
2 Adiantum capillus-veneris Majra (human habitat/exposed) 0.083 0.021 0.105 3.9
3 Adiantum capillus-veneris Shasthradhara (natural) habitat/shade) 0.16 0.008 0.205 20
4 Adiantum capillus-veneris Patel nagar human habitat/exposed) 0.045 0.012 0.058 3.7
5 Adiantum capillus-veneris Biharigarh (human habitat/exposed) 0.045 0.13 0.058 3.7
Fig. 2. 1.shashtradhara 2. Maldevta 3.Majra 4. Biharigarh 5. Patel nagar
Fig. 3. Graphical representation of chlorophyll a content estimated in plants of different locations at Doon valley
8499 Asian Journal of Science and Technology, Vol. 09, Issue, 08, pp.8497-8501, August,
2018

Fig. 4. Graphical representation of chlorophyll b content estimated in plants of different locations at Doon valley
Fig. 5. Graphical representation of Total chlorophyll content estimated in plants of different locations at Doon valley
Fig. 6. Graphical representation of chlorophyll a/b ratio estimated in plants of different locations at Doon valley
8500 Asian Journal of Science and Technology, Vol. 09, Issue, 08, pp.8497-8501, August,
2018

Environmental Pollutant can easily interact with essential plant
metabolic processes leading to defect in Chl a and b
production or function and leading to impairment in whole
photosynthetic activity & as a result affecting plant fitness.
Low concentrations of chlorophyll can directly limit
photosynthesis and hence growth and yield. The Adiantum
capillus-veneris L. collected from the moist places were
having large and thick leaves as compared to those growing
near human habitat as also seen in Fig 2.
Conclusion
From the present study it can be concluded that variation in
chlorophyll can be observed in ferns of same species from
shady places to moist and damp places. Since the habitat
where ferns are reported are more under threat due to human
stress may be habitat fragmentation, human interference in the
forest area etc.The major factor responsible is environmental
pollution which has deleterious effects on the production of
chlorophyll. Further study has to be undertaken by analysis of
different factor such as plant biomass of plant collected from
different source, variation in the size of the leaves, changes in
the stomatal structure etc.As it is well known that chlorophyll
or chlorophyll derivatives can be utilized as photodynamic
agent in Tumor and cancer therapy. (Brandis et al., 2006) this
aspect can also be experimentally carried out in case of
Adiantum capillus-veneris.
Acknowledgment
We are greatly thankful to the Vice-chancellor Dr P.P. Dhyani
of Shri Guru Ram Rai university for providing support and
Head of Department Dr Kunal Kishore of Life sciencies for
providing research facilities.
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