1. Presented by:
Debarati Chakraborty
M.Phil,2nd year
School Of Oceanographic
Studies,
Jadavpur University
Under supervision of :
Dr. Punarbasu Chaudhuri, Faculty
Department of Environmental Science
University of Calcutta
Dr. Sugata Hazra ,Director
School of Oceanographic Studies
Jadavpur University
Under supervision of :
Dr. Punarbasu Chaudhuri, Faculty
Department of Environmental Science
University of Calcutta
Dr. Sugata Hazra ,Director
School of Oceanographic Studies
Jadavpur University

2. Graph showing gradual increase in heavy metal pollution since Industrial Revolution.
J.O. Nriagu, “History of Global Metal Pollution,” Science,Vol.272
Primary sources are the burning of fossil fuels, mining and smelting of metalliferous ores,
automobiles, agrochemicals municipal wastes & sewage (Velez &Montoro,
1998;Conacher, et al.,1993). The most common heavy metals found at contaminated
sites, in order of abundance are Pb, Cr, As, Zn, Cd, Cu & Hg. USEPA, Report(1996).
Debarati Chakraborty,J.U

3. Plant exposure to Environmental stresses (high salinity, high light intensity,
heavy metal toxicity, temperature extremes, drought, herbicide treatment
etc.)
Disturbed redox homeostasis balance between the production of reactive
oxygen species(ROS)[singlet oxygen(1O2),superoxide anion(O2-), hydrogen
peroxide H2O2,hydroxyl radical(OH-)[via the Haber-Weiss and Fenton
reactions(K.J.Dietz et al.1999;A.Polle et al.2002)].
Often resulting in oxidative damage(Hernandez et al. 1999).
Mangroves , a complex intertidal ecosystem possess a remarkable capacity to
retain heavy metals(Dwivedi et al.1983).
Debarati Chakraborty,J.U

4. Occurrence:
Primarily along sheltered
tropical & sub-tropical coastal
regions within 300 of the
equator(Alongi 2002,Upadhyay et al.2002)
Mangroves occur in 2 distinct bio-geographical
region:
1. The Indo-West Pacific(IWP)
including Asia ,Australia &
Eastern coast of Africa.
2. The Atlantic-East Pacific
(AEP) covering America &
western coast of Africa.
Covers about 6,768 km2 constituting
7% of world mangroves(Krishnamurthy
1987)
Of which ,70% is on East coast & 12% is
on West coast.
Debarati Chakraborty,J.U

5. •Mangroves survives extremely hostile & inhospitable conditions(high salinity,
tidal extremes, high temperature & muddy anaerobic soil conditions)
[Kathiresan & Ramesh 1991].
•Mangroves and their associates have high antioxidant potential [Kathiresan et al.2013].
•Mangroves can act as a long term sink for heavy metals because of their
precipitation with sulphides during diagenetic reactions (Huerta-Diaz,Morse,1992).
•Endophytes hidden within them can also have contributed towards relieving HM
stress (Redman et al.2002; Rodriguez et al. 2004; Swarthout et al.2009;Tian et al.2008).
•A novel application of endophytes in the area of phytoremediation has been reported in
many reviews (Ma et al. 2011).
•Metal bioavailability is a function not only of their total concentration but also
of physico-chemical (e.g. pH, Eh, organic matter, clay content)& biological
(e.g. biosorption, bioaccumulation & solubilization) factors (Berthelin et al.,1995).
Debarati Chakraborty, J.U

6. Aims & Objectives
• To identify Mangal population having heavy metal polluted zones in West Bengal.
•To analyze the amount of Stress Enzymes namely SOD,POD,CAT( for
assessing
antioxidant potential) of five mangal species and their endophytes.
•To assess the photoactive state of mangals in heavy metal polluted zones by
measuring chlorophyll content.
•To gather information regarding various physico-chemical parameters which
may affect bioavailability of heavy metals of the sites.
•Establishing the role of enzymatic potential of the concerned Mangroves
conferring them heavy metal tolerance.
Debarati Chakraborty,J.U

7. Map showing location of sampling sites
Debarati Chakraborty,J.U

8. Acanthus ilicifolius Sonneratia caseolaris Excoecaria agallocha
Sonneratia apetala Avicennia officinalis
Methodology- Samples collected
Debarati Chakraborty,J.U

9. Biochemical parameters-
•Chlorophyll Content ( Chla ,Chlb, Total Chlorophyll, Chla /Chlb )by Arnon’s
Method(1949)& application of Porra’s Correction(1991).
• Protein Content (Folin-Ciocalceteau method,1940).
•Peroxidase content(Cippolini 1998) and Superoxide dismutase content ( Giannapolitis
& Ries,1977 with modifications of Chowdhury & Chowdhuri,1985).
Physico-chemical parameters –
• pH, Conductivity, oxidation reduction potential of soil samples.
•Percentage of organic carbon and Percentage of organic matter.
•Available phosphorus
Debarati Chakraborty,J.U

10. Chlorophyll content Calculation Formula
(D.I. Arnon (1949) Plant Physiol. 24: 1)
Chla(mg g-1)= [(12.7 × A663) - (2.6 × A645)] × ml acetone/mg leaf tissue
Chl b(mg g-1)= [(22.9 × A645) -(4.68 ×A663)] × ml acetone/mg leaf tissue
Total Chl = [Chla + Chlb ]A
Total chlorophyll pigment value, although flawed by interferences by other
chlorophylls, phaeo-pigments etc. is the only value that remains fairly independent
of chlorophyll methodology. Therefore, it is the only measurement that provides
historical consistency.
Corrections(Porra et al. 1989; Porra 1991)
Chl a/bT = 0.593 +0.459·(Chl a/bA)+0.229·(Chl a/bA)2
[Chl a + b]T = 0.895[Chla + Chlb]A
Ref: Standard Methods for the Examination of Water and Wastewater.17th Edition. APHA. 1989.
Debarati Chakraborty,J.U

11. Acid digestion protocol
(Santra et al.2009)
0.5gm sample was soaked overnight in
5ml nitric acid and next day the samples
were heated upto 600 c for two hours and
precautions are taken so that the samples
don’t be dried.
The tubes were cooled and sulphuric acid and perchloric acid were
added to the samples in 3:2 ratio and was heated for 4 hrs.until dense
white fumes were obtained along with white ppt.Finally volume
makeup was done with double distilled water and filtrate was obtained
with Whatman filter paper no.42.
Debarati Chakraborty,J.U

12. Avicennia officinalis correlation matrix(Pb)
*=95% confidence interval,**=99% confidence interval
Debarati Chakraborty,J.U

13. Avicennia officinalis correlation matrix (Cd)
% *=95% confidence interval,**=99% confidence interval
Debarati Chakraborty,J.U

14. Sonneratia apetala correlation matrix(Cd)
*=95% confidence interval,**=99% confidence interval
Debarati Chakraborty,J.U

15. Sonneratia apetala correlation matrix(Pb)
*=95% confidence interval,**=99% confidence interval
Debarati Chakraborty,J.U

16. Excoecaria agallocha correlation matrix(Cd)
*=95% confidence interval,**=99% confidence interval
Debarati Chakraborty,J.U

17. Excoecaria agallocha correlation matrix(Pb)
*=95% confidence interval,**=99% confidence interval
Debarati Chakraborty,J.U

18. Avicennia alba correlation matrix(Cd)
*=95% confidence interval,**=99% confidence interval
Debarati Chakraborty,J.U

19. Avicennia alba correlation matrix(Pb)
*=95% confidence interval,**=99% confidence interval
Debarati Chakraborty,J.U

20. Table showing correlations of various parameters with increase in
Heavy Metal(Pb & Cd) content
Parameters Pb content Cd content
Protein content ‘+’ve ‘+’ve
Peroxidase
content
‘+’ve ‘+’ve
Total Chlorophyll
content
‘-’ve ‘-’ve
Chla/Chlb ‘-’ve ‘-’ve
Organic matter ‘-’ve ‘-’ve
Available
phosphorus
‘-’ve(A.alba) ‘-’ve(A.alba)
Debarati Chakraborty,J.U

21. Excoecaria agallocha, leaf
endophytes
Acanthus ilicifolius, leaf
endophytes
Sonneretia apetala,
pneumatophore endophytes
Endophytes isolated from various parts of mangrove samples of Jharkhali
Debarati Chakraborty,J.U

22. Endophytes isolated from
Sonneratia apetala, leaf,Ghushighata
Endophytes isolated from
Sonneratia apetala, leaf, Jharkhali
Debarati Chakraborty,J.U

23. Future Scope:
1) Isolation of endophytes from plant samples showing high enzyme activity
(especially those collected from polluted sites) and their identification.
2) Obtaining enzyme data from isolated fungi & correlating fungal data with plant
data.
3) Obtaining total Phenolics & total Flavonoids data from leaf samples
4) Obtaining metal uptake data of various plant parts.
5) Antioxidant assay by DPPH(1,1-diphenyl-picrylhydrazyl) radical scavenging.
Debarati Chakraborty,J.U

24. •Physico-chemical Data courtesy: Harish Sharma, M.Sc, 2nd year, Dept.of Environmental
Science,University of Calcutta.
•Niharda, sample collection, Jharkhali.
•Akbarda, sample collection ,Shamshernagar.
•Somdeep Ghosh, research fellow, Dept.of Environmental Science,University of Calcutta.
•Madhurima Bakshi, research fellow, Dept.of Environmental Science,University of Calcutta.
•Kushal Banerjee,M.Sc, 1st year, Dept.of Environmental Science, University of Calcutta.
• Avra Chanda,Anirban Akhand, School of Oceanographic
Studies,Jadavpur University.
•Dr. Ashish Ghosh, Guest faculty,. School of Oceanographic Studies, Jadavpur University.
•Dr. Joydeep Mukherjee, Environmental Science Department, Jadavpur University.
Acknowledgement

25. References
•Mohammad Anwar Hossain, Pukclai Piyatida,Jaime A. Teixeira da Silva and Masayuki
Fujita(2012) MolecularMechanism of HeavyMetal Toxicity and Tolerance in Plants: Central Role
of Glutathione in Detoxification of Reactive Oxygen Species and Methylglyoxal and in HeavyMetal
Chelation.
•Zhen-Jian Lin, Guo-Jian Zhang, Tian-Jiao Zhu, Rui Liu, Hong-Juan Wei, and Qian-Qun
Gu(2009)Bioactive Cytochalasins from Aspergillus flavipes, an Endophytic Fungus Associated with
the Mangrove Plant Acanthus ilicifolius. Helvetica Chimica Acta – Vol. 92, 1538-1544.
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