Molecular Plant Breeding 2015, Vol.6, No.14, 1
-
8
7
kept in hot air oven (80-90ºC) for 7 days till the
constant weight comes. Each type of sample of equal
weight was completely immersed in different Cd
solutions (0, 50, 100 and 200 µM of CdCl
2
) for 24 h
with constant agitation at 37°C. After incubation
period was over, the plant samples were filtered
through Whatman 42 filter paper and the filtrates were
analyzed for Cd content through Atomic Absorption
Spectrophotometry (Spectra AAS 240 Agilent
Technologies, γ for Cd absorption-228.8 nm) .The
amount of metal bioadsorbed by the leaf sample was
calculated from the difference of Cd content before
and after immersion of plant material in respective
solutions (Pandey and Banerjee 2011).
All observations were recorded considering four
replications (n = 4).The statistical analysis was
performed by one-way ANOVA analysis taking P ≤
0.05. The data presented in the figures are as mean
value ±SE.
Detection of metal distribution in tissues.
Scanning Electron Microscopy (SEM) coupled with
Energy Dispersive Analysis for X-ray (EDAX) was
employed to monitor the cell surface variations with
different concentrations of Cd adsorption (Srivastava
and Thakur 2006). The distribution of metals in
different proportions of the cells and Cd binding sites
were detected by EDAX spectra of the biosorbent
from surface pictures of SEM (JSM 6700 F, Japan)
Detection of functional groups for biosorption
The different functional groups present in the cell wall
constituting moieties were done by Fourier Transform
Infrared Spectrometry (FTIR). For FTIR analysis
plant material was ground into fine powder and mixed
with Potassium Bromide (KBr, AR) in 1:1000 p/p. The
range of absorption spectra was between 400-4000
cm
-1
. In our experiment, three specific wavelengths
for probable changes within the major biomolecules as
carbohydrate (1200 cm
-1
- 1000 cm
-1
), lipid substances
(3000 cm
-1
to-2800 cm
-1
) and protein (1800- cm
-1
to
1500 cm
-1
) were considered. From the spectra
obtained the possible changes of IR absorption was
detected for specific functional groups as suggested
(Luo et al., 2010)
Acknowledgement
The corresponding author acknowledges the partial financial
support both from DST-PURSE programme activated to
University of Kalyani. The K. Das, Assistant Prof. in Botany,
acknowledges the FDP programme, UGC XI
th
Plan.
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