Role of Metallothionein and Phytochelatin in Combating Abiotic …
351
2 Materials and Methods
2.1 Plant Materials and Treatments
Brinjal [Solanum melongena L.] [Variety-BCO-1, Bidhan Chandra Krishi
Viswavidyalaya, West Bengal, India] seeds were used for this study. About 500,
800, and 1000 μM solution of copper chloride (CuCl 2 ) (SRL, Mumbai, India) were
used as treatment. LD 50 was measured and it was found to be 1100 mM for Cu
treatment . Hence all doses were taken lesser than the LD 50 value.
2.2 Evaluation of Stress
2.2.1 Estimation of Chlorophyll and Carotenoid Content
The amount of chlorophyll was measured according to the method of Lichtenthaler
(1987) with little modifications.
2.2.2 Lipid Peroxidation
The level of lipid peroxidation can be determined by measuring the amount of malondialdehyde (MDA). Method of Heath and Packer (1968) with little modifications
(Ghoshal et al. 2015) was used to measure the extent of lipid peroxidation. The
result was expressed as MDA equivalents in μM/g fresh weight of tissue.
2.3 Preparation of Plant Extract
Plant extracts were prepared according to Brolis et al. (1998) with little modifications
(Talukder et al. 2016).
2.4 Evaluation of Antioxidant Activity
2.4.1 Total Antioxidant Assay
This assay was performed according to Prieto et al. (1999).
351
2 Materials and Methods
2.1 Plant Materials and Treatments
Brinjal [Solanum melongena L.] [Variety-BCO-1, Bidhan Chandra Krishi
Viswavidyalaya, West Bengal, India] seeds were used for this study. About 500,
800, and 1000 μM solution of copper chloride (CuCl 2 ) (SRL, Mumbai, India) were
used as treatment. LD 50 was measured and it was found to be 1100 mM for Cu
treatment . Hence all doses were taken lesser than the LD 50 value.
2.2 Evaluation of Stress
2.2.1 Estimation of Chlorophyll and Carotenoid Content
The amount of chlorophyll was measured according to the method of Lichtenthaler
(1987) with little modifications.
2.2.2 Lipid Peroxidation
The level of lipid peroxidation can be determined by measuring the amount of malondialdehyde (MDA). Method of Heath and Packer (1968) with little modifications
(Ghoshal et al. 2015) was used to measure the extent of lipid peroxidation. The
result was expressed as MDA equivalents in μM/g fresh weight of tissue.
2.3 Preparation of Plant Extract
Plant extracts were prepared according to Brolis et al. (1998) with little modifications
(Talukder et al. 2016).
2.4 Evaluation of Antioxidant Activity
2.4.1 Total Antioxidant Assay
This assay was performed according to Prieto et al. (1999).