2.3.3. Chlorophyll Content

Fresh leaf samples (0.25 g) were randomly chosen from three plants per replicate and homogenized in 20 mL of acetone (80% *v*/*v*). The absorbance was measured using a spectrophotometer (JENWAY 6300, Staffordshire, UK) at 663 and 646 nm, and the chlorophyll content was determined using the following equations [21]:

$$\text{Chl a (mg g}^{-1} \text{ fw)} = 12.21 \times \text{A} \cdot \text{663} - 2.81 \times \text{A} \cdot \text{646}$$

$$\text{Chl b (mg g}^{-1} \text{ fw)} = 20.13 \times \text{A} \cdot \text{646} - 5.03 \times \text{A} \cdot \text{663}$$

$$\text{Total chlorophyll content (mg g}^{-1} \text{ fw)} = \text{Chl a} + \text{Chl b}$$
