Total soluble sugars:

Total soluble sugars were determined by Riazi et al., (1985). Fresh leaves (1 g) were grounded in 10 mL of 80% methanol. Centrifuged at 10,000rpm and filtered the extract and supernatant was used for the estimation of sugars. Plant extract 0.3mL was taken in test tubes and 3 ml of anthrone reagent was added to each test tube. The samples were placed for 30 minutes at room temperature. Optical density was recorded at 480 and 490 nm on a spectrophotometer. The concentration of soluble sugars was calculated from the standard curve developed with different concentrations of glucose.

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