Relative water contents (RWC) (%):

The relative water content will be calculated following the method of Jones and Turner (1978). Fresh leaf samples will be weighed (Fw), soaked in deionized water in the dark for 24 h to re-hydrate and their turgid weight (TW) recorded. After that, the samples will be oven-dried at 80°C for 48 h to get their …

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Soil pH:

Soil samples were used to measure the pH of the soil. Each replicate was subjected to pH analysis to determine the average acidic or alkaline nature of the soil. The pH meter was used which was standardized with buffers provided by the manufacturers that were of 4.01, 7.00, 10.00 standards. 5g soil was added with …

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Soil texture:

The hydrometer method was used to determine soil texture. Particle size determination of samples was done through this method. 1% solution of sodium Hexa Meta phosphate was made and 30g soil was added with this solution and left it for 24 hours. Dilution was done with distilled water in a 1L cylinder, using a mechanical …

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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 …

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Total flavonoids:

Flavonoids were determined by using the AlCl3 method. 1ml of the sample was taken in cuvette and 0.3ml of NaNO2 was added. Before adding the next chemical shaken well and then added 0.3ml of AlCl3. After mixing well, 2ml of NaOH was added. The optical density was taken at 510nm via a spectrophotometer.

Total phenolics:

Following Julkenen-Titto (1985), fresh leaf 0.5g was ground in 10ml of 80% of methanol. Centrifugation was done at 10,000rpm for 10 minutes.  Then 0.25ml of plant extract, along with 1.25ml of Folin-Phenol Ciocalteus reagent was mixed. The addition of 1.25ml of sodium carbonate was the last step. The total phenolics in leaf samples were anticipated …

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Total free amino acids:

Total free amino acids were determined using Hamilaton & Slyke (1943) method. 0.5g of plant material was ground in 10ml of phosphate buffer using pestle and mortar that was placed on an ice tub. The phosphate buffer used was made of two salts K2HPO4 and KH2PO4 and the buffer pH was maintained at 7.8. The …

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Total soluble protein contents:

The total soluble proteins were determined by following Bradford’s (1976) techniques. 0.5g of plant leaf was ground in 10ml of the buffer while the pestle and mortar were placed on an ice plate. The extract was subjected to centrifugation at 10,000rpm for 10 minutes, while the temperature was maintained at 4C. The supernatant after centrifugation …

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