Practical 1: Transverse Section of Nuxvomica Seed, Cinchona Bark and Rauwoflia Root.

Aim: a) To observe the transverse sections of Nuxvomica seed, Cinchona bark and Rauwolfia root b) To observe the longitudinal section of Cinchona bark under a microscope through staining chemical tests.

Apparatus and materials: Watch glass, knife, microscope, microscope glass, brush, nuxvomica seed, cinchona bark, rauwolfia root, nuxvomica powder, cinchona powder, distilled water, phloroglucinol solution, conc. HCl, dilute iodine solution, conc. sulphuric acid, alcoholic picric acid solution, Sudan red-III solution, dilute sodium hydroxide, dilute sulphuric acid, and nitric acid.

Procedures: 1. Nuxvomica seed was cut into small and thin section before they were placed in a watch glass. 2. The thin section was transferred onto a microscopic glass using a brush and the reagents that stated in the observation table below were added. 3. The transverse section was observed through the microscope and the observation was recorded. 4. Steps 1-3 were repeated on Cinchona bark and Rauwolfia root.

Observations: Transverse section of Nuxvomica seed No Reagents 1 1 drop of Phloroglucinol and 1 drop of conc. HCl Observation under microscope Characteristics Lignified epidermal trichomes

1 drop of dilute iodine solution and conc. Sulphuric acid is

Hemicelluloseendospermic wall (plasmodesmata)

Alcoholic picric acid solution

Aleurone grains present in the cells of endosperm

Sudan red-III solution

Oil globules present in the cells of endosperm

Nuxvomica powder 5 3-4 drops of phloroglucinol and 3-4 drops of conc. HCl (1:1) Trichomes: entire or fragments of lignified trichomes Endosperm: abundant fragments of endosperm cells whose walls are thickened

Transverse section of cinchona bark No Reagents 1 1 drop of dilute sodium hydroxide solution, warmed gently then washed with water and 1 drop of phloroglucinol and conc. HCl are added 2 1 drop of iodine solution Starch grains are observed Observation under microscope Characteristics Lignified phloem fibers

Dilute sulphuric acid solution

Calcium oxalate crystals

Cinchona powder 4 Dilute sodium hydroxide solution, 3-4 drops of phloroglucinol and 3-4 drops of conc. HCl Fibers: phloem fibers or blast fibers, numerous entire or in fragments, spindle shaped, yellowish, thick walled, strongly lignified Cork: typical thin

walled cork cells which appear reddish brown Starch grains: minute, both simple and compound

Longitudinal section of cinchona bark No Reagents 1 1 drop of dilute sodium hydroxide solution, warmed gently then washed with water and 1 drop of phloroglucinol and conc. HCl are added 2 1 drop of iodine solution Starch grains are observed Observation under microscope Characteristics Lignified phloem fibers

Dilute sulphuric acid solution

Calcium oxalate crystals

Transverse section of rauwolfia root No Reagents Observation under microscope Characteristics

1 drop of phloroglucinol and conc. HCl (1:1)

Lignified cork cells, xylem region and lower part of medullary rays.

1 drop of iodine solution

Starch grains are observed

Dilute sulphuric acid solution

Calcium oxalate crystals

2 drops of nitric acid and 1 drop of water

Presence of reserpine

Conclusion: It can be concluded that, for nuxvomica seed, the microscopic characteristics were observed under the microscope as followed:Test 1 (powder): Oil globules and lignified trichomes which is pink colour. Test 2: Hemicellulose endospermic wall which is red colour (probably due to iodine solution) Test 3: Aleurone grains which is yellow colour. Test 4: Oil globules which is red colour. For cinchona bark, the following microscopic characteristics were observed:Test 1: elongated form of pink fibers for longitudinal section, circular form of pink fibers for transverse section. Test 2: Starch grain which is blue in colour. Test 3: Calcium oxalate crystals and starch grain. Test 4: Cork cells, fibers, starch grain. For rauwolfia root, the following characteristics were observed under the microscope:Test 1: Xylem vessel and medullary rays and is in pink colour. Test 2: pith region and starch grain and red in colour( probably due to iodine solution). Test 3: calcium oxalate crystals. Test 4: medullary ray in slightly pink colour.(presence of reserpine).