11 PUC Biology lab manual a EXERCISE - 4: stan Aim: Staining of nucteic acid by acey carmine ora Principle: Acctocarmine or acetogreer iio acetoorcein, acid present in nucleic and present 1, 21 CA? the nucl ed conjugate nucleus to from deep Pomumements! “Onion bulb) -Acetocarmin«), blotting paper, slide, cover slips brush, necagnctooreein, blade and microscope ae Hed Procedure : + Peel off epidermis from the fleshy scaly leaf of an onion bulb and place it on a slide + Add a few drops of water with the help of a brush to avoid drying. + Take out a small piece out of the peel + Remove the water present in the peel with the help of blotting paper. * Add 2 drops of acetocarmine / acetoorcein oh the peel and heat gently on a spirit lamp. + Place a coverslip over the peel avoiding air bubbles and temove excess stain with the help of blotting paper. + Observe the cells under the microscope. Observation : Observe the shape of cell : Number of nuclei in acell: Position of nuclei in the cell: = les, scissors/ ABM tional Publications — Cytopiasm NUCLEAR STAINED PLANT CELLSlogy lab manual a EXERCISE - 3 ; im: To study polle: Aim: pollen tube growth on st ares igma ina pistil iple: Pollen ruibe as the outer othe, Sigma germ through the germ po er growth of intine. Poll: ed and produce ens Bern pore present in the exi lien tube comes out tissues of style to reach the ovule «xine and passes through the Requirements: Excised st tyles with stiema of (Retina et a2e/masize/ounilowsr/ eee suitable plant. Beaker, water, slides, coverslips, brush and acedle Cot finger), stain and microscope and needle, Cotton blue Procedure : + Take 2 beaker with water and boil, Place the cut styles with stigma in boiling water for 5-10 minutes to soften the tissues Take out a stigma and place on slide containing drop of water. Make an incision lengthwise through the style and stigma. Put a drop of cotton blue over it and allow to stand for 3-5 minutes . Wash the excess stain away with water or blotting paper. Add a drop of glycerin / water and place a cover slip on it. ji Press the cover slip gently with a needle. . If fail to observe pollen tubes mount another stigma. wing through the tissues of style and tubular mo’ ABM Educational Publications ne + POLLEN TUBE GROWTH ON STIGMA i ; LONGITUDINAL SECTION OF A FLOWER ‘SHOWING GROWTH OF POLLEN TUBE,oxy lab manual tb manuat —— Exine—— —__ Ungerininated a > pollen grains Vacucle— Generative cell — Germinated — Tube (vegetative) Pollen Grains intine Mature pollen grain cell Pollen tube — Pollen tube Male gamets < tube nucleus —}) Microscopic field sO GERMINATION OF POLLEN GRAINS Number of Total number of Total number of pollen | % of pollen germination observation pollen - N germinated - n = x 100 ollen | 1 25 15 48 x 100 = 60 | 2 | 3 | Average i tl ination = nX100/N.. Percentage of pollen germ:°) Three chambered ovary a) Tetralocular chambered ovat |] I] I e) Pentalocular : Five c I] eg: Hib angement of ovules s of the following types. || 5) Placentation : It |] with ir ary, i a) Marginal placentation - Placenta bearing he margin on placenta. I] ovules develop near |] eg:pe b) Axil placentation - Ovules arises from the | central axis of the ovary, eg: Hibiscu: || ¢) Parietal placentation - Ovules are arises ||" from the inner wall of the ovary, eg - Must || d) Free central placentation - ovules are ses from the center of the e) Basal placentation - Ovules are arises from the unilocular ovary, ¢g the base of sunflow "TS OF OVARY SHOWING LOCULES Y/—Pracen 0 Free central placentation Basal placentation TS. OF OVARY SHOWING PLACENTATION arginal placentation|H PUC Biology lab manag EXERCISE - 2 : POLLEN GERMINATION A Aim: Tc he percentage of polle la Requirements: Mature pollen s crose solution (10 08 c y Petridish, slide er slips, bru and ‘ Procedure: 1, Take a drop of 10% sucros ear slip an ene! e 2 ce the cover 3. After 10 minutes observe the sli 1s 4. Count total number of polle: tk punt the of pi grains that have germina\ Observation ) and caleulate the percentage of pollen grains germinat Tabuiate the observation (3 microscopic fields (a) Name of the plant used as pollen source : Vinca (p) Number of pollen grains ina microscopic field = N (c) Number of germinated pollen grains in the same field=n (a) percentage of pollen germination = nx 100/N %11 PUC Biology lab manual a EXERCISE - 4: stan Aim: Staining of nucteic acid by acey carmine ora Principle: Acctocarmine or acetogreer iio acetoorcein, acid present in nucleic and present 1, 21 CA? the nucl ed conjugate nucleus to from deep Pomumements! “Onion bulb) -Acetocarmin«), blotting paper, slide, cover slips brush, necagnctooreein, blade and microscope ae Hed Procedure : + Peel off epidermis from the fleshy scaly leaf of an onion bulb and place it on a slide + Add a few drops of water with the help of a brush to avoid drying. + Take out a small piece out of the peel + Remove the water present in the peel with the help of blotting paper. * Add 2 drops of acetocarmine / acetoorcein oh the peel and heat gently on a spirit lamp. + Place a coverslip over the peel avoiding air bubbles and temove excess stain with the help of blotting paper. + Observe the cells under the microscope. Observation : Observe the shape of cell : Number of nuclei in acell: Position of nuclei in the cell: = les, scissors/ ABM tional Publications — Cytopiasm NUCLEAR STAINED PLANT CELLSlogy lab manual a EXERCISE - 3 ; im: To study polle: Aim: pollen tube growth on st ares igma ina pistil iple: Pollen ruibe as the outer othe, Sigma germ through the germ po er growth of intine. Poll: ed and produce ens Bern pore present in the exi lien tube comes out tissues of style to reach the ovule «xine and passes through the Requirements: Excised st tyles with stiema of (Retina et a2e/masize/ounilowsr/ eee suitable plant. Beaker, water, slides, coverslips, brush and acedle Cot finger), stain and microscope and needle, Cotton blue Procedure : + Take 2 beaker with water and boil, Place the cut styles with stigma in boiling water for 5-10 minutes to soften the tissues Take out a stigma and place on slide containing drop of water. Make an incision lengthwise through the style and stigma. Put a drop of cotton blue over it and allow to stand for 3-5 minutes . Wash the excess stain away with water or blotting paper. Add a drop of glycerin / water and place a cover slip on it. ji Press the cover slip gently with a needle. . If fail to observe pollen tubes mount another stigma. wing through the tissues of style and tubular mo’ ABM Educational Publications ne + POLLEN TUBE GROWTH ON STIGMA i ; LONGITUDINAL SECTION OF A FLOWER ‘SHOWING GROWTH OF POLLEN TUBE,oxy lab manual tb manuat —— Exine—— —__ Ungerininated a > pollen grains Vacucle— Generative cell — Germinated — Tube (vegetative) Pollen Grains intine Mature pollen grain cell Pollen tube — Pollen tube Male gamets < tube nucleus —}) Microscopic field sO GERMINATION OF POLLEN GRAINS Number of Total number of Total number of pollen | % of pollen germination observation pollen - N germinated - n = x 100 ollen | 1 25 15 48 x 100 = 60 | 2 | 3 | Average i tl ination = nX100/N.. Percentage of pollen germ:H PUC Biology lab manag EXERCISE - 2 : POLLEN GERMINATION A Aim: Tc he percentage of polle la Requirements: Mature pollen s crose solution (10 08 c y Petridish, slide er slips, bru and ‘ Procedure: 1, Take a drop of 10% sucros ear slip an ene! e 2 ce the cover 3. After 10 minutes observe the sli 1s 4. Count total number of polle: tk punt the of pi grains that have germina\ Observation ) and caleulate the percentage of pollen grains germinat Tabuiate the observation (3 microscopic fields (a) Name of the plant used as pollen source : Vinca (p) Number of pollen grains ina microscopic field = N (c) Number of germinated pollen grains in the same field=n (a) percentage of pollen germination = nx 100/N %°) Three chambered ovary a) Tetralocular chambered ovat |] I] I e) Pentalocular : Five c I] eg: Hib angement of ovules s of the following types. || 5) Placentation : It |] with ir ary, i a) Marginal placentation - Placenta bearing he margin on placenta. I] ovules develop near |] eg:pe b) Axil placentation - Ovules arises from the | central axis of the ovary, eg: Hibiscu: || ¢) Parietal placentation - Ovules are arises ||" from the inner wall of the ovary, eg - Must || d) Free central placentation - ovules are ses from the center of the e) Basal placentation - Ovules are arises from the unilocular ovary, ¢g the base of sunflow "TS OF OVARY SHOWING LOCULES Y/—Pracen 0 Free central placentation Basal placentation TS. OF OVARY SHOWING PLACENTATION arginal placentation|EXE) Svat) Abia aati a RCISE - 1: 1: REPRODUCTIVE PARTS OF A FLOWER commonly udy the rep: eproductive par fe site of sexual reproduction Corolla, androecium and Whorls are Androecium and ? Commonly available Rowers (Hibiscus Requirements : Lathyrus, Crotaloria, Pea, Onion, Helianthus, Cucumi dissect roscupe, hand lens, needles, scalpel, Procedure : 1) Observe the typical flower and identify androecium sol ee ae 2) Carefully take out stamens and carpels. \ 3) Take a thin T/S of ovary and mount the section on a \ slide and examine under desection microscope, record Agee We wpa the number of locules and type of placentation. ~ ovary 4) Locules are the chambers of the ovary containing \ ovules. f- rete STAMEN pISTI, \ 4