ONE PAPER CHROMED ceAtAY LAB — PRINCIPLES IMPORTANCE OF PHOTOSYNTHESIS | Photosynthesis is the process of trapping light ‘energy with the aid of chlorophyll to combine ‘water and carbon dioxide to form carbohydrate. ‘The importance of photosynthesis to life on ‘earth is great. Photosynthesis 1. provides the energy, stored in the form of chemical bonds, in food, i 2. is responsible for the energy stored in coal and petroleum, ri 3. largely provides the oxygen in the ar. ‘The scale on which photosynthesis functions is enormous. In one year, it has been estimated that photosynthesis = = takes in 128 billion tonnes of carbon dioxide = takes in 52 billion tonnes of water ~ gives off $2 billion tonnes of oxygen = manufactures 87 billion tonnes of carbohy- drate ‘All the green parts of a plant cary out photo- symivess in daylight hours, but leaves ae the | principal photosynthetic organs. ‘The Role of Starch Starch is an important storage material in plants. Starch is made up of many glucose molecules joined together. During periods when light energy is insuf cient, such as at night, and during the long win- ters in many parts of Canada, plants can stay alive by using the energy stored in their starch molecules. By converting starch to a 3-carbon compound called phosphoglyceraldehyde, the plant obtains the energy released in the con- version reaction, Photosynthesis — A Laboratory View For many years, the commonly stated equation for photosynthesis has been: Take water + caibon desde —"®™\—y cartohydate + one Horo” “gucore! Chloroplast: Site of Photosynthesis The reactions of photosyihess tke place within the celular organelle called the chloroplast Each leat cell usualy contains about forty chiovo- plass. In certain plans, there may be as many as 500 000/mm* of leaf surface. Under the light microscope, the chloroplast is oval in shape and green in colour. Under the electron microscope, however, chloroplasts can be seen to be packed with ‘membranes, called lamellae. These membranes are folded over in places to form stacks of coin- like structures. These stacks are visible under the light microscope as “tlle green specks.”” The early microscope workers called these "specks" grains or grana. They are still called this today. Each “coin” in the stack is called a thylakoid. Chlorophyll Photosynthesis starts wit the absorption of light by chlorophy/l. Chlorophyll s a green pigment found in the grana of the chloroplast. Why Are Most Leaves Green Under White Light? Ordinarily visible light consists of a mixture of different wavelengths — red, orange, yellow, green, blue, and violet. These light waves con- tain variable amounts of energy. Red light is the longest wavelength that can be seen by humans and contains the least energy; violet is the short- est that can be seen by us and contains the most energy. The graph en page 2 indicates what hap- pens when ordinary “white” light falls on a chlorophyll molecule: Investigating Other Photosynthetic Pigments Light is captured in chloroplasts by groups of pigments. Different pigments absorb light en- ergy at different wave lengths. A, There are several different kinds of chloro- phylls. Two of the most common chlorophylls are chlorophyll a and chlorophyll b. All photosynthetic cells contain chlorophyll a, Most photosynthetic cells also contain a sec- ‘ond chlorophyll (chlorophyll b) and a repre- sentative of another group of pigments called the carotenoids,Oo 2m 40 6 8 (percent of maximum absorption) GEM 1 - ABOAITON SrecrtuM of VISIBLE LIGMT AY CHLOROPHYLL ENRICHMENT THE CHLOROPHYLL MOLECULE. The key molecule involved in the’ process of photosynthesis is the green pigment chlorophyll. ‘The structure of chlorophyll is similar to hemo- globin in red blood cells and the cytochrome mol- ecules involved in cellular respiration. Instead of having a central atom of iron, however, as in hemoglobin and cytochrome, the chlorophyll has a central atom of magnesium. octrome ——_Chorophy Chlorophyll b differs from chlorophyll a in that it replaces a CH, group with a CHO group (at the location indicated). Chiorophyll (CathnO.NMg) +One of the carotenoids inthe higher plans i plants is | beta-carotene. When a beta-carotene molecule | is spit in two, it forms two molecules of vitamin THREE The oxidation of vitamin A produces a pig- ' ‘ment called retinene, Retinene is the pigment involved in vision. ste o-daci-cie Hy CH CH, ce cH, - Vitamin k= > x j cidact-ci chor oe ow ry : met bor oror oer oi (Sy 4 re Cy, oH, = cmc on = CH, ‘The red, blue, lavender, and purple colours of various plant parts are due to water-soluble pigments called the anthocyanins. ‘Anthocyanins are found dissolved in the cy- toplasm and are never present in plastids. These pigments occur commonly in the petals of flow- ers. ‘Any food present moves into the body of the tree for storage. All that remains in the cell cavities of the leaf is a few oil globules and erystals, and fa small number of yellow structures. It is these yellow structures that give the leaf its yellow col- oar. FALL COLOUR Yellow Golden & Bronze Orange-yellow Golden-yellow Yellow, Orange, Red ‘Orange, Red Red Yellow, Purple Purple-red to Violet-purple Deep Purple: \f more sugar is in the leaf than can be transferred to the body of the tree, the accumulation of sugar results in the production of red pigments (called anthocyanins). The colours produced depend on ‘weather conditions. The best colour condition is @ warm, sunny day (for sugar production) followed by a coo! night temperature (below 7°C). This pre~ ‘vents sugars from being transported down to the branches and roots. ‘The shades of fall colour vary with the tree in- volved: TREE ‘Ginkgo or maidenhair (Ginkgo biloba) Tulip tree (Liriodendron tulipifera) Norway maple (Acer platanoides) |White birch (Betula papyrifera) Linden (Tilia cordata) American beech (Fagus grandi- fala [European beech (Fagus sylvatica) Yellow wood (Cladass lutea) [Eastern redbud (Ceris canadensis) ‘Sugar maple (Acer saccharum) Sumac (Rhus typhina) Red Oak (Quercus borealis) Red maple (Acer rubrum) Black gum (Nyssa sylvatica) Sree ‘gum (Liquidambar styraci- ua) Pin oak (Quercus Palustris) Flowering dogwood (Cornus flor- ial Hawthorn (Crataegus phaenopy- um) [Mountain ash (Sorbus aucuparia) [Green ash (Fraxinus pennsylvan- ica) White oak (Quercus aba) | white ash (Fraxinus american) _c Glossary of NeW TEAMS Four js — putting together): the process by which ‘Photosynthesis (rom Greek phos — light and synthesis — ul mthesize carbohy- chlorophyll-containing cells in green plants use the energy of light to synthesize carbohy- rates from carbon dioxide and water. - 1 a chlorophyll-bearing organelle chloroplast (from Greek chloros — green and plastos — formed): a chlorophyl-bearing orga Aamg athe cytoplasm of many plans, lamellae throm Latin lamina — plate): layers; thin sheets. ; ‘grana (from Latin granum — seed): a structure within the chloroplast containing the photosyn- thetic pigments, s thylakoid throm Greek thylaco — sac or pouch): a saclike membranous structure found in the chloroplast. | 5 chlorophyll (from Greek chloros — green and phyllon — leaf): the green pigment of plant cells. «carotenoids (rom Latin carota — carrot and Greek eidos — likeness): a group of pigments that includes the carotenes (yellows, oranges) and the xanthophylls (yellow); accessory pigments in photosynthesis of various plant pars. fetinene (from Latin retina — light sensitive membrane lining the inner eyeball and the Greek ‘ene — having a double bond): pigment involved in vision. anthocyanins (om Greek anthos — flower and kyanos — dark blue): a group of pigments found in various plant parts. plastids (ftom Greek plastos — formed): relatively large bodies found in the cytoplasm of plant cells that function in photosynthesis, food storage and the lke. flammable (from Latin flamma — flame): capable of burning with great rapidity; easily ignited. toxic (rom Latin toxicum — poison): poisonous. pulverized (rom Latin pulvis — dust): to crush to a powder or dust. extract (from Latin extrahere — to draw out: to obtain by chemical ot mechanical action. fluorescence {from Latin fluere — to flow and escence — becoming: the emission of electro- ‘magnetic radiation resulting from the absorption of radiation and lasting only as long as the stimulating radiation is continued, Chromatography (irom Greek Khroma — colour and graphein — write): the analysis of a sub- stance by sending it in liquid form slowly through some material such as paper. The sub- >) stance is broken into its constituent parts which are distinguished by different colours or similar colours found at different locations on the paper. ‘migration (from Latin migrare — to move): the act of moving from one place to another. solvent (fom Latin solvere — to loosen): a liquid capable of dissolving another substance, legend ({rom Latin legere — to read): an unauthenticated story from early times, preserved by tradition.Five @ yellow 0 a 6 80 {percent of maximum absorption) GRAEAAL —- ABSUETION SPECTUM OF VISIBLE LIGHT AM AN wknow Nl FAMILA OF PIOMENTS . 2 0 o & (percent of maximum absorption) GAAP HD WODRETON SPECTRUM OF VISGLE LIGHT PA THE CAROTENOIDSLABORATORY ACTIVITY | To Extract Various Photosynthetic i Pigments 1. Obtain 5 g spinach leaves and remove the leaves’ midribs. Tear the leaves into small, confett-size pieces. Put the leaf tearings into a clean mortar. 2. Add a pinch of sand to the leaf tearings. 3. Add 5 ml of acetone and grind the leaf tear- ings for several minutes. Caution: Acetone is extremely flammable and | toxic. Avoid breathing the vapours. Do not use a source of flame. 4, Filter the pulverized leaf tissues and acetone solution into a test tube, The solution in the test tube is the chlorophyll extract. 5. Examine the chlorophyll extract in direct light. ENRICHMENT FLUORESCENCE Dissolved chlorophyll is green in colour since the chlorophyll absorbs all but the green wavelengths of light. When the green solution is illuminated by bright light, the colour changes to red. This phe- ‘nomenon is called fluorescence. tis caused by the absorption and retransmission of light by chloro- phyll, When the light “excites” the chlorophyll | molecules, high-energy electrons become dis- lodged and the chlorophyll acquires a positive charge. | Some of the high-energy electrons return to the chlorophyll immediately. The “extra energy" is ven off as heat or light (at the wave length cor- responding to red). yaLABORATORY ACTIVITY ©: To Identify Photosynthetic Pigments A technique known as paper chromatography can be used to separate mixtures of chemical substances. These substances are separated on the basis oftheir different rates of migration with a flowing solvent on special paper. Inthis activity four pigments will be detected. The four pigments and their colours are: chlorophyll a . blue-green chlorophyll b . yellow-green carotenes .. orange-red xanthophylls -selight yellow 1. Cut a strip of chromatography paper long enough to reach almost the length of a large test tube. Handle the paper with forceps at the edges only. Why? 2, Insert a bent paper clip into the bottom of a cork that fits the mouth ofthe large tet tube. Suspend the paper strip on the hook and test the length and fit before proceeding further. Taper the bottom edge of the paper so that the point nearly touches the bottom of the test tube. Be absolutely sure that the strip hangs straight and does not touch the sides. Your setup should look as follows: 3. Remove the paper from the test tube. 4, Using a micropipette, capillary pipette or dropper, place as small a drop as possible of the chlorophyll extract 3 to 5 mm from the pointed end. Let the spot dry. Reapply the extract at least three times. 5, Place sufficient solvent (9 parts petroleum ‘ether and 1 part acetone) into the test tube toreach the tip ofthe paper, but not enough | to touch the pigment spot, : Caution: Acetone and petroleum ether are ex- ‘tremely flammable, Do ot use a source of flame. ' ‘Avoid breathing the vapours, SPuEW 6. Insert the strip of paper back into the test tube, Fit the cork tightly into the tube. Let the tube stand vertically in a test tube rack. 7. The solvent will move up the paper strip car- tying successively dissolved pigments in so- lution. The most soluble pigment will ravel farthest. When the solvent front is within 1 cm of the top of the strip, remove the strip ‘and mark the position of the front, Let the strip dry. 8. Mark the centre of the original dot of chlo- rophyll extract. Mark the centre of each pig- ment. Measure the distance travelled by each pigment. Measure the total distance travelled by the solvent front. Calculate the migration rate (R, for each pigment. The migration rate (Ry) of each pigment can bbe calculated using the following formula: p, = sistance pigment travelled " ™ ‘distance solvent travelled 9. Make a copy of the following table and fill in the information. : ENRICHMENT FALL COLOURS Many of our native peoples possess a legend re- garding the fall colours. The legend tells how the fall colours were created by “*hunters-in-the-sky"” ‘who killed the ‘Great Bear."” The blood dripping from the wounds onto the forests changed many tees to red. Other tees were tured yellow by the fat that splattered from the Keitle as the hunters cooked the meat. ‘Actually, the fall colour change stems from chemical processes which take place in the tree during its preparation for winter. Within the ces, the chlorophy/l is broken down into various com- ponents.LENE MEMENG EXTRACTION RES Dy Puartst(i) REF = BL guement@) RE = oe = 40 memen(S) Rs me = FE ae women ruecment (4) REF dee = 4 INA. Del OF QUVERT (AVERT FA) ORIN Lene UGE = GERANWM . ,bik, | avarosrnrwes vn va OL peewe eHomsyi-THEsis ; Le IN WHAT CELUULAL ORGANELLE 10% PHOTSMNTHESIS OCCA: DECLINE 1% RPeReANCE 4. GWE 9 Whortinur Une of PHOTOSNTESS. Yo IF UEAVA FRE We HAPINN PHOTOCHMTHETIC. 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