PART 1. PLANT ANATOMY Y a oI BEL Pee eee: Pee Dy ype MyChapter 1. General structure of higher plants The angiosperms, which represent the most recently evolved group of plants, form the main part of the natural and cultivated vegetation on the earth. The general structures of a flower-bearing seed plant, starting with the seed, are outlined below. shoot opex Lg SSE leat primordia The seed contains an embryo plant, SS oxillary buds enveloped and f f= protected by a seed coat, and is supplied with a source of stored food. The plant embryo contains a minute axis with two poles—the root growing point and the shoot growing point. On the minute axis the cotyledons or seed leaves occur laterally. The food required for the germinating plantlet may be stored in the cotyledons or in a special tissue, the endosperm. Under suitable growing conditions the seed cortex —1 germinates and a young plant or seedling emerges. The seedling grows, extends its roots into the soil and its shoot ATH epidermis (stem and leaves) phloem root apex into the atmosphere. Wee cop The growth of the shoots and roots is due to the formation A The principal organs and tissues of the body of seed plants of new cells by B Cross section of stem C cross section of root internode —— node shoot ground line root branch roots meristematic (divided) tissues of the growing points, followed by growth and differentiation of these cells. When the plant attains adult size, flowers are formed. After pollination (transfer of pollen grains from the stamens to the style) and fertilization, fruit, containing seeds, develops, thus completing the life cycle. Some plants die shortly after seed set (annual plants), others (perennials) continue to grow for many years and become shrubs or trees.THE PLANT ORGANS The plant organs, as the organs of animals, are composed of tissues (groups of cells which carry on specific activities). The cells of plant tissues are small compartment possessing living material, the protoplasm enclosed by a cell wall. All metabolic processes take place in the cells. The roots anchor the plant in the soil, take up water and mineral salts from it, and in many cases store food. The shoot consists of stem and leaves. The leaves produce food by photosynthesis and give off water vapour by transpiration. The stem supports the leaves and has the role of conducting water and mineral salts from the roots to the leaves and synthesized organic substances from the leaves to regions of growth or storage. At the tips of the shoots and roots the apical meristems are situated. The cells that make up these meristems divide, grow, differentiate, and thus cause the extension growth of the plant. INTERNAL STRUCTURE OF THE YOUNG ROOT The epidermis is a protective tissue and consists of a single layer of densely packed cells. Below the epidermis occurs a relatively thick region, the cortex. The cortex is composed chiefly of structurally unspecialized cells, parenchyma cells, with large intercellular spaces. The innermost layer of the cortex is a single row of cells, the endodermis. The central region of the root is termed the vascular cylinder. It consists of the water-conducting tissue, the xylem, and the food-conducting tissue, the phloem. Between the vascular tissues (xylem and phloem) and the endodermis occurs a layer of unspecialized parenchyma cells, the pericycle, which originate from the same group of meristematic cells as the xylem and phloem. The pericycle which retains meristematic properties gives rise to lateral roots. INTERNAL STRUCTURE OF YOUNG SHOOT The zone between the epidermis and the vascular tissue is termed the cortex. Inside the cortex there is the vascular cylinder, which contains the vascular tissues. In the young stem portions the vascular tissues are generally organized in strands called vascular bundles. Each bundle contains xylem facing the centre of the stem and phloem outside. In the gymnosperms and dicotyledons the vascular bundles are arranged in a circle around a pith, which is usually parenchymatous, whereas in monocotyledons they are generally scattered throughout the stem cross section. INTERNAL STRUCTURE OF LEAVES The external and internal structure of the leaf is correlated to its role in photosynthesis (the synthesis of organic compounds using light as a source of the necessary energy) and transpiration (the loss of water as vapour). The leaf is flat and thin, thus enabling the solar rays to penetrate into all its cells. The high surface to volume ratio also enables successful gas exchange. The veins seen in the leaf blade (the expanded part of the leaf) contain vascular tissues. In the parenchymatous tissue (the mesophyll) occurring between the upper and lower epidermis of the leaf two zones can be distinguished: the upper—palisade parenchyma—consisting of elongated cells, and the lower—spongy parenchyma—consisting of irregularly shaped cells with large intercellular spaces. In the epidermis the stomata occur, which serve for gas exchange between the leaf tissues and the atmosphere.Chapter 2. The Plant Cell CELL AND CYCLOSIS Plants are composed of countless number of cells. Depending on its functions there are several types of cells in the plants. From the improved microtechnical and microscopical methods (e.g. light microscopy, transmission and scanning elektronmicroscopy) came our actual knowledge of cell composition. The eucaryotic cells (plants, fungi) have many distinc internal parts called organelles. The plant cells have some different features comparing with animal cells. The most importants are the cellulose containing cell wall, plastids and vacuoles. The main components of the ground tissue of plant organs are the parenchyma cells. These are structurally simple, and usually characterized by membrane isodiametric shape, thin extensible cell wall, a large central vacuole and a thin Dictyosome layer of peripheral cytoplasm. Parenchyma cells are the site of different metabolic _ activities and sometimes retain ability to become [= meristematic (divided) cells. A cell is not a static thing. Movements are going on within the cell all the time so long as it remains alive. The most conspicious of these movements is that referred to as streaming or cyclosis in which the organelles move in relation to one another in streams. This movement involves the mass movement of whole regions of the cytoplasm. This allows the movement of organelles, dissolved and colloidal elements of cytoplasm. There are two types of cyclosis: rotation - movement around a big, central vacuole, and circulation - movement around a lot of small vacuoles. The rotation can be observed e.g. in the leaf of Elodea sp. The circulation can be seen mainly in the hair-cells. /Nuclear. membrane. Nucleus ae * ek Chromatin yceslas The plant cell Object: Elodea canadensis (canadian pond weed) (or Vallisneria spiralis) leaf Method: Mount a leaf from close to the tip of an Elodea plant in a drop of water on a slide. Use the same water in which the plant is living (not distilled water!). Cover it with coverslip. Observation: Observe at first with the low-power objective (10x), then with the high- power objective (40x). The leaf consists of only two layers of cells in thickness, except the midrib. The cells are rectangular. Note the cell walls, the nucleus (large, gray body, round or elongate, near the cell wall), vacuoles (which are clear spaces containing the cell sap), cytoplasm (usually appears granular) and the green chloroplasts.In some cells the plastids are moving. They are being carried by streaming cytoplasm. The cells which show cyclosis best are long cells lying immediately over the midrib. The shock of detaching the leaf can cause the temporary cease of the protoplasmic movement, but at room temperature it should begin again within five minutes. Draw: Draw one Elodea leaf cell, label the parts of the cell and the direction of the movement with arrays. PLASMOLYSIS The plasmamembrane of the plant cell is differentially permeable. Water passes | cytoplasm through easily in both directions but the passage of many dissolved materials is restricted or prevented. Some kinds of molecules are held against a diffusion pressure gradient by active adsorption in a living cell. If a cell is immersed in a solution <=yewole that has a higher solute concentration than of that of the cell sap, water diffuses outward. Such loss of water by living cells is called plasmolysis. If plasmolyzed cells are immersed in a solution whose concentration is less than that of the cell sap, the cells regain their turgor, water molecules diffuse rN inward. The type of plasmolysis depending on the viscosity of the cytoplasm can be concave or convex. In the first case, the layer of cytoplasm remains in contact with the cell wall in several points, so that the vacuole, because of the loss of the fluids, forms concave indentations. If the layer of cytoplasm detaches itself from the cell wall completely, however, and the vacuole becomes round, this is convex plasmolysis. Concave plasmolysis, which can be maintained over long periods of time, occurs when cells are placed in a solution of bivalent kations (e.g. Ca”’), because these ions dehydrate the cytoplasm and thus make this firm. A solution of an univalent kation also gives rise to concave plasmolysis, but this soon turns into convex plasmolysis. This is because these ions increase the imbibition of the cytoplasm, which then assumes the smallest possible surface area. Object: Mnium sp. leaflet Method and observation: Mount a small leaflet of Mnium in a drop of water on a slide. Look at it with microscope. Place a drop of 10 % CaCl, solution at the edge of the coverslip, and draw it under by placing a folded edge of blotting paper at the opposite side of coverslip. Do not soak up all the salt solution. Wait five minutes then look at it. The cell content will now be seen to have shrunk away from contact with the cell wall. The space between the cytoplasm and cell wall is filled with the plasmolyzing solution in which the cells are immersed. Repeat it with KNO; solution (on another sample!). Lift the cover-glass and remount the cells in pure water. The cells can rapidly de-plasmolyze. Draw: The different plasmolysis types. nucleus, nucleolus: -+-—cell wall plasmalemma: chloroplasts tonoplast Diagram of a living cell from an Elodea leaf THE ANTHOCYANINS The anthocyanins are well-known red and blue pigments, which usually are dissolved in the cell-sap. They are flavan-derivatives. Apart from a few exceptions of uncertain nature they occur in plants as glycosides. Anthocyanin-aglykones (sugar-free anthocyanins) are called anthocyanidins.Object: Ligustrum vulgare fruit mesocarp cells Method: Tease out a small portion of the soft tissue of the fresh berry with needles into a drop of water on a slide. Cover it with cover-glass and tap on it gently with the point of needle to separate the cells. Add 0.1 N HCI solution to the cells (see the method in examination of plasmolysis) and observe the process of colour changing. Repeat it with 0.1 N NaOH solution! Draw: The mesocarp cells and label the color change. INULIN Inulin is a polysaccharide which occurs in the storage organs of many species. This is built of mainly fructose-units. Object: Dahlia variabilis root tuber Method: sections of tubers, which have been previously steeped for some days in 95 % ethanol. Mount in glycerine. Observation: The tissue consist of parenchyma cells. Here and there will be seen (particularly around vascular bundles) large rugged masses, with rounded outlines, formed of inulin, which is Sphaerocrystals of inulin in cells of dissolved in the cell sap during life but is a Dahlia tuber precipitated by concentrated alcohol in which it is insoluble. Draw: The inulin crystals in the cells. FATS, OILS These are a particularly important class of ergastic substances. Oils and fats are glycerides of certain organic acids. The fats being solid at ordinary temperature, the oils are liquids. These are the storage forms of lipids being present in every cell, at least in small amounts. They are found frequently in liquid form as oil droplets, but sometimes are stored in elaioplasts, a kind of storage plastids. Object: Ricinus communis (castor oil plant) seed endosperm Method: Make a thin section of the endosperm of the seed. Oils and fats may be identified by a reddish colour when they are treated with Sudan III. reagent. Draw: The section. Label the place of the oil-droplets, inside the cell. ‘TANNINS Tannins are a heterogenous group of phenol-derivatives widely distributed in the plant body. They appear as coarsely or finely granular masses or as bodies of various sizes colored yellow, red or brown. They are abundant in leaves, in vascular tissues, in unripe fruits, in seed coats. They occur in the cytoplasm and the vacuole and may impregnate the walls. They may be present in many cells of a tissue or in isolated cells (idioblasts). Object: Rosa canina (rose) stem Method: Identify the tannin idioblasts in the stem cross section of Rosa canina. Test for them with the reagents: 12% potassium-cromate (K2Cr207) solution or 3% ferric- chloride (FeCl) solution. Draw: Draw the cross section. Label the place of tannins and the colour of this with the different reagents.PLASTIDS The plastids are as distinctive a feature of the plant cell as the wall and the vacuole. The types of plastids are chloroplasts, which contain the green pigment chlorophyll, the chromoplast, which contain pigments other than chlorophyll, the colourless leucoplast, the various plastids that function in food storage (elaioplast, amiloplasts) and the immature proplastids. CHLOROPLASTS The source of energy available to organism living on Earth is light energy. This is transformed into chemical energy through photosynthesis. This conversion involves the synthesis of carbohydrate from carbon-dioxide (found in the atmosphere). The over-all process of photosynthesis can be written in chemical terms: 6 CO, + 6 H20 > CeHi20¢ + 6 Or This biological conversion of energy occurs in green plants. The site of photosynthesis in the plant cell is the chloroplast. The most distinctive chemical components of it are the pigments: chlorophylls and carotenoids. Light used in photosynthesis is absorbed by them. Object: chloroform extract of green leaves Method: Drop 25 l extract with micropipette 1.5 cm far from the bottom edge of the TLC (thin layer chromatography)-plate. Take it to the mixture of benzene:izo- propanol:water (10:1:1drop). Wait 45 minutes and look at it! Draw: The TLC-plate with the colour and place of the different pigments. The inner structure of chloroplast AMYLOPLASTS * 56 bast On These are plastids filled with starch. ki 9 Amyloplasts have specialized as organelles for the accumulation of starch in storage cells. Next to cellulose, starch is the most abundant carbohydrate in the plant world. During photosynthesis starch is formed in chloroplasts. Later it is hydrolized, transferred and resynthetized as storage strach in amyloplasts. Starch consist of two kinds of molecules: amylopetin and amylose. These both are built of glucose units joined by a.-1,4 linkages. (Cellulose also consists of glucose units, but these are B-linked.) Amylose is unbranched to give a spiral chain of 300-1000 units, but amylopectin has i Different types of starch grains: concentric also 1,6-linkages producing a (Triticum-a, Phaseolus-c), excentric (Solanum-b), branched molecule. simple, half-compound (Solanum), compound Starch grains are varied in shape. (Avena-e)Look at the types of starch grains in figure. Amyloplasts can be seen under polarisation microscope showing the shape of a maltese cross, the arms of which are separated by blue and yellow sectors. This arises because the starch molecules in these plastids are arranged in concentric layers. Objects: Solanum tuberosum (potato) tuber, Triticum aestivum (wheat) caryopsis, Phaseolus vulgaris (bean) seed, Zea mays (maize) caryopsis, Avena sativa (oat) caryopsis Method: Scrape a little material from a cut surface into a drop iodine dissolved in potassium-iodide (I-KJ) solution on a slide. Observation: Observe the shape and the type of the different starch grains. See the slides also with polarized light. Draw: The different types of grains. Try to decide the type! Label the colour! PROTEINOPLASTS - ALEURON GRAINS These are leucoplasts with proteins in them. Proteins are stored in the plants as solid protein bodies or aleurone grains, mainly in the seeds and fruits of many species. Protein bodies are enclosed in a unit membrane. Object: Ricinus communis (castor oil plant) seed endosperm Method: Make a thin section of the endosperm of the seed. Dissolve the fats in 95 % ethanol. Stain with iodine and mount in glycerine. Observation: The cells contain numerous aleurone grains, which are spherical bodies, each with a delicate mebrane on its own, enclosing a hexagonal crystalline body, the crystalloid, which is a protein, and a smaller round body, the globoid which consist of a double Aleurone grains in an : . endosperm cell of Ricinus phosphate of calcium and magnesium. These compound conuronie grains are special forms of reserve food vacuole sometimes found in oily seeds. Draw: The aleuron grains. SSOIGSOISE YQEOSES Object: Triticum vulgare (wheat) caryopsis Method: transverse section of the wheat grain, stained with iodine and mounted in glycerine. Observation: The central part of the wheat grain contains only strach as a reserve, but there is a layer of cubical cells, just under the protective coat of the grain, which stains yellow in iodine and contains protein grains. This is known as aleuron layer. Draw: The aleuron layer. CRYSTALS ‘ Crystals usually develop in vacuoles. They are often defined as excretory products, but possibly some of their elements (e.g. calcium) is recycled. Calcium-oxalate is most prominently represented among crystals in plants. These crystals assume several different forms. They appear as raphides (bundles of needles), elongated columnar crystals, prisms, druses (spheroidal aggregates of prismatic crystals) or as crystal sand. Appearance and location of crystals may be specific and useful in taxonomic classification. Crystals can be found sometimes in special crystal-containing cells: idioblasts. ’ Distinction can be taken between Ca(COO), and CaCO; crystals with the reaction with HCI or H2SO,.Reactions: Ca(COO),+ 2 HCl = CaCl2 + (COOH). Ca(COO) + H2SO4 = CaSO, + (COOH)2 CaCO ; + 2 HCI = CaCl, + H20 + CO2 Object: Allium cepa (onion) bulb epidermis, Agave americana (agave) leaf L.S., Vanilla planifolia (vanilla) leaf T.S., Opuntia sp. cladodium T.S., Ficus elastica (ficus) leaf T.S. Method: Make a section of the organ. Treat it with HCl and an other sample with H)SO4. NK Nat Allium cepa columnar Opuntia sp. rosetta are 1, " Ficus e. cystolith gave a. raphide The different types of the crystals CELL WALL A young cell is surrounded by a single wall called the primary wall. This is thin and elastic. As growth ceases, a secondary wall is laid down between the cytoplasm CNT Ame membrane and the primary wall. This is most often thick and : rigid. The primary walls of two cells are joined by a common layer, the middle lamella. This layer functions as an intercellular matrix, which holds groups of cells together. The middle lamella and the primary cell wall are composed of cellulose, hemicellulose and pectins. The microfibrils which consist of a lot of paralelly oriented cellulose chains The structure of thickened cell wall are embedded in a ground substance of cell wall, the matrix. Other carbohydrate polymers of noncellulosic character are found here. These are known collectively as hemicelluloses and pectin substances. In the secondary cell wall a new component, lignin, appears. Secondary wallChemical components of the cell wall: Cellulose: Cellulose is composed of B-glucose units, linked to each other through 1-4 linkages. A macromolecule comprises 3000-10000 units. Object: Gossypium hirsutum (cotton) Method: Mount some hair in a drop of water on a slide. Treat it with Schweitzer’s- reagent (Cu(OH)2 in concentrated NH,OH). Observation: The cellulose is dissolved in Schweitzer’s-reagent. Draw: The hairs before and after the reaction. Hemicelluloses: These are part of the matrix, mainly polymers of pentoses situated between the cellulose microfibrils. Pectins: Pectin substances are macromolecules that consist of essentially galacturonic acid units linked by a-glycosidic 1,4 linkages. In each case several 4 cotton hair before and after the hundred units are involved. treatment with Schweitzer- Lignin: The woody material, lignin, is a very reagent important structural substance of plants and is universally distributed in the plant kingdom from the mosses upward. In vascular plants it is usually found in the xylem , the individual elements of which have cell walls incrustated with lignin. It is a highly polymeric substance in which phenyl-propane units are linked to form a three-dimensional network. The substitution patterns of p-coumaric- acid, ferulic-acid and sinapinic-acid can be recognized in this phenyl-propane residues. However, it is not the acids but the corresponding alcohols, which are incorporated into lignin. The relative amount of these three components can vary greatly, depending on the nature and age of the plant. Object: Rosa canina (rose) stem Method: Make some cross section of the rose stem. Treat it with one drop alcoholic floroglucin and one drop cc. HCI (Wiesner-reaction). Observation: The cell wall containing lignin become cherry-red. Draw: The section. Label the different part. Pits Certain portions of the cell wall remain thin even as the secondary wall is formed and these parts consist of primary wall material. These areas, which are of variable in shape, are called pits. The pits are areas through which substances pass from cell to cell. Two principal types are recognized: simple pits and bordered pits. Bordered pits are found in cells which assist the transport of water. The structure of a bordered pit 10THICKENINGS The secondary cell wall remains thick in different parts of the plant cells. CENTRIFUGAL pollen grains, spores oT CENTRIPETAL equal thickenings unequal thickenings - the whole cell wall eg. scalariform, | e.g. pitted, helical, annular reticulated Pyrus achras (pear) stone | Ricinus communis (castor | Pinus silvestris cells oil plant) macerated stem | (pine) wood radial section - a part of the cell wall endodermis, collenchyma Centrifugal cell wall thickenings parenchyma cells a 4 yp VAN" ya QB = Z = ; = == war Different types of centripetal cell wall thickeningsChapter 3. The Plant Tissues The term tissue denotes a group of cells of a similar kind and/or with similar functions. In the case of flowering plants, we distinguish between two main groups of tissues: meristematic and mature tissue. MERISTEMATIC TISSUES Meristems are tissues in which cells actively divide. According to the position they are divided to apical meristems, intercalary meristems (which are found between mature tissues) and lateral meristems (which are situated parallelly to the circumference of the organ in which they are found). According to the origin they are divided in to primary meristems (the cells of which develop directly from the embryonic cells) and secondary meristems (developing from mature tissues). Apical meristems are found at the tips of the roots and shoots. They form the three primary meristematic tissues (primary meristems): protoderm (differentiating into epidermis), ground meristem (differentiating into pith and cortex), and procambium (differentiating into vascular tissues - xylem and phloem). Object: Allium cepa (onion) root apex T.S. (cross section) and L.S. (longitudinal section) Observation, draw: Draw the sections and label the meristematic tissues and the different parts of the root tip. low-power plan showing tissue arrangement. x 10 region of elongation mel prominent nucleolus growing point or apical eristeen | — Notes: 1. The cells are small and cubical with large, prominent nuclei, and they are arranged in vertical rows 2. Seven stages of cell division, numbered in sequence. are pointed out. Stage 7 shows two young daughter cells. the result of cell division The regions of the root tip root cap See al Note: Outer celts at A breaking down ate replaced by those at 8. which have been produced by the apical meristem. This is a continuous process 12GROUND TISSUES PARENCHYMA The most abundant tissue in almost all major parts of higher plants. The cells are more or less spherical. The parenchyma cells often store different types of ergastic substances, e.g. starch, fatty oil or protein crystals. Object:Ficaria verna (lesser celandine) root bulbs T.S. Observation, draw: Draw the reserving starch in the parenchymatous tissue. vascular cylinder Ficaria verna bulb T.S. reserving parenchyma CHLORENCHYMA These parenchyma cells contain chloroplasts. AERENCHYMA. This is a type of parenchyma with extensive connected air spaces. Object: Nymphaea alba (white water-lily) petiole T.S. Observation, draw: Draw the aerenchyma. SECRETORY TISSUES Mature parenchymatous tissues may have a well-developed system of intercellular spaces. The development of these is either schizogenous or lysigenous. Schizogenous spaces are formed by splitting apart of the cell walls at the middle lamella of the contiguous cells. In schizogenous glands a ring or lining of intact cells, the epithelium, surrounds the cavity. Lysigenous spaces are formed by the disintegration (lysis) of entire cells. These cavities can store biologically active substances. Objects: Citrus limon (lemon) fruit, Pinus silvestris (pine) wood T.S., Coriandrum sativum (coriander) stem T.S. Observation, draw: Lysigenous oil cavities of Citrus, schizogenous intercellul of Coriandrum stem, and schizogenous resin ducts with epithelium of Pinus. COC woodray — tracheids resinduct wood parenchyma, I Se ), GID 5 SJ ) = 3 ik < globule of resin secretory cells A Lysigenous oil-cavity of Citrus B Schizogenous resin duct of a pine 13 = We ars ———— MI MIANIMININAI ATASUPPORTING TISSUES OF THE PLANTS Plants owe teir stability partly to the turgor pressure in the parenchyma cells. Flowering plants, however, possess special tissues which provide further rigidity. These tissues are made up either of living cells (collenchyma) or of cells that are already dead (sclerenchyma). Stone cells lignified wall primory cell wall and middle lomella ~amellar collenchyma Different types of supporting tissues COLLENCHYMA _Collenchyma cells have living protoplasts and partly thickened cell wall. The thickened cell walls provide the necessary rigidity, while the unthickened walls allow the exchange of material. The size and the shape varies. There are several types depending on the position of cell wall thickening (.g. angular, lamellar, lacunar). When the thickening is concentrated at the corners of the longitudinal walls and the middle of each wall remains unthickened, we refer to angular collenchyma. In the case of lamellar collenchyma, the cell walls running parallel to the surface of the organ are thickened, while those perpendicular to it appear thin. If intercellular spaces occur between the walll thickenings, the tissue is referred to as lacunate. There are transitory stages between various types of collenchyma. SCLERENCHYMA. Sclerenchyma cells collenchynna have thick walls, normally impregnated with lignin. hlorenchyma The lignin increases the strength of the cell wall, but reduces its elasticity and permeability to water. These are usually dead sclerenchyma cells consisting either of isodiametric stone cells or of long — sclerenchyma fibers. patency Object: Daucus carota (carrot) stem T.S. Observation, draw: The T.S., label the parenchyma, collenchyma, Different types of ground tissues in Daucus c. stem T.S. _ chlorenchyma. ~ 14