5. Move several sections into the second well that contains a few drops of Toluidine Blue.
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1 HAIGLER PROTOCOL FOR VIEWING CROSS SECTIONS OF PLANTS Materials (per student team): Plants Double-edged razor blades (split into two as demonstrated by the teacher) Masking tape for razor blade handle Spot Plate Carrot slices Knife Paint brush (small with some bristles removed) Bottle of distilled water Glass slides Microscope Toluidine Blue Stain Procedure: 1. Collect plant material to be observed, immersing cut stems in water. 2. Place a small section of the stem of a plant on a glass cutting plate or slide. Using a very sharp side of a double-edged razor blade, cut at a slight slant and take as thin a section as possible (0.5 1 mm). See Diagram For softer tissue, make a small slit in a slice of carrot to hold a small piece of plant stem. Insert the plant tissue in the carrot. Using a very sharp razor blade, take a very thin section of the plant at an angle. Attention: Keep the razor wet at all times by dipping it in water before each cut. 4. Immerse the sections in distilled water in the first well of the spot plate. See Diagram II. Student 1 Water TB Water Water Diagram II 1
2 5. Move several sections into the second well that contains a few drops of Toluidine Blue. 6. After 30 seconds, move the tissue to the third well to rinse and follow with the fourth well for a final rinse. Hint: Times may need to be adjusted slightly for different plants. 7. With a paintbrush, lift the section out of water and place it, fiat side down, on the center of the slide. Never use forceps to handle the sections. The water surrounding the section should allow it to adhere to the slide. Water should not drip from the slide; blot off extra water if needed. 8. Move the condenser downward to allow room for the inverted section. Place the slide, SECTION SIDE DOWN, on the microscope stage so that the adhering section is in the center of the stage and positioned over the center of the condenser. See Diagram III. Diagram III 9. Focus on the plant section while looking through the slide. Do this on the lowest magnification objective available. Start with the objective positioned high above the slide and slowly move it toward the specimen to find the focus. TAKE CARE not to ram the objective into the slide. 10. The cellular anatomy should come into focus very clearly. Different magnifications can be used to observe different levels of detail. (If bubbles appear, they can be minimized by carefully removing and reapplying the section. Do not confuse bubbles with cells!) Refer to Table I to identify cell types and tissues. 11. If the slide begins to dry out, add a small amount of water around the section. Place a small drop of water to one side of the section and then placing tissue or filter paper on the other side of the section, drawing the water across the section. See Diagram IV. Diagram IV 2
3 Table I Tissue Element or Structure Xylem Phloem Schlerenchyma Collenchyma Parenchyma Cellulose, Starch DNA Color Developed by Toluidine Blue Green or Blue-green Red Blue-green, sometimes green Red-Purple Red-Purple Unstained Blue 3
4 Elaboration: The table below shows plant adaptations observed at the microscopic level. Descriptors, images and functions are given in addition to modifications observed in specified environments. Xerophytic plants have adaptations for dry climates, mesophytic plants grow in moderate climates (not extreme) and hydrophytic plants grow in water environments. Description of Plant Structure Cuticle: waxy coating on the surface of leaves and stems Epidermal Layer: outer layer of cells covering leaves, stems and roots Trichomes: small hair-like structures on leaves and stems Anatomical Adaptations in Plants Image Function Modification of Structure Protects structure Thick cuticle to prevent water from water loss and loss sometimes present serves as defense Thick cuticle on top of floating (bacteria and viruses) leaves water rolls off Protects inner tissue; regulates gas and water exchange; produces cuticle Traps moisture; can give a silver coloration that reflects light Multiple layers in leaves with many chloroplasts (sunny locations); same plant can have reduced layers in a shaded leaf Abundant Present Adaptation Environment Xerophytic Hydrophytic Xerophytic Xerophytic Hydrophytic Mesophytic Stoma: (also stomata) microscopic pores in the epidermal layer surrounded by guard cells Air spaces: intercellular spaces between mesophyll or other parenchyma cells Allows gas exchange between the environment and the interior of the plant Holds gases and water vapor for gas exchange and buoyancy Sunken; often closed and on the bottom side of the leaf On top of leaf; increased and open most of the time Large spaces Large spaces Xerophytic Hydrophytic Xerophytic (succulent) Hydrophytic Collenchyma cells: plant cells with thickened, uniform walls Helps stems in soggy or windy conditions to maintain strength Abundant just inside the epidermis Xerophytic Hydrophytic 4
5 Name What Does the Plant Image Tell? Finding the Environment from Anatomical Structures For each microscopic image provided, describe anatomical characteristics that provide clues on the plant s environment. Analyze the information and derive the climate where the plant would grow (xerophytic, mesophytic or hydrophytic). Unknown Characteristics Environment #1 #2 #3 #4 5
6
7 Teacher Key for Unknowns 1. Typical monocot: corn (mesophytic) 2. Dicot: Oleander (xerophytic: thick cuticle, many layered epidermis, succulent with space for water, sunken stomata surrounded by trichomes to hold moisture) 3. Monocot: Cypress Plant (hydrophytic: thick cuticle, layers of collenchyma to provide support, air spaces for buoyancy) 4. Dicot: Water Lily (hydrophytic: air spaces for buoyancy and stoma with guard cells on the top side) 7
8 References for Anatomical Adaptations in Plants Cuticle: clivia cuticle jpg 804x556 - jpeg k - jpg images.xoila.com Air spaces: intercellular space below k - jpg vcbio.science.ru.nl Trichome: Aerenchyma with lignified "inner hairs" (Nymphaea spec.): section through the leaf stalk. Staining with phloroglucin-hcl. (photo by W. KASPRIK). Collenchyma: pharamcytimes.wordpress.com/2009/04/page/2/ Epidermis: 1 upper epidermis k - jpg vcbio.science.ru.nl Epidermis: k - jpg - botit.botany.wisc.edu/.../images/epidermis.jpg Image may be subject to copyright. Below is the image at: botit.botany.wisc.edu/courses/botany_130/anat... Corn cross section: Hydrophytic plant: 8
9 Elaborate Teacher Section Where s Home? Background Information: Students apply information from previous activities to look for patterns and decide where plants live. They should build an understanding of plant adaptations as they move through the unit. Purpose: The purpose of this activity is to compare adaptive characteristics of plants at the tissue level to determine where they live. Objectives: The student will derive the habitat of a plant by generalizing from patterns. Age Appropriateness: Grades 9-10 Time Requirements: 20minutes Materials: Student Sheets Transparencies of leaf cross sections Preparation and Procedure: 1. Copy student sheets. 2. Have students compare the examples provided to typical examples on the student sheet to determine the habitat of each. Questions to Ask: 1. Look at each example that shows an adaptation. How does it look different from the typical plant? 2. What could be the purpose for the change? 3. If structure relates to function, how do you think the structure could help the plant survive? 4. Infer where the plant lives. 9
10 Elaborate Student Section WHERE S HOME? Purpose: The purpose of this activity is to compare adaptive characteristics of plants at the tissue level to determine where they live. Materials: Illustrations of plant microscopy slides Procedure: 1. Observe the examples given in the illustrations. Compare the typical leaves to examples that have special adaptations. 2. Look for overall patterns and predict where each example lives. Select and answer, and justify your response. 10
11 PLANT CROSS SECTIONS. 1. The example provided is a typical dicot leaf. What are some clues that help you to identify this cross-section? 2. Observe the cross-section below. The adaptations indicate the plant grows in a a. desert b. rainforest c. lake d. tundra Justify your answer. 11
12 3. Where would this plant grow? a. forest b. desert c. rainforest d. pond Justify your answer. 12
13 4. The cross-section below is a typical monocot leaf. What are some clues that help you to identify this cross-section? 5. Look at the cross-section below. In what climate would the plant grow? a. arid b. rainforest c. pond d. tundra Justify your answer. 13
14 TEACHER ANSWER SHEET PLANT CROSS SECTIONS TRANSPARENCIES 1. The example provided is a typical dicot leaf. What are some clues that help you to identify this cross-section?. Response: Top and bottom are evident (palisade and spongy mesophyll) Vein in the middle indicates a vein Pattern of the vascular bundle is not a monkey face 2. Observe the cross-section below. The adaptations indicate the plant grows in a e. desert f. rainforest g. lake h. tundra Response: Lake The plant has air pockets to float and stomata on top rather than on the bottom. 14
15 3. Where would this plant grow? e. forest f. desert g. rainforest h. pond Justify your answer Response: Desert The plant has multiple epidermal layers on top for protection with a thick cuticle. The spaces within show it is a succulent. The stomata are within special enclosed structures that prevent water loss from the stomata. Trichomes (hairs) are abundant to keep water near the surface. 15
16 4. The cross-section below is a typical monocot leaf. What are some clues that help you to identify this cross-section? Response: Top and bottom look the same (if it were turned upside down, you could not tell) Veins are looking at you (parallel is often observed) Veins are monkey faces 5. Look at the cross-section below. In what climate would the plant grow? e. arid f. rainforest g. pond h. tundra Justify your answer. Response: arid The leaf rolls to prevent water loss and the trichomes help to hold moisture. Also, the leaf has a thick cuticle. 16
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