2 CROP SCIENCE LAB MANUAL LABORATORY MANUAL CROP SCIENCE AGR499 Prepared by: Ts. Dr. SITIKHADIJAH ABDUL KARIM FPA (JASIN) 2023 Universiti Teknologi MARA Scanned with CamScannerCROP SCIENCE ae LAB MANUAL anos, Lab1 PLANT STRUCTURE AND PHYSIOLOGY Introduction The roots, stems, and leaves make up the three primary organs of the vascular plant. These develop from meristems, regions of cells that still have the capacity for cell division. In vascular plants, the primary roles of the roots, stems, and leaves are reflected in their structures. The huge surface area of roots is an adaptation that makes it easier for plants to absorb water. The cellular changes in stems that support the leaves in the air are the result of evolution. The large surface area of leaves is an ‘adaptation for the photosynthetic tissues to absorb sunlight. Vascular tissue connects all of these structures and carries the photoassimilates throughout the entire plant. There are two distinct leaf morphologies that serve as excellent illustrations of how structure and function interact. So-called C3 plants make up the majority of all plants. ‘The majority of the chlorophyll-containing cells in these plants’ tissues are where the carbon fixation reaction of photosynthesis takes place. Contrarily, C4 plants only perform carbon fixation into the Calvin cycle in specialised cells that are close to the vascular bundles and shielded as far away from the leaf surface as possible to avoid contact with oxygen, as contact with oxygen reduces total photosynthetic yield. ‘Sugarcane and maize are two common examples of C4 plants. Mesophyl cell layers are tightly packed, protecting bundle sheath cells from 02 entering the leaf from the outside, Objectives. 1. To identify and compare the anatomy and morphology of different plant cells and tissues. 2. To know various plant structures of C3 and C4 plants. Materials Plant, Eosin, Prepared slaid (Cross-section of root), Cover slip, Clean slip Filter paper, Scalpel, Forceps, Beaker, Tiles, Microscopes. Procedure A. Experiment 4. Stems 41, Obtain plant A with leaves and root. 2. Remove soil from the plant 2023 Universiti Teknologi MARA Scanned with CamScannerCROP SCIENCE rea ers B= 3. Slice off thin section of the stem and leaf on the tiles 4, Put a few drops of eosin on the thin sections for 5-10 minutes 5. Transfer carefully the stem on the slaids and put a few drops of distilled water. 6. Observe under the miscroscope. 7. Draw your observation under low and high magnification. (See if you can distinguish xylem from phloem cells. In the cross section, look for the different tissue types. In the cross section also look for vascular bundles and cells that divide xylem and phloem). 2. Leaves 1. Cuta thin slice of cross section of the the C3 and C4 leaves 2. Observe the C3 and C4 leaves under the microscope. 3. Draw and label your observation. 3. Root 1, Observe the prepared slides provided (root) (Label your drawing with each celltissue type and don't forget to write on your drawing the name of the plant, the magnification, organ, and the type of section (cross section vs, longitudinal etc). 2023 Universiti Teknologi MARA, Scanned with CamScannerCROP SCIENCE Bea LAB MANUAL Ese LAB2 DISTRIBUTION OF STOMATA IN THE UPPER AND LOWER SURFACES OF LEAVES Introduction Plants can have between 1,400 and 40,000 stomata per square centimetre of leaf, which is referred to as stomatal density. The numbers and location of stomata have a significant impact on how quickly gases are exchanged and how much water a leaf loses. For instance, we may hypothesized that the rate of transpiration increases as the number of stomata increases. Stomatal frequency is determined by counting the number of stomata in the microscope field of view (after we calculate the area of view). ‘The majority of stomata are found in leaves, although they can also be found on stems, petioles, sepals, and petals, Stomatal index can be used to determine how many stomata are present in a given leaf. The ratio of total stomata to total epidermal cells in a given unit area is known as the stomatal index. Various plants exhibit varying ‘stomatal indices. Additionally, the number changes under various environmental conditions as well as from plant to plant. Objectives 1. To count and compare the density of stomata on upper and lower epidermises of the same leaf with relation to transpiration. 2. To compare the distribution of stomata of different plants, with relation to transpiration Materials Clear ni amish, Slides and cover-slip, Forceps, microscope, mature leaves. Procedure 1. Students will be given leaves from different plant. 2. Make a replica of the leaf surface using clear nail varnish by spreading a thin layer of nail varnish over the surface of the leaf using the brush in the bottle on the upper and lower surfaces of the leaf Allow to dry for 15-20 minutes Peel off the replica of the leaf surface using the forcep and the knife Place the film of nail varnish on slide and add cover slip. Examine under the microscope and count the number of stoma in a given view and repeat 3 times to obtain a mean value Determine the area of the field of view by measuring the diameter of the lens. Calculate the area of view using the formula 12, where r is the radius and 1 2023 Universiti Teknologi MARA N gmae Scanned with CamScannerCROP SCIENCE ee TAB MANUAL pee 33.142 8. Calculate the number of stomata per square centimeter 9. Tabulate the results and compare. Questions: 1. What is the density of stomata on the upper and lower surfaces of a leaf? 2. How many percent of the leaf surface is open pore? Stomatal Spidermtal cells Opening an Radially ar Closing Eaiuiage merits Ledaact Guard cells e8 Soliulode microfibrils (b) Closed! Spldertal cette 2023 Universiti Teknologi MARA, Scanned with CamScannerCROP SCIENCE iietiaar LAB MANUAL. a ie LABS TRANSPIRATIONAL PULL DOES WATER TRANSPORT IN CELERY XYLEM Introduction Water is lost from plants through stomata through a process called transpiration. Most plants’ transpiration is a passive process, mostly governed by soil moisture levels and ambient humidity. Only 1% of the water that transpires through a plant is utilised for growth, Transpiration is also employed to prevent tissues from overheating by transferring nutrients from the earth into the roots and delivering them to the plant's numerous cells. From the roots to the stem and leaves, the xylem transports water and minerals. ‘Several factors come together to cause water to travel up a plant stem. Due to the cohesive and polar characteristics of water molecules and their attraction (adhesion) to xylem cell walls, some movement is caused by capillarity, the ascent of water up a thin tubule cell (xylem). Root pressure, which is a result of active transport in the roots, ‘adds additional force. Active transport moves the required ions up the xylem and into the root when water content rises. The tension-cohesion (transpiration pull) force that results from water vapour loss through stomata is an additional force. Water from xylem veinlets replaces water that escapes to the atmosphere in the leaf mesophyll. Water is drawn up the xylem by cohesion as a result of this constant loss and replacement of water. Water lost from the leaves through transpiration is continuously replaced by water in leaf tissue, a “pulling” of water molecules all the way from the roots and up to the stem. Water molecules are held together by hydrogen bonds and cling to xylem cell walls. The rate of transpiration might change depending on the atmosphere. The rate of transpiration is measured as the amount of water lost/ square meter! minute. Because water evaporates through the many stomata on the leaf surface, the rate of transpiration is directly related to the surface area. To arrive at the rate of transpiration, therefore, you must calculate the leaf surface area of each plant: Because most stomata are found in the lower epidermis, you will determine that surface area. 2023 Universiti Teknologi MARA Scanned with CamScannerCROP SCIENCE Give LAB MANUAL Ege Experiment 1 Objective 1. To study the effect of wind and light factors on transpiration in celery stalks, Materials Gelery stalks Eosin dye (red) Gooseneck lamp Fan Scalpel Tiles Tissue Ruler Procedure 4. Cut 1 cm from the bottom end of each celery stalk as it remains submerged in water. Cut the celery stalks at the same height. 2. Label flask A, B, and C which contain 200 ml eosin dye. 3. Quickly place the cut end of each celery stalk into the flasks. 4. Place flask A on a normal room temperature, flask B in front of the running fan and flask C in front of gooseneck lamp. 5. Note the time and allow all the celery stalks to stand for less than 5 minutes in their designated environmental conditions. 6. Stop the experiment once any of the eosin reach at the top end of the celery stalk. and take the time. 7. Remove the celery stalks from their flask. Rinse excess dye from the stalks under running water and place them on a tissue. 8. Use a metric ruler and scalpel to cut 1 cm segments from the bottom end of the stalks. 9. After each cut is made, examine the cut end of the stalk for the presence of eosin dye in the xylem tissue positioned along the outer edge of the stalk. 10.Continue cutting 1 cm segments from the each stalk until the dye begin to fade in the xylem. 11. Tabulate the distance the dye traveled up of each stalk. 2023 Univers Feknologi MARA Scanned with CamScannercrop science ay ee LAB MANUAL. een Experiment 2 Materials Celery Leaves Graph paper Objective 41) To calculate the rate of transpiration in celery leaves. Procedure 1. Lay the celery leaves to be measured on a 1-cm grid and trace their outlines. 2. Count the number of square centimeters. Estimate the area of the partial squares. (Here's a simple method for this estimate: Count a partial square if it is at least half covered by the leaf; do not count partial squares that are less than half covered.) 3. Do not include the area of the stem (petiole) in your calculations. 2023 Universiti Teknologi MARA, Scanned with CamScannerCROP SCIENCE ee I LAB MANUAL reo Lab 4 ‘SEED GERMINATION USING HYDROPHONIC SYSTEM Introduction Without utilising soil, hydroponics is a technique for growing plants with mineral nutrient solutions. With their roots exclusively in the mineral fertiliser solution or in an inert media like perlite, gravel, or mineral wool, terrestrial plants can be grown. As inorganic ions in water, vital mineral nutrients are absorbed by plants. Soil serves as a storehouse for mineral nutrients under natural circumstances, but it is not necessary for plant growth in and of itself. Plant roots can absorb mineral nutrients from the soil when they dissolve in water. The development of an embryonic plant inside a seed is known as germination, which Produces the seedling. The seed of a higher plant is a litle packet that develops in a fruit or cone as a result of the union of the sex cells from the male and female. Every completely grown seed has an embryo inside of it, and most plant species also have ‘some food reserves that are stored inside the seed coat, Some plants produce varied numbers of empty seeds, also known as seeds without embryos, which never germinate. The majority of seeds go through a phase of quiescence during which there is no active growth. During this time, the seed can be moved safely and/or endure unfavourable weather conditions until conditions are favourable for growth. Quiescent seeds are ripe seeds that do not germinate because they are subject to external environmental conditions that prevent the initiation of metabolic processes and cell growth. Under favorable conditions, the seed begins to germinate, and the embryonic tissues resume growth, developing towards a seedling. Plant nutrients are dissolved in the water used in hydroponics and are mostly in inorganic and ionic form. Primary among the dissolved cations (positively-charged ions) are Ca** (calcium), Mg” (magnesium), and K* (potassium); the major nutrient anions in nutrient solutions are NOs" (nitrate), SO. (sulfate), and HzPO. (dihydrogen phosphate).. Commonly-used chemicals for the macronutrients include potassium nitrate, calcium nitrate, potassium phosphate, and magnesium sulfate. Various micronutrients are typically added to hydroponic solutions to supply essential elements; among them are Fe (iron), Mn (manganese), Cu (copper), Zn (zinc), B (boron), Cl (chlorine), and Ni (nickel). Chelating agents are sometimes used to keep Fe soluble. Plants will change the composition of the nutrient solutions upon contact by depleting specific nutrients more rapidly than others, removing water from the solution, and altering the pH by excretion of either acidity or alkalinity. 2023 Universiti Teknologi MARA, Scanned with CamScannerCROP SCIENCE iene LAB MANUAL io Objectives: To observe the seed germination using hydrophonic system Materials Styrofoam box with lid (as nutrient tank) Water spinach seeds (or any available seeds) fom » 2023 Universit Teknologi MARA Scanned with CamScannerCROP SCIENCE Gime LAB MANUAL, agent and B liquid fertilizer (can be obtained at any plant nursery) Methods. Stage 4 4) Pour 2L of tap water in the container (about 1 inch from the top level). 2) If you are using plastic drinking cups as the media pots, make holes at the bottom of the cups for the threads. 3) Cut out a hole on the Styrofoam lid with the same size of the media pot opening, Prepare about 6 holes for one nutrient tank. Make sure the keep about 10 cm space in between the holes to provide enough space for the plant growth, 4) Put the threads in the media pot and pull unti the threads are outside the pot holes. Make sure that the holes are fully covered to avoid mosquito. 5) Place the pots in the nutrient tank filled with tap water and then position them in ‘StyroFoam holes. 6) Place one wet planting sponge or adequate cocopeat into each pots. Then germinate 2 seeds into each pots. 7) Leave the hydrophonic system and observe the seed germination every day. Stage 2 4. After seeds have been germinated, remove the nutrient tank lid. Add 10m! A and 10 mB fertilizer into the nutrient tank. Stir using a spatula to mix it well. 2. Place back the lid. Place the tank at area with good light exposure. 3. Harvest the plant after 3/4 weeks. 2023 u Scanned with CamScannertear Heaven CROP SCIENCE a LAB MANUAL Net Pot Thread AB MIX Nutrient tank alaeperlik Figure 1.0 : HYDROPHONIC SYSTEM 2023 Universiti Teknologi MARA 2 Scanned with CamScannerCROP SCIENCE LAB MANUAL Egos LAB 5: PHOTOSYNTHESIS AND RESPIRATION Introduction Plants and certain other organisms transform solar energy into chemical energy in the form of sugars through a process called photosynthesis. Cellular respiration uses the chemical energy in sugar to meet the energetic requirements of plants, whereas photosynthesis uses the energy of the sun to make sugar. However, not al of the sugar that a plant produces is consumed during respiration; some is utiised to build new plant biomass, such as roots, leaves, stems, wood, and bark. Plant biomass temporarily stores carbon, but it wll eventually degrade, burn, or be consumed and ‘metabolised, retuming the carbon to the atmosphere, The plant will take carbon dioxide from the water as it photosynthesizes, lowering ‘carbon dioxide concentrations in the water and raising the pH of the solution if itis Placed in water that contains dissolved gases. When a plant or seed is placed in water and begins to breathe, carbon dioxide is released into the water, lowering the PH of the solution. Therefore, by keeping an eye on the pH fluctuations in a solution, ‘you may tell ifthe plant or seed in the solution is photosynthesizing (pH rises) or respiring (pH decreases). We can use pH as an indirect indicator of photosynthesis rates since carbon dioxide uptake is one way to gauge the rate of photosynthesis, and changes in carbon dioxide concentrations result in pH changes. When the pH is basic (and carbon ioxide concentrations are low), bromothymol blue is blue; when the pH is acidic (and carbon dioxide concentrations are high), itis yellow. When the solution is basic, it is blue; when it is neutral or acidic, it is greenish yellow. Objectives 1. To observe the effect of photosynthesis and respiration process on the amount of carbon dioxide in water. Materials aquatic plant Elodea bromothymol blue straw tubes tube caps gooseneck lamp petri dish jars and two jar covers ‘germinating mung bean (Phaseolus aureus) seeds strainer 2023 Universiti Teknologi MARA, Scanned with CamScannerLAB MANUAL ee CROP SCIENCE a vasemacan Methods (I) Photosynthesis 1. Place about 75 ml of bromothymol blue into a 125 ml Erlenmeyer flask (the triangular glass containers at your work bench), 2. Observe the color of the solution. Introduce carbon dioxide into the solution. By using a straw, slowly blow CO, into the solution until it just tus yellow. 3. Compare the color of the solution to the printout showing bromothymol blue at various pHs. Find the color on the printout that best matches the color of your solution. If your color is between those shown, use the pH value that is intermediate between them. ‘4. Record the color of the solution before and after CO, treatment as well as the pH for each of the tubes in Table 1. When comparing or describing colors, be sure to hold the tubes in front of a white background and be as detailed as possible when describing the color (e.g., use terms lke "greenish-biue" or "yellow-green. 5. Pour the solution into three screw cap tubes, dividing it evenly between them. Obtain two 2-inch pieces of Elodea from the open dish ("Light Elodea”). Place them in one of the tubes, and cap it. Obtain two 2-inch pieces of Elodea from the covered dish, place them in a second tube covered with aluminum foil (to prevent the entry of light), and ap it. In both tubes, make sure the plant is completely submerged in the solution. Cap the remaining tube — it will serve as your control. 6. Place the tubes upright on the lab bench approximately 8-10 inches from the lamp. Ensure the light hits all three tubes equally and from the side, not from the top. Turn ‘on the lamp. While you allow the experiment to run, set up the respiration experiment described on the next page. 7. Allow the plants to sit undisturbed for 30 minutes, and then carefully remove the Elodea plants from the tubes. Determine the color of the solution against a white background, record them in Table 1, and compare the colors to the bromothymol blue printout to determine pH. Table 1: Tube Initial color | initial pH | Finalcolor| Final pH Elodea - ight Elodea - dark No Elodea 2023 Universiti Teknologi MARA Scanned with CamScanner(CROP SCIENCE LAB MANUAL (Wl) Respiration 1. Obtain two jars and two jar covers. One of the jars should be partially filed with germinating mung bean (Phaseolus aureus) seeds and the other should be empty. 2. Rinse out both jars thoroughly (without pouring out the seeds) to ensure that each has a fresh supply of air then fill each jar with equal amounts of bromothymol blue to just above the top of the seeds. Record the initial color and estimated pH of the solutions in Table 2. 3. Cover the jars and make sure that the two jars are in similar conditions. Allow the jars undisturbed for 30 minutes. 4. Filter the seeds out of the jars by pouring the solution through the provided strainer into a separate container. To control all your variables, do the same for the solution with no seeds (even though there is nothing to filter out). Compare the color of the solution in each of the jars against a white background. Record the final color and estimated pH of the solutions in Table 2. Table 2: Container | Initial color | Initial pH | Final color Final pH With seeds No seeds Peas 12023 Universiti Teknologi MARA Scanned with CamScannerCROP SCIENCE LAB MANUAL, Lab Report Guidelines Reports should be typed or neatly hand written (Depends on lecturers’ decision). Kindly refer the report format that is included in this manual. All the reports can be compiled and submitted at one time or be submitted separately a week after each lab session (Depends on lecturer's decision). Lab report must be prepared and submitted indivually. However, lab session will be carried out in groups. 2023 Universiti Teknologi MARA Scanned with CamScannerLogo UiTM (PROGRAM) SUBJECT CODE: ‘TITLE OF EXPERIMENT: Name: Student ID: LECTURER NAME: DATE OF SUBMISSION: Scanned with CamScannerAttention: 2. 3. 4. 5. Lab report must be submitted a week after the laboratory session or be compiled altogether and submitted at Week 10 (Depends on lecturer's decision). Submit lab report by individual. ‘The report should be submitted to the lecturer only not to anyone else. Contents of the report should follow as the examples given to you. Overall practical part carries 20% of your assessments. (Practical participation and lab report) Lab report must include these: 1) Introduction (including the objectives) 2) Materials and Methods 3) Results and Discussions 4) Conclusions 1) Introduction Objective: 1), 2, Introduction: ‘+ Itstates the objective of the experiment and provides the reader with background to the experiment. ‘© State the title of your report clearly and concisely, in or two sentences. ‘© Example: The purpose of this experiment was to identify Must Have: 1) Purpose of the experiment 2) Important background and/or theory. May includ 1) Deseript ‘*Note: The experiment is already finished. Use the past tense and passive sentences. n of specialized equipment. 3) Materials and Methods ‘* Materials: Can usually be a simple list, but make sure it is accurate and complete. ‘+ Methods: -Describes the process in chronological order. = Using clear numbering structure, explain all steps in order they actually happened, not as they were supposed to happen. Scanned with CamScanner4) Results and Discussions * Results: Graphics need to be clear, easily read, and well labeled. Discussions: -The most important part of your report. Explain, Analyse, Interpret. Compare expected results with those obtained. Analyze experimental error, Explain your results in terms of theoretical issues. Relate results to your experimental objective (s). 5) Conclusion © Canbe very short in most undergraduate laboratories. © Simply state what you have known. Must have: 1) State what's known. 2) Justify statement. May include: Prepared By: 1) Suggest further research. Dr. Siti Khadijah Abdul Karim Senior Lecturer Scanned with CamScanner