Measuring Transpiration

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Transpiration is the evaporation of water from plants, which occurs mostly through the stomata. The amount of water lost can be significant: a medium-sized tree can lose 50 to 100 gallons of water on a single sunny day. Water forms a long, continuous column in the tracheids and vessels of the xylem; this column goes from the roots through the stem to the mesophyll of the leaves. This continuous column allows water to move from the soil to the air. As water evaporates through the open stomata of the leaves, water moves out of the veins of the leaf into the mesophyll cells and surrounding airspaces to replace the lost water. This movement pulls water from the stem xylem out into the leaf veins to replace the lost leaf water, and the pull in the stem is translated all the way down to the roots, where water is being pulled in from the surrounding soil. The force of the evaporating water is transmitted to the roots because of the cohesion of water molecules to each other throughout the plant, so this description of water movement in plants is called the transpiration-cohesion hypothesis. When a plant wilts, the movement of water from the soil isnt sufficient to keep up with the evaporation of water from the leaves, so water is pulled from the living cells, as well as from the xylem. The strength that the water in the cells provided the plant through turgor pressure is thus lost, and the plant collapses. When you look at the surface of a leaf, you see the pattern of the veins that distributes the water throughout the mesophyll. This pattern is called the venation. In a dicot the venation is generally reticulate or net- like and somewhat irregular. In a monocot the venation is often parallel, with branches from major veins coming off at right angles, so the venation is very regular. In this lab you are going to (1) examine the components of leaf structure and anatomy that are important in transpiration, (2) determine the transpiration rate for your coleus plants, and (3) look for correlations between leaf area and transpiration rate.

1. Examine the venation of a coleus leaf under the dissecting microscope. Is it parallel or reticulate? Also look at both the top and bottom sides of the leaf. Can you see the stomata? Cut 2 small bits from the leaf. Place one right-side up on a microscope slide in a drop of water and cover with a cover slip. Place the other with the bottom side up on a slide in a drop of water and cover with a cover slip. Examine both slides under the compound microscope, focusing up and down to find the epidermis. Identify the guard cells of the stomata. You might also try using clear nail polish to obtain an epidermal peel of each leaf surface that will allow you to observe the stomata more easily. Estimate the density of stomata by using the known area of your field of view and counting the number of stomata within that field of view. Do the two sides of the leaf differ in the number and/or density of stomata? Which side has more? Note also the shape of the other epidermal cells. Does this shape differ between the two sides? 2. Now examine a prepared cross-section of a dicot leaf under the compound microscope. Think about what you would expect to see if you took a cross-section of your coleus leaf. Would you expect the veins to be in cross-section, longitudinal section, both, or neither? Identify the upper and lower epidermis, palisade mesophyll, spongy mesophyll, and xylem

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and the phloem in the slide of the dicot leaf cross-section; notice the position of the vascular bundle in the mesophyll. Can you identify different branching orders of veins? Locate the stomata in cross section. 3. We are going to determine the transpiration rate for the coleus plants on which you studied apical dominance. Choose 2 coleus plants that are similar in size. If there are multiple stems per pot, cut away all but the largest stem. Remove branches that are close to the soil from the remaining stem. Also remove buds or branches that are beginning to expandyou want to have a single stem that doesnt have too many leaves and branches. Remove app. of the remaining mature leaves from 1 plant. This will facilitate measurement of leaf area. Count the number of leaves on each plant and measure and record the length of each leaf blade. Your TA will give you a general equation that relates leaf length to leaf area in these plants. Use this to estimate the total leaf area for each plant. If you did not have this equation to start with, think of how you might estimate it1. Water each pot until the soil is saturated (water runs freely from the pot). Let the water drain until it has stopped dripping water. Wrap the pot in clingwrap, making a tight seal around the base of the stem but leaving the shoot free. This will prevent evaporation from the soil. Cover the clingwrap with a layer of aluminum foil. Weigh the wrapped plant on the balance in the lab, recording the weight and the time. Repeat for the second plant. Label the plants with your group name and a plant treatment name (A for lots of leaves, B for fewer leaves). Put the plants in the greenhouse. After 1 to 2 days, re-weigh the plants and record the time. There is a balance in the greenhouse for this purpose. Using the fact that 1 gm water has a volume of 1 mL, figure out how much water the coleus plant has lost in all (= total water loss) and what the rate was (water loss per hour and per day). How much water would the plant lose in a week? Did the two plants differ in how much water was lost? Using your estimates of total leaf area, estimate the water loss per unit leaf area for each plant. Compare the data from you lab group to that of other groups in your lab. Would you expect them to have similar or different results? Why?

One way to estimate this is to relate weight of a piece of paper to the area of the paper; you can then make paper tracings of leaves, weigh them, and use that weight to determine what your area was (i.e., if 1 piece of paper that is 50 cm 2 weighs 2 g, then if you trace some leaves, cut them out and weigh them, and find you have 1 g of paper leaf tracings, you can estimate that your leaf area was 25 cm 2 .

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