Kingdom Plantae: Vascular Seed Plants; Stems and Roots

Objectives: Identify the structure and function of the major plant cell and tissue types Differentiate between the structures and functions of the vascular tissue Describe the organization of tissues and cells in a plant organ and relate this organization to the function of the organ Identify the areas of primary and secondary growth in a typical plant stem and root Relate the meristems to the type of growth expected in that plant region Describe the major differences that distinguish the roots of typical monocots and typical dicots Introduction and Background: The vascular plants (as a loose grouping) have been evolutionarily successful for hundreds of millions of years. Their success, in part, is due to their successful adaptation/evolution to a terrestrial environment. Although we see many adaptations to a terrestrial environment in the avascular plants, the vascular plants take it to another level. The terrestrial habitat presents many challenges that aquatic algae did not have to deal with. In particular are the issues of (not an exhaustive list): 1) Lack of support for the plant body (water is a decent support medium) 2) How to get water from the ground and transport it to the upper reaches of the plant 3) How to transport photosynthetic products to the non-photosynthetic parts of the plant 4) How to disperse pollen and seed (not dealt with in this lab per se) The vegetative plan organs we will focus on in this lab are the results of evolutions and adaptations to the terrestrial lifestyle. Specifically, we will focus on three organs: stems, roots, and leaves. These organs have many cell and tissue types in common, and we will begin by considering the cell types and tissue types. By the time you are looking at slides, you should be familiar with the cells and tissues. (That is a hint to make the lab go smoother!) Plant Tissue Types (a brief overview): This is not meant to be an exhaustive primer on plant cell and tissue types, but instead to point out some of the more common cell and tissue types. (To get a more complete description and list, consult your textbook or chat with your instructor). Epidermal cells are typically flattened, rectangular cells that line portions of the plant body. Specialized epidermal cell extensions include trichomes and root hairs and a specialized epidermal cell type you will see on the leaves are the guard cells of the stomata. Most epidermal cells have a waxy cuticle covering to help prevent water loss through the cells and the organs they line. Primary Meristematic tissues are found in the growing regions of the plant. These tissues consist of small actively dividing cells (Remember the mitosis lab?). These cells produce the primary tissue types (epidermal, ground, and vascular) of these growing regions. In general, we can think of the primary meristem as lengthening these plant parts. (Hint: look in the stem and root tips for the primary meristems) Lateral Meristematic tissues are also found in growing regions of the plant. These tissue types (vascular and cork cambium for instance) are largely responsible for the thickening (secondary growth) of the plant parts they are found in. (Hint: Look on the sides of stems and roots (in cross-section) for these tissues). We will also compare the plant organs of monocots and dicots. Although this is no longer a systematically significant distinction, it is still important to be familiar with the two basic body types. There are obvious differences in basic structure and organization in these two body types and these provide an easy, functional way to compare plants. For more information about monocots and dicots consult your text or your instructor.

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General terms to define and structures to identify: (you dont have to write out the definitions, but you should be familiar with them) stems, shoot system, meristems, primary growth, secondary growth, dermal tissue system, vascular tissue system, ground tissue system, lignin, parenchyma, collenchyma, sclerenchyma, herbaceous, woody, primary xylem, primary phloem, vascular cambium. Part 1: Structure of the Dicot and Monocot Stem Use illustrations and guides of the herbaceous dicot and monocot stems to help you examine and label the slides provided Slides: Basswood (Tilia americana)-woody dicot stem Identify the following: periderm, cortex, primary and secondary phloem, primary and secondary xylem, phloem ray, xylem ray, vascular cambium Various woody trunks will be available for you to examine the external structures. Use the Secondary Stem Model to assist you as you identify the following structures: periderm, bark, phloem, vascular cambium, secondary xylem, xylem ray, heartwood, transverse surface, tangential surface, radial surface Part 2: Live Specimens 1. Obtain a herbaceous dicot plant of the pea family that contains both intact roots and stems (You will use this plant for both the stem and root section of this lab). Perform a cross section of the stem and apply the Toludine blue and Phloroglucinol staining techniques as described from the plant anatomy lab. Draw your observations and label as many of the following structures as possible from your section: epidermis, cortex, pith, vascular bundle, phloem (sieve tube members, companion cells), xylem, vascular cambium. Also, answer the following questions in the Results section of your lab notebook. a. b. c. d. e. 2. How would you describe the arrangement of the vascular bundles in your stem section? Based on your section and your observations, how might the arrangement of the cortex and pith relate to its function in the plant stem? When examining the vascular bundles or your section, how does the placement of collenchyma cells differ from that of parenchyma cells? Is the vascular cambium only found in the vascular bundles? Explain and hypothesize a reason for your observation. Is there a cork cambium visible in your section? Explain.

Obtain a herbaceous monocot plant that contains both intact roots and stems (You will use this plant for both the stem and root section of this lab). Perform a cross section of the stem and apply the Toludine blue and Phloroglucinol staining techniques. Draw your observations and label as many of the following structures as possible from your section: epidermis, vascular bundles, phloem (sieve tube members, companion cells), xylem, vascular cambium. Also, answer the following questions in the Results section of your lab notebook. a. b. c. Can you distinguish cortex and pith in your monocot section? Explain. How would you describe the arrangement of the vascular bundles in your stem section? The vascular bundles of monocots (and some dicots) are considered closed which means that after the cambial cells mature, the potential for further growth within the bundle is lost. How might you infer this based on the bundle arrangement in your section and in comparison to your dicot stem which has open bundles?

Part 3: Deciduous twig external features 1. Various deciduous twigs will be available for you to examine the external structures. Use the Botany Photo Atlas and your text to assist you as you identify the following structures. Draw your observations and label: terminal bud, lateral bud, terminal bud scale scars, leaf scars, vascular bundle scars, lenticels

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a. b. c. How many years old is your twig (based on the cutting)? Explain your reasoning. Based on your observation, what could be a possible function of the lenticels? Where on your twig would you expect meristematic growth to occur during the growing season? Explain your reasoning.

Part 3: Water conduction in a stem Background: You have learned in lecture that water moves through a vascular system by mass movement and not by diffusion. This experiment will provide evidence to support the mass movement hypothesis. 1. Set-up: Obtain two Red Top grass seedlings from the germination tray. With a sharp razor blade, completely remove the root system of one of the seedlings. Place both seedlings on a glass slide in a lengthwise position parallel with each other. Make sure the two seedlings are aligned with their stems in the same position and location. Use grease or vasaline to construct a barrier perpendicular to the seedlings along the base of their stems. Place 2-3 drops of Saranin on the root systems of the seedlings. Make sure your grease/vasaline barrier prevents the Safranin from oozing onto the stem portion. Obtain a coverslip and place it over the root system of the seedlings. a. What do you hypothesize will happen in your experiment? Make a prediction and write it down your hypothesis explanation. Set your experiment aside for 30-60 minutes, occasionally observing the movement of the Safranin. Describe what you observe in your Results section of your lab notebook.

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Part 4:

Structure of the Dicot and Monocot Root

Use illustrations and guides of the herbaceous dicot and monocot roots to help you examine and label the slides provided Slides: Root Tip (l.s.)-monocot or dicot Identify the following: root cap, apical meristem, zone of elongation, epidermis, root hairs Basswood root (Tilia spp.) woody dicot root Identify the following: secondary xylem and phloem, vascular cambium

Part 5: Live Specimens 1. Re-examine the herbaceous dicot plant of the pea family you obtained from your stem analysis. Perform a cross section of the root and apply the Toludine blue and Phloroglucinol staining techniques as described from the plant anatomy lab. Draw your observations and label as many of the following structures as possible from your section: epidermis, cortex, endodermis, stele, pericycle, xylem, phloem, and vascular cambium. Also, answer the following questions in the Results section of your lab notebook. a. b. How would you describe the arrangement of the vascular bundles in your root section?

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How does this arrangement compare to that of the stem section? Describe the similarities and differences in detail. Re-examine the herbaceous monocot plant you obtained from your stem analysis. Perform a cross section of the root and apply the Toludine blue and Phloroglucinol staining techniques as described from the plant anatomy lab. Draw your observations and label as many of the following structures as possible from your section: epidermis, cortex, endodermis, stele, pericycle, xylem, phloem, and vascular cambium. Also, answer the following questions in the Results section of your lab notebook

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a. How does the monocot root section compare to the dicot root section? Describe the key similarities and differences. Part 6: External structures of Roots Root hairs: Root hairs are elongate epidermal cells which vastly increase the surface area and thus the absorption capability of roots. As roots age and secondary tissue is produced, they no longer function. Therefore only the younger root branches have root hairs and are involved in water and nutrient absorption. If Red top grass seedlings are available, make a wet mount of 2-3 germinating seeds and observe. After an initial observation, stain the mount with acetocarmine. Observe the relative denseness of the cells and cytoplasm in various regions of the root. Try to distinguish among the meristematic region, zone of elongation, and zone of maturation. Note especially the root cap cells. Examine a fresh root system in lab and make a drawing of a typical root hair in the Results section of your lab notebook. Label all recognizable cell types and tissues. Part 7: Internal structures of Roots While stem branches are superficial in origin, root branches are derived from meristematic tissue inside the stele of the root. The point of derivation and pattern of development can be observed by sectioning roots of bean seedlings. Extract a bean seedling and carefully wash and clean the roots. Make a series of sections through a branching region. Select the better sections using a dissecting microscope and stain with acetocarmine in a small watch glass. Make a drawing of what you observe and any distinguishable cell types and tissues in the Results of your lab notebook. Finally make a careful observation and drawing of the transitional region between a typical stem and root anatomy. Describe characteristics that can be used to distinguish roots and stems. Questions to review and answer in Discussion section of lab notebook: 1. In the cross-section of the stem, which tissue type was more noticeable: xylem or phloem? Why do you think this is so? What types of cells provide support for an herbaceous stem? 2. Where did you typically find the primary meristematic tissue of the stem? What does this illustrate about the primary growth of a stem? In your Water Conduction experiment, how did the Safranin behave between the cut and the uncut seedling? Can you explain your results? Looking at your cross section of your bean seedling stem and root, what differences did you see between the two sections? Were you able to find all the same tissue and cell types? Explain. In the cross-section of the root, which tissue type was more noticeable: xylem or phloem? Why do you think this is so? Were the differences between monocot and dicot cross section greater for the root section or stem section? How do you explain this observation? What is the function of the endodermis of the root? Why is the endodermis important to living life on the land? Root hairs tend to wilt rapidly when removed from a moist environment. How does this apparent weakness relate to their function? Why dont stems and leaves have a similar problem? Do roots show secondary growth? Explain why or why not.

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5 Conclusion: Write a paragraph of two in the Conclusion section of your lab notebook about the adaptation of stems and roots as evolutionary keys for land plant colonization. Also describe advantages and disadvantages the differences between monocots and dicots might have in stem and root structure. Finally, draw conclusions on the experiments performed during this lab and how the results aided you in your analysis or stem and root function.