Name: ____________________ Chapter 11: Protists Revised: Spring 2007 Learning Objectives: Gain an appreciation of the diversity of organisms considered to be “protists.” Describe differences between the major kingdoms of protists. Understand some of the different methods used by protists for obtaining nutrients and for moving about in the world. Explain the key role that protists play in supporting life in aquatic communities, as well as sustaining human life on the planet. Understand what information is important for classifying organisms. Contents: Introduction ............................................................................................................... 146 Ecological Importance of Protists ............................................................................. 147 Exercise 1: Protozoa ................................................................................................. 148 Ciliated Protozoa ........................................................................................... 148 Paramecium ....................................................................................... 148 Stentor................................................................................................ 150 Amoeboid Protozoa ....................................................................................... 151 Exercise 2: Algae ...................................................................................................... 152 Green Algae ................................................................................................... 153 Chlamydomonas ................................................................................ 153 Volvox ................................................................................................ 154 Spirogyra ........................................................................................... 154 Ulva ................................................................................................... 155 Diatoms.......................................................................................................... 155 Dinoflagellates ............................................................................................... 156 Brown Algae .................................................................................................. 157 Red Algae ...................................................................................................... 157 Euglenoid Algae ............................................................................................ 158 Exercise 3: End of Lab Questions ............................................................................ 159 Exercise 4: Protist Review Worksheet ..................................................................... 160 protists: 145 INTRODUCTION Protists are eukaryotes, but beyond that, they are difficult to classify. The group of organisms that fit into the “protist” category has more than 7 kingdoms and 10,000 species. About the only thing they all have truly in common is that they are all eukaryotes. They also are all similar in having a less complex organismal organization than the fungus, plants, or animals. To make more sense of this catch-all group, it is divided into three major groups. These groups are: 1. The protozoa - single celled animal-like protists. 2. The algae - plant-like protists. 3. Slime molds - fungus-like protists. Note that these are also informal group names, like “protist,” and not official classification taxa. You already studied protists a bit during the Diversity Lab at the start of this semester. You will continue to study protists here and learn about some of the formal Kingdom and Phylum names that may (or may not) remain in place. However, in this lab, we will restrict our further study to the protozoa and algae, and we will not be viewing examples of slime molds. Why is it that we have so much trouble classifying these organisms? You tried to classify many of them during the Diversity Lab, but may remember that it wasn’t easy. The quick answer as to why it is so difficult is because we have only recently developed the appropriate methods to classify them properly! You see, years ago, people came up with a classification scheme based on readily observable organizational features. Before the 1900s, we classified all living things as either plant or animal. Then we observed fungus more closely and realized that they were too different from plants to be grouped with them. As our microscopes improved, we learned about microscopically-small organisms, and tackled grouping them as well; this was when the name “protists” arose. By the mid-1900s, we had learned that there were different types of cells, and by the later 1900s we determined that there were different kinds of prokaryotic organisms (thus, Domain Archaea and Domain Bacteria were named). However, observable organismal characteristics are not the only ones we can now use. We can now directly compare the DNA from different organisms to decipher their relatedness. The more similar the genetic code between two species, the more closely related they are. It turns out that not all organisms that look similar are closely related! You can probably relate to this notion if you think about the fact that while some human siblings look similar, others don’t look at all alike—and yet they are still siblings and very closely related. It is the genetic code of the organisms that provides the key to understanding how related different organisms truly are. We have only learned how to obtain the genetic code of organisms quickly and easily in the last 15 years with the international effort made on the Human Genome Project. Therefore, now we have a lot of new data to look at and re-evaluate organismal relatedness. This new information is going to help us shape our taxonomic classification, and make it more accurate. Until that time when we know exactly how many kingdoms lie within the informal grouping called the protists, we will have to just do our best to try to understand these organisms. protists: 146 Ecological Importance of Protists While responsible for causing many dreaded human diseases (malaria, sleeping sickness, and giardiasis to list a few), protozoa also benefit humans tremendously. Protozoa are important nutrient recyclers in natural ecosystems. Without organisms like bacteria, fungi and protozoa, soils would quickly become depleted of nutrients making plant growth impossible. All organisms on earth depend on healthy plant populations for survival. Protozoa also play a key role in wastewater treatment plants by consuming enormous quantities of bacteria, and are important components of food chains. Algae is much more than simply "seaweed" or "pond scum"--it is a vital component of our biosphere, and the survival of humans would be jeopardized without it. We often hear of the rain forests as being important in making oxygen, and they are important in many ways, but most of the oxygen we actually breathe is made by algae in the ocean. Indeed, algae produced more than 50% of the oxygen molecules that your cells consumed during this laboratory! Algae also form the base of aquatic food chains. This algae is primarily microscopic, and is called phytoplankton. Without algae in lakes, rivers, and oceans, there could be no life in these environments since all aquatic life either directly feeds on algae, or feeds on organisms that feed on algae. Humans usually eat from the top of the food pyramid. Without phytoplankton in the oceans, salmon, tuna, cod, shrimp, lobsters and a myriad of other seafood products would not be available to us. An Aquatic Food Chain 1000 pounds of algae (phytoplankton). 100 pounds of zooplankton. protists: 10 pounds of small fish. 147 1 pound of large fish for dinner. Exercise 1: The Protozoa Protozoa are single-celled, heterotrophic organisms. Although most are single celled, that one cell is still able to carry out all the functions necessary for life: ingesting and digesting food, expelling waste, and obtaining oxygen. Protozoa live just about anywhere that is moist, on both land and sea, but in this lab we will primarily be looking at protozoa that live in fresh water environments. Protozoa that live in fresh water face a problem; compared to the surrounding water the high concentration of solute in their cells creates an osmotic gradient. Which way do you think the water moves in this gradient? Protozoa respond to this problem with a contractile vacuole. The contractile vacuole is a cellular structure that constantly pumps water out of the cell. There are so many different protozoa, it’s not easy to divide them up into species. The current approach is for taxonomists to classify protozoa according to their method of movement, and it is the mode of locomotion on which we will base our study of the protists today. Contractile vacuole in a paramecium. Note the array of channels that carry water to the central vacuole. Ciliated Protozoa Ciliates are protozoa that move and obtain food by means of short, hair-like structures called cilia. It is thought that they fit into Kingdom Alveolata. A cilium (singular) extends from the cell membrane and is composed of bundles of microtubules. In most ciliates, cilia cover the entire cell surface. Protozoa use their cilia for two purposes: 1. Locomotion 2. Feeding. The cilia sweep smaller protists, bacteria, and non-living organic matter into an oral groove on the cell surface. At the far end of the oral groove, a food vacuole forms. Ciliates occupy a variety of habitats including freshwater, soil and as symbiotes in the guts of invertebrates. Study of Paramecium - Prepare a wet mount of living Paramecium. Add methyl cellulose to your wet mount to slow the Paramecium down. View using 10x magnification. Paramecium will be gray or tan and will move quickly in a spiraling manner. Find a Paramecium that cannot move and observe this individual under 40x magnification. protists: 148 Questions about Paramecium. Do the cilia beat in a synchronized pattern, or does each just beat on its own? ________________ Explain why Paramecium’s body spirals as it moves through the water? ____________________________________________________________________________________ ____________________________________________________________________________________ ____________________________________________________________________________________ Look for contractile vacuoles within each cell. What collects within the contractile vacuoles of the cell? __________________________________________ Why are contractile vacuoles are needed by these organisms (hint: think in terms of osmotic gradients)? ____________________________________________________________________________________ ____________________________________________________________________________________ ____________________________________________________________________________________ Please explain what might happen to Paramecium if the contractile vacuoles stopped functioning? ____________________________________________________________________________________ ____________________________________________________________________________________ ____________________________________________________________________________________ The following two graphs show how many times Paramecium’s vacuole might contract in pond waters of varying saltiness. One graph is correct and the other is not. Circle the correct graph. Explain your choice on the next page. Circle the correct graph. 6 6 5 5 Graph A Contractions 4 per minute. Graph B Contractions 4 per minute. 3 3 2 2 1 1 0 1 2 3 0 4 1 Less salty More salty Saltiness of pond water. protists: 2 3 4 Less salty More salty Saltiness of pond water. 149 Did you circle Graph A or Graph B on the previous page? ______________________________ Please explain your choice. _____________________________________________________________ ____________________________________________________________________________________ ____________________________________________________________________________________ ____________________________________________________________________________________ Sketch a Paramecium in the space below the way it looks to you. Label your sketch. Study of Stentor: Prepare a wet mount of living Stentor. Add methyl cellulose to your wet mount to slow the movements of these very large organisms. You will be able to see these using only your scanning power objective. Once you find one, increase your magnification to answer the questions below. Food vacuoles should be apparent in these organisms. Stentor Are the cilia the same all over the cell, or are they more dense in some areas. Explain. ________________________________________________________ _________________________________________________________________________________ Sketch a Stentor in the space below the way it looks to you. Label your sketch. protists: 150 Amoeboid Protozoa Amoeboid protozoa are a large group that includes the amoebas. Amoebas move using pseudopodia. Pseudopodia are protoplasmic extensions that pull the organism in a particular direction. Amoebas also use these "false feet" to capture food through a type of endocytosis known as phagocytosis. Most freshwater environments include harmless species of amoeba, and it is one of these we will be observing today. In addition to slow moving streams and ponds, Amoeba species also inhabit soil and saltwater and a few even live as parasites in the digestive systems of humans and other animals. Study of Amoeba - Make a wet mount of Amoeba. Using your scanning objective, scan your slide for a grayish, irregular-shaped object. Reducing your light intensity may help you find this organism. Don’t be fooled by dead vegetation, which is yellowish brown, the amoebas you are looking for are more grayish black. If after a few minutes you have still not located an amoeba, rinse your slide and try another drop. Persistence is the key to finding these organisms! Note the irregular shape, the slow movement (streaming) of the protoplasm, and the formation of pseudopodia. List 2 functions of pseudopodia __________________________________________________________ ____________________________________________________________________________________ Do you see any food vacuoles? If so, describe their appearance. ______________________ ___________________________________________________________________________ Do you think it is likely that amoeba require contractile vacuoles? Please explain your answer. ___________________________________________________________________ __________________________________________________________________________ __________________________________________________________________________ Sketch an Amoeba in the space below the way it looks to you. Label your sketch. protists: 151 Exercise 2: The Algae Algae are autotrophic (photosynthetic) organisms. Like the protozoa, some algae are single-celled. Unlike the protozoa however, many algae are multicellular and may be quite large. Most algae live in water, but they may also be found in any place that is moist at least part of the time. Algae are around us all the time and can be found on shady brick walls, the fur of some mammals, in swimming pools, birdbaths, flowerpots, and gutters on the roof. Over 30,000 species of algae have been described by Phycologists (Phycology is the study of algae). Like the protozoa, taxonomists need a way to divide up all these species. The way they do it is by color and by how complicated the algae is. Divisions of algae by color. Different species of algae use different types and mixes of chlorophyll and this leads to their being different colors. Based on color, there are four main groups of algae. They are: A. Green algae B. Brown algae C. Golden-brown algae D. Red algae Look at the seaweed the next time you’re at the beach. The four colors are usually pretty easy to find. Divisions of algae by complexity. In addition to being different colors, some algae are more complex than others. Algae can be divided into the following four categories based on the complexity of the organism: A. Unicellular algae Consist of only one cell living on its own. B. Colonial algae Consists of a colony or cluster of more or less identical cells. Each cell could live on its own, but they choose to live together. C. Filamentous algae Consists of a long chain of more or less identical cells. Again, each cell could live on its own, but they choose to live together. D. Multicellular algae These are diverse, since they may be very simple (where all the cells look similar) or they may be more complex (and have specialized cells). Large seaweed, for example, has special cells that form rootlike holdfasts to hold on to rocks. protists: 152 Groupings of algae by genetic comparisons into Kingdoms A. Kingdom Euglenozoa These algae are motile (using flagella) and can either eat like a protozoan or carry out photosynthesis. B. Kingdom Stramenopila This group contains the brown algae, the diatoms and other groups. C. Kingdom Rhodophyta This group contains the red algae. D. Kingdom Alveolata Although we saw this kingdom back in the protozoa section as the group that contains ciliates, it also contains the dinoflagellates. E. No Green Algae! The green algae have been temporarily re-assigned into the Kingdom Plantae as Division Chlorophyta. These organisms may stay in with the plants, or may return to the group of protists as their own kingdom. Question: Which of these products contains algae? Answer: They all do! Green Algae Chlamydomonas Chlamydomonas is algae, but it has an eyespot to detect light and flagella to swim toward the light. It’s a good example of how it is hard to fit protists into categories. Each cell also has a granule of stored starch and two flagella. Examine a living specimen or prepared slide of Chlamydomonas beneath your microscope. Note their small size. Identify flagella and the starch granule if possible. To which of the 4 color categories does Chlamydomonas belong? __________________________ To which of the 4 complexity categories does Chlamydomonas belong? __________________________ protists: 153 Volvox Each of the cells in Volvox also has a flagella. Flagella are beat by individual cells in synchrony with other cells. This allows the whole Volvox to move through the water in an orderly manner. Prepare a wet mount of Volvox. If available, use a depression slide for Volvox so it is not crushed by the coverslip. Note the daughter (reproductive) colonies that may be seen inside some Volvox. Volvox To which of the 4 color categories does Volvox belong? __________________________ To which of the 4 complexity categories does Volvox belong? __________________________ Sketch a Volvox in the space below the way it looks to you. Label your sketch. Spirogyra Spirogyra is what we may think of as "pond scum", since it forms thick mats on the surface of ponds when conditions are right. Spirogyra takes the form of thread-like strands called filaments. Each filament is composed of many cells linked end to end. Spirogyra gets its name from the fact that the chloroplasts inside the cells are arranged in distinctive spirals. Prepare a wet mount of Spirogyra and examine beneath the microscope. Note the characteristic spiral-shaped chloroplasts. Look for a nucleus within each cell of the filament. The nucleus is suspended in strands of cytoplasm, much like a spider might be found sitting in the center of a web. Spirogyra Note the filamentous shape of the algae. Are the filaments branched? _________________ Can you see cell walls separating the individual cells of the filament? _________________ To which of the 4 color categories does Spirogyra belong? __________________________ To which of the 4 complexity categories does Spirogyra belong? __________________________ protists: 154 Sketch Spirogyra below and label any visible features. Ulva An abundant marine species of algae is "sea lettuce" or Ulva. The body or thallus of Ulva includes a tiny root-like holdfast that anchors the algae onto rocks or other substrate on which it is growing. The thallus of Ulva is only several cell layers thick. Observe the living specimens of Ulva on display in the lab. To which of the 4 color categories does Ulva belong? __________________________ To which of the 4 complexity categories does Ulva belong? __________________________ Diatoms Diatoms usually have very small cells and are found in abundance in both terrestrial, marine, and freshwater environments. The defining characteristic of diatoms is the presence of their glass-like cell walls or frustules composed of silica. The walls are arranged in overlapping halves, much like candy box or petri dish fits together. Electron microscope picture of a diatom's frustule. An incredibly complex shell for a creature that is only one cell! a Once the organism dies, the cell inside decomposes but the glasslike frustule remains intact for hundreds or thousands of years. Over thousands of years significant deposits of frustules may accumulate on the ocean floor. These deposits may then be mined to provide diatomaceous earth. Humans use this material in a wide array of products, including paints, cement, fertilizers, toothpaste, polishes, and cleaners. Diatomaceous earth also kills snails and slugs. Examine the diatomaceous earth on display in lab. Examine prepared slides of diatoms or, if available, prepare a wet mount and examine sediments for both living and dead diatoms. Note the many shapes. To which of the 4 complexity categories do Diatoms belong? ________________ protists: 155 Sketch a few diatoms below, in the area between the photos of diatoms: To which of the 4 color categories do Diatoms belong? ________________ To which of the 4 complexity categories do Diatoms belong? ________________ Dinoflagellates Dinoflagellates are unicellular algae that move with two flagella. Because of the positioning of their flagella, they tend to whirl about in the water. The prefix “dino-” means whirling, so they are named according to the way they move. Although dinoflagellates move, they are also autotrophic. They are also algae because they have very interesting cell walls made up of many plates like armor. Their two flagella have to stick out through grooves in their cell plates. Observe the living specimens of Peridinium available in the lab. To which of the 4 complexity categories does Peridinium belong? __________________________ protists: 156 Brown Algae Laminaria and Fucus are types of kelp. Kelps are typically dark brown in color and are only found in marine environments. They generally prefer cooler oceans over warm. Each individual kelp is composed of a basal holdfast, which anchors the organism, blades which look and act like leaves, and a stalk-like stipe. Keep in mind though, that kelp are not plants because they lack true stems, leaves and roots. Distributed along the blade may be several or many air bladders that help to keep the blade buoyant. A thick growth of kelp can create an underwater forest along many temperate shores. Kelp forest are extremely important ecologically because they create an environment where marine mammals, fish, and invertebrates can hide, raise their young, and seek protection from storms. Observe living specimens of kelp on display. Make a sketch in the space below. Include and label on your sketch: Holdfast, blade, and stipe. If time allows, make a wet mount of a small section of the blade, and draw the cells you see below as well. To which of the 4 color categories does kelp belong? __________________________ To which of the 4 complexity categories does kelp belong? __________________________ Red Algae Callithamnion is a filamentous algae of the type that can often be found floating in the surf along New England shores. A commercially important species related to Callithamnion is Irish Moss, which is a source of the carageenan used as a thickener in many food products. You may also have the opportunity to see Chondrus crispus, which is a larger multicellular organism. Observe the living specimens of Callithamnion on display in the lab. Note the filamentous shape of the algae. Are the filaments branched? _________________ Can you see cell walls separating the individual cells of the filament? _________________ To which of the 4 color categories does Callithamnion belong? __________________________ protists: 157 To which of the 4 complexity categories does Callithamnion belong? __________________________ Sketch Callithamnion in the space below the way it looks to you. Label your sketch. Euglenoid Algae Study of Euglena - Euglena are freshwater protists that exhibit characteristics of both protozoa and algae. Algal characteristics include the ability to photosynthesize and store starch. Protozoan characteristics include the lack of a cell wall, and the ability to ingest food. Some species of Euglena are very tolerant of polluted waters, and act as water quality indicators for humans when expensive pollution monitoring equipment is not available. Prepare a wet mount of Euglena. Methyl cellulose will help slow these fast-moving organisms. Reduce the light with the iris diaphragm and work up to your 40x objective. Find an individual that cannot move and note the movement of the flagellum. Look for the red stigma or “eyespot” near the flagellum. This structure allows the organism to sense different light intensities. Note the very flexible external membrane or pellicle, as well as green coloration of the organism that indicates the presence of chloroplasts. Why is a structure such as the stigma important for photosynthetic organisms like the Euglena? ____________________________________________________________________________________ Do you think Euglena require contractile vacuoles? Explain. _______________________ ____________________________________________________________________________________ Chlamydomonas is a lot like Euglena in many ways. There is another organism we have seen today that also shares some similarity to Euglena. What other organism has traits in common with Euglena and what are the traits? ___________________________________________________________________________________ ___________________________________________________________________________________ ___________________________________________________________________________________ To which of the 4 complexity categories does Euglena belong? __________________________ Note that you were not asked to put Euglena into a color category… that is because this organism is NOT a green algae, but is in a different phylum, the Euglenoids. Sketch a Euglena in the space below the way it looks to you. Label your sketch. protists: 158 Exercise 3: End of Lab Questions You may have to consult other sources of information to correctly answer some questions. 1. Approximately 100,000 species of protists have been identified and classified. Most taxonomists, however, believe that this number is low and that many thousands more await discovery by humans. Explain why you think it is so hard to find new species of protists. 2. List functions of the following: Pseudopodia, flagella, chloroplast, stigma, holdfast, cilia, food vacuole, contractile vacuole. 3. Suppose that as the result of extensive pollution to the ocean, kelp forests are killed. Explain how this would affect not only kelp populations, but also many other forms of marine life. 4. Why are protozoa not classified into the animal kingdom? List 2 reasons. 5. Why are algae not classified into the plant kingdom? List 2 reasons. 6. What is agar? Carrageenan? Algin? 7. List 3 ways an amoeba is similar to a human. 8. List 3 ways humans and Paramecium differ. 9. Protozoa are: Heterotrophic or Autotrophic? Define your answer. 10. On your first day on the job as Naturalist at Cape Cod National Seashore, an angry visitor approaches you complaining about all the "good fer nuthin seaweed" littering the beach. In a paragraph, explain to the visitor 5 ways humans benefit from algae. 11. If both protozoa and most bacteria are single-celled and heterotrophic, why are they classified into separate kingdoms? Explain. 12. Paramecium possesses unique organelles called trichocysts. See if you can find out what these are and their function. 13. Suppose that as a result of a genetic mutation a particular freshwater protozoan loses the use of its contractile vacuole. Is this organism likely to survive, reproduce, and pass this characteristic on to its offspring? Explain. protists: 159 Exercise 4: Protist Review Worksheet protists Protozoa Brown Algae Dinoflagellate Multicellular Euglenoids UniColonial FilaMulticellular mentous cellular Diatoms UniUniUnicellular cellular cellular Red Algae Green Algae Flagellates Ciliates O r g a n i s m Amoebas G r o u p i n g Algae MultiUniUniUniMulticellular cellular cellular cellular cellular no example given Organisms you may have seen today: protists: Spirogyra Chlamydomonas Euglena Stentor Chondus Crispus 160 Amoeba Fucus or Laminaria Paramecium Callithamnion Volvox Ulva Diatoms Peridinium Put each organism you saw into an organism box above to organize all that you viewed! protists: 161