Handout- Lab 2 Division: Gymnomycota Scientific name: Physarum polycephalum (Genus species)  Common name: Slime mold Slime molds are intriguing creatures; they have characteristics of both fungi and animals. A slime mold produces fruiting bodies and spores typical of fungi but it also exists as a free-living mass of protoplasm or a plasmodium that changes form as it creeps along, engulfing bacteria and other organic matter like a giant amoeba. The plasmodium moves across the agar surface in a fan shape, secreting enzymes and ingesting its food. Within this fan-shaped area is a network of veins that carry the fluid protoplasm. The protoplasm moves in one direction for about a minute, then reverses its flow. This protoplasmic streaming is probably most associated with metabolism but also causes movement of the slime mold from place to place. The plasmodium is unicellular and multi-nucleate and is bounded by a cell membrane that is often enclosed in a slime sheath, which is shed as the plasmodium moves, leaving a white slime track in its path.  Slime molds occur in cool, moist, shady places like crevices of decaying wood or under leaf litter. Generally the plasmodia are easily spotted because they are a bright yellow, orange, or red. After a period of feeding and growth, the plasmodium moves into a drier, more exposed location and develops fruiting structures called sporangia, which are stalked balls or cups. If conditions become too dry before the slime mold is ready to fruit, the plasmodium will convert into a hardened resistant mass of tissue called a sclerotium, which upon return of favorable conditions, converts to free-flowing plasmodium again.  TO DO: 1. To view protoplasmic streaming, place the plate carefully on the stage of the COMPOUND scope and using the 10x objective carefully focus on a slime mold vein. DO NOT TOUCH THE OBJECTIVE TO THE PLATE.  a. Time the protoplasmic streaming and watch the flow reverse.  b. Speculate on why the flow reverses.  2. Is the slime mold pathogenic?? Why or why not?        Division: Mastigomycota  Class: Oomycetes Fungi in this division are water or soil inhabitants and produce asexual sporangia containing zoospores. Zoospores have one or more flagella with which they swim. The Oomycetes also produce thick-walled sexual oospores that act as the overwintering stage because they survive unfavorable conditions.  Several of the Oomycetes (Pythium spp. are examples) are saprophytic or parasitic on plants and commonly cause damping-off and root rot diseases of trees. Phytophthora infestans caused the historical potato late blight of Ireland in the 1800s.  Saprolegnia sp. is a common freshwater fungus and the main genus of water molds responsible for fungal infection of fish and their eggs. The asexual sporangia of this genus are elongate and look a lot like hyphae so they are difficult to see on a culture plate. TO DO: 1.  Make a slide from the Oomycete (Saprolegnia sp.) plate. Look for big round oogonia (the female sexual reproductive structure) and the oospheres inside. When fertilized by the male antheridia, the oospheres will be released as oospores.   2.  Familiarize yourself with the photos of Phytophthora sp.  Be able to distinguish the sporangia from the oospores.     Division: Amastigomycota  Subdivision: Zygomycotina Scientific name: Zygorhynchus sp. (Genus sp.)  Members of this genus are commonly found in soil. The asexual fruiting structure is the sporangium formed on a single stalk, the sporangiophore. The sporangium contains many small sporangiospores that are released when the sporangium wall disintegrates. Zygorhynchus is homothallic which means only 1 mating type is required to produce the sexual fruiting structure, the zygospore. The zygospore of Zygorhynchus is produced from the union of 2 gametangia, which are the structures containing male and female or + and - gametes, from the same mycelium. The zygospore is large and orange with a very warty surface.  TO DO: 1. Carefully place the plate on the stage of the COMPOUND scope and using the 4x and 10x objectives observe the fruiting structures. DO NOT LET THE OBJECTIVE HIT THE AGAR SURFACE.  2. Make a slide of the mycelia and spores.  a. Are the mycelia septate or aseptate?  b. Find and draw the sporangia.  c. Find and draw a zygospore. Note the 2 arms or suspensor cells holding the zygospore.  d. Can you tell the difference between a sporangium and a zygospore? What are the differences?  Division: Amastigomycota  Subdivision: Zygomycotina Scientific name: Phycomyces sp. (Genus sp.)  This genus produces stalked sporangia as asexual fruiting bodies.  Phycomyces is heterothallic, meaning it requires both a + and a - type of mycelium for compatible sexual reproduction and production ofzygospores. These plates were made by placing + sporangiophores on one side of the plate and - sporangiophores on the other side. Mycelia or hyphae grew from each side and where they met in the center of the plate, zygospores developed.  TO DO: 1. Using the dissecting scope, look at the asexual sporangia produced on sporangiophores. Draw a diagram.  2. Using the dissecting scope, look at the zygospores produced in the center of the plate where the + and - mycelia have crossed. Note the distinctive appendages on the Phycomyces zygospores.  Division: Amastigomycota  Subdivision: Deuteromycotina or Fungi Imperfecti Scientific name: Alternaria sp. (Genus sp.)  Alternaria may cause leaf spots on some plants. The fungus is usually saprophytic and may cause hay fever-like symptoms.  Alternaria produces chains of dark conidia on conidiophores.  TO DO: 1. Look at the plate under the dissecting scope and view the chains of black spores.  2. Make a slide and observe the muriform septation within each spore. Draw a diagram.  3. What type of spores does this fungus produce?  Division: Amastigomycota  Subdivision: Deuteromycotina or Fungi Imperfecti Scientific name: Fusarium sp. (Genus sp.)  Fusarium is commonly found in the soil. It is parasitic on many higher plants including trees.  The fruiting structure is the conidiophore, which produces macroconidia and microconidia.  TO DO: 1. Make a slide from the plate and look for macro and micro conidia. Draw diagrams.  2. Look at the mycelia on the slides. Is it septate oraseptate?  Division: Amastigomycota  Subdivision: Deuteromycotina or Fungi Imperfecti Scientific name: Aspergillus sp. (Genus sp.)  This fungus is found from the arctic to the tropics and feeds on an enormous variety of substances. It is commonly a problem as a lab and kitchen contaminant.  Aspergillus produces conidiophores and conidia, which is the fruiting stage most often found.  The perfect (sexual) stage of this fungus is Eurotium sp., which produces cleistothecia and ascospores.  TO DO: 1. Look at the plate under the dissecting scope. Observe the conidiophores of Aspergillus growing up from the agar and the round cleistothecia of Eurotium growing on the agar surface.  2. Make a slide of both fruiting bodies. Observe the conidia and conidiophores. Look at a crushed cleistothecium and observe the ascospores produced inside. The asci or sacs that contain the ascospores have already disintegrated.  3. To which subdivision does Eurotium sp. belong?  Division: Amastigomycota  Subdivision: Deuteromycotina or Fungi Imperfecti Scientific name: Phomopsis sp. (Genus sp.)  This fungus causes disease of many plants, including trees. You will meet it again in later labs.  Phomopsis produces pycnidia within which form 2 types of conidia:  Alpha conidia Beta conidia  TO DO:  1. Make a slide from some of the orange-yellow spore tendril coming from a pycnidium on the plate.  2. Look for the 2 types of conidia and draw and label them appropriately.  Division: Amastigomycota  Subdivision: Deuteromycotina or Fungi Imperfecti Scientific name: Sirococcus sp. (Genus sp.)  This fungus causes butternut canker.  Other members of the genus cause diseases of conifers.  You will meet it again in later labs.  Sirococcus produces pycnidia within which form 2-celled conidia:    TO DO:  1. Make a slide from some of the yellow spore mass coming from a pycnidium on the plate.  2. Look at the conidia under the compound scope and draw and label them appropriately.  Division: Amastigomycota  Subdivision: Fungi Imperfecti Scientific name: Leptographium sp. or Verticicladiella sp. (Genus sp.)  This fungus commonly causes a blue stain of conifer wood. It is one of the asexual stages of Ophiostoma sp. (Ceratocystis sp.). Leptographium produces conidiophores with branching ends on which the conidia are produced.  TO DO: 1. View the plate and conidiophores under the dissecting scope.  2. Make a slide of the conidiophores and observe under the compound scope. Draw a conidiophore. Can you see any conidia?  3. What is the other name for the subdivision to which this fungus belongs?  4. Using Chapter 4 of your text Principles of Forest Pathology, determine to which subclass this fungus belongs.  Division: Amastigomycota  Subdivision: Deuteromycotina or Fungi Imperfecti Scientific name: Pesotum sp. or Graphium sp. (Genus sp.)  This is another asexual stage of Ophiostoma sp. (Ceratocystis sp.). This fungus produces a special conidiophore called a synnema or coremium, which is a thick, black, multiple-stranded stalk upon which conidia are produced.  TO DO: 1. View the plate under the dissecting scope and observe the synnemata (plural).  2. Make a slide of a synnema and compare it to the conidiophore of Leptographium. How do they differ? Draw a diagram.  3. What type of spores does this fungus produce?  Division: Amastigomycota  Subdivision: Ascomycotina Scientific name: Ceratocystis sp. or Ophiostoma sp. (Genus sp.)  This fungus cause blue stain of conifers and vascular wilt diseases in certain hardwoods such as oak and maple. It produces long-necked black perithecia, which produce ascospores.  TO DO: 1. Observe the plate under the dissecting scope and view the perithecia.  2. Make a slide and observe the perithecia and ascospores. Draw the perithecium. If you made the slide carefully, you will see the small round ascospores coming out of the top of the long neck of the perithecium.  3. Also you may find endoconidia on the slide. This is another asexual stage of Ceratocystis. Endoconidia are rectangular spores produced in endoconidiophores:  Draw some endoconidia being released from an endoconidiophore.  Division: Amastigomycota  Subdivision: Ascomycotina Scientific name: Sordaria sp. (Genus sp.)  This fungus is commonly found growing on dung. Sordaria produces perithecia, which contain asci and ascospores.  TO DO: 1. View the perithecia on the plate under the dissecting scope.  2. Make a slide of a perithecium and observe the asci containing large dark-colored spores. How many spores do you see in each ascus? What type of spores are they?  3. What is the most likely function of this fungus?  4. Using Chapter 4 of your text Principles of Forest Pathology, determine the order and family to which this fungus belongs.  Division: Amastigomycota  Subdivision: Basidiomycotina Scientific name: Schizophyllum commune (Genus species)  This is a common wood rotting fungus that produces a gilled basidiocarp which produces basidiospores. The gills may be facing up in the plates if the plates were stored upside-down.  TO DO: 1. Compare the lab culture to the natural formation on the wood sample. Note the gills (spore producing surfaces) on both basidiocarps.  2. What type of spores are produced?  Division: Amastigomycota  Subdivision: Basidiomycotina Scientific name: Agaricus sp. (Genus sp.)  This is the common edible mushroom sold in grocery stores.  TO DO: 1. Look for the parts of the mushroom. Draw a diagram and label the cap, stipe (stem), and gills.  2. Make a slide of a thin section of a gill and look for the basidia and basidiospores. Draw a diagram of a basidium and label the sterigma and basidiospores.  3. How many basidiospores are typically produced per basidium?  Symptomology I  Can you find the:  1. Leaf spot?  2. Canker?  3. Gall?  4. Tip blight?  Symptomology II  Can you find the:  1. Marginal chlorosis?  2. Interveinal necrosis?  3. Purple stippling?  4. Marginal necrosis?  5. Tip necrosis?