FUNGAL DIVERSITY AND LIFE CYCLE

INTRODUCTION: The fungal functional type is comprised of sessile heterotrophs with cell walls. Rather than ingesting food as animals do, fungal organisms absorb food across the cell wall. The assemblage of organisms termed fungi are classified into two general categories. First, are the true fungi (Kingdom Fungi) which evolved from motile, aquatic protozoa that are also ancestors to the animal kingdom. True fungi first evolved as the chytrids (Phylum Chytridiomycota) who produce an enlarged globular cell from which numerous filaments grow into the food source. Chytrids produce motile spores and gametes, and the vegetative cells are ceonocytic (many nuclei float around in one big cell). Chytrids gave rise to the Zygomycetes (Phylum Zygomycota), which produce no motile cells but form ceonocytic hyphae. From the Zygomycetes, the advanced fungi arose and in time formed the Phylum Dikaryomycota. Organisms in the Dikaryomycota produce hyphae comprised of individual nucleated cells separated by walls (septate hyphae) with each cell having two haploid nuclei in what is called an N + N configuration. The two main groups of dikaryotic fungi are the ascomycetes (sac-forming fungi) and the basidiomycetes (club-forming fungi). The majority of fungi affecting humans are ascomycetes and basidiomycetes.
The second category of fungal organisms is the pseudofungi, made up of various unrelated protista groups. The pseudofungi were formerly classified into the catch-all kingdom Protista but have recently been reclassified into more-specific kingdoms that reflect genetic relationships. Important pseudofungi are the Oomycetes (egg fungi and water molds), and the slime molds. Oomycetes are closely related to the stramenopilous algae - the brown algae, golden-brown algae and diatoms of the Kingdom Stramenopila. The close relationship between the oomycetes and the brown algae is evident in that both have cellulose walls, and they share the same type of flagella. Oomycetes descend from algae that lost their chloroplasts, and hence have adopted a heterotrophic life form. Oomycetes also produce filamentous hyphae to better absorb nutrients from the food source. Because they have no relationship with the chytrids, it is clear that the oomycetes and chytridiomycetes independently evolved the mycelial life form.
Slime molds evolved from various ancient protozoa and have little genetic affinity to other fungi or algae groups. They are animal-like in that they ingest food early in the life-cycle, but are fungal-like in that they produce walled sporangia and spores.
In today’s lab, you will have a chance to examine the diversity of both false and true fungi. The lab will begin with the false fungi and then follow an evolutionary sequence through the true fungi. Many specimens illustrate important reproductive phases in the life-cycles of these organisms. You should examine these carefully because reproductive features are important to understanding and distinguishing the various groups of fungi.
Although not required, we urge you to draw and label the specimens you examine. Our experience has been that drawings with appropriate labels are the best way to learn the features emphasized in lab and lecture. Drawings are also an excellent study tool to refresh your memory just prior to the exam
PART I: THE PSEUDOFUNGI
A) Slime molds (Kingdoms Myxomycota and Dictyosteliomycota): Slime molds are largely saprophytic and are typically found on decaying wood in moist forests. During the vegetative phase of the life cycle (see figure 16-6 on page 353 of your text), they begin life as independent amoebae, ingesting microscopic bits of organic debris. The free-living amoebae eventually swarm together to produce a multicellular blob called a plasmodium. After a while, the plasmodium forms cellulose walls around the nuclei and produces sporangia (or fructifications). The sporangia release large numbers of air-borne spores which germinate in the presence of water to form free-living amoebae, thus completing the life cycle. Slime molds defy simple categorization. They are animal-like in that they ingest food during the amoeboid- and plasmodial-phase. They are plant-like in their formation of cell walls and sporangia during reproduction. Because of the cell-walls formed during the reproductive phase they are considered fungal in nature. There are two major types of slime molds:
A) the Myxomycota are the acellular, or true plasmodial slime modes: The plasmodium stage of this group is made up of large blobs of ceonocytic protoplasm with many nuclei inside.
B) the Dictyosteliomycota are the cellular clime molds, where the plasmodium is made of individual cells separated by membranes (but not cell walls).
Observations on Display: Fruiting bodies of slime molds are readily found in Ontario forests in the autumn. Some will be on display for you to examine, along with a diagram of the life cycle.
B) Oomycetes: the water molds, or egg fungi (Kingdom Stramenopila)
Oomycetes are characterized by the formation of large egg-bearing cells on the tips of specialized hyphae termed oogonia (see figure 17-4 on page 374 of your text). Large, non-motile eggs form inside the oogonia and are fertilized by male-like hyphae termed antheridia. The antheridia grow into the egg and deposit the male gametes, which then fuse with the egg to form a zygote, termed the oospore. The oospore undergoes mitosis and forms a sporangia. The spores that are produced disperse to infect leaves, seedlings, fish and dead organisms.
Oomycetes are important saprophytes in aquatic habitats. In terrestrial habitats, they are generally parasitic. Important diseases caused by water molds are downy mildews (Peronospora), potato late blight (Phytophthora infestans), and damping off disease (Pythium spp.). We will examine three species, Achlya, a saprophytic water mold; Albugo candida, the white rust of mustard plants and Phytophthora infestans.
Examine the following cultures with the dissecting microscope:
1. Achlya whole mounts: Achlya is a water mold that grows on organic debris in lakes and rivers. It forms floating mycelia mats arising from the food material, and in these mats sexual reproduction occurs. On your bench are prepared slides for you to examine with the light microscope. Focus on the blue-green material in the center of the slide. This is a hyphal mat with oogonia.
Observe the large conspicuous oogonia within the mycelium. Inside the oogonia you can see zygotes (oospores), that will later divide to form sporangia. Examine the oogonia closely to see if additional hyphae are attached to it. These would be antheridia, which present the sperm nucleus to the egg cells
within the oogonia. Note the properties of the vegetative hyphae. Do you see cross walls, or are the cells continous within a filament?
2. Phytophthora infestans: This organism causes potato late blight, one of the worst crop diseases in the history of humanity. In the 1840’s, Phytophthora infestans was introduced to Europe from Peru. It rapidly spread across the continent, destroying much of the potato crop. In Ireland, peasants were particularly dependent on the potato for survival, and the crop losses in 1845-1848 killed millions of Irish and forced the migration of millions more to North America. In continental Europe, the loss of the potato crop led to widespread economic failure and social revolt. Many of the radical worker movements that would later influence world history, such as Marxism-Leninism, arose in the wake of the food crisis caused by the Phytophthora outbreak.
Examine the Phytophthora slide prepared fresh from a culture stained with cotton blue. Identify the round oogonia in the slide and examine the hyphae for cross walls between the cells. Are there any oospores within the Oogonia? Can you see any antheridial hyphae attached to the oogonia? If you do, show other students in your group.
3. Albugo candida (White Rust of Mustards): Albugo infects plants of the mustard family, forming white pustules on the leaves. These pustules are comprised of many asexual conidiospores bursting through the plants epidermis. Inside the plant, fungal hyphae form oogonia and antheridia, which will mate and form oospores. The oospores develop sporangia which disperse genetically-distinct spores. The two prepared slides show the extent of the infection by the parasitic white rust fungus.
Examine the prepared cross section showing Albugo conidiaspores bursting through the epidermis of a mustard fruit. Note the strings of conidiospores forming under the epidermis. The bulge formed by the mass of conidia produce the rust pustule. Upon rupturing, the conidiospores are released on the wind to start a series of new infections.
Examine the prepared slide of the Albugo sexual organs (oogonia and antheridia). The oogonia are evident as dense, red-staining circles among the cells of the leaf tissue.
Notes and Drawings:
PART II: THE TRUE FUNGI (KINGDOM FUNGI)
We have assembled a range of specimens from the Kingdom Fungi, and they are arranged for you to examine beginning with the most primitive (the Chytrids) and then progressing through the Zygomycetes to the Ascomycetes and Basidiomycetes.
A. CHYTRIDIOMYCOTA (The Chytrids):
Chytrids are mainly aquatic or parasitic. They are important decomposers of pollen, dead insects and seeds that fall into ponds and rivers. Others are parasites of algae, higher fungi, mosquitoes, rotifers, and water molds. The general body plan is to form a large, central ceonocytic globule that either directly invades the body of the host, or produces diminutive hyphae termed rhizomycelium, which invade the surface cells of the host. The rhizomycelium grows into the food source and absorbs nutrients across the chitenous cell wall.
Chytrids are difficult to maintain in culture and have to be baited from natural sources. We have purchased slides showing a common parasitic chytrid, Synchytrium, invading a plant host, and have prepared a fresh culture of chytrids on snake skin. Chytrids are important decomposers of animal epithelial tissue in aquatic habitats. To capture them, we have placed strips of snake skin in pond water. Chytrid spores swim to the snake skin, and then germinate on the skin, forming a simple globular body with rhizomycelium.
Examine:
Synchytrium: Examine the prepared slide of Synchytrium that has infected either leaves or potato tubers. Note the simple globular body (the sorus) of the chytrid embedded in the tissue of the plant. Some of the globules may have matured into sporangia, and you may see spores inside.
Chytrids on snakeskin: In the dissecting scope, you can see the rhizomycelia extending from the snake skin, along with many protozoa. Note the pinhead-like cells within the mycelia. These are either the central globules from which multiple filaments of rhizomycelium arise, or they are sporangia.
In the light microscope, examine the globular cells and the hyphae growing away from them. The globular cell and rhizomycelium form the basic body plan of the saprophytic chytrids.
You will also see many tiny creatures swimming about the mycelium. Some of these may be zoospores released from sporangia. Examine the mycelium for any evidence of sporangia. Sporangia will be apparent as they will have only one hyphae attached to them.
B. ZYGOMYCOTA:
The Zygomycetes are important saprophytes, including species that are major decomposers of dung and food. Members of this group have a zygotic lifecycle (see figure 15-11 on page 316 of your text). The gametes are non-motile, and are born on the tips of specialized, fertile hyphae termed gametangia. The gametangia contain many haploid nuclei. In zygomycetes, the sexual act consists of two fertile hyphae growing towards each other. As they approach each other the ends of the hyphae form gametangia. The gametangia come in contact, after which the end walls disintegrate, releasing the haploid gamete nuclei into one common space. Pairs of haploid nuclei then fuse, creating many diploid nuclei.


The nucleate cell formed from the residuals of the two gametangia is termed a zygosporangium. Zygosporangia typically develop a thick wall that protects the diploid nuclei from harsh conditions, forming a many nucleate (ceonocytic) resting cell. Zygosporangia germinate when the diploid nuclei undergo meiosis to produce many new haploid nuclei. The haploid nuclei are walled off into distinct spores, which are released from a dispersal sporangium that grows out of the zygosporangium.
The most commonly encountered zygomycetes are the bread molds, which are important saprophytes that grow on carbohydrate-rich foods, including bread. The mycelium formed on the surface of the bread is a cottony mass that is initially white but soon darkens as the mycelium forms asexual sporanagia. Large numbers of mitospores are released, allowing the fungus to quickly spread.
Examine the following, first under the dissecting scope and then with the light microscope:
1. Zygorrhynchus moelleri: This mold growing on an agar plate shows the major stages of a typical Zygomycete life cycle. First examine the culture under the dissecting microscope, and then take a very small scraping of the agar with a dissecting needle or knife. Place this on a microscope slide, stain with the cotton blue stain on your bench, and examine at medium power with both phase contrast and normal visible light.
With the dissecting scope, examine the cottony matrix of the mycelium, and the sporangia rising above it, forming dark spheres on elongated stalks. These are mostly mitosporangia used in asexual reproduction. You may be able to see zygosporangia mixed in amongst the mycelium. They will be dark, barrel-shaped granules on the surface of the agar.
With the light microscope, observe the slide of agar, note the clear tubular nature of the hyphae and the absence of cross walls. Next observe any mitosporangia you may have opened while pressing down the cover slip.
Finally, observe the zygosporangia. These are dark, barrel-shaped structures with rough walls. Note the hyphae attached to the zygosprangia. These are the stalks of the gametangia, and are termed suspensors.
2. Rhizopus stolonifera: This is the common bread-mold, a regular feature of most pantries. We have provided you with a Petri plate of Rhizopus to examine. Note the following with the dissecting scope under high power:
Examine the mycelium and sporangia. You may be able to see elongated, horizontal hyphae connecting the sporangial stalks. Rhizopus spreads by these elongated hyphae, termed stolons (after the strawberry runners of the same name). Where stolons settle onto a food source, they produce anchoring hyphae that penetrate the food. Sporangia form above this contact point. This habit allows for rapid spread of Rhizopus over a loaf of bread. Typically, Zygomycetes reproduce asexually by mitospores when conditions are good, allowing for rapid spread over a new food source. They switch to sexual reproduction when the food is exhausted and conditions deteriorate.
3. Ungulate dung: We may display some moose or cow feces which may show a range of dung-zygomycetes, possibly including the hat-throwing fungus Pilobus. If anything of interest appears to be present, examine it with the dissecting scope and note the nature of the sporangia.


C. THE DIKARYOMYCOTA
The dikaryomycota were formerly classified as the phyla Ascomycota and Basiodiomycota (for example, see chapter 15 in your text), but recent advances in the systematic understanding have led to the merging of these two groups in a single phylum of higher fungi, the Dikaryomycota, with the ascomycetes and basiodiomycetes being separated into subphyla termed Ascomycotina and Basiodiomycotina. We will focus on these two groups.
The common feature of these groups is the formation of dikaryotic hyphae. The dikaryon arises when the protoplast of haploid hyphae fuse (they undergo plasmogamy). The nuclei do not initially fuse, and the resulting mycelium is made up of cells that are dikaryotic, or in the N + N state. Fusion of the two nuclei occurs in the fruiting body of the fungus, forming a diploid cell that immediately undergoes meiosis and mitosis to produce four to eight spores. The spores are released from sporangia formed in the fruiting body. In the ascomycetes, eight spores are released from sacs termed asci (singular is ascus, from the Latin word for sac). In the basiodiomycetes, the spores are formed on the end of a club-like sporangia termed a basidium (from the Latin word for club). The fruiting bodies of each fungus are termed an ascocarp (ascoma in your text), and the basidiocarp (basidioma in your text). The mushroom cap is a basidiocarp.
We have a number of specimens for you to examine today from each subphylum.
1. Subphyllum Ascomycotina
a. Unicellular forms: the yeasts: These are single-celled fungi that typically live within the food medium. Most are saprophytic, although some can become parasitic. The yeast fungus Candida albicans is an important pathogen in humans, forming diaper-rash, vaginal and urethral tract infections, and the potentially deadly sexually-transmitted disease candidiasis. The common yeast Saccharomyces cerviseae is the yeast of baking, brewing and enology (wine making). This yeast is preferred in fermentations as it rapidly grows, produces pleasant as opposed to noxious or toxic waste-products, and is tolerant of high (>10%) concentrations of ethanol.
Saccharomyces cereviseae: The common brewers yeast is growing on agar. Take a very small portion off the culture and smear into a drop of water on a microscope slide. Add a cover slip, and examine at low and then high power with the compound scope.
Look for budding cells amongst the large numbers of indistinct single yeast cells. These are apparent from the blob-like cellular extensions, termed buds that arise from mature yeast cells. Rather than simply dividing in two as most algae and plant cells do, yeasts divide by extruding protoplasm into a bud. This extrusion is then encapsulated in a wall and split off to form a new independent cell.
Occasionally, you may see some yeast forming asci: Yeasts live in both a haploid and diploid state. When conditions are harsh, two diploid yeast nuclei merge to form a zygote, which then undergoes meiosis to produce a four celled sac, the ascus. The ascus splits open to release the four cells, which then bud to start a new population of yeast cells. You may be able to see four-celled asci floating among the many cells in your slide. If so, show your classmates.

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b. Filamentous Ascomycetes: Multicellular ascomycetes produce hyphae and mycelium, and form ascocarps. Three types of ascocarps are produced by these fungi, cleistothecia (enclosed spheres), perithecia (vase-like) and apothecia (cup shaped). You should examine examples of each.
b.1 Cleistothecial species:
i. Powdery mildew (Uncinula spp.): Powdery mildews are common pathogenic fungi that infect leaves, forming a powdery mycelium on the surface. Powdery mildews reproduce asexually by forming chains of spores (conidiospores) on special hyphae termed conidophores. During sexual reproduction, they form a simple enclosed ascocarp, the cleistothecia. Cleistothecia are completely enclosed, with no opening for the developing spores to escape. When mature, the ascocarp wall ruptures, allowing enclosed asci with their ascospores to spill out and disperse. Often, cleistothecia have barbs and hooks, which can help disperse the entire ascocarp by clinging onto the fur of passing animals.
Examine the following:
Dissecting scope: Scan across the leaf infected with Uncinula to note the powdery mycelium, with chains of conidia rising above it. Periodically, you will see a pepper grain-like object with multiple elongated hooks attached. This is the cleistothecia.
Light microscope: Scrape some cleistothecia onto a microscope slide and cover gently with a cover slip. Examine at low power. Next press of the cover slip to rupture the ascocarp and release the spores inside.
ii. Powdery mildew on leaves: We also have specimens of unknown powdery mildews collected on leaves from around Toronto. Examine these under the dissecting scope for cleistothecia and conidia.
b.2 Perithecial species: The perithecium is a vase-shaped ascocarp with a narrow, open neck. Inside are multiple asci with spores. When mature, the asci protrude from the neck of the perithecia and forcibly eject the spores into the air. Sordaria is a dung saprophyte that is closely related to Neurospora, the fungus that has become one of the leading model organisms in genetic research.
Examine:
Dissecting scope: Sordaria is growing on agar plates, and the perithecia can be seen as dark pepper-like grains mixed in a mass of conidia-forming hyphae. Examine the perithecia closely and note the pear-like shape of the ascocarp. Are any asci protruding from the perithecia?
Light microscope: Scoop some perithecia onto a slide, cover and examine at low-to medium power. Gently push on the cover slip to squash open the perithecia. Note any football-shaped spores and asci that emerge.
b.3. Apothecial species: The apothecium is an ascocarp where the asci are directly exposed to the air in a cup, dome or invaginated surface. The fungi are commonly called the saucer, or cup fungi, and they include many beautiful, brightly colored forest species. The delectable morel is an apothecial ascocarp. To demonstrate the apothecium structure and form, we have a set of prepared slides and live specimens from a number of species.

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i. Bispora centrina (Yellow-fairy cups – live specimens): These are wood decomposing fungi that form small, brightly yellow cup shaped apothecium. Examine the stick with the fairly cups closely. You may take the stick to you bench to examine with a dissecting scope. The asci are formed on the inner surface of the cup. Return the stick to the display area when finished as we have only a few specimens.
ii. Peziza (prepared slides): Peziza is a cup fungus that grows on wood and is similar in form to Bispora. A life cycle of Peziza is shown in Fig. 15-14 on page 318 of your textbook.
Examine the prepared cross sections of the Peziza ascocarp under medium power with your light microscope. You will note the sac-like structure of the ascus with 8 haploid spores inside. These arose from meiosis and one subsequent round of mitosis. Note the zone of fertile tissue where the asci form. Below this are fattened vegetative cells that form the support structure of the ascocarp.
b.4 Ascomycetes of special note
i. Claviceps purpurea (Ergot of Rye)
We have display specimens of rye shoots infected with Ergot, caused by the perithecia-forming ascomycete Claviceps purpurea. Claviveps is an example of an endo-parasite, a fungus that grows within the stem and leaves of grasses. The fungus retards growth, but does not kill the host plant. In many instances, toxins produced by the fungus deter herbivory, and so the grass host can actually show superior performance relative to a non-infected plant that is eaten. In the case of ergot, the toxin produced is lysergic acid amide, from which the hallucinogenic drug lysergic acid diethyamide (LSD) was derived.
Examine the infected rye and note the grain heads with enlarged, dark-colored protrusions extending out from the grass stalk. These are sclerotia, which occur where the fungus has completely infected a developing grain and replaced the grain with a tight mat of interwoven mycelium. As the growing season ends, the sclerotia fall to the ground and overwinter. In the spring, they produce perithecia and in turn, large numbers of spores that infect the new rye crop.
Sclerotia break free and mix with the rye grain at harvest. People eating rye contaminated with ergot sclerotia experience severe poisoning, called ergotism. Symptoms include wild hallucinations coupled with extreme burning sensations in the extremities. Constriction of minor veins is common, leading to limbs dying and falling off. The pain is severe, and a typical victim would scream in agony while madly hallucinating. Before modern science explained the cause, people in the past would interpret the symptoms as an attack of demons, and in regions affected by ergot outbreaks, the citizens often turned to extreme religious practices to exorcise the devil. Throughout history, witch hunts, new religious movements, and mass hysteria have been attributed to ergot outbreaks.
Today, ergot poisoning is rare, and rye grain is routinely screened to filter out the larger sclerotia. Sclerotia are now intentionally grown as a source of drugs to control internal bleeding, migraine headaches, and to alter mental states in psychiatric patients.
ii. Peach Brown Rot (Monilinia fructicola): Many ascomycetes are severe pathogens of fruit crops. One of the worst is Peach Brown rot, which stunts trees and destroys mature peaches, apricots, cherries and related fruit. Infected trees form cankers on the twigs and leaves. Conidia erupting from the cankers are dispersed to infect other trees by asexual means. Fruits are infected as they near maturity.

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After infection, lesions develop on the fruit and it prematurely rots, falls to the ground and dries to a mummified carcass. Peach mummies are completely infected with the mycelium and in this form, the fungus will overwinter. In the spring, the fungus in the mummy form apothecia, from which spores will be released in huge numbers to infect new trees.
Examine the Monilinia cultures on agar with the dissecting microscope and note the lemon-shaped conidiospores arising from the mycelium. You can examine these more closely by taking a small piece of agar and preparing it on a slide for examination with the light microscope.
iii. Penecillium: Many ascomycetes are saprophytes that infect food and building materials in the home. Some are also sources of important drugs, while other produce powerful carcinogens. Penicillium is one of the most common molds in the household pantry, where it infects bread, fruits and milk products. Penicllium species are also important in making strong-flavored cheeses such rouquefort, gorgonzola, chamenbert, brie and Danish Blue. The blue-green color is actually the reproductive conidia of the Penicillium mold. Pennicilium is also the source of penicillin, the antibiotic that prevents wall synthesis in gram negative bacteria.
Examine the culture with the dissecting scope and note the green-blue broom-like conidiospore masses rising above the mycelium. These masses give Penicillium molds their characteristic color. Take a sample and prepare a microscope slide of it. Examine the conidiophores with conidia under the compound microscope. Note the broom-like structure of the spore-bearing mass.
We also have some blue-cheese on display. Examine the Pennicilium colony through the dissecting scope and try to identify the sporangia.
iv. Aspergillus: Aspergillus species are common black-colored molds in the household environment. They are frequently found on bread, drywall, and grains. Many species produce aflotoxins, which are powerful carcinogens of the liver found in stored grains, peanuts and cereals, including corn flakes. It is unwise to eat foods contaminated with wild Aspergillus species as they likely contain aflotoxins. (For example, never eat wild peanuts, or musty old grain). Beneficial Aspergillus spp. are used to produce soy sauce, miso (fermented soy paste), and to ferment rice in an early step in sake production.
Examine with a dissecting scope the culture on the agar plate and note the fan-shaped mass of conidia arising above the mycelium. Next, examine a piece of the mycelium to see the bulbous conidiophore. The dark masses of conidia give this fungus its particular color and shape.
Take a small chunk of infected agar and prepare a slide of the sporangia for the light microscope. Examine the swollen top of the condiophores and the attached fan-shaped array of conidia.
2. The subphylum Basidiomycotina
The most familiar fungi are the basidiomycetes. The fruiting bodies of the basidiomycetes (the basidiocarp) are the recognizable features of species of mushrooms, toadstools, coral fungi, shelf fungi and tooth fungi. In each, the main body lives underground or in wood as a dispersed mycelium. Although all basidiomycetes reproduce by forming spores on club-shaped basidia, there are actually two main groups: the homobasidiomycetes and the heterbasidiomycetes. The homobasidiomycetes produce one type of spore, the basidiospore. The heterobasidiomycetes produce two types of spores

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during the sexual life cycle. We will focus on the homobasidiomycete life cycle as exemplified by the common food mushroom, Agaricus campestris. Heterobasidiomycetes are the pathogenic rusts and smuts.
a. Basidiomycete yeast (Rhodotorula ruba): Some basidiomycetes also have evolved the unicellular life form. A common basidiomycete yeast is the red yeast, Rhodotorula ruba, a contaminant of bathroom curtains, tile and grout. The pink scum in filthy bathtubs and showers is caused by Rhodotorula. (You may remember the battle between the Cat-in-the-Hat and pink bathroom scum).
Examine the red yeast culture on display. If time permits, you may prepare a microscope slide of the cells from the agar culture. Examine them for budding and basidia, which are distinguished by an elongated shape and horn-like points on one end of the cell.
b. Heterobasidiomycetes: The heterobasidiomycetes include the rust diseases of grasses, and smut diseases of maize. Other members of this group are wood decomposers such as the jelly fungi. We may have a jelly fungus in the wild mushroom display.
Examine the specimens of grasses infected with wheat rust (Puccinia graminis). Note the rust-colored pustules forming on the blades of the grass. These are where asexual spores are formed to allow for continued infection of healthy plants during the summer. Black pustules appear in the late-summer. These are where teliospores are formed. Teliospores are overwintering spores that form basidiospores in the spring.
If available, examine any corn smut (Ustilago maydis) that may be on display. Corn smuts attack developing corn kernels and produce large, grey-colored smutballs that are filled with dark spores. Immature smutballs are served as a delicacy in Latin American cuisines.
Rusts and smuts are virulent parasites of grain crops, with the potential to wipe out the production of an entire region in any given year. The primary means of preventing infestation is to breed crop cultivars that are resistant to rust infections. The rusts eventually evolve new ways to infect the cultivar, so government agencies are continuously breeding resistance into varieties to stay ahead of the rust capacity to re-evolve virulence. Should breeding efforts fall behind (for example, via cost-cutting measures by governments and agribusiness), major rust outbreaks could result, ruining grain crops and causing food prices to sky-rocket.
Notes:

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c. Homobasidiomycetes: the Mushrooms
We have fresh specimens of the common store-bought mushroom for examination, along with cultures of the inky cap mushroom, and a range of wild mushrooms from southern Ontario. To aid in examining fine detail, we have prepared slides showing cross sections of mushroom caps for you to examine under the light microscope. A detailed diagram of the life-cycle of the mushroom is presented in Figure 15-19 of your textbook (page 321).
c.1. The common food mushroom Agricus bisporus:
Examine a) the mycelium of the spawn blocks on display, b) the young button mushrooms and c) mature-spore producing mushrooms from the collection of fresh mushrooms provided.
i. Mycelial stage: sample mycelia of the mushroom spawn that is available. Stain with cotton blue and view with the light microscope under both normal light and phase contrast. Find some isolated hyphae and examine this under high power. Note the septate nature of the hyphae. This is one of the diagnostic features of the basidiomycetes.
Each cell contains two haploid nuclei in the N + N configuration. A key feature of Basidiomycetes is the presence of clamp connections, which form after cell division in order to keep the N + N dikaryotic configuration intact (see figure 15-21 in your text). Clamp connections may be visible along the end walls of the hyphal cells, forming bulges or loops around the septate wall.
ii. The mushroom button stage: The basidiocarp forms from tightly woven mycelia. Initially, the basidiocarp form a button, or egg stage. Cut open a button and examine a) the immature stalk (or stipe), b) the young, white to pink gills, and c) the developing cap which extends down over the stipe. With a razor blade, cut a thin slice of a gill, and look at the slice with the light microscope. Stain the gill with Melzer’s blue. You may be able to see developing basidia with miniature spores.
iii. The mature mushroom: Note the features of the basidiocarp structure. The main parts are a) a well developed stalk, termed the stipe. The cap, termed the pileus, and the gills, where the spore bearing tissues occur. The gills have turn chocolate brown as millions of spores mature. Cut a thin slide of the mature gill and examine for spores and horned basidia. Stain with the Melzers stain placed on your bench. You should be able to isolate one or two good basidia from the mass of tissue.
iv. Prepared slide of the mushroom cap: Examine under the light microscope the cross sections prepared of an Agaricus pileaus. The cross sections of gills clearly demonstrate the club shaped basidia arising from the zone of fertile tissue (the hymenium). Examine the basidia under high power and note how the spores are attached to the horn-like basidial tips (the sterigmata). The spores appear as party balloons taped to a club.
v. Spore prints: As the spores mature, they are released and drift into the air below the cap. If the cap is place over paper, the spores cannot disperse and settle onto the paper to form a print of the mushroom gills. Spore prints have been prepared for you to examine. Spore prints are often used to identify particular mushrooms, and they are an easy way to verify the color of the spores.

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c.2. The Inky-cap mushroom, Coprinus cinereus. We have displays of the Inky-cap mushroom for you to examine. The cultures show how the mushroom arises from the mycelium. Prepared slides are also available if you wish to examine the gill structure and the basidia of Corpinus.
The cap self-digests upon maturity forming a purple-ink colored mass of goo.
c.3. The oyster mushroom Pleurotus ostreatus: Oyster mushrooms are delightful edible mushrooms that grow on logs and decaying stumps. They produce white spores on short gills, and exhibit a large, fan-shaped pileus with a short stipe. They are now commonly cultivated and are available in many supermarkets.
Examine the oyster mushrooms for morphological structure, then thin slice a gill and examine for the basidia and white spores under the microscope.
c.4. The chanterelle (Chanterellus cineareus): Chanterelles are a wonderful delicacy that is prized for its gentle, buttery flavor. Unlike the gilled mushrooms, chanterelles form their basidia on gently folded tissue underneath a wavy pileus. Assuming we have these available, take a small piece of the pileaus of a chanterelle and examine for basidia and spores. What color are the spores?
c.6. The wild mushroom display: We have collected a variety of wild mushrooms for you to examine for variation in form. Examine each carefully, paying attention to the pileus, the stipe (if present) and where the spore bearing tissues are located. In many of the samples, gills are not present. Instead, the spores are produced on elongated tubes that form pores on the underside of the pileus, on teeth-like protrusions that hang below the pileus, on coral-like prongs, or on rumpled folds of tissue that resemble elbow skin. These traits distinguish the major families of mushroom-forming species. You may take some of the specimens and examine them under the dissecting scope in order to better see the pores, teeth or prongs of the spore-bearing tissue.
PART IV: LICHENS
Lichens are structures formed by close symbiotic relationships between an algae and a fungus. Both green and blue-green algae can serve as the algal symbiont, while the fungus is typically an ascomycete. Because the sexual stage of the lichen that is visible is that of the fungal partner (the mycobiont), the lichen is typically named after the fungus.
In the symbiosis, the algae provide carbohydrates from photosynthesis while the fungus shelters the algae and gathers water and nutrients. Lichens can completely desiccate with no harm to the organisms inside. Upon wetting they rapidly rehydrate and resume activity. This ability allows them to live in extremely harsh surfaces, such as the branches and trunks of trees, the sides of rocks, and bare ground in deserts. In the boreal zone, lichens are important ground covers on bare soil, fallen branches and the surface of rocks. They also are common as epiphytes on trees. In general, they grow extremely slow, reflecting the harsh conditions of the habitats they live in.
Lichens come in three general categories based on morphology. Examine the specimens displayed in the lab room.

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A. Foliose lichens: Lichens that exhibit a leafy shape are termed the foliose lichens. These are common in wetter habitats, for example, forest interiors in eastern Canada. Foliose lichens are important epiphytes growing on branches of standing trees.
B. Fructicose lichens: These lichens are shrubby in appearance, with many narrow, highly branched stem-like structures. Fructicose lichens are common in the boreal forest, forming dense ground covers in spruce forests. When dry, they are extremely flammable and can be used as a fire starter. They also help wildfires spread and thus contribute to some of the severe forest fires that occur every summer in Canada. Fructicose lichens are common on the sides of trees, and often hang from the bra nches in dense growths termed Witch’s hair, or Old Man’s Beard.
C. Crustose lichens: Crustose lichens occur in the most extreme terrestrial environments where life is possible. They grown on the sides of rocks, buildings and on bare soil, and are common in arid and polar deserts, including the dry valleys of Antarctica. Crustose lichens form brilliant yellow, orange, red and yellow-green colors on the rock, and are some of the most beautiful features in what are otherwise barren landscapes.
Study Guide: You should be familiar with the major categories of fungi and the names of the common species of yeast, store mushrooms, and major disease organisms displayed in the lab. You need to know the terms presented in bold font, and should recognize an organism well enough to classify it to phylum or where relevant, to subphylum. Know and understand the distinguishing characteristics of the major phyla presented in lab, as well as that of the ascomycetes and basidiomycetes.