DIVISION EUMYCOTA ‘Most fungi belong to this group. Their thallus organization differs in that they do not possess plasmodium or pseudoplasmodium but are unicellular or filamentous. They may have diverse evolutionary organs and relationships however; they do possess certain features of organization, nutrition and physiology as follows: 1. THALLUS ORGANIZATION: the thallus of a fungus is generally based on a branched filament. In most fungi, the thallus is differentiated into a vegetative part which absorbs nutrients and their reproductive parts, Such thalli are described as eucarpic. However, in some the thallus does not show this differentiation and following a phase of vegetative growth, the entire thallus becomes converted into propagules. Thalli of this kind are called holpearpie and are found in certain parasitic fungi in which the thallus lives inside the host cells of some unicellular free floating organisms. The unicellular type of thallus is typical of yeast\and yeast like fungi e.g. saccharomyces and sporobolomyces. Some fungi especially animal pathogens can exist either in the filamentous or unicellular phase, this phenomenon is called dimorphism. Changes from the filamentous to unicellular form can be brought about by manipulating the environmental conditions (composition of the medium and CO; concentration). The vegetative filaments of fungi are termed hyphae and the collective term for a thallus made up of hyphae is called mycelium. ‘An important distinguishing feature between groups, of fungi, is the presence or absence of cross walls or septa in the hyphae. Certain groups of fungi however, generally have non septate hyphae. Oomycetes, Zygomycetes, Basidiomycetes and Deuteromycetes generally have septate hyphae. In non septate forms the mycelium contains numerous nuclei which are not separated from each other by cross walls but lie in a common mass of cytoplasm. Such a condition is called Coenoeytic. In septate forms however, the hyphal segments may contain one, two or more nuclei. When the nuclei are genetically identical as in a mycelium derived from a single uninucleate spore, the mycelium is said to be homokaryotic but where a cell or mycelium contains nuclei of different genotypes possible as a result of mutation, itis said to be heterokaryotic. A special condition is found in the mycelium of certain Basidiomycetes where each cell may contain two genetically distinct haploid nuclei, this condition is called dikaryotic. In contrast, mycelia with segments containing single, haploid and genetically identical nuclei are called monokaryotic. 2. WALL STRUCTURE: The filament owes its form to the turgor of the protoplast it contains and to the rigidity of the wall. The change in form of the fungal thallus as it undergoes development is caused by changes in shape and proportion of fungal cells, which are closely linked with changes in the wall structure. There are also correlations between the chemical structure of fungal cell walls and taxonomy. LAC ne inne TY OFCoMO ONY Page 14Chemical analysis of cell walls which have been cleaned free of cytoplasm by physical and chemical techniques shows that they contain 80 — 90 % polysaccharides with most of the remainder consisting of protein and lipid. The skeletal material of the cell wall consists in most forms of spirally oriented layers of chitin, cellulose or sometime glucans. Chitin is however, the most usual component but cellulose is present in oomycetes cell walls together with glucans. Occasionally, chitin and cellulose are found together in some members of the ascomycetes. A characteristic feature of oomycetes cell walls is the presence of the amino acid hydroxyproline. The microfibrillar components are embedded in a matrix of other substances. Protein is an important component some of this may represent enzymes closely bound to wall components. The presence of enzymes as an integral part of the wall explains why the wall fragments are not inert but demonstrate branched activity. ‘The walls of yeast however, have distinctive chemical properties being composed of mannan - glucan complex (MGCx). 3. INTERNAL STRUCTURE: The cells of Eumycota are eukaryotic and apart from chloroplast contain many of the familiar organelles characteristic of eukaryotes. The nucleus is surrounded by a double membrane which is continuous with endoplasmic reticulum. ‘There are rmumerous pores on the nuclear envelope which may allow for interchange of materials between nucleus and cytoplasm when mitotic nuclear division occurs, the nuclear membrane does not always break down in most other organisms but may construct in the middle to separate the two sister nuclei. This process is called karyochorisis. Mitochondria are of diverse shape but often elongate. They are sufficiently large in many fungi to be seen by light microscope and move about rapidly within the fungal protoplast. The endoplasmic reticulum may be smooth or rough i.e. with the ribosome concerned with protein synthesis attached to the endoplasmic reticulum. Cytoplasmic microtubules have been widely reported in fungal cells and maybe concerned with protoplasmic movement and the maintenance of cell shape. In many non-fungal eukaryotic cells, a Golgi apparatus consisting of stacks of folded membranes called dietyosomes, functioning in secretion have been reported but they are of comparatively rare occurrence in fungal cells, they have however, been reported in the oomycetes. Other characteristic cytoplasmic inclusions are lipid droplets and glycogen, a typical fungal storage product. Lipid and glycogen are especially abundant in mature cells, food storage structures and spores. The surface of fungal protoplast is the plasmalemma a typical membrane consisting of lipoprotein. Where vacuoles are present, they are surrounded by a similar membrane called the tonoplast. 4, HYPHAL GROWTH: In most cases is apical and as the hyphae extend there must obviously, be rapid synthesis of new wall components and membrane to provide for the increase in the plasmalemma. The driving force which thrust the growing apex forward is the turgor of the protoplast possibly generated by the increasing vacuolation of the hyphae which is distal to the apex. The fine structural changes which accompany the growth of hyphae show minor variations Luck tat Cnentanrevorcvaasnsnesnciaey Page 15in different groups of fungi. The dictyosomes which are possibly the equivalent of the Golgi apparatus of other organisms are differentiated into a pole proximal to the nucleus or endoplasmic reticulum a distal pole which faces away from the nucleus towards the plasmalemma. The dictyosomes are in a state of dynamic equilibrium, receiving membrane material derived from the endoplasmic reticulum on the proximal face and giving rise to secretory vesicles, migrate towards the hyphal apex directly or may coalesce to form into a larger vesicle. Ribosomes are scarce in the vicinity of the apex. At the hyphal apex vesicles fuse with the plasma membrane and release their contents into the wall region. It is presumed that the secretory vesicles contain polysaccharides needed in wall construction and possible materials for the plasma membrane and enzymes. Various research works have shown that the incorporation of new wall material is confined to the very tips of the growing hyphae. It is estimated ‘that about 10,000 vesicles is required to support 1 mm of growth for a hypha 5 mm in diameter growing at arate of 1 mnyhr. It is believed that a specialized organelle, chitosome, is involved in fungi having chitinous cell walls e.g. in species of Zygomycetes. It is also reported that in septate fungi a dark apical body, the spitzenkérper (German for pointed body), has been reported whieh is probably associated with apical vesicles and hyphal tip growth. This body disappears when growth ceases and reappears when growth resumes. The forces which cause the secretory vesicles to move forward are probably due to electrophoresis and it has been claimed that an electric current may account for the movement of the vesicles. SEPTA IN FUNGAL CELLS ARE OF THREE KINDS: i, Where septa do not contain a pore, septa of this type, are rare in vegetative hyphae. ii, Where itis a simple transverse plate Lying at right angles to the axis of the hyphae and is, usually perforated. The movement of cytoplasmic organelles such as the mitochondria and nuclei occur freely through the pores of ascomycetes and deuteromycetes. iii, Where it is more complex, where surrounding the central pore of the septa is a curved structure of wall material which is often thickened to form a barrel shaped or cylindrical outline, septa of this type called Dolipore Septa e.g. in Basidiomycetes excluding the nest and smut fungi > cell wall QS cell wall 2 * nuclei (a) septate hypha (b) nonseptate hypha ceca ov un Ouanaantmnarvorananaonsesncen Page 16Septate vs. non-septate hyphae When you see a mushroom, you are looking at a tiny part of the whole fungus. Mushrooms are the fruiting bodies, the reproductive structure, for some types of fungi. The rest of the fungi is a body of fine threads weaving through the substrate and slowly digesting nutrients. While not all fungi form mushrooms, most do form a network of hyphae, tube-like structures that allow the fungus to search out and absorb new food sources. Non-septate hyphae are generally single-cell organisms. Hyphae growth and structures ‘A fungus starts from a spore and the initial hypha grows out from that germ. The first hypha grows out, extending at the tip, or apex, and then begins to branch out into richer areas of food, forming a body of hyphae, the mycelium. The hyphae exude digestive enzymes and absorb nutrients, As the mature fungus exhausts its food supply, it cannibalizes old hyphae and expands. Hyphae form more branches in areas that are richer in nutrients. Depending on the type of fungus, hyphae can be one large multi-nucleated cell, when they are called non-septate hyphae, or can have dividers between the individual cells, when they are called sepate hyphee, Septate hyphae Septate hyphae have dividers between the cells, called septa (singular septum). The septa have openings called pores between the cells, to allow the flow of cytoplasm and nutrients throughout the mycelium. Although the septa separate the cells, in some hyphae the cellular components, including the nucleus, can fit through the pores. When new cells bud at the apex of the hypha, a septum docs not form immediately. As the new cell matures, the cell wall grows down into the cytoplasm, forming the septum. Members of the classes Basidiomycetes and Ascomycetes form septate hyphae. Non-septate hyphae Non-septate hyphae, also known as aseptate or coenocytic hyphae form one long cell with many nuclei. They are the more primitive form of hyphae; species with septate hyphae diverged from a common ancestor with coenocytic hyphae. Most fungi with coenocytic hyphae belong to the class Zygomycetes. While they do not form septa between nuclei, they do form a septum at branch points that connect one filament to another, preventing the entire network from being compromised if one hypha is injured. Comparing hyphal structures Coenocytic hyphae allow nutrients to move quickly throughout the filament because the cytoplasm is continuous, without any dividers to slow transport. On the other hand, if a coenocytic hypha is ruptured, the entire filament will die because nothing keeps the cytoplasm from leaking out. Septate hyphae can completely close the septa if they are injured, preserving the integrity of the rest of the filament. The septa also provide increased structural stability for the hyphae. al Larnaca den nro cove seancmioce Page 175. CYTOSKELETON: consists primarily of microtubules and microfilament with associated protein. They are found at the hyphal tips and are reported to be involved in the growth of the hyphae. The specialized role is reported to be in providing a framework for directing vesicles to the tip or actively moving the vesicles as occurs in non-fungal cells where they have been shown. to play a main role in moving nuclei and other organelles. This represents a 2" theory for the movement of the vesicles to the hyphal tip. 6. DIFFERENTIATION OF HYPHAL APEX As hyphal growth takes place at the apex differentiation takes place behind it. This is evident by the formation of septa in septate fungi-and vacuole. It is true that the extreme tip is viscoelastic so that it flows outwards and backwards as new components is added to the tip then the wall rigidifies progressively by the formation of extra bonds behind the tip. At the apex there is hydrolysis of glycogen to sugar which is used in building the cell wall. ‘The vigorous streaming out of cytoplasm and the formation of vacuoles behind the apex are important in pushing material forward to the growing apex. The importance of perforations in the septa in allowing translocation in nutrients is also evident. Differentiation of hyphae also occurs in many parasite and fungi. The tip of the hypha which penetrates the host may form a specialized adhesive structure. Within the host cell specialised absorptive organs called haustoria are formed e.g. in Downey mildew, powdery mildew, rust. Increasing wall crase-linking Vesicles (V) derived fiom a decreasing actin cytoskeleton Golgi body (G) are transported to the apex, pethaps by Microtubule (M) mediated systems the actin meshwork at the apex is thought to provide structural support where the wall is thinnest and where there is little or no cross-linking of the wall polymers. Behind the —— = : '\ | extreme tip the wall is Diagrammatic representation of the organisation of wall | progressively rigidified by cross- rowth at the hyphal tip. Only half of the tip is shown. _| linking of wall polymers. if Representation of the steady-state model of hyphal tip growth, in which the wall is envisaged as being viscoelastic. New wall polymers synthesised at the extreme tip are suggested to flow | outwards and backwards as new components are added at the extending tip. The decreasing thickness of the arrows behind the tip signifies progressively reduced flow as the wall components become cross-linked. [Based on a diagram in Wessels, 1990.) ————— errorcnsuansmecvon Page 18 scrim or ah7. BRANCHING OF THE COLONY Branching occurs by the development of new species as the colony grows and synthesizes new protoplasm. Branches seldom develop at the tip but at a considerable distance behind it. Most often they arise immediately behind the septa. Branching is initiated by the development of new apices from the previously mature hyphal wall resulting from a thinning and softening of wall structure. The softened wall balloons out as a result of turgor of the protoplast and extends as a new apex with an organization resembling that of a main axis. o © The duplication cycle of Basidiobolus ranarum, a fungus that grows as hyphae with complete, unperforated septa on agar plates. (@) - (b) An apical cell extends and synthesises the protoplasm, drawing forwards during growth. (© When the protoplasmic volume attains a critical size the nucleus divides and a septum is formed. (@ - ©) The new apical cell grows on to repeat the process; the sub-apical cell forms a branch, and the protoplasm migrates into this, forming another apical cell 8. AGGREGATION OF HYPHAE Most macroscopic fungal structures are formed by hyphal aggregation and give rise to mycelial strands which produce differentiated structures. (@ RHIZOMORPHS this resulted from a relatively undifferentiated hyphae common in basidiomycetes e.g. Amellaria Spp, a major root rot pathogen. The strands form most readily when the mycelium has developed a food base; a robust leading hyphae extend from the food base and branch at fairly wide intervals to form finer laterals most of which grow ovary from the parent hyphae. A few however may form an acute angle and tend to ‘grow parailel to the parent. Mycelia strands are capable of translocating materials in both directions. Certain fungi however have highly differentiated aggregates of hyphae with a well-developed apical meristem. The main part of the rhizomorph is fairly uniform in thickness and is differentiated into zones. An outer cortex of melanized cells often pigmented in an extracellular matrix. A medulla of thinner walled parallel hyphae and a central channel where the medulla has broken down serving a role in gaseous exchange or diffusion. Rhizomorphs branch by producing new multicellular apices behind the tip. They may spread underground from one root system to another however; they need to be attached to a food base because the growth depends on translocated nutrients. = ‘ecru. ase ceasnasnuseciy Page 19Breathing pore Soil surface Diagrammatic representation of a rhizomorph of Armillaria mellea 5 Zit Melanized rind’ oxygen conducting // Soft Apex camel (b) SCLEROTIA these are specialized hyphal bodies involved in dormant survival. They are thick walled food reserves. They are found similarly amongst plant pathogens e.g. Sclerotium Spp. Sclerotia develops initially by repeated localized hyphal. As the slerotium initially expands and mature the outer hyphae form a rind while the interior of the structure differentiates into a tissue like cortex of thick walled melanized cells and a central medulla consisting of hyphae with substantial nutrients storage reserves of glycogen, lipids and trehalose. Slerotium can survive for considerable periods and ‘germinate in suitable conditions either by producing hyphae or sexual fruiting body. Rasen | Diagrammatic representation of cut sclerotium cross section of Athelia rolfsii (© MYCORRHIZA these are a specialized mycelial aggregate that surrounds like a sheath to the roots of trees. These trees may grow in fertile soils and its roots are covered by these sheaths of fungal cells. The external layer of fungal tissue replaces root hairs as @ system of absorption of nutrients. (@ REPRODUCTIVE STRUCTURE hyphae may become aggregated together to form fruiting bodies bearing spores of various kinds e.g. ascomycetes, basidiomycetes and deuteromycetes. In the ascomycetes, the sexually produced spores formed by nuclear fusion and meiosis are called ascospores and are enclosed in an aggregation of hyphae called the ascocarp. In the basidiomycetes fruiting bodies e.g. mushroom, the aggregation of hyphae is called the basidiocarp. ——————— a rarnneaneeneteesss==—-—= nnn scr. pn Cnn mores nomena Page 20(© INFECTION STRUCTURES in fungal parasites infection of the host is often preceded by production of a specialized pre-penetration structure. The simplest of these are terminal swellings called appressorium if they occur as germ tubes, or hyphopodia, if they develop on short Jateral branches of hyphae, These pre-penetration structures serve to anchor the fungus to the host surface usually by secretion of mucilaginous matrix. The penetration process is achieved by a narrow hypha termed the penetration peg which develops beneath the pre- penetration structure. u Uredospore GT Germ tube A Appressorium Gc v ssc Stomatal guard and cell Sub-stomatal Vesicle Sub-stomatal cavity 1H Infection Hypha HMC Haustorial Mother cell H Haustorium Pp Infection Peg 2 Appressorium Necrotic tissue PRE-PENETRATION STRUCTURES OF PLANT PATHOGENS Larrea or cova mmecioey Page 21