Introduction: Fungi can be divided into two basic morphological forms, yeasts and hyphae.

Dimorphism is the condition where by a fungus can exhibit either the yeast form or the hyphal form, depending on growth conditions. Yeasts are common unicellular fungi which reproduce asexually by budding or fission, especially in situations where efficient penetration of the substratum is not required, for example on plant surfaces or in the digestive tracts of animals. But, hyphae are multicellular fungi which reproduce asexually and/or sexually. Most fungi are composed of microscopic filaments which branch eventually to form a network of hyphae, called a mycelium. Each hypha is essentially a tube containing protoplasm surrounded by a rigid wall. Depending upon the species, a continuous and uninterrupted mass may be formed by the protoplasm running the length of the branching hyphae, or may be interrupted at intervals by cross-walls called septa. Hyphae are divided by septa into individual discrete cells or interconnected hyphae compartments. Hyphae exhibit apical growth and elongate at their tips. At least in theory, hyphae are capable of growing indefinitely, provided that environmental conditions remain favorable for growth. The environment eventually limits or restricts their growth. Hyphae may initially develop from a short, immature hypha called a germ-tube that emerges from a germinating spore. However, fungi belonging to the Chytridiomycota exist as either single round cells or primitively branched chains of cells.

Objective: Experiment 1: Experiment 2: Observation of growth of fungal spore and yeast. Isolation of soil by direct plate method.

Experiment 1:Material: Fusarium oxysporum, Rhodotorula species, Potato dextrose agar (PDA)

(Oxoid, England), slide glasses, cover slips, dropping bottle (distilled water), scalpel (surgical knife) and microscope. Procedure: A. Germination of conidia 1. The dish was observed for 21 hr of incubation from the bottom of dish, that is, reverse the dish and put on the stage of microscope. 2. The germinated conidia were focused by following the instruction of compound microscope. 3. The position of observation was remembered and the dish was removed from stage once the germinated conidia are found. The dish was opened and agar is cut in size for cover slip by scalpel. Then it was put on the slide glass and sealed with a cover slip. 4. The germinated conidia were observed (up to 40 objective lens) and the way of conidia germinates was drawn. The length of germ tube is estimated and comparing with the length of conidia. 5. As the same manner, the dishes after 2 and 12 hours of incubation were observed and the way of conidia germinates were drawn. B. Budding of yeast cells 1. The dish was observed from the bottom of dish, that is, reverse the dish and put on the stage of microscope.

2. The budding yeast cells was focused by following the instruction of compound microscope. 3. The position of observation was remembered and the dish was removed from stage once the budding yeast cells have been found. The dish was opened and agar was cut in size for cover slip by scalpel. Then it was put on the slide glass and sealed with a cover slip. 4. The budding of yeast cells were observed (up to 40 objective lens) and the way of yeast cells bud was drawn. The position where the budding appears most frequently on the yeast cells was detected.

Experiment 2:Material: Tobacco field soils from several different places, aluminum foil, balance, spoon, marker, Rose Bengal agar (Martin & Johnson) and microscope. Procedure: 1. 0.01g of soil sample was spread on Rose Bengal agar medium in the

sterile disposable Petri dish (90mm). 2. 3. Each soil was examined with replicates. The sample was incubated in room temperature (25-27C) on the

bench. 4. The sample was then observed under the microscope.

Discussion: 1. Binary fission is a relatively simple type of cell division: the cell elongates, replicates its chromosome and separates the newly formed DNA molecules so there is one chromosome in each half of the cell. A septum is then formed at mid cell, dividing the parent cell into two progeny cells, each having its own chromosome and a complement of other cellular constituents. Fission yeast cells are rod shaped and divide by medial fission. The division cycle is quite rapid, with a generation time of Schizosaccharomyces pombe between 2 and 4 hours. It's easy and inexpensive to grow fission yeast and manipulate the cells in the laboratory. Fission yeast life cycle could be vegetative or mitotic growth and meiotic cycle. For the mitotic cycle, in exponential phase, G2 phase is particularly long, with G1, S phase and M phase each taking about 10% of the division time. Generally fission yeast is a haploid, with diploid growth only a transient stage in sexual differentiation. However, in the laboratory, diploids can be recovered and grown in the vegetative cycle. The central events of cell reproduction are chromosome duplication, which takes place in S (Synthetic) phase, followed by chromosome segregation and mitosis and cytokinesis, which is collectively called M phase. G1 is the gap between M and S phases, and G2 is the gap between S and M phases. In the budding yeast, the G1 phase is particularly extended, and cytokinesis does not happen until a new S phase is launched. Fission yeast governs mitosis by mechanisms that are similar to those in multicellular animals. It normally proliferates in a haploid state. While for the meiotic cycle, the specialized meiotic cycle that results after two cells conjugate, forming a transient diploid or zygote that proceeds through meiosis to produce four spores. When starved, cells of opposite mating types fuse to form a diploid zygote that immediately enters meiosis to generate four haploid spores. When conditions improve, these spores germinate to produce proliferating haploid cells. Hence, the fission yeast is obviously different from budding yeast

which new individuals are produced as buds. When the buds reach a certain size, they are released as self-sufficient individuals. 2. (I) Soil plating method: The soil-plate method was designed by Warcup to isolate fungi from soil and to study the ecological distribution of various species of fungi. Often it is most convenient to place materials that are of interest directly on a nutrient agar medium. This technique is simple and encourage rapidly spreading moulds at the expense of other fungi but is widely used which is requiring the placing of small bits of the substance on the surface of the agar or the pouring of melted but cooled agar over the fragments. After a few days' incubation mould colonies appear on the surface, and can be transferred into pure culture. The amount of material placed in the dish varies, depending upon how heavily it is infected with mould spores. This technique is commonly used in soil studies, requiring only a pinch of soil, evenly dispersed over the surface of the agar. It is difficult to apply any rules here because a small amount of material may yield a different set of moulds from a large amount. If the amount of material is large, the result may be a combination of a plating technique and a moist chamber. Martin's Rose Bengal Medium is a good choice for direct plating as the rose bengal dye and antibiotics in the medium slow down the colony growth and keep the colonies from growing together at beginning.

(II) Soil dilution method: In this technique, a set of serial dilutions is made, a sample of each is placed into a liquefied agar medium, and the medium poured into a petri dish. The agar solidifies with the fungal cells locked inside of the agar. 1 gram of the material to be studied is ground up and dispersed in 9 ml of sterile water. One ml of this solution is transferred to a second tube containing 9 ml of sterile water, resulting in a 0.01 dilution of the spore mass in the original material. The process is repeated to yield dilutions of 0.001, 0.0001, and 0.00001 or even further if necessary. A 1-ml portion from each dilution is pipetted to a separate petri dish, and cooled, melted agar medium poured over it. The plate should be moved gently on the table in a figure-of-eight motion to effect proper dispersion. Alternatively, the solution can be put on the surface of solidified medium and spread evenly throughout. After a few days' incubation, colonies will appear in varying densities, depending upon the amount of dilution from the original material. The number of spores present in the original sample can be calculated roughly by selecting the plates showing 40100 colonies and writing down the colony count. With this formula the following calculation can be performed:

The accuracy of this technique is low when only one plate is counted. There are numerous contributing factors, including improper dispersion of spores during dilution, failure to break up spore masses, or mutual inhibition of growth by certain fungi. Greater accuracy is attained by doing several plates at the most desirable dilution, perhaps ten or more. Dilutions are frequently used in studies on soil fungi. Although the technique has been criticized as not reflecting the "true" soil flora, it is probably the most commonly used

method in this area of study. The technique does allow scientists to compare one soil with another on a statistical basis.