Isolation and Culture of Slime Molds

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  • Topic: Fungus, Spore, Chytridiomycota
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  • Published : February 17, 2013
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Isolation and Culture of Zoosporic Fungi using Baiting Technique -------------------------------------------------

Rillera DP , Talibsao K , Talucod AC and Tan P
Department of Biological Sciences, College of Science, University of Santo Tomas, España Street, Manila 108

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Zoosporic fungi belonging to class Oomycetes of Kingdom Chromista and Phylum Chytridiomycota of Kingdom Eumycota, were isolated and cultured in this experiment. Baiting technique was conducted using 6 baits: pollen grains, snake skin, animal dung, shrimp exoskeleton and cockroach,. The morphological and reproductive characterization was also conducted on the isolated zoosporic fungi. -------------------------------------------------

Introduction
Zoospores are motile, asexual spores having one or more flagella. They may be haploid or diploid and are formed inside a sporangium, discharged from the sporangium to become a free swimming stage, and after the motile stage , the zoospore encysts in a suitable substrate or host. ( Lange & Olson, 1983). Baiting technique is a method that allows further growth of fungal mycelia on prepared baits. Major groups of fungi isolated using baiting techniques are those from Kingdom Chromista, particularly in the Class Oomycetes. The Oomycetes or “egg fungi”, also called water molds were previously grouped with fungi based on their filamentous growth and the presence of coenocytic hyphae, one of the morphological traits and characteristic of fungi. However, their life cycle, unlike that of many fungi, includes a diploid, asexually or sexually reproducing phase. (Madigan, et al., 2009). Oomycetes differ from fungi in other fundamental ways as well. The cell walls of oomycetes are typically made of cellulose, not chitin, as for fungi, and they have flagellated cells, which are lacking in all but a few fungi. (Madigan, et al, 2009) Zoospores are produced by almost all Oomycetes. They are biflagellate bodies, having one whiplash flagellum directed backward and another tinsel type flagellum directed forward. The flagella are attached anteriorly or laterally. (Sharma, 2011). Interestingly, zoosporic fungi belonging to Phylum Chytridiomycota of Kingdom Eumycota are also commonly isolated using the baiting technique. Chytridiomycetes, or chytrids, are the earliest diverging lineage of fungi with about 100 genera consisiting of approximately 1000 species. Their name refers to the structure of fruiting body (“little pot”) which contains zoospores. Cell wall contains chitin like other true fungi. However they differ from other fungi, in producing motile, flagellated spores called zoospores, a possible remnant of their adaptation to aquatic environment, mostly freshwater and moist soils, where they are commonly found. (Madigan, et al., 2009). According to JW Deacon & G Saxena (1997), Chytridiomycetes are very common as saprotrophs, facultative parasites and obligate parasites in moist soil and freshwater habitats. They depend on their zoospores for dispersal and site-selection. In fact, this is how the chytridiomycota can be detected - by placing baits such as plant seedlings, pollen grains, insect exoskeletons etc. in water or wet soil, because the zoospores encyst on these materials and then give rise to microscopic colonies. This study was designed with two to three objectives. The first was to assess the possible sources of fungi growing on the prepared baits, second is to discuss how these zoosporic fungi reach their substrates and thirdly, to characterize the fungal isolates based on their baits. The ultimate goal of this study is to isolate and culture zoosproic fungi from different baits like dead insects, dungs, and pollen grains. Morphological and reproductive characterization will also be conducted on the isolated fungi.

Methodology

A. Isolation and culture of zoosporic fungi from pollen and...
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