International Research Journal of Biological Sciences ___________________________________ ISSN 2278-3202Vol. 2(6), 31-34, June (2013) Int. Res. J. Biological Sci. International Science Congress Association 31 Isolation and Characterization of Some Indian HyphochytriomycetesDubey Manish Kumar and Upadhyay R.S. Laboratory of Mycopathology and Microbial Technology, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi- 221 005, Uttar Pradesh, INDIAAvailable online at: www.isca.in Received 19th March 2013, revised 8th April 2013, accepted 5th May 2013Abstract In the present study, three species of Hyphochytriomycetes were isolated, identified and described. They are Rhizidiomyces hirsutus Karling, Rhizidiomyces apophysatus Zopf and Rhizidiomyces bulbosus Karling. Among them, Rhizidiomyces bulbosus Karling species is being reported for the first time in India. Keywords: Hyphochytriomycetes, Rhizidiomyces and Rhizidiomyces bulbosus Karling. Introduction Hyphochytriomycetes (Hyphochytriales) is a small group of chytrid-like organisms that contain about 23 known species. They are strikingly similar in morphology and development to many of the true chytrids, but are distinguishable from them by the presence of single anterior tinsel-type flagellum on their zoospores. Fuller included them in the class Hyphochytriomycetes of phylum Hyphochytriomycota. According to Berbee and Taylor the Hyphochytriomycota, Labyrinthulomycota and Oomycota belong to the Kingdom Stramenopila. Hyphochytriomycota, consisting of a single order Hyphochytriales, has been classified into three families, namely Anisolpidiaceae, Rhizidiomycetaceae and Hyphochytriaceae on the basis of their thallus structure4,5,2,6. The Hyphochytriomycota are a small group of little known fungi with almost no economic importance; but molecular and ultrastructural evidences place them together with the biagellate heterokont organisms such as Oomycota and Chromistan algae. In India the study of Hyphochytriomycetes started as early as 1935, when Chaudhuri and Kochhar reported Rhizidiomyces apophysatus from the oogonia of Achlya klebsiana. Few years later, Mundkur reported the same species on A. klebsiana. However, the real start of research on Hyphochytriomycetes in India should be credited to J.S. Karling, a mycologist of USA who visited India in 1963 and described 6 species of Hyphochytriomycetes10. The purpose of this study was to throw light on description and information pertaining to habitats, substrates, and geographical locations of the following Hyphochytriomycetes isolated from fresh water and soil sources. Three species of Rhizidiomyces were isolated and illustrated in this paper. The detail of their life history, with particular emphasis on their delepmental stages on soild media is described. Rhizidiomyces bulbosus Karling species is newly recorded in India. Material and Methods Isolation: Baiting technique11,12 was used for the recovery of Hyphochytriomycetes. Samples of water and soil were collected at random and taken to the laboratory. Each sample was divided into triplicates, which were introduced in separate Petri dishes and flooded with 40 mL of sterile deionized water. Each triplicate was baited with chitin (purified shrimp exoskeleton) and keratin (purified snake skin). All triplicates were incubated at ambient room temperature for two weeks. The baits were periodically examined under a microscope for about two weeks and when the isolates became visible on the baits, they were transferred to 1/4YpSs (yeast extract peptone soluble starch) agar medium containing 300 ppm penicillin G and 300 ppm streptomycin sulfate. All isolates were obtained in to pure form by a series of regular subculture carried out on 1/4YpSs agar. Stock culture of all the isolates was maintained on 1/4YpSs agar slants. The cultures were stored at 10°C and subcultured after every three months onto fresh media. All specimens were deposited at the Laboratory of Mycopathology and Microbial Technology, Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi, India. Observation and identification: Thallus morphological features and developmental pattern of the isolates were examined using a light microscope on 1/4YpSs agar, PYG broth and on baits. The isolates were examined by light microscopy to assess range and variation in thallus structural features, including sporangial shape and size, discharge apparatus, number of discharge pores/ tubes, type of zoospores discharge, flagellation of the zoospore, possession of apophysis and morphology of rhizoidal system. The type of habitat and substrata from which each culture was originally isolated were analyzed. Identification and characterization of isolates were made with the help of Sparrow's `Aquatic Phycomycetes11 and Karling's `Chytridiomycetarum Iconographia'13 and other relevant taxonomic literatures.