Uniflagellate zoosporic "fungi" (=Chytridiomycota and the zoosporic protista Hyphochytriomycota) are common inhabitants of soil. However, at what scale differences in their spatial distribution can be detected is poorly known. The first objective of this study was to assess the association of organismal distribution and frequency with two microhabitats: moss-covered and exposed forest soils, at four macroscopically similar but spatially separate sites in the Blue Ridge and Allegheny Mountains of Virginia. The second objective was to provide statistically either acceptance or denial of inferences derived from sampling regimes involving a more limited number of samples. To evaluate the scale where distributional differences may occur within a site, protocols involved four collection regimes and random point and linear transect sampling. Chytrid frequency on thalli of two moss genera was greatest in the soil surrounding and under the moss rhizoids. Random point sampling methods suggested differences in zoosporic fungal frequency between moss-covered soil and the exposed soil adjacent to mosses, as well as between two moss taxa. Linear transect sampling methods also suggested differences in zoosporic fungal frequencies between moss-covered soil and soil proximal to mosses. However, statistical analysis of random point samples using a goodness-of-fit test demonstrated that there was no significant difference in frequency of zoosporic fungi from moss-covered soil and exposed soil proximal to mosses. More importantly, there was a significant difference in the frequency of ubiquitous and common zoosporic fungal species between different moss/soil complexes. This study demonstrates that differences in chytrid distribution can be detected at a microscale while at a larger scale, similarity in frequency and distribution was found.
Mass culture of algae for the production of biofuels is a developing technology designed to offset the depletion of fossil fuel reserves. However, large scale culture of algae in open ponds can be challenging because of incidences of infestation with algal parasites. Without knowledge of the identity of the specific parasite and how to control these pests, algal-based biofuel production will be limited. We have characterized a eukaryotic parasite of Scenedesmus dimorphus growing in outdoor ponds used for biofuel production. We demonstrated that as the genomic DNA of parasite FD01 increases, the concentration of S. dimorphus cells decreases; consequently, this is a highly destructive pathogen. Techniques for culture of the parasite and host were developed, and the endoparasite was identified as the Aphelidea, Amoeboaphelidium protococcarum. Phylogenetic analysis of ribosomal sequences revealed that parasite FD01 placed within the recently described Cryptomycota, a poorly known phylum based on two species of Rozella and environmental samples. Transmission electron microscopy demonstrated that aplanospores of the parasite produced filose pseudopodia, which contained fine fibers the diameter of actin microfilaments. Multiple lipid globules clustered and were associated with microbodies, mitochondria and a membrane cisternae, an arrangement characteristic of the microbody-lipid globule complex of chytrid zoospores. After encystment and attachment to the host cells, the parasite injected its protoplast into the host between the host cell wall and plasma membrane. At maturity the unwalled parasite occupied the entire host cell. After cleavage of the protoplast into aplanospores, a vacuole and lipids remained in the host cell. Amoeboaphelidium protococcarum isolate FD01 is characteristic of the original description of this species and is different from strain X-5 recently characterized. Our results help put a face on the Cryptomycota, revealing that the phylum is more diverse than previously understood and include some of the Aphelidea as well as Rozella species and potentially Microsporidia.
AbstractChytridiales is an order of zoosporic fungi currently comprising species representing 19 genera. Although morphologically and genetically diverse, these taxa have in common a zoospore with a suite of ultrastructural characters unique among Chytridiomycota. However, multiple states have been reported for almost every character that defines the Chytridiales zoospore. Two zoospore types have been recognized, each corresponding to a family. Here we examine zoospore ultrastructure of 52 isolates in Chytridiales and assess states for six characters to hypothesize evolutionary trends, using parsimony ancestral state reconstruction for evolutionary analysis. Based on suites of character states, we describe four additional zoospore types in Chytridiales. Five of the six characters ([i] location of the nucleus, [ii] morphology of the kinetosome-associated structure, [iii] complexity of the microtubular root, [iv] microbody-lipid globule complex cisterna structure and [v] thickness of the flagellar plug) revealed ancestral and derived states. The sixth character, structure of the paracrystalline inclusion, did not resolve ancestral and derived states. In each of the lineages within Chytridiales, the evolutionary trend appears to have been from a more complex zoospore to a less complex zoospore with reduced features. As we isolate and analyze additional taxa, we discover new ultrastructural character states that assist in taxon delineation and phylogenetic interpretation.KeywordschytridChytridiaceaeChytriomycetaceaesamplingsystematicstaxonomy AcknowledgmentsThis study was supported by the National Science Foundation through MRI DEB-0500766 and REVSYS DEB-00949305. We sincerely appreciate JE Longcore (Univ Maine) for supplying isolates for this study and WJ Davis (Univ Alabama) for discussions about chytrid evolution.
