Neocallimastix californiae O’Malley, Theodorou & Solomon, sp. nov.
Index Fungorum number: IF 551675, Facesoffungi number: FoF 02061, Fig. 3
Etymology – The specific epithet refers to the state of California where the fungus was isolated.
Holotype – Neocallimastix sp. G1 (O’Malley Lab, University of California, Santa Barbara, NCBI Taxon ID: 1550276), JMRC:SF:12176.
An obligate anaerobic fungus isolated from the feaces of a goat (Capra aegagrushircus) housed at the Santa Barbara Zoo (www.sbzoo.org) in 2013. The species is monocentric and has a determinate (finite) life cycle. The fungus exhibits endogenous zoosporangial development (i.e., the encysted zoospore retains the nucleus). The encysted zoospore germinates to form a rhizoidal system and a single typically sphaerical zoosporangium (≥ 120μm diam.) that on maturity liberates ≥ 100 zoospores. The rhizoidal system is devoid of nuclei (as seen under DAPI staining) and is highly branched and tapering. The zoosporangium is typically attached to the rhizoidal system via one main rhizoid or sporangiophore. A septum is often visible in mature zoosporangia, separating the zoosporangium from the sporangiophore. Free swimming zoospores are typically sphaerical (ca. 10μm diam.) and the species is characterized by the presence of ca. 16 or more posteriorly directed flagella that are in length up to 3–fold the diam. of the zoospore. When swimming the flagella beat together as if they were a single flagellum and thus propel the zoospore forward in a spiral or helical motion.
The reference culture is maintained by continual passage at the University of California, Santa Barbara (G1, JMRC:SF:12176, holotype), and under cryopreservation in repositories at the O’Malley Lab, University of California, Santa Barbara, and University of Jena and Leibniz Institute for Natural Product Research and Infection Biology, Jena, Germany (Jena Microbial Resource Collection JMRC: SF: 012176 – ex-type). Fixed glutaraldehyde preparations are also kept by the O’Malley Lab.
The internal transcribed spacer regions of the ribosomal RNA were amplified with primers JB206/JB205 (Tuckwell et al. 2005). Phylogenetic analysis of the ITS1 regions of several cultured anaerobic fungal specimens spanning all eight known genera and partial 28 s reads, firmly place G1 in the genus Neocallimastix as a distinct, previously unclassified sister species to established cultures such as N. frontalis (Figs. 1, and 2). The ~190 Mbp genome has also been sequenced by the US Department of Energy’s Joint Genome Institute (JGI) to reveal that G1 is a polyploid organism. The genome will be made available at Mycocosm in 2016 (http://genome.jgi.doe.gov/programs/fungi/index.jsf).

Fig. 1 Molecular phylogeny generated by maximum likelihood analysis of ITS1 sequence data from the Neocallimastigomycota. Representative species from all known eight genera (indicated) are shown. Bootstrap values above 50 % are indicated above each branch. Ex-types (reference strains) are bolded and new isolates are indicated in blue.

Fig. 2 Molecular phylogeny generated by maximum likelihood analysis of partial large subunit (28S) ribosomal DNA sequence data from the Neocallimastigomycota. Bootstrap values above 50 % are indicated above each branch. New isolates are indicated in with a filled shape.

Fig. 3 Neocallimastix californiae (holotype) a Sphearical zoospores with multiple flagella which are splayed out b Multiple sporangia, demonstrating the predominantly sphearical to ovoid structure c Goat.