Phylogeny of Hirsutella species (Ophiocordycipitaceae) from the USA: remedying the paucity of Hirsutella sequence data

Abstract

Hirsutella (Ophiocordycipitaceae: Hypocreales) is a genus of insect, mite, and nematode pathogens with an asexual morph, which generally produce a mucilaginous cluster of one or several conidia on phialides that are basally subulate and taper to a fine neck. The generic name Hirsutella has been proposed for suppression in favour of Ophiocordyceps as a consequence of the ending of dual nomenclature for different morphs of pleomorphic fungi in 2011. Though the generic name is well established, geographically dispersed, and speciose, exceptionally few sequences are available in online databases. We examined 46 isolates of 23 Hirsutella species from the USA, curated by the USDA-ARS Collection of Entomopathogenic Fungal Cultures (ARSEF Culture Collection), that previously had not been molecularly characterized and produced a phylogeny of these organisms; we included previously published Hirsutella and Ophiocordyceps taxa. In producing the largest phylogeny of Hirsutella isolates so far, we provide: (1) context for discussing previously-hypothesized relationships; (2) evidence for revisions as taxonomic transitions move forward; and (3) available molecular data to be incorporated into further evolutionary studies of Ophiocordycipitaceae.

A new entomopathogenic fungus, Ophiocordyceps ponerus sp. nov., from China

“…A total of 45 taxa were selected to represent the morphological and ecological diversity of Ophiocordyceps, including outgroup taxon Colletotrichum gloeosporioides, which is classified within Glomerellaceae (Sung et al, 2007). Axenic mycelia (0.05-0.1g) of tested fungi which needed to extract molecular data were harvested from PDA plates and transferred into 1.5 ml eppendorf tubes for genomic DNA extraction and PCR amplification, which were carried out as previously described (White et al, 1990;Rehner et al, 1994;Rehner et al, 2005;Sung et al, 2007;van den Brink et al, 2012;Simmons et al, 2015). Sequences from four nuclear loci, including the small subunit ribosomal RNA (SSU), the transcription elongation factor-1 alpha (TEF), the largest and second largest subunits of RNA polymerase ІІ (RPB2) and the first and the internal transcribed spacers (ITS1-5.8S rDNA-ITS2 region, ITS) were used for phylogenetic analyses.…”

“…To conform to Article 59 of the International Code of Botanical Nomenclature, some mycologists have proposed suppressing the use of some asexual names proposed for taxa in this 'O. sphecocephala clade', including Hymenostilbe, in favor of Ophiocordyceps as the genus name for the entire clade (Sung et al, 2007, Kepler et al, 2013, Quandt et al, 2014, Simmons et al, 2015, Spatafora et al, 2015. In light of these recommendations by respected researchers of these fungi, we describe the fungus represented by GZUIFR-2012xch03 as Ophiocordyceps ponerus.…”

The complete mitochondrial genome of fungal endosymbiont, Ophiocordycipitaceae sp., isolated from Ricania speculum (Hemiptera: Ricaniidae)

“…Phylogenetic trees present that our species is clustered with Hirsutella and Ophiocordyceps spp. (Figure 1; Simmons et al 2015). While rRNA of this mitogenome displays that 85.94% identity against Tolypocladium inflatum as a best hit, which is incongruent with phylogenetic tree.…”

Isolasi dan Identifikasi Entomopatogen Hirsutella citriformis (Speare) dan Potensi Miselianya sebagai Sumber Inokulum untuk Pengendalian Wereng Cokelat (Nilaparvata lugens Stål.)

“…Kuantitas DNA dicek dengan spektrofotometer dan kualitasnya dengan elektroforesis. Amplifikasi urutan DNA ITS dilakukan dengan teknik PCR mengikuti prosedur yang dikembangkan oleh Simmons et al (2015). Komponen reagen PCR dengan total volume 25 μl dalam tabung PCR (0,2 ml) yang terdiri atas larutan penyangga PCR 10× sebanyak 2,5 μl; MgCl 2 1,5 mM; d NTP 0,2 mM; marka ITS1 dan ITS4 masing-masing 0,5 μM; Taq polimerase DNA 1U; ekstrak DNA 0,2 ng.…”