Molecular characterization and evaluation of plant litter-associated fungi from the spring ‘grazing corridor’ of a sheep herd vulnerable to alveld disease

Abstract

This study sample and identify species of fungi on withered vegetation in the spring 'grazing corridor' from infields to Narthecium bogs for a sheep herd almost chronically vulnerable to phototoxic disease. Hepatogen photosensitizing disorders like alveld attack grazing sheep, especially lambs, in various parts of the world. It has been hypothesized that saponin metabolites in the monocotyledonous plant Narthecium ossifragum causes the disorder in Norway, however, this has not been verified. Thus, the search for other causal agents or saponin cofactors (the cofactor hypothesis) has been intensified, and endophytic poisonous fungi associated with dead N. ossifragum leaves and grasses are among the prime suspects. The fungal diversity was targeted by obtaining axenic cultures from surface-sterilized plant material, with subsequent DNA isolation, PCR, and sequencing of the ITS nrDNA region. The taxonomic affinities of the obtained sequences were thereafter explored by similarity searches against the public access sequence database EMBL/GenBank/DDBJ. Among the spectrum of identified taxa were representatives of Cladosporium, Fusarium, Penicillium, and Trichoderma, all of which are well known as potential producers of toxins. A possible involvement of these toxic species in the etiology of alveld is evaluated and discussed.

Mollisiaceae: An overlooked lineage of diverse endophytes

“…(direct submission) Root tips of Pinus sylvestris Finland 3.46.4A KM068442 — — — — Sarjala et al. (direct submission) Withered plant material Norway A4-3 AM231338 — — — — Mysterud et al. (2007) Wood Antarctica AB45 FJ235978 — — — — Arenz & Blanchette (2009) Myotis septentrionalis wing USA APA-2013 clone LJ76MstMg10w KF212280 — — — — Johnson et al.…”

The secrets of Cryptomyces maximus (Rhytismataceae): ecology and distribution in the Nordic countries (Norden), and a morphological and ontogenetic update

“…DNA was extracted from fresh herbarium samples using a cetyltrimethyl ammonium bromide (CTAB) extraction SURWRFRO 0XUUD\ 7KRPSVRQ ZLWK VRPH PRGL¿-cations described in detail in Mysterud et al (2007). PCR DPSOL¿FDWLRQ ZDV WDUJHWHG WR DPSOLI\ WKH LQWHUQDO WUDQscribed spacer (ITS) region of nuclear ribosomal DNA, using the primer pair ITS5/ITS4 (White et al 1990) and Illustra PuReTaq Ready-To-Go PCR beads (GE Healthcare, Buckinghamshire, UK).…”

Morphological update on Calycina marina (Pezizellaceae, Helotiales, Leotiomycetes), a new combination for Laetinaevia marina

“…We extracted DNA of Laetinaevia marina voucher specimens and the culture isolate using a modified cetyltrimethyl ammonium bromide (CTAB) extraction protocol (Murray and Thompson 1980) and amplified the target loci according to Mysterud et al (2007) and Johnson et al (2009) with slight modifications. PCR amplifications were performed on a PTC-0200 DNA engine (MJ Research, Waltham, MA, USA) and GeneAmp PCR System 9700 (Applied Biosystems, Foster City, CA, USA) using the primer pairs ITS5-ITS4 (White et al 1990), LR0R-LR5 (Vilgalys andHester 1990, Rehner andSamuels 1994), NS1-NS24 (White et al 1990, Gargas andTaylor 1992) and RPB1Ac-RPB1Cr (Stiller andHall 1997, Matheny et al 2002, both with unpublished modifications by Giho Sung).…”