Optimization of protein and cellulase secretion in Neocallimastix frontalis EB188

Tsai, K.-P.; Calza, R. E.

doi:10.1007/bf00205036

Cited by 5 publications

(1 citation statement)

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“…The feasibility of using a slot-blot detection system would require 25-100 ng target DNA to be loaded (see Figs 4 and 5). Using the method of DNA extraction from the zoospores of anaerobic gut fungi described by Tsai & Calza (1993), the yield of DNA per zoospore was reported to be 2n2 pg. The anaerobic gut fungal population is about 10& zoospores per ml of rumen contents.…”

Section: Its1 Slot-blot Analysis Shows Potential For Exploitationmentioning

confidence: 99%

Identification and characterization of anaerobic gut fungi using molecular methodologies based on ribosomal ITS1 and 18S rRNA The GenBank accession numbers for the sequences determined in this work are given in Methods.

et al. 2000

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The gut fungi are an unusual group of zoosporic fungi occupying a unique ecological niche, the anaerobic environment of the rumen. They exhibit two basic forms, with nuclear migration throughout the hyphal mass for polycentric species and with concentration of nuclear material in a zoosporangium for monocentric species. Differentiation between isolates of these fungi is difficult using conventional techniques. In this study, DNA-based methodologies were used to examine the relationships within and between two genera of monocentric gut fungi gathered from various geographical locations and host animals. The ribosomal ITS1 sequence from 16 mono-and 4 polycentric isolates was PCR-amplified and sequenced ; the sequences obtained were aligned with published sequences and phylogenetic analyses were performed. These analyses clearly differentiate between the two genera and reflect the previously published physiological conclusions that Neocallimastix spp. constitute a more closely related genus than the relatively divergent genus Piromyces. The analyses place two type species N. frontalis and N. hurleyensis together but, contrary to a recent suggestion in the literature, place them apart from the other agreed species N. patriciarum. In situ hybridization and slot-blotting were investigated as potential methods for detection of and differentiation between monocentric gut fungi. DNA slot-blot analysis using ribosomal sequences is able to differentiate between gut fungal genera and thus has considerable potential for use in ecological studies of these organisms.

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Section: Its1 Slot-blot Analysis Shows Potential For Exploitationmentioning

confidence: 99%