Application of a yeast method for DNA extraction associated with database interrogations for the characterization of various filamentous fungi from diseased hop

Abstract

Filamentous fungi were collected from diseased hop (Humulus lupulus; L.) and DNA was prepared from 19 isolates. The critical step of cell lysis was carried out using zymolyase for fungal cell wall digestion. DNA was successfully prepared for all isolates and allowed ribosomal DNA region amplifications by PCR. The amplicons were sequenced and sequences were compared with nucleotides databases. These analyses allowed identification of each fungus and gave a precise view of the ecosystem. Seven different genera were found among the 19 isolates. A phylogenetic tree was constructed and revealed the diversity of the hop plant environment.

Gene expression in Fusarium graminearum grown on plant cell wall

“…Several dicotyledons including Arabidopsis, tobacco, tomato and soybean were described as potential hosts of this fungus (Urban et al, 2002). Furthermore, diseased hop (Humulus lupulus) plants were repeatedly sources of virulent Fusarium species (Hatsch et al, 2002;Phalip et al, 2004).…”

A histone deacetylase-dependent screen in yeast

“…Both β-2,6 and β-1,3 linked glucans provide strength to the cell wall, forming a microfibrillar network. Digestion of the yeast cell wall by lytic enzymes, such as zymolyase, is necessary for many experimental procedures, including spheroplasting, immunofluorescence, transformation, and protein purification 43–45. The main enzymatic activities of zymolyase are β-1,3 glucanase and β-1,3 glucan laminaripentao-hydrolase, which hydrolyze glucose polymers at β-1,3 glucan linkages, 46 leading to cell wall digestion.…”

Identification and characterization of filamentous fungi isolated from fermentation starters for Hong Qu glutinous rice wine brewing

“…PCR reactions were performed in 50 μl of 1.5 mM MgCl 2 , 0.2 mM dNTPs, 0.5 μM of each primer, 0.025 U of Taq polymerase (Promega Corp., USA) and 10 30 ng of fungal DNA. Reactions were performed on a MyCycler thermal cycler (BIO RAD, Hercules, USA) using a program described by Phalip et al (2004).…”