Molecular Identification Of Fungi Research Articles

Penapisan Kapang Laut Penghasil Senyawa Sitotoksik dari Beberapa Perairan di Indonesia

ABSTRACT Marine-derived fungi have proven to be a rich source of cytotoxic compounds for the development of new anti cancer drugs. The aims of this research were to: 1) screen cytotoxic activity of marine fungi from Indonesian waters, 2) indentify marine fungus that produced that produced the most active cytotoxic compound and 3) investigate inhibition concentration 50 (IC50) value of cytotoxic compound. The fungi were isolated from marine organism collected from Wakatobi Marine National Park-South East Sulawesi, Binuangeun Beach-Banten, Manado watersNorth Sulawesi and Kepulauan Seribu Marine National Park-Jakarta. Liquid cultures of the fungi were carried out in Malt Extract Broth and Soluble Starch Soytone medium for 4 weeks at 27–28oC without shaking. Molecular identification of fungus was conducted through PCR amplificatin using primers of ITS1 and ITS4 primer. Cytotoxic activity of the extract was tested by using MTT (3-(4.4-dimethylthiazol-2-yl)-2.5-diphenyl-tetrazolium bromide) method. The MTT test showed that MFW39 strain exhibited the strongest cytotoxic activity. Molecular identification revealed that MFW39 marine fungus was similar to Emericella nidulans with precent identity of 99%. Mycelium and broth extract of MFW39 fungus inhibited the growth of T47D cell with IC 50 values of 21.9 and 169.3 µg/mL, respectively. Further research will be focus on to the strain of MFW39 marine fungi.

Towards standardization of the description and publication of next‐generation sequencing datasets of fungal communities

Towards standardization of the description and publication of next‐generation sequencing datasets of fungal communities

Conspecificity of DAOM 197198, the model arbuscular mycorrhizal fungus, with Glomus irregulare: molecular evidence with three protein-encoding genes

Ribosomal nuclear genes are routinely utilized in the molecular identification of fungi. The variation in the multiple copies of these genes within each Glomeromycota strain and species reduces their usefulness for molecular characterization of arbuscular mycorrhizal fungi. To explore the potential of molecular tools for the identification of Glomus species, a multi-gene analysis approach was undertaken. Three protein-encoding genes were tested, namely elogation factor 1-α (765 bp), V-H+-ATPase VHA5 (1468 bp), and F0F1-ATPase β-subunit (621 bp). The latter is newly reported for the Glomeromycetes. Eleven species, including the type-specimen of Glomus irregulare Blaszk., Wubet, Renker & Buscot, a reference strain of G. intraradices N.C. Schenck & G.S. Sm. (DAOM 225240), and five strains of Glomus sp. formerly identified as G. intraradices, were analysed. These genes did not show polymorphisms within strains, and results indicated a close relationship between molecular identification and morphological characterization. Species with closely related spore morphological features, G. aggregatum N.C. Schenck & G.S. Sm., G. diaphanum Morton & Walker, G. irregulare , and Glomus sp. DAOM 197198, showed more than 99% nucleotide similarity, while the morphologically distinct species, G. cerebriforme McGee, G. clarum T.H. Nicolson & N.C. Schenck, G. claroideum N.C. Schenck & G.S. Sm., G. custos C. Cano & Dalpé, G. mosseae (T.H. Nicolson & Gerd.) Gerd. & Trappe, and G. proliferum Dalpé & Declerck, showed less than 97% similarity for at least one gene. A 100% molecular similarity for all three genes was found between G. irregulare and Glomus sp. DAOM 197198, confirming the new identity of the model arbuscular mycorrhizal fungus. Similarity thresholds for identification by DNA sequencing are discussed.

An open source software package for automated extraction of ITS1 and ITS2 from fungal ITS sequences for use in high-throughput community assays and molecular ecology

We introduce an open source software utility to extract the highly variable ITS1 and ITS2 subregions from fungal nuclear ITS sequences, the region of choice for environmental sampling and molecular identification of fungi. Inclusion of parts of the neighbouring, very conserved, ribosomal genes in the sequence identification process regularly leads to distorted results. The utility is available for UNIX-type operating systems, including MacOS X, and processes about 1000 sequences per minute.

Molecular identification of fungi isolated from stem tissue of Upland cotton (Gossypium hirsutum)

Molecular techniques such as restriction fragment length polymorphism (RFLP) analysis, random amplification of polymorphic DNA (RAPD) fingerprinting, and DNA sequencing and database comparison, were employed to identify fungi isolated from field-grown cotton plants (Gossypium hirsutum L.). DNA fragments of between 510 and 590 bp, representing the two rDNA (rDNA) internal transcribed spacers (ITS1 and ITS2) and the intervening 5.8S rRNA gene, were amplified from the fungi with eukaryotic consensus primers. Subsequent digestion with the restriction endonucleases AluI, CfoI, HaeIII, HinfI and HpaII enabled the allotment of all 57 isolates to 13 different groups. Restriction analysis was supported by RAPD–PCR analysis of multiple isolates and rDNA sequencing of representative fungi from each group. Sequence alignment and comparison with rDNA sequences of other fungi available in GenBank allowed for putative identification of three different taxa of Fusarium, two taxa each of Cladosporium, Diaporthe and Nectria, and one taxon each of Alternaria, Ampelomyces, Bartalinia, Phaeosphaeria and Rhizoctonia. Many of the stem-colonising fungi identified in this study are either pathogenic on cotton or have elsewhere been found to act as biocontrol agents.

Molecular identification of fungi causing rot of building timbers

Molecular identification of fungi causing rot of building timbers