Application of molecular techniques for identification of fungal communities colonising paper material

被引:99
|
作者
Michaelsen, Astrid
Pinzari, Flavia
Ripka, Katrin
Lubitz, Werner
Pinar, Guadalupe
机构
[1] Univ Vienna, Dept Med Chem, A-1090 Vienna, Austria
[2] Ist Cent Patol Libro, Biol Lab, I-00184 Rome, Italy
基金
奥地利科学基金会;
关键词
objects of art; DGGE; internal spacer regions; PCR; fungal communities;
D O I
10.1016/j.ibiod.2006.06.019
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Archives and libraries all over the world suffer from biodeterioration of writings caused by microorganisms, especially fungi. With traditionally used culture-dependent methods, only a small amount of effectively colonising organisms is detected. Restoration and maintenance of written cultural heritage is therefore problematic due to incomplete knowledge of the deterioration agents. In the present study, culture-independent molecular methods were applied to identify fungal communities colonising paper samples of different composition and age. Nucleic-acid-based strategies targeting the internally transcribed spacer (ITS) regions, which are nested in the nuclear rDNA repeats, were selected to investigate the fungal diversity on paper. The ITS regions possess a high variation among taxonomically distinct fungal species and even within the species. With this aim, several molecular biological methods were optimised for working with paper materials. Here, we introduce a DNA extraction protocol, which allowed the direct extraction of PCR-amplifiable DNA from samples derived from different kinds of paper. The DNA extracts were used to amplify either the ITS1 or ITS2 region by using different fungi-specific primer sets. The ITS-aniplified regions were subsequently analysed by denaturing gradient gel electrophoresis (DGGE). Conditions for DGGE analysis, gradient, voltage, and running time, were established to accurately discriminate different fungal species in complex communities. Pure fungal strains were used to constitute a marker for further comparative investigations of historic papers. (c) 2006 Elsevier Ltd. All rights reserved.
引用
收藏
页码:133 / 141
页数:9
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