Sugiyama Junta, Kiyuna Tomohiko, Nishijima Miyuki, An Kwang-Deuk, Nagatsuka Yuka, Tazato Nozomi, Handa Yutaka, Hata-Tomita Junko, Sato Yoshinori, Kigawa Rika, Sano Chie
TechnoSuruga Laboratory Co. Ltd., Chiba Branch Office.
TechnoSuruga Laboratory Co. Ltd.
J Gen Appl Microbiol. 2017 May 12;63(2):63-113. doi: 10.2323/jgam.2017.01.007. Epub 2017 Mar 24.
Microbial outbreaks and related biodeterioration problems have affected the 1300-year-old multicolor (polychrome) mural paintings of the special historic sites Takamatsuzuka Tumulus (TT) and Kitora Tumulus (KT). Those of TT are designated as a national treasure. The microbiomes of these tumuli, both located in Asuka village, Nara, Japan, are critically reviewed as the central subject of this report. Using culture-dependent methods (conventional isolation and cultivation), we conducted polyphasic studies of the these microbial communities and identified the major microbial colonizers (Fusarium spp., Trichoderma spp., Penicillium spp., dark Acremonium spp., novel Candida yeast spp., Bacillus spp., Ochrobactrum spp., Stenotrophomonas tumulicola, and a few actinobacterial genera) and noteworthy microbial members (Kendrickiella phycomyces, Cephalotrichum verrucisporum (≡Doratomyces verrucisporus), Sagenomella striatispora, Sagenomella griseoviridis, two novel Cladophialophora spp., Burgoa anomala, one novel species Prototheca tumulicola, five novel Gluconacetobacter spp., three novel Bordetella spp., and one novel genus and species Krasilnikoviella muralis) involved in the biodeterioration of mural paintings, plaster walls, and stone chamber interiors. In addition, we generated microbial community data from TT and KT samples using culture-independent methods (molecular biological methods, including PCR-DGGE, clone libraries, and pyrosequence analysis). These data are comprehensively presented, in contrast to those derived from culture-dependent methods. Furthermore, the microbial communities detected using both methods are analytically compared, and, as a result, the complementary roles of these methods and approaches are highlighted. In related contexts, knowledge of similar biodeterioration problems affecting other prehistoric cave paintings, mainly at Lascaux in France and Altamira in Spain, are referred to and commented upon. Based on substrate preferences (or ecological grouping) and mapping (plotting detection sites of isolates), we speculate on the possible origins and invasion routes whereby the major microbial colonizers invaded the TT stone chamber interior. Finally, concluding remarks, lessons, and future perspectives based on our microbiological surveys of these ancient tumuli, and similar treasures outside of Japan, are briefly presented. A list of the microbial taxa that have been identified and fully or briefly described by us as known and novel taxa for TT and KT isolates since 2008 is presented in Supplementary Materials.
微生物爆发及相关生物劣化问题已对高松冢古坟（TT）和吉野里古坟（KT）这两处拥有1300年历史的多色壁画造成了影响。高松冢古坟的壁画被指定为国宝。本报告重点审视了位于日本奈良飞鸟村的这两处古坟的微生物群落。我们采用依赖培养的方法（传统分离培养法），对这些微生物群落进行了多相研究，确定了主要的微生物定殖者（镰刀菌属、木霉属、青霉属、暗色顶孢霉属、新型念珠菌酵母属、芽孢杆菌属、慢生根瘤菌属、土居寡养单胞菌，以及少数放线菌属）和值得关注的微生物成员（藻状肯德里克氏菌、疣孢头孢霉（≡疣孢刀孢霉）、条纹孢萨氏霉、灰绿萨氏霉、两个新型枝孢瓶霉属物种、异常布尔戈霉、一个新型物种土居原壁菌、五个新型葡糖醋杆菌属物种、三个新型博德特氏菌属物种，以及一个新型属和物种壁画克拉西尼科夫氏菌），它们参与了壁画、石膏墙和石室内部的生物劣化过程。此外，我们还使用非依赖培养的方法（分子生物学方法，包括PCR - DGGE、克隆文库和焦磷酸测序分析）从高松冢古坟和吉野里古坟的样本中获取了微生物群落数据。与依赖培养方法获得的数据相比，这些数据得到了全面呈现。此外，对两种方法检测到的微生物群落进行了分析比较，结果突出了这些方法和途径的互补作用。在相关背景下，还提及并评论了影响其他史前洞穴壁画（主要是法国的拉斯科洞穴壁画和西班牙的阿尔塔米拉洞穴壁画）的类似生物劣化问题的相关知识。基于底物偏好（或生态分组）以及绘图（绘制分离株的检测位点），我们推测了主要微生物定殖者侵入高松冢古坟石室内部的可能来源和入侵途径。最后，简要介绍了基于我们对这些古代古坟以及日本境外类似宝藏的微生物学调查得出的结论、经验教训和未来展望。自2008年以来，我们已鉴定并完整或简要描述的高松冢古坟和吉野里古坟分离株已知和新的微生物分类单元列表见补充材料。
