绿背山雀人工巢箱真菌多样性差异分析

真菌是巢穴微生物的重要组成部分，与鸟类的生存、繁殖和环境适应息息相关。本研究通过悬挂人工巢箱招引绿背山雀(Parus monticolus)入住，基于内转录间隔区(ITS)测序技术，对绿背山雀繁殖成功巢箱与筑巢未产卵巢箱真菌群落的组成差异进行探究。结果显示，在门水平上，子囊菌门(Ascomycota，98.81%)是繁殖成功巢箱内微生物的主要菌门；子囊菌门(85.59%)和担子菌门(Basidiomycota，8.33%)是筑巢未产卵巢箱内微生物的主要菌门。在属水平上，繁殖成功巢箱的优势属为拟单宽皿菌属(Phialemoniopsis，83.04%)、曲霉属(Aspergillus，4.75%)、子囊菌属(Arthroderma，4.29%)和柄帚霉属(Scopulariopsis，1.78%)；筑巢未产卵巢箱的优势属为拟单宽皿菌属(36.06%)、曲霉属(14.53%)、青霉属(Penicilliu，6.22%)、单端孢霉属(Trichothecium，5.80%)、德巴利酵母菌属(Debaryomyces，1.67%)和蝶孔耳属(Papiliotrema，1.09%)。Alpha多样性分析表明，筑巢未产卵巢箱中真菌的多样性和丰富度均显著高于繁殖成功巢箱(P < 0.05)；Beta多样性分析表明，繁殖成功巢和筑巢未产卵巢箱之间的真菌群落存在显著差异；LEfSe分析共检测到19个具有统计学差异的生物标记物，繁殖成功巢箱和筑巢未产卵巢箱的显著生物标志物分布在子囊菌门和担子菌门中，两种巢箱的标记物种存在显著差异。整体来说，与繁殖成功巢相比，筑巢未产卵巢内分布有更多的潜在病原菌。

Analysis of Fungal Differences in Artificial Nest Boxes of Parus monticolus

objectives] Fungi are important components of nest microbiota, which are closely related to the survival, reproduction, and environmental adaptation of birds. However, most of the research on the relationship between birds and microbiota is mainly focused on intestinal microbes, but few studies focused on the micro-ecological environment in the nest. So, we want to investigate the fungal community diversity in the nest of the Green-backed Tit (Parus monticolus). Methods] From April to June 2020, in Aha Lake National Wetland Park in Guiyang, Guizhou Province, the Green-back Tits were attracted by hanging artificial nesting boxes. All nests were inspected once a week in the early breeding period, and once every 2 days after the emergence of nest materials. Microbial samples were collected from successful breeding nests and the nests without laying eggs. Based on internal transcribed spacer (ITS) sequencing technology, the differences in fungal community composition between successful breeding nests and the nests without laying eggs of Green-backed Tit were investigated. Results] At the phylum level, the results showed that Ascomycota (98.81%) was the main phyla of successful breeding nests, Ascomycota (85.59%) and Basidiomycota (8.33%) were the main phyla of the nests without laying eggs (Fig. 2a). At the genus level, the dominant genera of successful breeding nests were Phialemoniopsis (83.04%), Aspergillus (4.75%), Arthroderma (4.29%) and Scopulariopsis (1.78%), and the dominant genera of the nests without laying eggs were Phialemoniopsis (36.06%), Aspergillus (14.53%), Penicilliu (6.22%), Trichothecium (5.80%), Debaryomyces (1.67%) and Papiliotrema (1.09%) (Fig. 2b). Alpha diversity analysis showed that the diversity and richness of fungi in the nests without laying eggs were significantly higher than that in successful breeding nests (P < 0.05) (Fig. 3). Beta diversity analysis showed that there were significant differences in fungal communities between successful breeding nests and the nests without laying eggs (Fig. 4). A total of 19 biomarkers with statistical differences were detected by LEfSe (Linear discriminant analysis effect size) analysis. The significant biomarkers of successful breeding nests and the nests without laying eggs were distributed in Ascomycota and Basidiomycota, and there were significant differences between these two biomarker species (Fig. 5). Conclusion] In general, by comparing the differences in fungal composition in successful breeding nests and the nests without laying eggs, we find that birds may affect the composition structure of microbiota in the nests during breeding, showing more potential pathogens in the nests without laying eggs than in successful breeding nests.