Agriculture and Agri-Food Canada Saskatoon Research Centre, Saskatoon, SK, Canada.
Department of Veterinary Microbiology, University of Saskatchewan, Saskatoon, SK, Canada.
New Phytol. 2014 Apr;202(2):542-553. doi: 10.1111/nph.12693. Epub 2014 Jan 21.
In order to address the hypothesis that seeds from ecologically and geographically diverse plants harbor characteristic epiphytic microbiota, we characterized the bacterial and fungal microbiota associated with Triticum and Brassica seed surfaces. The total microbial complement was determined by amplification and sequencing of a fragment of chaperonin 60 (cpn60). Specific microorganisms were quantified by qPCR. Bacteria and fungi corresponding to operational taxonomic units (OTU) that were identified in the sequencing study were isolated and their interactions examined. A total of 5477 OTU were observed from seed washes. Neither total epiphytic bacterial load nor community richness/evenness was significantly different between the seed types; 578 OTU were shared among all samples at a variety of abundances. Hierarchical clustering revealed that 203 were significantly different in abundance on Triticum seeds compared with Brassica. Microorganisms isolated from seeds showed 99-100% identity between the cpn60 sequences of the isolates and the OTU sequences from this shared microbiome. Bacterial strains identified as Pantoea agglomerans had antagonistic properties toward one of the fungal isolates (Alternaria sp.), providing a possible explanation for their reciprocal abundances on both Triticum and Brassica seeds. cpn60 enabled the simultaneous profiling of bacterial and fungal microbiota and revealed a core seed-associated microbiota shared between diverse plant genera.
为了验证假设,即来自生态和地理上多样化的植物的种子携带特征性的附生微生物群,我们对小麦和油菜种子表面相关的细菌和真菌微生物群进行了特征描述。通过扩增和测序伴侣蛋白 60(cpn60)的片段来确定总微生物组成。通过 qPCR 对特定微生物进行定量。从种子冲洗液中分离出与测序研究中鉴定的分类操作单元(OTU)相对应的细菌和真菌,并对其相互作用进行了研究。从种子冲洗液中观察到了 5477 个 OTU。种子的总附生细菌负荷或群落丰富度/均匀度在两种种子类型之间没有显著差异;578 个 OTU 在各种丰度下存在于所有样本中。层次聚类显示,与油菜相比,在小麦种子上有 203 个 OTU 的丰度存在显著差异。从种子中分离出的微生物在其 cp60 序列与该共享微生物组的 OTU 序列之间具有 99-100%的同一性。鉴定为成团泛菌的细菌菌株对一种真菌分离株(链格孢属)具有拮抗特性,这为它们在小麦和油菜种子上的相对丰度提供了可能的解释。cpn60 能够同时对细菌和真菌微生物组进行分析,并揭示了不同植物属之间共享的核心种子相关微生物组。