Application of denaturing gradient gel electrophoresis for analysing the gut microflora of Lumbricus rubellus Hoffmeister under different feeding conditions
Application of denaturing gradient gel electrophoresis for analysing the gut microflora of Lumbricus rubellus Hoffmeister under different feeding conditions
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DOI：
10.1017/s0007485308006056
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发表时间：
2008-06-01
影响因子：
1.9
通讯作者：
Insam, H.
中科院分区：
文献类型：
--
作者：
Knapp, B. A.;Seeber, J.;Insam, H.
蚯蚓在土壤生态系统中起着至关重要的作用，因为它影响有机质的分解和养分的循环。通过摄入有机和矿物质的混合物，各种细菌和真菌被带到蚯蚓的肠道。为了更好地了解红曲霉与摄取的微生物之间的相互作用，本研究试图揭示饮食是否影响红曲霉肠道微生物区系的组成，或者它的肠道是否拥有本土的、物种特有的微生物群。建立了红曲霉的饲养实验，在野外采集个体，转移到气候室，用不同质量的食物来源(矮灌木凋落物、草凋落物和马粪)饲养6周。从蚯蚓的肠道、食物来源和周围土壤中提取DNA，并用聚合酶链式反应-变性梯度凝胶电泳法(PCR-DGGE)进一步分析。我们能够证明，肠道微生物区系受到摄取的食物来源的强烈影响，与周围土壤的微生物区系有很大不同。对细菌DGGE指纹优势条带的测序显示，所有肠道样本中都有强烈的Y-变形杆菌存在，与食物来源无关。在肠道中找不到一种对饮食变化有很强抵抗力的特定微生物群。
The earthworm, Lumbricus rubellus, plays an essential role in soil ecosystems as it affects organic matter decomposition and nutrient cycling. By ingesting a Mixture of organic and mineral material, a variety of bacteria and fungi are carried to the intestinal tract of the earthworm. To get a better understanding of the interactions between L. rubellus and the microorganisms ingested, this study tried to reveal if the diet affects the composition of the gut microflora of L. rubellus or if its intestinal tract hosts an indigenous, species-specific microbiota. A feeding experiment with L. rubellus was set up; individuals were collected in the field, transferred to a climate chamber and fed with food sources of different quality (dwarf shrub litter, grass litter or horse dung) for six weeks. DNA was extracted from the guts of the earthworms, as well as from the food sources and the surrounding soil, and further analysed by a molecular fingerprinting method, PCR-DGGE (Polymerase Chain Reaction - Denaturing Gradient Get Electrophoresis). We were able to demonstrate that the gut microbiota was strongly influenced by the food source ingested and was considerably different to that of the surrounding soil. Sequencing of dominant bands of the bacterial DGGE fingerprints revealed a strong occurrence of y-Proteobacteria in all gut samples, independent of the food source. A specific microflora in the intestinal tract of L. rubellus, robust against diet changes, could not be found.