Analysis of the Dynamics of Bacterial Communities in the Rhizosphere of the Chrysanthemum via Denaturing Gradient Gel Electrophoresis and Substrate Utilization Patterns
Analysis of the Dynamics of Bacterial Communities in the Rhizosphere of the Chrysanthemum via Denaturing Gradient Gel Electrophoresis and Substrate Utilization Patterns
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DOI：
10.1128/aem.64.12.4950-4957.1998
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发表时间：
1998-12
影响因子：
4.4
通讯作者：
B. M. Duineveld;A. Rosado;J. V. van Elsas;J. A. van Veen
中科院分区：
文献类型：
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作者：
B. M. Duineveld;A. Rosado;J. V. van Elsas;J. A. van Veen
为了更好地了解菊花根际细菌群落的时空动态，采用了两种互补的方法：一种是基于16S rRNA基因的聚合酶链式反应(16S RRNA)结合变性梯度凝胶电泳(DGGE)的分子细菌群落分析方法；另一种是琼脂平板法，其中使用了11种单一碳源利用试验。DGGE图谱表明，直接从根际DNA提取的细菌群落在菊花发育的根部基本稳定，随着时间的推移或不同年龄的根部之间的变化很小。这些模式也与从大量土壤样本中获得的DNA提取液相似。用稀释板上的微生物菌落作为目标DNA的来源，得到的DGGE图谱与直接提取的DNA图谱不同。此外，这些模式在植物重复之间也表现出差异，但在重复平板之间也是如此。单一碳源利用试验的结果表明，根尖根际细菌群落的代谢谱在植物生长过程中没有发生实质性的变化。这表明通过根尖的存在来选择性地发展特定的细菌种群。另一方面，在植物生长过程中，根基根际细菌群落的代谢谱也发生了变化。通过8个单一碳源利用试验，观察到植物发育阶段对能够在这些碳源上生长的细菌数量有显著影响。
ABSTRACT In order to gain a better understanding of the spatial and temporal dynamics of bacterial communities of the rhizosphere of the chrysanthemum, two complementary methods were used: a molecular bacterial community profiling method, i.e., 16S rRNA gene-based PCR followed by denaturing gradient gel electrophoresis (DGGE), and an agar plate method in which 11 sole-carbon-source utilization tests were used. The DGGE patterns showed that the bacterial communities as determined from direct rhizosphere DNA extracts were largely stable along developing roots of the chrysanthemum, with very little change over time or between root parts of different ages. The patterns were also similar to those produced with DNA extracts obtained from bulk soil samples. The DGGE patterns obtained by using microbial colonies from dilution plates as the source of target DNA were different from those found with the direct DNA extracts. Moreover, these patterns showed differences among plant replicates but also among replicate plates. Results obtained with the sole-carbon-source utilization tests indicated that the metabolic profile of the bacterial communities in the rhizosphere of the root tip did not change substantially during plant growth. This suggests selective development of specific bacterial populations by the presence of a root tip. On the other hand, the metabolic profile of bacterial communities in the rhizosphere of the root base changed during plant growth. With eight sole-carbon-source utilization tests, a significant effect of the development stage of the plant on the number of bacteria which were able to grow on these carbon sources was observed.