A novel method for sampling bacteria on plant root and soil surfaces at the microhabitat scale
A novel method for sampling bacteria on plant root and soil surfaces at the microhabitat scale
复制标题
DOI：
10.1016/j.mimet.2008.04.013
复制
发表时间：
2008-09-01
影响因子：
2.2
通讯作者：
Hirsch, Penny R.
中科院分区：
文献类型：
--
作者：
Dennis, Paul G.;Miller, Anthony J.;Hirsch, Penny R.
本研究报告了第一种在空间尺度上近似微生境取样细菌的方法。该方法的核心是使用具有已知表面积（0.013 mm(2)）的激光切割尖端的钨棒。用解剖显微镜观察暴露的植物根系或土壤表面，用微机械臂引导微采样棒到采样点。粘附在采样尖上的细菌随后被回收用于微生物学分析。这种方法从根表面去除细菌的效率与使用传统的洗涤技术从解剖的根段中回收细菌的效率相似。然而，由于微采样尖端的表面积是已知的，新方法的优点是消除了由于对采样的根或土壤表面估计不准确而导致的细菌密度估计不准确。当用于调查根际细菌的空间分布时，新技术揭示了与现有方法一致的趋势，同时提供了在更小的空间尺度上获得关于群落结构的额外信息。这种新方法的空间尺度比其他涉及抽拭的采样方法小约1000倍。这种新技术代表了在微生境尺度上促进微生物生态研究的重要方法步骤。爱思唯尔B.V.版权所有
This study reports the first method for sampling bacteria at a spatial scale approximating a microhabitat At the core of this method is the use of tungsten rods with laser-cut tips of known surface area (0.013 mm(2)). Exposed plant root or soil surfaces were viewed with a dissecting microscope and micro-sampling rods were guided to sample sites using a micro-manipulator. Bacteria that adhered to the sampling tips were then recovered for microbiological analyses. The efficiency of this method for removing bacteria from root surfaces was similar to that with which bacteria are recovered from dissected root segments using the conventional technique of washing. However, as the surface area of the micro-sampling tips was known, the new method has the advantage of eliminating inaccuracy in estimates of bacteria[ densities due to inaccurate estimation of the root or soil surface sampled. When used to investigate spatial distributions of rhizoplane bacteria, the new technique revealed trends that were consistent with those reported with existing methods, while providing access to additional information about community structure at a much smaller spatial scale. The spatial scale of this new method is ca. 1000-times smaller than other sampling methods involving swabbing. This novel technique represents an important methodological step facilitating microbial ecological investigations at a microhabitat scale. Crown Copyright (C) 2008 Published by Elsevier B.V. All rights reserved.