Resolving colocalization of bacteria and metal(loid)s on plant root surfaces by combining fluorescence in situ hybridization (FISH) with multiple-energy micro-focused X-ray fluorescence (ME μXRF).
Resolving colocalization of bacteria and metal(loid)s on plant root surfaces by combining fluorescence in situ hybridization (FISH) with multiple-energy micro-focused X-ray fluorescence (ME μXRF).
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DOI：
10.1016/j.mimet.2016.09.018
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发表时间：
2016-12
影响因子：
2.2
通讯作者：
Maier, Raina M.
中科院分区：
文献类型：
--
作者：
Honeker, Linnea K.;Root, Robert A.;Chorover, Jon;Maier, Raina M.
金属(固体)--人类活动造成的环境污染是一个全球性问题。要了解污染物的去向，需要阐明影响金属(Loid)形态的生物和非生物因素，从分子到领域。需要改进的方法来评估微尺度的过程，如发生在植物组织、微生物细胞和金属(LOID)之间的生物地球化学界面的过程。在这里，我们提出了一种先进的方法，将荧光原位杂交(FISH)和同步加速器多能量微聚焦X射线荧光微探针成像(MEμXRF)相结合，以检测细菌和金属(LOID)S在植物根面上的共存。使用SytoBC核酸染色和FISH探针将细菌显示在一个小的根部切片上，目的是针对特定的细菌和特定的组(阿尔法蛋白细菌、伽马蛋白细菌或放线细菌)。用ME-μ-XRF分析了同一根区As和Fe的元素分布和金属(LOID)形态。FISH和MEμXRF图像使用ImageJ软件进行比对，以关联微生物和地球化学结果。共定位定量分析结果显示，AS与根面氧化铁菌斑共定位的比例(重叠率为0.072.49±0.19)显著高于与细菌(0.072±0.052)的共定率(p<；0.05)。在与金属(Loid)S共定位的细菌中，以耐金属著称的放线杆菌与As和Fe的相关性高于甲蛋白细菌和伽马蛋白细菌。这种方法展示了如何结合这些微技术可以扩展我们对根、金属(Loid)S和微生物之间微观相互作用的理解，这些信息应该导致改进金属(Loid)形态和命运的机制模型。
Metal(loid)-contamination of the environment due to anthropogenic activities is a global problem. Understanding the fate of contaminants requires elucidation of biotic and abiotic factors that influence metal(loid) speciation from molecular to field scales. Improved methods are needed to assess micro-scale processes, such as those occurring at biogeochemical interfaces between plant tissues, microbial cells, and metal(loid)s. Here we present an advanced method that combines fluorescence in situ hybridization (FISH) with synchrotron-based multiple-energy micro-focused x-ray fluorescence microprobe imaging (ME μXRF) to examine colocalization of bacteria and metal(loid)s on root surfaces of plants used to phytostabilize metalliferous mine tailings. Bacteria were visualized on a small root section using SytoBC nucleic acid stain and FISH probes targeting the domain Bacteria and a specific group (Alphaproteobacteria, Gammaproteobacteria, or Actinobacteria). The same root region was then analyzed for elemental distribution and metal(loid) speciation of As and Fe using ME μXRF. The FISH and ME μXRF images were aligned using ImageJ software to correlate microbiological and geochemical results. Results from quantitative analysis of colocalization show a significantly higher fraction of As colocalized with Fe-oxide plaques on the root surfaces (fraction of overlap 0.49 ± 0.19) than to bacteria (0.072 ± 0.052) (p < 0.05). Of the bacteria that colocalized with metal(loid)s, Actinobacteria, known for their metal tolerance, had a higher correlation with both As and Fe than Alphaproteobacteria or Gammaproteobacteria. This method demonstrates how coupling these micro-techniques can expand our understanding of micro-scale interactions between roots, metal(loid)s and microbes, information that should lead to improved mechanistic models of metal(loid) speciation and fate.
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DOI：
10.1038/ismej.2012.8
发表时间：
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期刊：
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影响因子：
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影响因子：
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影响因子：
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DOI：
10.1016/j.scitotenv.2016.04.168
发表时间：
2016-09-15
期刊：
The Science of the total environment
影响因子：
--
作者：
Gil-Loaiza J;White SA;Root RA;Solís-Dominguez FA;Hammond CM;Chorover J;Maier RM
通讯作者：
Maier RM