Single-cell Raman spectral profiles of Pseudomonas fluorescens SBW25 reflects in vitro and in planta metabolic history
Single-cell Raman spectral profiles of Pseudomonas fluorescens SBW25 reflects in vitro and in planta metabolic history
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DOI：
10.1007/s00248-006-9138-5
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发表时间：
2007-04-01
影响因子：
3.6
通讯作者：
Spiers, Andrew J.
中科院分区：
文献类型：
--
作者：
Huang, Wei E.;Bailey, Mark J.;Spiers, Andrew J.
单细胞拉曼显微光谱具有报道细菌全细胞化学组成的潜力，反映了代谢状态以及生长历史。这种潜力已经通过对从土壤和植物相关细菌荧光假单胞菌SBW 25获得的拉曼光谱轮廓（RSP）的判别函数分析得到证明，所述细菌在体外使用限定培养基生长，并且在植物中使用3个月大的甜菜（甜菜变种，甜菜Roberta）。SBW 25体外RSP数据显示，生长在不同氨基酸、糖、TCA循环中间体、丰富的King's B和来自甜菜植物圈的培养基上的那些细胞之间存在显著差异。拉曼分析也能够跟踪SBW 25在一段时间内缺乏碳的转变，并且SBW 25在植物中的RSP数据也显示出在从土壤和根际回收的细菌之间具有显著差异的变化。SBW 25全细胞的化学组成，因此，生长和代谢的历史，可以解释通过共分析在体外和在植物RSP数据。SBW 25恢复从植物圈被认为是更相似SBW 25在体外生长的Fru或Asp，而不是葡萄糖或精氨酸，和很不一样，从碳饥饿。这表明SBW 25在植物圈中的生长通常既不受碳分解代谢物的抑制，也不受碳限制。这些发现表明，单细胞RSP的分析可以区分具有不同代谢历史或从其自然环境的不同部分恢复后的细菌的同基因群体。此外，拉曼分析还能够提供生物学相关的生化推断，然后可以对其进行测试，以揭示区分在复杂环境中生长并暴露于不同条件的细菌的机制基础（生物化学-代谢-遗传）。
Single-cell Raman microspectroscopy has the potential to report on the whole-cell chemical composition of bacteria, reflecting metabolic status as well as growth history. This potential has been demonstrated through the discriminant functional analysis of Raman spectral profiles (RSP) obtained from the soil and plant-associated bacterium Pseudomonas fluorescens SBW25, grown in vitro using defined media, and in planta using 3-month-old sugar beets (Beta vulgaris var. Roberta). SBW25 in vitro RSP data showed significant variation between those cells grown on different amino acids, sugars, TCA cycle intermediates, rich King's B, and culture media derived from the sugar beet phytosphere. Raman analysis was also able to follow the transition of SBW25 starved of carbon over a period of days, and SBW25 in planta RSP data also showed variation with significant differences between bacteria recovered from soil and the rhizosphere. SBW25 whole-cell chemical composition, and therefore growth and metabolic history, could be interpreted by coanalyzing in vitro and in planta RSP data. SBW25 recovered from the phytosphere was found to be more similar to SBW25 grown in vitro on Fru or Asp, rather than on Glc or Arg, and quite dissimilar to that resulting from carbon starvation. This suggests that SBW25 growth in the phytosphere is generally neither carbon-catabolite-repressed nor carbon-limited. These findings demonstrate that the analysis of single-cell RSP can differentiate between isogenic populations of bacteria with different metabolic histories or after recovery from different parts of their natural environment. In addition, Raman analysis is also capable of providing biologically relevant biochemical inferences, which might then be tested to uncover the mechanistic basis (biochemical-metabolic-genetic) differentiating bacteria growing in complex environments and exposed to different conditions.