Enumeration of actively respiring bacteria in soil using the CTC-SYBR green I double-staining method
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- Ryuda Noriko
- Division of Soil Environment, Faculty of Agriculture, Saga University:The United Graduate School of Agricultural Sciences, Kagoshima University
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- Wang Xiaodan
- Division of Soil Environment, Faculty of Agriculture, Saga University
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- Ueno Daisuke
- Division of Soil Environment, Faculty of Agriculture, Saga University
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- Inoue Koichi
- Division of Soil Environment, Faculty of Agriculture, Saga University
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- Someya Takashi
- Division of Soil Environment, Faculty of Agriculture, Saga University
Bibliographic Information
- Other Title
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- CTC-SYBR Green I二重蛍光染色法による土壌中の呼吸活性陽性菌の定量
- CTC-SYBR Green 1二重蛍光染色法による土壌中の呼吸活性陽性菌の定量
- CTC SYBR Green 1ニジュウ ケイコウ センショクホウ ニ ヨル ドジョウ チュウ ノ コキュウ カッセイ ヨウセイキン ノ テイリョウ
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Abstract
The tetrazolium salt 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) has been employed for the detection and enumeration of respiratory-active bacteria in environmental samples. In this method, CTC was reduced by a respiratory chain to form CTC-formazan crystals, which were deposited intracellularly and were enumerated under an epifluorescence microscope. We evaluated a CTC-SYBR Green I double-staining (CTC-SG) method, by which both CTC-formazan particles and whole cells could be observed by CTC and SG, respectively. Pure culture studies of several Gram-positive and -negative bacteria revealed that one or more CTC-formazan crystals were observed in a single bacterial cell and extracellularly. Consequently, the conventional CTC method gave counts 1.2-1.6 times higher than the CTC-SG method, suggesting overestimation by the CTC method. A starved culture of E coil showed the presence of viable but nonculturable cells, as detected by the CTC-SG method after two weeks of incubation. The counts of metabolically active bacteria in upland and paddy soils by the CTC-SG method were 4.5×10^8-1.4×10^9 cells g^<-1> dry soil, which were as much as 10-12% of the total bacterial count and 1.5-5.2 times higher than the plate counts. These results suggest that the CTC-SG method gives more accurate counts of respiratory-active bacteria in soil samples than the conventional CTC method.
Journal
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- Soil Microorganisms
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Soil Microorganisms 64 (1), 18-24, 2010
Japanese Society of Soil Microbiology
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Details 詳細情報について
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- CRID
- 1390001204253062656
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- NII Article ID
- 110009468711
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- NII Book ID
- AN00352796
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- ISSN
- 21896518
- 09122184
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- NDL BIB ID
- 10673236
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- Text Lang
- ja
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- Data Source
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- JaLC
- IRDB
- NDL
- CiNii Articles
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- Abstract License Flag
- Disallowed