【引用本文】 李强, 黄雅丹, 何若雪, 等. 岩溶水体惰性有机碳含量及其存在机理[J]. 岩矿测试, 2018, 37(5): 475-478. doi: 10.15898/j.cnki.11-2131/td.201807250088
LI Qiang , HUANG Ya-dan , HE Ruo-xue , et al. The Concentration of Recalcitrant Dissolved Organic Carbon in the Karst Hydrosphere and Their Existing Mechanism[J]. Rock and Mineral Analysis, 2018, 37(5): 475-478. doi: 10.15898/j.cnki.11-2131/td.201807250088

岩溶水体惰性有机碳含量及其存在机理

1. 自然资源部广西岩溶动力学重点实验室, 中国地质科学院岩溶地质研究所, 广西 桂林 541004;

2. 联合国教科文组织国际岩溶研究中心, 广西 桂林 541004;

3. 桂林医学院研究生学院, 广西 桂林 541004

收稿日期: 2018-07-25  修回日期: 2018-08-26 

基金项目: 广西自然科学基金杰出青年基金项目“岩溶生物地球化学”(2015GXNSFGA139010);中国地质科学院项目“岩溶水体碳固定微型生物群落特征及其碳汇效应”(YYWF201505);中国地质调查局子项目“人为干预增加岩溶碳汇试验示范及碳汇效应综合评价”(DD20160305-05)

作者简介: 李强,研究员,主要从事岩溶生物地球化学研究。E-mail:liqiang@karst.ac.cn。。

The Concentration of Recalcitrant Dissolved Organic Carbon in the Karst Hydrosphere and Their Existing Mechanism

1. Key Laboratory of Karst Dynamics, MNR & GZAR, Institute of Karst Geology, Chinese Academy of Geological Sciences, Guilin 541004, China;

2. International Research Center on Karst under the Auspices of UNESCO, Guilin 541004, China;

3. Graduate School of Guilin Medical University, Guilin 541004, China

Received Date: 2018-07-25
Revised Date: 2018-08-26

摘要:BACKGROUND:The accepted paradigm is that recalcitrant dissolved organic carbon (RDOC) concentrations in the ocean are about 42 μmol/L. However, the concentrations of RDOC in the karst river systems remain unidentified and there is little evidence about these compounds unavailable to microbial degradation.
OBJECTIVES:To reveal the existence of RDOC and its concentrations in the karst river systems, water samples were collected at different depths (0 m, 5 m and 10 m) from three typical sites (city-river section, reservoir area and outflow area) in dammed Liu River in 2006.
METHODS:Water samples (3 L) were pre-filtered in-situ using 3 μm filter membranes, and then filtered through 0.22 μm poresize filter membranes. The 0.22 filters were used for DNA extraction following the manufacturer's instructions of the Power Water DNA Isolation Kit (Mobio Laboratories, Inc., Carlsbad, CA, USA). The real-time PCR assay of the 16S rRNA gene, as a marker of phytoplankton-bacteria, was carried out in a volume of 25 μL. The assay mixture contained 12.5 μL Green-2-Go qPCR Mastermix (Sangon Biotech Co., Ltd, China), 1 μL of primer (10 μmol/μL), 9.5 μL of distilled water, and 1 μL of template DNA (at 5 ng/μL). Thermal cycling conditions for 16S rRNA gene were as follows:an initial cycle of 95℃ for 3 min, 39 cycles of 95℃ for 60 s, 56℃ for 60 s, 72℃ for 60 s, and 72℃ for 5 min. The primers for 16S rRNA gene were designed as 338f (5'-CCTACGGGAGGCAGCAG-3') and 518r (5'-ATTACCGCGGCTGCTGG-3'). Thermal cycling, fluorescent data collection, and data analysis were carried out with CFX96 TouchTM Real-Time PCR Detection System (Bio-Rad, USA) according to instruction manual. The relationship between Ct and log starting concentration was linear (R2>0.99). The amplification efficiency was 90%-105%. The filtrates after through 3 μm filter membranes were transferred to sterile glass bottles. The bottles were incubated in the dark room at 30℃ for 180 days. Dissolved organic carbon (DOC) concentrations of all water samples were determined at 30 days interval on a total organic carbon analyzer (Multi N/C®3100) at the Institute of Karst Geology, CAGS. The obtained data were performed for correlation analyses using the Pearson correlation method (one-tailed t test) with significance defined as P<0.05 by using SPSS 13.0 software for Windows XP (IBM, Armonk, NY, USA).
RESULTS:About 24.63 μmol/L RDOC is stored in the dammed Liu River systems, supporting the hypothesis that they are resistant to microbial degradation and the dilution limits dissolved organic carbon utilization.
CONCLUSIONS:These findings demonstrate the mechanism for the long-term storage of RDOC in karst hydrosphere, which has hitherto been largely ignored. In any case, the advanced techniques (Fourier-transform ion cyclotron resonance mass spectrometry) can provide new evidence to test the initial hypotheses for existence of RDOC in the deep karst reservoir waters.

关键词: karst hydrosphere, dissolved organic carbon, resistant to microbial degradation, dilution limiting dissolved organic carbon utilization

The Concentration of Recalcitrant Dissolved Organic Carbon in the Karst Hydrosphere and Their Existing Mechanism

KEY WORDS: karst hydrosphere, dissolved organic carbon, resistant to microbial degradation, dilution limiting dissolved organic carbon utilization

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岩溶水体惰性有机碳含量及其存在机理

李强, 黄雅丹, 何若雪, 于奭, 宋昂, 曹建华