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书书书 2018 年 9 月 September 2018 岩矿测试 ROCK AND MINERAL ANALYSIS Vol. 37,No. 5 475 -478 收稿日期 2018 -07 -25; 修回日期 2018 -08 -26; 接受日期 2018 -08 -28 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 ; 中国地质科学院基本科研 业务费项目 “岩溶水体碳固定微型生物群落特征及其碳汇效应” YYWF201505 ; 中国地质调查局地质调查 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。 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. LI Qiang,HUANG Ya- dan,HE Ruo- xue,et al. The Concentration of Recalcitrant Dissolved Organic Carbon in the Karst Hydrosphere and Its 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 李强1, 2,黄雅丹3,何若雪1, 2,于奭1, 2,宋昂1, 2,曹建华1, 2 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 通常认为全球超过 1000 m 的海洋深处稳定存 在着不超过 42 μmol/L 的溶解有机碳, 并被认为是 微生物难以利用的惰性溶解有机碳[1 -2 ]。这为进一 步评估水圈溶解有机碳的稳定性提供了研究依 据 [3 ]。河流不但是陆地水的主体、 水圈的重要组成 部分, 而且还是海洋 - 陆地物质和能量交换的主要 通道。近年来,河流的自然性质和作用过程不断受 到人文活动的影响, 特别是河流筑坝引发的“水库 效应” [4 -5 ]。然而 “水库效应” 下岩溶区筑坝河流中 究竟存在着多少惰性溶解有机碳以及惰性溶解有机 碳存在的机理仍不清楚。 2016 年以来, 作者以水化学类型为 HCO3- Ca 型 [6 ]的广西柳江梯级筑坝河流为研究区 图 1 , 采 集不同层位 表层水、 5 m 和 10 m 的水样, 利用荧 光定量 PCR 技术 [7 ]和改进的惰性有机碳测试方 法 [8 ], 定量评估岩溶筑坝河流水体惰性溶解有机碳 含量及其形成机制。 室内培养实验结果显示 随着培养时间的延长, 水体中的溶解有机碳浓度趋于稳定, 约为 24. 63 μmol/L, 远低于海洋惰性溶解有机碳浓度 图 2 。 同时, 不同取样条件下的水样经过 180 天培养后, 细 菌丰度与溶解有机碳浓度显著正相关 r 0. 658, p 0. 038 表 1 , 从而说明此时的溶解有机碳不再 被微生物利用, 并成为微生物生长的限制因子。而 0 ~150 天培养时间段内溶解有机碳浓度的波动, 在 于岩溶水体中存在着大量化能自养微生物, 进而造 成大 量 暗 碳 non - photosynthetic/dark inorganic carbon incorporation 的固定 [9 ]。因此, 在培养后期, 随着化学能量的消耗殆尽, 化能自养微生物对水体 溶解有机碳的影响可以忽略不计。从而说明, 岩溶 筑坝河流水体中可以存在着约为 24. 63 μmol/L 并 且难以被微生物利用的惰性溶解有机碳。 那么, 岩溶筑坝河流水体中约为 24. 63 μmol/L 的惰性溶解有机碳如何存在呢 本文假设岩溶筑坝 河流水体中存在三种浓度梯度的溶解有机碳 DOC 。这三种浓度梯度的溶解有机碳在红线以 下表示不可以被微生物利用 灰色部分 , 即惰性溶 解有机碳, 而只有高于红线部分的溶解有机碳可以 表 1研究区水样细菌丰度与不同培养时间段溶解有机碳浓度的 Pearson 相关分析 Table 1Correlative analysis of bacteria abundance and dissolved organic carbon concentration 项目0 天30 天60 天90 天120 天150 天180 天 相关系数 r0. 394 0. 3240.3220.228-0.0530.2400.658* 注 * 表示 p 在 0.05 水平 单侧 上显著相关。 574 ChaoXing A橡胶坝拦截区; B红花水库区; C红花水库泄洪区。 图 1广西柳江梯级筑坝河流采样点 Fig. 1Sampling location in dammed Liu River,Guangxi 图 2不同培养时间段溶解有机碳浓度 Fig. 2Dissolved organic carbon concentration in different incubating phase 被微生物利用 图 3 。因此, 当溶解有机碳浓度增 加到 2. 5 倍和 4 倍时, 能够被微生物利用的溶解有 图 3 “稀释假说” 下的溶解有机碳利用速率 Fig. 3Expected increase in dissolved organic carbon availability with increasing dissolved organic carbon enrichment 机碳比例快速增加, 即从 14. 33 增加到 73. 28。 从而说明当岩溶筑坝河流水体的溶解有机碳浓度低 674 第 5 期 岩矿测试 http ∥www. ykcs. ac. cn 2018 年 ChaoXing 于微生物可以利用的阈值 24. 63 μmol/L 时, 岩溶 区筑坝河流特别是水库底层可以保存大量不易被微 生物利用的惰性溶解有机碳, 并与海洋惰性溶解有 机碳存在的 “稀释假说” 相吻合 [10 ]。此外, 岩溶筑坝 河流中还存在着部分自身难以被微生物利用的溶解 有机碳, 即 “生物惰性有机碳” 进而造成其积累并成 为岩溶区筑坝河流特别是水库底层溶解有机碳库的 重要组成部分 [2 -3 ]。 为此, 通过室内模拟培养实验, 本研究认为岩溶 区筑坝河流特别是水库底层存在着约为 24. 63 μmol/L 的惰性溶解有机碳, 并且基于“稀释假说” 和 “生物惰性假说” 的共同理论而存在, 从而为进一 步评估岩溶水圈的碳汇潜力提供了理论依据。为进 一步验证岩溶水体惰性溶解有机碳的稳定性, 今后 还可以利用傅里叶变换离子回旋共振质谱对大型岩 溶水库底层水体与培养实验的溶解有机碳分子结构 进行进一步的比较分析。 参考文献 [ 1]Hansell D A. Recalcitrant dissolved organic carbon frac- tions[J] . Annual Review of Marine Science, 2013, 5 421 -445. [ 2]Arrieta J M, Mayol E, Hansman R L, et al. Dilution limits dissolved organic carbon utilization in the deep ocean [ J] . Science, 2015, 348 331 -333. [ 3]Zhang C L, Dang H Y, Azam F, et al. Evolving paradigms in biological carbon cycling in the ocean[J] . National Science Review, 2018, 5 4 481 -499. [ 4]Peng X, Liu C Q, Wang B L, et al. The impact of damming on geochemical behavior of dissolved inorganic carbon in a karst river[J] . Chinese Science Bulletin, 2014, 59 19 2348 -2355. [ 5]Cui G, Li X, Li Q, et al. Damming effects on dissolved inorganic carbon in different kinds of reservoirs in Jialing River, Southwest China[J] . Acta Geochimica, 2017, 36 4 581 -597. [ 6]原雅琼, 何师意, 于奭, 等. 柳江流域柳州断面水化学特 征及无机碳汇通量分析[ J] . 环境科学, 2015, 36 7 2437 -2445. Yuan YQ,HeSY,YuS,etal. Hydrochemical characteristics and the dissolved inorganic carbon flux in Liuzhou Section of Liujiang Basin[J] . Environmental Science, 2015, 36 7 2437 -2445. [ 7]Li Q, Song A, Peng W J, et al. Contribution of aerobic anoxygenic phototrophic bacteria to total organic carbon pool in aquatic system of subtropical karst catchments, SouthwestChina Evidencefromhydrochemicaland microbiological study[J] . FEMS Microbiology Ecology, 2017, 93 6 1 -8. [ 8]Mermillod- Blondin F, Simon L, Maazouzi C, et al. Dyna- mics ofdissolvedorganiccarbon DOC through stormwater basins designed for groundwater recharge in urban area Assessment of retention efficiency[ J] . Water Research, 2015, 8127 -37. [ 9]Casamayor E O, Llirs M, Picazo A, et al. Contribution of deep dark fixation processes to overall CO2incorporation and large vertical changes of microbial populations in stratified karstic lakes[J] . Aquatic Sciences, 2012, 74 61 -75. [ 10]Jiao N, Legendre L, Robinson C, et al. Comment on “Dilution limits dissolved organic carbon utilization in the deep ocean” [ J] . Science, 2015, 3501483 -1483. The Concentration of Recalcitrant Dissolved Organic Carbon in the Karst Hydrosphere and Its Existing Mechanism LI Qiang1, 2,HUANG Ya- dan3,HE Ruo- xue1, 2,YU Shi1, 2,SONG Ang1, 2,CAO Jian- hua1, 2 1. Key Laboratory of Karst Dynamics, Ministry of Natural Resources, 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 HIGHLIGHTS 1 24. 63 μmol/L recalcitrant dissolved organic carbon is stored in the karst river systems. 2 Recalcitrant dissolved organic carbon is resistant to microbial degradation and the dilution limits dissolved organic carbon utilization in the karst river systems. 774 第 5 期李强, 等 岩溶水体惰性有机碳含量及其存在机理第 37 卷 ChaoXing ABSTRACT BACKGROUND It is generally accepted that recalcitrant dissolved organic carbon RDOC concentrations in the ocean are in the region of 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,by collecting water samples at different depths 0 m, 5 m and 10 m from three typical sites city- river section,reservoir area and outflow area in the dammed Liu River in 2006. S Water samples 3 Lwere pre- filtered in- situ using 3 μm filter membranes,and then filtered through 0. 22 μm pore- size 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 95C 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 designated as 338f 5 - CCTACGGGAGGCAGCAG - 3and 518r 5 - ATTACCGCGGCTGCTGG - 3 . Thermal cycling,fluorescent data collection, and data analysis were carried out with CFX96 Touch Real- time PCR Detection System Bio- Rad,USA according to the instruction manual. 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 was used for correlation analyses using the Pearson correlation 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 The relationship between Ctand log starting concentration is linear R2> 0. 99 . The amplification efficiencies are 90 -105. 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. The advanced techniques Fourier- trans Ion Cyclotron Resonance Mass Spectrometry provide new evidence to test the initial hypotheses for existence of RDOC in the deep karst reservoir waters. KEY WORDS karst hydrosphere; dissolved organic carbon; resistant to microbial degradation; dilution limiting dissolved organic carbon utilization 874 第 5 期 岩矿测试 http ∥www. ykcs. ac. cn 2018 年 ChaoXing
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