Difference study on microbial community structure and diversity of groundwater in different coastal zones
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摘要:
研究目的 海水入侵已成为全球沿海地区面临的重大环境地质问题及研究热点，探究海水入侵区地下水微生物群落特征可以对海水入侵的管理与防治起到支撑作用。
研究方法 本研究选取黄渤海地区两个典型海岸带（烟台龙口西海岸及青岛大沽河海岸），基于高通量测序方法对当地地下水微生物进行16S rDNA分析，对比海水入侵程度不同的海岸带地下水微生物群落结构与多样性特征差异。
研究结果 大沽河研究区相比龙口研究区地下水受海水入侵的影响更为严重，地下水TDS含量在1.06~3.19 g/L，以Na–Cl–HCO3和Na–Cl型水为主，而龙口研究区地下水TDS相对较低，以Ca–Na–Cl–HCO3型水为主。Alpha多样性指数表明，龙口研究区地下水微生物的均匀度和丰富度随着海水入侵程度的增强而减小，而大沽河研究区地下水微生物多样性变化复杂。龙口研究区三个监测点地下水微生物群落结构相似，而大沽河研究区的微生物群落结构差异较大。龙口研究区标志微生物为伯克霍尔德氏菌目（Burkholderiales），丛毛单胞菌科（Comamonadaceae）和噬氢菌属（Hydrogenophaga）；大沽河研究区标志微生物较少，在阈值St≥4.0时标志微生物仅为变形菌门（Proteobacteria）。龙口研究区地下水TOC和DO与微生物丰度呈现正相关关系；而大沽河研究区中，Na+、Cl–、SO42–等指标与微生物丰度呈显著负相关关系，这与当地受海水入侵时间较长有关。
结论 本研究揭示了在不同程度的海水入侵影响下地下水微生物与环境的响应特征，并表现出了不同的标志微生物，表明微生物指标可作为识别海水入侵的新型有效方法。
Abstract:This paper is the result of environmental geological survey engineering.
Objective Seawater intrusion has become a major environmental and geological problem as well as research hotspot in coastal areas all over the world. To explore the characteristics of groundwater microbial communities in coastal areas can play a fundamental role in the management and prevention of seawater intrusion.
Methods This study took two typical costal zones in Yellow Sea and Bohai Sea such as Longkou west bank and Dagu river coast. The microbial 16S rDNA of groundwater was analyzed by high–throughput sequencing method, to explore the diversity and structure characteristics of groundwater microbial communitiesin different areas.
Results The groundwater in Dagu river study area was more seriously affected by seawater intrusion than that in Longkou. In Dagu river study area, the TDS concentration of groundwater was 1.06−3.19 g/L, mainly Na–Cl–HCO3 and Na–Cl type water. In Longkou study area, the TDS of groundwater was relatively low, mainly Ca–Na–Cl–HCO3 type water. The Alpha diversity index showed that the microbial uniformity and richness of groundwater decreased with the increasing seawater intrusion in Longkou, while the variation of microbial diversity of groundwater in Dagu river study area was complex. The groundwater microbial community structures of three monitoring wells in Longkou study area were similar, but the groundwater microbial community structures in Dagu river study area were different. The representative microorganisms of groundwater in Longkou were Burkholderiales, Comamonadaceae and Hydrogenophaga. In Dagu river study area, the representative microorganisms of groundwater are a little, only Proteobacteria was identified at the threshold St≥4.0. TOC and DO were positively correlated with bacterial abundance in groundwater in Longkou study area. In the Dagu river study area, Na+, Cl–, SO42– and other indicators were negatively correlated with bacterial abundance in groundwater, which was related to the long period of seawater intrusion.
Conclusions This study revealed the response characteristics of groundwater microorganisms and the environment under the influence of seawater intrusion at different degrees, and showed different representative microorganisms, indicating that microbial indicators can be used as a new and effective method to identify seawater intrusion.
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表 1 龙口和大沽河研究区地下水样品物化指标
Table 1. Physico–chemical parameters of groundwater samples of Longkou and Dagu river study areas
物化指标 龙口 大沽河 DZ1 DZ2 DZ3 DG1 DG2 DG3 GL/m –3.31 –2.91 –1.89 –0.52 –0.40 –0.69 T/℃ 16.51 17.08 16.60 18.97 18.74 19.50 pH 7.62 7.30 7.35 7.61 7.68 7.54 TOC/(mg/L) 1.70 1.80 1.70 1.80 2.50 2.80 EC/(mS/cm) 1.37 1.82 2.52 1.45 4.13 4.40 TDS/(g/L) 0.68 0.91 1.26 1.06 3.05 3.19 ORP/mV 13.00 21.00 6.00 72.00 49.00 –59.00 DO/(mg/L) 5.28 4.38 4.40 6.28 6.05 2.10 Cl–/(mg/L) 173.08 311.79 604.26 279.15 1314.62 1408.37 HCO3–/(mg/L) 246.01 289.27 376.42 311.71 616.20 787.62 NO3–/(mg/L) 286.01 98.34 153.07 1.08 1.14 32.73 SO42–/(mg/L) 114.09 178.80 167.22 135.74 299.03 269.80 Ca2+/(mg/L) 198.20 208.62 284.48 52.02 50.43 75.18 Na+/(mg/L) 81.00 106.37 174.50 188.03 886.81 676.60 Mg2+/(mg/L) 22.00 26.74 34.90 40.07 67.59 91.07 K+/(mg/L) 2.10 3.08 3.24 23.40 48.04 45.84 注：GL—潜水地下水位；TOC—总有机碳；EC—电导率；TDS—总溶解固体；ORP—氧化还原电位；DO—溶解氧。 
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表 2 龙口与大沽河研究区地下水样品的微生物群落Alpha多样性指标
Table 2. Microbial community Alpha diversity indices for the groundwater samples of Longkou and Dagu river study areas
多样性指标 龙口 大沽河 DZ1 DZ2 DZ3 DG1 DG2 DG3 Shannon 4.49 4.75 3.46 4.54 4.48 4.20 Chao1 706 573 701 728 338 572 Observed species 239 200 179 218 131 190 PD 17.0 17.0 14.2 20.6 9.4 15.9
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表 3 龙口和大沽河研究区地下水微生物丰度与环境因子回归分析
Table 3. Regression analysis of microbial richness and environmental factors in Longkou and Dagu river study areas
物化指标 龙口 大沽河 回归方程 R2 P 回归方程 R2 P GL y=−0.00654x+0.69245 0.079 0.463 y=7.91×10−6x²−0.0075x+1.126 0.658 < 0.05* T y=18.7x−0.0181 0.004 0.864 y=17.8x0.0118 0.044 0.589 pH y=0.00159x+6.6094 0.099 0.410 y=3.83×10−6x²−0.0037x+8.425 0.817 < 0.01** TOC y=3.98×10−5x1.72 0.530 < 0.05* y=−2.25×10−5x²+0.0203x−2.0486 0.256 0.411 EC y=−6.0932x+5080.9 0.097 0.415 y=−0.04561x²+32.549x−1479.7 0.908 < 0.01** TDS y=−3.1274x+2578.8 0.107 0.391 y=−0.03122x²+21.672x−597.56 0.905 < 0.01** ORP y=−0.00475x²+4.9775x−1286.9 0.097 0.415 y=0.00399x²−3.4867x−714.53 0.642 < 0.05* DO y=0.01107x−0.91302 0.507 < 0.05* y=0.00036x²−0.2680x+52.943 0.902 < 0.01** Cl– y=−2.0873x+1454.8 0.087 0.442 y=−3.5751x+2545.5 0.792 < 0.01** HCO3– y=−0.7111x+673.06 0.111 0.380 y=−0.00886x²+6.966x−650.11 0.835 < 0.01** NO3– y=0.86755x−276.62 0.102 0.401 y=−0.0009x²+0.811x−159.45 0.648 < 0.05* SO42– y=−0.4688x+395.58 0.224 0.198 y=−0.7276x+567.62 0.887 < 0.01** Ca2+ y=35468x−0.807 0.047 0.576 y=−0.00088x²+0.7918x−100.75 0.554 0.089 Na+ y=−0.3520x+306.59 0.053 0.550 y=−2.4546x+1671.8 0.949 < 0.01** Mg2+ y=−0.0731x+65.035 0.089 0.435 y=−0.00142x²+1.1802x−151 0.762 < 0.05* K+ y=5528x−1.2 0.164 0.280 y=−0.00044x²+0.3235x−9.36 0.847 < 0.01** 注：**表示显著相关（P<0.01）；*表示相关（P<0.05）。
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