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1 土壤 (Soils), 2016, 48(1): DOI: /j.cnki.tr 石墨烯施用后对土壤酶活性及土壤微生物群落的影响 1,2 1* (1 ( ) ) ( mg/kg) mg/kg AWCD mg/kg DGGE 3 3 (Graphene) sp 2 [1], [1] [2-3] [4] [5] [6 8] [9] t [10] [5 6] [11] [12 17] [17] [10] mg/l [10] Biolog DNA [18 20] h ( >99%) 0.8 ~ 1.2 nm ( ) * (yteng@issas.ac.cn) (1989 ) lnli@issas.ac.cn

2 2 48 [21] ( 1) ph (g/kg) 表 1 供试土壤的基本理化性质 Table 1 Physico-chemical properties of soil tested (g/kg) (mg/kg) (mg/kg) (cmol/kg) 图 1 Fig. 1 氧化石墨烯和石墨烯的扫描电镜图片 SEM images of graphene and graphene oxide kg 100 mg/kg mg/kg 60% mg/kg [22 23] g 2 mm 40 ml 5 ml 0.3% 120 r/min 30 min 5 ml 1.5 mol/l 25 ml mol/l 5 g 2 mm 50 ml 1 ml 15 min 10 ml 10% 20 ml (ph 6.7) h 37 1 ml 50 ml 9 ml 4 ml 3 ml 20 min 578 nm Biolog-ECO [17] Biolog-ECO min 10 g 0.85% NaCl 250 r/min 10 min µl Biolog-ECO h 12 h ( quant BioTek ) 590 nm (average well color development, AWCD) 1.5 MPBIO Fast DNA SPIN Kit for Soil 56 DNA DNA GC-clamp 515F 907R CGCCCGCCGCGCG CGGCGGGCGGGGCGGGGGCACGGGGGGGTGCC AGCMGCCGCGG CCGTCAATTCMTTTR AGTTT 150 ng PCR DGGE (C.B.S. Scientific Company Inc USA) 45% ~ 80% 8% TAE 80 V 16 h SYBR Green I

3 1 3 (Generay TM Biotechnology) 30 min (Bio-Rad) GC-clamp 515F 907R PCR peasy Cloning KitT3(TransGen Biotech) NCBI BLAST mg/kg mg/kg [24] mg/kg - - [20] 3 [24 25] (GO-100 GO mg/kg mg/kg G-100 G mg/kg mg/kg ) 图 2 潮土脲酶 (A) 红壤脲酶(B) 潮土过氧化氢酶(C) 红壤过氧化氢酶(D) 相对于对照的活性变化 Fig. 2 Relative activities of ureases and catalases in Fluvo-aquic soil and red soil treated with graphene and graphene oxide compared with soil controls [26 27] [24] ( 2) Ahmed [10]

4 4 48 [6] [16] 56 [24] DNA [10] AWCD 12 h 96 h [28 29] 100 mg/kg mg/kg AWCD [31] Fig. 3 图 3 培养 56 天后潮土和红壤各处理 AWCD 值随时间的变化 (A. 潮土 B. 红壤 ) Variations of average well color development (AWCD) in two soil types treated with graphene and graphene oxide Akhavan Ghaderi [17] AWCD Tong [18] Shrestha [24] mg/kg Jin [30] Shrestha Jin 2.3 PCR-DGGE 16S rrna ( 4) (>99%) AWCD DGGE [19] mg/kg 3 3 Tong [18]

5 1 5 Fig. 4 图 4 DGGE 条带的系统发育树 Phylogenetic tree of DGGE bands and their nearest neighbors in GenBank mg/kg [24] [30] [31] [34] [11,33] Lanphere [34] [14] mg/kg 3 56 [1] Geim A K. Graphene: Status and prospects[j]. Science, 2009, 324: [2] Geim A K, Novoselov K S. The rise of graphene[j]. Nature Materials, 2007, 6: [3] Li D, Müller M B, Gilje S, et al. Processable aqueous dispersions of graphene nanosheets[j]. Nature Nanotechnology, 2008, 3: [4] Zhang L, Xia J, Zhao Q, et al. Functional graphene oxide as a nanocarrier for controlled loading and targeted delivery of mixed anticancer drugs[j]. Small, 2010, 6:

6 6 48 [5] Paek S M, Yoo E, Honma I. Enhanced cyclic performance and lithium storage capacity of SnO 2 /graphene nanoporous electrodes with three dimensionally delaminated flexible structure[j]. Nano Letter, 2009, 9: [6] Pan Y, Sahoo N G, Li L. The application of graphene oxide in drug delivery[j]. Expert Opinion Drug Deliver, 2012, 9: [7] Yang K, Li Y, Tan X, et al. Behavior and toxicity of graphene and its functionalized derivatives in biological systems[j]. Small, 2012, 9: [8] Srivastava G, Talat M, Raghubanshi H, et al. Cicer alphagalactosidase immobilization onto functionalized graphene nanosheets using response surface method and its applications[j]. Food Chemistry, 2014, 142: [9],,,. [J]., 2014, 23: [10] Ahmed F, Rodrigues D F. Investigation of acute effects of graphene oxide on wastewater microbial community: A case study[j]. Journal of Hazard Mater, 2013, 256: [11] Dinesh R, Anandaraj M, Srinivasan V, et al. Engineered nanoparticles in the soil and their potential implications to microbial activity[j]. Geoderma, 2012, : [12],,. [J]., 2015, 42(1): 5 15 [13],, 旸,. [J]., 2014, 59(20): [14] Begum P, Ikhtiari R, Fugetsu B. Graphene phytotoxicity in the seedling stage of cabbage, tomato, red spinach, and lettuce[j]. Carbon, 2011, 49: [15] Zhang X, Yin J, Peng C, et al. Distribution and biocompatibility studies of graphene oxide in mice after intravenous administration[j]. Carbon, 2011, 49(3): [16] Lammel T, Boisseaux P, Fernández-Cruz M L, et al. Internalization and cytotoxicity of graphene oxide and carboxyl graphene nanoplatelets in the human hepatocellular carcinoma cell line Hep G2[J]. Particle and Fibre Toxicology, 2013, 10(1): 27 [17] Akhavan O, Ghaderi E. Toxicity of graphene and graphene oxide nanowalls against bacteria[j]. ACS Nano, 2010, 4(10): [18] Tong Z H, Bischoff M, Nies L, et al. Impact of Fullerene (C60) on a Soil Microbial Community[J]. Environ. Sci. Technol., 2007, 41: [19],,. [M]. :, 2008 [20],,. [J]., 2006, 43(6): [21] Kang S, Herzberg M, Rodrigues D F, et al. Antibacterial effects of carbon nanotubes: Size does matter[j]. Langmuir, 2008, 24: [22] Klaine S J, Alvarez P J, Batley G E, et al. Materials in the environment: Behavior, fate, bioavailability, and effects[j]. Environ. Toxicol. Chem., 2008, 27: [23] Stone V, Nowack B, Baun A, et al. Nano-materials for environmental studies: Classification, reference material issues and strategies for physico-chemical characterization[j]. Sci. Total Environ., 2010, 408: [24] Shrestha B, Martinezb V A, Cox S B, et al. An evaluation of the impact of multiwalled carbon nanotubes on Soil microbial community structure and functioning[j]. Journal of Hazardous Materials, 2013, 261: [25],,,. [J]., 2012, 7(2): [26],,,. [J]., 2014, 46(4): [27],,,. [J]., 2014, 46(1): [28] Wei X l, Ge Z Q. Effect of graphene oxide on conformation and activity of catalase[j]. Carbon, 2013, 60: [29],,,. [J]., 2012, 23(3): [30] Jin L X, Son Y H, DeForest J L, et al. Single-walled carbon nanotubes alter soil microbial community composition[j]. Science of the Total Environment, 2014, : [31] Tao X Q, Lu G N, Liu J P, et al. Rapid degradation of phenanthrene by using sphingomonas sp. GY2B immobilized in calcium alginate gel beads[j]. Int. J. Environ. Res. Public Health, 2009, 6: [32],,. Sphingomonas sp. DX-T3-03 [J]., 2010, 37(10): [33],. [J]., 2014, 25( 9): [34] Lanphere J D, Luth C J, Walker S L. Effects of Solution Chemistry on the Transport of Graphene Oxide in Saturated Porous Media[J]. Environmental Science & Technology, 2013, 47 (9):

7 1 7 Effects of Graphene on Soil Enzyme Activities and Microbial Communities LI Lina 1, 2, TENG Ying 1*, REN Wenjie 1, LI Zhengao 1, LUO Yongming 1 (1 Key Laboratory of Soil Environment and Pollution Remediation, Institute of Soil Science, Chinese Academy of Sciences, Nanjing , China; 2 University of Chinese Academy of Sciences, Beijing , China) Abstract: In this study, soils were treated with 0, 100 and g/kg pristine graphene and graphene oxide for up to 56 days, respectively. Treatment effects on urease and catalase activity, microbial communities were evaluated. Microbial community sizes were evaluated by using both Biolog-eco plate and DGGE method. The results showed that the introduction of graphene, as either pristine graphene or graphene oxide, had little impact on urease and catalase activities after cultivation for 0, 7, 14 and 56 days. AWCD values of red soil and fluvo-aquic soil treated with graphene and graphene oxides showed no differences as compared with control. Only DGGE results showed a little changes of bacterium genera in fluvo-aquic soil treated with graphene oxide. Key words: Graphene; Soil enzyme activities; Microbial community

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