石油污染对植物生长毒性效应的影响研究
|
摘要
随着石油开采和使用量的增加,大量的石油及其产品进入环境,不可避免地对环境造成了污染,给生物和人类带来了严重危害。石油类污染物进入土壤后,可引起土壤理化性质的变化、改变土壤微生物区系、可渗入地下并污染地下水等一系列问题,所以如何修复石油污染土壤成为人们关注的焦点。而植物修复技术以其投入低、操作性强、适应性广、无二次污染,可在清除土壤污染的同时,清除污染土壤周围的大气和水体中的污染物等优点,正逐渐成为未来石油污染治理的一个重要研究方向。
本试验以黑麦草Lolium perenne、高羊茅Festuca arundinacea、披碱草Elymus dahuricus、紫花苜蓿Medicago sativa、沙打旺Astragalus adsurgens和翅碱蓬S.hetroptera Kitag这6种植物为研究对象,通过盆栽模拟试验,研究0%、1%、5%和10%4个不同浓度石油污染土壤处理对这6种植物生长生理的影响,全面监测和评价石油污染物胁迫对这6种受试植物的毒性效应,及种植前后土壤含油率和生物指标的变化情况,得出的主要结论如下：
1)紫花苜蓿和沙打旺在盐碱化与石油污染双重胁迫的土壤中发芽率低,各浓度石油处理中的翅碱蓬在生长14d左右死亡率高,均不适宜作为修复植物进行进一步的研究。故本研究选取黑麦草、高羊茅和披碱草为主要的研究植物进行修复后的植物生理和土壤生化指标分析。
2)随着土壤石油污染浓度的增加,在试验末期三种植物的根生长和株高均随着土壤石油污染浓度的升高而降低,说明土壤石油污染会对这三种植物的根生长和株高产生抑制作用,根生长和株高都可作为指示土壤受石油污染情况的植物指标进行研究。
3)植株叶片中超氧化物歧化酶(SOD)与过氧化物酶(POD)的活性都随着土壤石油污染浓度的升高而增大,其活性变化与石油污染浓度具有一定的相关性；植株叶片中丙二醛(MDA)含量也随土壤中石油烃浓度的升高而增大。说明SOD、POD活性和MDA含量等植物酶指标都能较早、较敏感地指示环境的变化。
4)石油污染对土壤脱氢酶、多酚氧化酶和脂肪酶这三种土壤酶活性的影响多表现为促进作用,土壤脱氢酶和多酚氧化酶的活性随着石油污染浓度的升高呈上升趋势,这两种土壤酶可以用来指示土壤受石油污染的程度。
5)石油污染在一定范围内可以刺激土壤微生物活动,且原油中部分成分可作为土壤微生物生长所需的C源和能量来源,使得石油烃的相对降解率(相对空白土)随着土壤石油污染浓度的升高而增大。与原油的组分相比,种植过受试植物的石油烃土壤,其土壤中饱和烃、非烃、胶质和沥青质的含量与供试土壤原油中各含量的百分比相比显著降低,而芳烃的百分比含量则增加。
With the increase of petroleum extraction and usage, a large amount of petroleum and its products were entry into the environment which inevitably polluted the environment and seriously harmed creatures and human. How to repair petroleum-contaminated soil has been attracting considerable public attention for petroleum may lead to a series of problems such as changing physicochemical property and microflora of soils, penetrating into underground and then polluting groundwater and so on. The phytoremediation technique of petroleum-contaminated soil is becoming an important research area for the advantage of low cost, high maneuverability, wide adaptability, removing or degrading pollutant in soil, atmosphere and water at the same time, no secondary contamination and so on.
6 plants (Lolium perenne., Festuca arundinacea, Elymus dahuricus, Medicago sativa, Astragalus adsurgens, S. hetroptera Kitag) planted in contaminated soils were observed at 4 pollution concentration levels (a serial petroleum concentration of 0,1%,5% and 10% by dry weight) in greenhouse pot experiments. Analyze seeding growth and serials biochemical indexes of soil, in order to identify remediation plants from these field crops and provide theoretical basis for the phytoremediation of petroleum-contaminated soil. The main results were as follows:
1) The results showed that these seeds significantly differentresponses to petroleum pollution. Compared with those planted in clean soils, the germination of most seeds planted in the contaminated soils was obviously inhibited. Medicago sativa L., Astragalus adsurgens Pall., S. hetroptera Kitag were the ornamental affected most adversely. Fortunately, strong endurance and high seed germination were observed for the other seeds under 4 different levels of petroleum pollution, thus displaying the potential of remedying petroleum contaminated soils. The responses of seeding growth and so on of Lolium perenne L.,Festuca arundinacea and Elymus dahuricus Turcz. to petroleum contamination were emphatically studied in the next experiments.
2) During the seedling growth initial stage, results show that the variation of oil concentration of soil has different effects on each plant growth. Crude oil restrains growth of them when its content is high. All treatments have less effect on seed germination, individual height and fresh weight of Lolium perenne L., Festuca arundinacea and Elymus dahuricus Turcz.. They can be fit to grow on the crude oil-contaminated soils by itself and with the biggest experimental potential to phytoremediation of oil-contaminated soils in this experiment.
3) With the increase of the concentrations of petroleum, the activities of superoxide dismutase (SOD) and the activities of peroxidase (POD) in all leaves were increased during stages, the contents of malondialdehyd (MDA) were significantly increased too. The activities of antioxidant enzymes in maize leaves were significantly negative correlated to the plant height. The activities of superoxide dismutase (SOD), the activities of peroxidase (POD) and the contents of malondialdehyd (MDA) were more sensitively changes in the environment.
4) The existence of petroleum pollution had mostly stimulated the activity of three kinds of soil enzymes (dehydrogenase, polyphenoloxidase and lipase in soils). The activity of dehydrogenase and polyphenoloxidase in soils were increased with the increase of the the concentrations of petroleum, and the activity of them were also more sensitively changes in the environment.
5) The phytoremediation of petroleum-contaminated soil harm obviously for growth indexes, seed quality of the three seeds, which can promote on the bacteria growth in the soil. With the increase of the concentrations of petroleum, the degradation rate was increased.
本试验以黑麦草Lolium perenne、高羊茅Festuca arundinacea、披碱草Elymus dahuricus、紫花苜蓿Medicago sativa、沙打旺Astragalus adsurgens和翅碱蓬S.hetroptera Kitag这6种植物为研究对象,通过盆栽模拟试验,研究0%、1%、5%和10%4个不同浓度石油污染土壤处理对这6种植物生长生理的影响,全面监测和评价石油污染物胁迫对这6种受试植物的毒性效应,及种植前后土壤含油率和生物指标的变化情况,得出的主要结论如下：
1)紫花苜蓿和沙打旺在盐碱化与石油污染双重胁迫的土壤中发芽率低,各浓度石油处理中的翅碱蓬在生长14d左右死亡率高,均不适宜作为修复植物进行进一步的研究。故本研究选取黑麦草、高羊茅和披碱草为主要的研究植物进行修复后的植物生理和土壤生化指标分析。
2)随着土壤石油污染浓度的增加,在试验末期三种植物的根生长和株高均随着土壤石油污染浓度的升高而降低,说明土壤石油污染会对这三种植物的根生长和株高产生抑制作用,根生长和株高都可作为指示土壤受石油污染情况的植物指标进行研究。
3)植株叶片中超氧化物歧化酶(SOD)与过氧化物酶(POD)的活性都随着土壤石油污染浓度的升高而增大,其活性变化与石油污染浓度具有一定的相关性；植株叶片中丙二醛(MDA)含量也随土壤中石油烃浓度的升高而增大。说明SOD、POD活性和MDA含量等植物酶指标都能较早、较敏感地指示环境的变化。
4)石油污染对土壤脱氢酶、多酚氧化酶和脂肪酶这三种土壤酶活性的影响多表现为促进作用,土壤脱氢酶和多酚氧化酶的活性随着石油污染浓度的升高呈上升趋势,这两种土壤酶可以用来指示土壤受石油污染的程度。
5)石油污染在一定范围内可以刺激土壤微生物活动,且原油中部分成分可作为土壤微生物生长所需的C源和能量来源,使得石油烃的相对降解率(相对空白土)随着土壤石油污染浓度的升高而增大。与原油的组分相比,种植过受试植物的石油烃土壤,其土壤中饱和烃、非烃、胶质和沥青质的含量与供试土壤原油中各含量的百分比相比显著降低,而芳烃的百分比含量则增加。
With the increase of petroleum extraction and usage, a large amount of petroleum and its products were entry into the environment which inevitably polluted the environment and seriously harmed creatures and human. How to repair petroleum-contaminated soil has been attracting considerable public attention for petroleum may lead to a series of problems such as changing physicochemical property and microflora of soils, penetrating into underground and then polluting groundwater and so on. The phytoremediation technique of petroleum-contaminated soil is becoming an important research area for the advantage of low cost, high maneuverability, wide adaptability, removing or degrading pollutant in soil, atmosphere and water at the same time, no secondary contamination and so on.
6 plants (Lolium perenne., Festuca arundinacea, Elymus dahuricus, Medicago sativa, Astragalus adsurgens, S. hetroptera Kitag) planted in contaminated soils were observed at 4 pollution concentration levels (a serial petroleum concentration of 0,1%,5% and 10% by dry weight) in greenhouse pot experiments. Analyze seeding growth and serials biochemical indexes of soil, in order to identify remediation plants from these field crops and provide theoretical basis for the phytoremediation of petroleum-contaminated soil. The main results were as follows:
1) The results showed that these seeds significantly differentresponses to petroleum pollution. Compared with those planted in clean soils, the germination of most seeds planted in the contaminated soils was obviously inhibited. Medicago sativa L., Astragalus adsurgens Pall., S. hetroptera Kitag were the ornamental affected most adversely. Fortunately, strong endurance and high seed germination were observed for the other seeds under 4 different levels of petroleum pollution, thus displaying the potential of remedying petroleum contaminated soils. The responses of seeding growth and so on of Lolium perenne L.,Festuca arundinacea and Elymus dahuricus Turcz. to petroleum contamination were emphatically studied in the next experiments.
2) During the seedling growth initial stage, results show that the variation of oil concentration of soil has different effects on each plant growth. Crude oil restrains growth of them when its content is high. All treatments have less effect on seed germination, individual height and fresh weight of Lolium perenne L., Festuca arundinacea and Elymus dahuricus Turcz.. They can be fit to grow on the crude oil-contaminated soils by itself and with the biggest experimental potential to phytoremediation of oil-contaminated soils in this experiment.
3) With the increase of the concentrations of petroleum, the activities of superoxide dismutase (SOD) and the activities of peroxidase (POD) in all leaves were increased during stages, the contents of malondialdehyd (MDA) were significantly increased too. The activities of antioxidant enzymes in maize leaves were significantly negative correlated to the plant height. The activities of superoxide dismutase (SOD), the activities of peroxidase (POD) and the contents of malondialdehyd (MDA) were more sensitively changes in the environment.
4) The existence of petroleum pollution had mostly stimulated the activity of three kinds of soil enzymes (dehydrogenase, polyphenoloxidase and lipase in soils). The activity of dehydrogenase and polyphenoloxidase in soils were increased with the increase of the the concentrations of petroleum, and the activity of them were also more sensitively changes in the environment.
5) The phytoremediation of petroleum-contaminated soil harm obviously for growth indexes, seed quality of the three seeds, which can promote on the bacteria growth in the soil. With the increase of the concentrations of petroleum, the degradation rate was increased.
引文
[1]Ding KQ, Luo YM, Sun TH, Li PJ. Bioremediation of soil contaminated with petroleum using forced-aeration composting [J]. Pedophile,2002,12(2):145-150.
[2]Gogoi BK, Dutta NN, Go swami P, Krishna Mohani TR.A case study of bioremediation of petroleum-hydrocarbon contaminated soil at a crude oil spill site [J]. Advances in Environmental Research 2003,7(4):767-782.
[3]刘五星,骆永明,滕应.石油污染土壤的生态风险评价和生物修复Ⅰ.一株具有乳化石油能力的细菌分离鉴定[J].土壤学报,2006,43(3)：461-466.
[4]张乃明,段永蒽,毛昆明.土壤环境保护[M].北京：中国农业科学技术出版社,2002.
[5]卜淑君.石油化学工业固体废物治理[M].北京：中国环境科学出版社,1992,12.
[6]任华峰,单德臣,李淑芹.石油污染土壤微生物修复技术的研究进展[J].东北农业大学学报,2004,35(3)：373-376.
[7]陈鹤建.原油在土壤中的渗透及降解规律[J].油气田环境保护,2000,10(4)：14-15.
[8]吕志萍,程龙飞.石油污染土壤中石油含量对玉米的影响[J].油气田环境保护,2001,11(1)：36-37.
[9]孟紫强.环境毒理学[M].北京：中国环境科学出版社,2003,163.
[10]袁红莉,杨金水,王占生.降解石油微生物菌种的筛选及降解特性[J].中国环境科学,2003,23(2)：157-161.
[11]孙铁布,李培军,周启星.土壤污染形成机理与修复技术[M].北京：科学出版社,2005,86-87.
[12]S Hayat, Iqbal Ahmad, ZM Azam. Effect of long term application of oil refinery waste water on soil health with special reference to microbiological characteristics [J]. Bioresearch Technology,2002,84:159-163.
[13]李永涛,吴启堂.土壤污染治理方法研究[J].农业环境保护,1997,16(3)：118-122.
[14]任华峰,单德臣,李淑芹.石油污染土壤微生物修复技术的研究进展[J].东北农业大学学报,2004,35(3)：373-376.
[15]钱署强,刘铮.污染土壤修复技术介绍[J].化工进展,2000(4)：10-12.
[16]陆秀君,郭书海,孙清.石油污染土壤的修复技术研究现状及展望[J].沈阳农业大学学报,2003,34(1)：63-67.
[17]L Zhang, P Somasundaran, V Ososkov etal. Flotation of hydrophobic contaminants from soil [J]. Colloids and Surfaces,2001,117:235-246.
[18]于晓丽.落地原油对土壤污染及治理技术[J].农业环境与发展,2000,17(3)：28-29.
[19]吕荣湖,王嘉麟,陈琴仪.热碱水溶液清洗-气浮分离对原油污染土壤的处理[J].油气田环境保护,1994,4(3)：3-8.
[20]谢飞,吴芳云,刘建刚.洗涤法处理含油土壤的研究[J].油气田环境保护,1997,7(1)：5-9.
[21]童蕾,魏昌华,赵中一.表面预处理+热洗涤法处理含油污泥的方法研究[J].安徽化 工,2005,137(5)：45-47.
[22]K西莫查.净化污染土壤的新的有效技术[J].国外金属矿选矿,2001(2)：43-45.
[23]Maria S,Irena B,Sergey O,et al. Effect of bio-surfactants on crude oil description and mobilization in a soil system [J]. Environment International,2005,31:155-161.
[24]N Vasudevan, P Rajaram. Bioremediation of oil sludge-contaminated soil [J]. Environment International,2001,26:409-411.
[25]罗孝俊,杨卫东,党志.有机物污染的土壤治理方法及研究进展[J].矿物岩石地球化学通报,2000,19(2)：130-135.
[26]段云霞,隋红,韩振为.生物通风修复石油污染土壤的研究进展[J].环境保护科学,2003,29(12)：25-27.
[27]刘其友,赵东风,张云波.石油污染土壤生物修复技术的研究进展[J].油气田环境保护,2005,15(4)：14-16.
[28]徐向阳,冯孝善.五氯酚(PCP)污染土壤厌氧生物修复技术的初步研究[J].应用生态学报,2001,12(3)：439-442.
[29]VENOSA AD, SUIDANMT, WRENNBA, et al. Bioremediation of experimental oil spill on the shoreline of Delaware Bay [J]. Environmental Science and Technology,1996,30(5): 1764-1775.
[30]罗孝俊,杨卫东,党志.有机物污染的土壤治理方法及研究进展[J].矿物岩石地球化学通报,2000,19(2)：130-135.
[31]Cunningham S D, ANDERSON T A, Schwab A P, etal. Phytoremediation of soils contaminated with organic pollutants [J]. Advances in Agronomy,1996,56 (1):55-114.
[32]Jordahl J L, FOSTER L, Schnoor J L, etal. Effect of hybrid poplar trees on microbial populations important to hazardous waste bioremediation [J]. Environmental Toxicology and Chemistry,1997,16 (6):1318-1321.
[33]赵玉霞,杨珂.石油污染土壤修复技术研究综述[J].环境科技,2009,22(1)：60-63.
[34]沈铁孟,黄国强,李凌.石油污染土壤生物通风修复及其强化技术[J].环境污染治理技术与设备,2002,3(7)：67-69.
[35]丁克强,郑昭佩,孙铁珩.石油污染土壤的生物降解研究[J].生态学杂志,2001,20(4)：16-18.
[36]Ayotamuno M J, Kogbara R B, Ogaji S O T, etal.Bioremediation of a crude-oil polluted agricultural-soil at Port Harcourt, Nigeria [J]. App 1 Energy,2006,83 (11):1249-1257.
[37]叶淑红,丁鸣,马达.微生物修复辽东湾油污染湿地研究[J].环境科学,2005,26(5)：143-146.
[38]高彦征,凌婉婷,朱利中.黑麦草对多环芳烃污染土壤的修复作用及机制[J].农业环境科学学报,2005,24(3)：498-502.
[39]宋玉芳,许华夏,任丽萍.两种植物条件下土壤中矿物油和多环芳烃PAHs)的生物修复研究[J].应用生态学报,2001,12(1)：108-112.
[40]Muratova A Y, Turkovskaya O V, Hubner T, et al. Studies of the efficacy of alfalfa and reed in the phytoremediation of hydrocarbon polluted soil [J]. App 1 Bioremediation,2003, 39(6):599-605.
[41]MerklN, Schultze-kraf t R, Infante C. Assessment of tropical grasses and legumes for phytoremediation of petroleum contaminated soils [J]. Water Air Soil Pollution,2005, 165(1-4):195-209.
[42]张福锁,曹一平.根际动态过程与植物营养[J].土壤学报,1992,29(3)：239-250.
[43]Anderson T A, Guthrie E A, Walton BT. Bioremediation in the rhizophere, plant roots and associated microbes clean contaminated soil [J]. Environ. Sci. Technol.,1993,27:2630-2636.
[44]Anderson T A. Bioremediation through Rhizosphere Technology [M]. Washington:DC Amer Chemi Soci,1993,71-157.
[45]丁克强,骆永明,刘世亮.黑麦草对土壤中苯并[a]芘动态变化的影响[J].土壤学报,2004,41(3)：348-353.
[46]Zhang X X, Cheng S P, Zhu C J et al. Microbial PAH-degradation in soil:Degradation pathways and contributing factors[J]. Pedo-sphere,2006,12(2):151-156.
[47]欧阳威,刘红,于勇勇.高羊茅对微生物强化修复石油污染土壤影响的研究[J].环境污染治理技术与设备,2006,7(1)：94-971.
[48]李方敏,姚金龙,王琼山.修复石油污染土壤的植物筛选[J].中国农学通报,2006,22(9)：429-4311.
[49]孙东平,胡凌燕,周伶俐.高效石油降解微生物堆制法处理油污土壤[J].环境科学与技术,2007,30(6)：86-891.
[50]蔺昕,李培军,台培东.石油污染土壤植物-微生物修复研究进展[J].生态学杂志,2006,25(1)：93-1001.
[51]易筱筠,党志,石林.有机污染物污染土壤的植物修复[J].农业环境保护,2002,21(5)：477-479.
[52]Alkorta I, Garbisu C. Phytore mediation of organic contaminants in soils [J]. Bioresearch Techno,2001,79(3):273-276.
[53]唐世荣,黄昌勇,朱祖祥.利用植物修复污染土壤研究进展[J].环境科学进展,1996,4(6)：10-16.
[54]凌婉婷,朱利中,高彦征.植物根对土壤中PAHs的吸收及预测[J].生态学报,2005,25(9)：2320-2325.
[55]焦杏春,陈素华,沈伟然.水稻根系对多环芳烃的吸着与吸收[J].环境科学,2006,27(4)：760-764.
[56]Parrish Z D, White J C, Isleyen M, et al. Accumulation of weathered polycyclic aromatic hydrocarbons (PAHs) by plant and earthworm species [J]. Chemosphere,2006,64(4): 609-618.
[57]郜红建,蒋新,常江.根分泌物在污染土壤生物修复中的作用[J].生态学杂志,2004,23(4)：135-139.
[58]Liste H, Felgentreu D. Crop growth, cultivable bacteria, and degradation of petrol hydrocarbons (PHCs) in a long-term contaminated field soil [J]. App l Soil Ecol,2006, 31(1-2):43-52.
[59]陈嫣,李广贺,张旭.石油污染土壤植物根际微生态环境与降解效应[J].清华大学学报(自然科学版),2005,45(6)：784-787.
[60]Kirk J L, Klironomos J N, Lee H, et al. The effects of perennial ryegrass and alfalfa on microbial abundance and diversity in petroleum contaminated soil. Environ Pollute, 2005,133 (3):455-465.
[61]Parrish Z D,Banks M K,Schwab A P. Effect of root death and decay on dissipation of polycyclic aromatic hydrocarbons in the chemosphere of yellow sweet clover and tall fescue[J]. Environ Qua 1,2005,34 (1):207-216.
[62]陈静,王学军,胡俊栋.腐殖酸对土壤持留多环芳烃的影响[J].农业环境科学学报,2005,24(5)：849-853.
[63]Del Arco J P, Franca F P. Influence of oil contamination levels on hydrocarbon biodegradation in sandy sediment [J]. Environ Pollute,2001,112(3):515-519.
[64]Trinidad P V O, Sacral L G, Rizzo A C L, etal. Biro mediation of a weathered and a recently oil-contaminated soil from Brazil:a comparison study [J]. Chemosphere,2005,58 (4): 515-522.
[65]Lin Qianxin, Mendelssohn I A.The combined effects of phytoremediation and biodegradation in enhancing habitat restoration and oil degradation of petroleum contaminated wetlands [J]. Ecol Engineering,1998,10 (3):263-274.
[66]宋玉芳,孙铁珩,许华夏.表面活性剂TW-80对土壤中多环芳烃生物降解的影响[J].应用生态学报,1999,10(2)：230-232.
[67]Kechavarzi C, Pettersson K, Leeds Harrison P, etal. Root establishment of perennial ryegrass (L. perenne) in diesel contaminated subsurface soil layers [J]. Environ Pollute,2007,145 (1): 68-74.
[68]Merkl N, Schultze kraft R, Infante C. Phytore mediation in the tropics-influence of heavy crude oil on root morphological characteristics of grained [J]. Environ Pollute,2005,138 (1): 86-91.
[69]Singh O V, Jain R K. Phytore mediation of toxic aromatic pollutants from soil [J]. App l Micro b Bio techno,2003,63(2):128-135.
[70]欧阳威,刘红,于勇勇.高羊茅对微生物强化修复石油污染土壤影响的研究[J].环境污染治理技术与设备,2006,7(1)：94-97.
[71]Huang Xiaodong, El AlawiY, Gurska J, et al. A m ulti2 process phytore mediation system for decontamination of persistent total petroleum hydrocarbons (TPHs) from soils [J]. J Microchemist,2005,81 (1):139-147.
[72]Roy S, Labelle S, Mehta P, etal. Phytore mediation of heavy metal and PAHs contaminated brown field sites [J].Plant Soil,2005,272 (1-2):277-290.
[73]丁克强,骆永明.苜蓿修复重金属铜和有机物苯并[a]芘复合污染土壤的研究[J].农业环境科学学报,2005,24(4)：766-770.
[74]沈铁孟,黄国强,李凌.石油污染土壤生物通风修复及其强化技术[J].环境污染治理技术与设备,2002,3(7)：67-69.
[75]LIX, Feng Y, Sawatsky N. Importance of soil water relations in assessing the endpoint of bioremediation soils:1.Plant growth [J]. Plant and soil,1997,192:219-226.
[76]孙铁布,李培军,周启星.土壤污染形成机理与修复技术[M].北京：科学出版社,2005,8-87.
[77]丁克强,郑昭佩,孙铁晰.石油污染土壤的生物降解研究明[J].生态学杂志,2001,20(4)：16-18.
[78]Ayotamuno, MJ, Kogbara, RB, O gaji, SOT, et al. Bioremediation of a crude-oil polluted agricultural-soil at Port Harcourt, Nigeria [J]. Applied Energy,2006,83(11):1249-1257.
[79]叶淑红,丁鸣,马达.微生物修复辽东湾油污染湿地研究[J].环境科学,2005,26(5)：143-146.
[80]何良菊,魏德洲,张维庆.土壤微生物处理石油污染的研究[J].环境科学进展,1999,7(3)：110-115.
[81]任磊,黄廷林.石油污染土壤的生物修复技术[J].安全与环境学报,2001,1(1-2)：50-54.
[82]丁克强,骆永明.生物修复石油污染土壤[J].土壤,2001,4：179-184.
[83]高彦征,凌婉婷,朱利中.黑麦草对多环芳烃污染土壤的修复作用及机制[J].农业环境科学学报,2005,24(3)：498-502.
[84]宋玉芳,许华夏,任丽萍.两种植物条件下土壤中矿物油和多环芳烃PAHs)的生物修复研究[J].应用生态学报,2001,12(1)：105-112.
[85]Muratova, A Y, Turkovskaya, O V, Hubner, T, etal. Studies of the efficacy of alfalfa and reed In the Phytoremediation of hydrocarbon-Polluted soil [J].Applied Biochemistry and Microbiology,2003,39(6):599-605.
[86]Merkl, Schultze-kraft, R, Infante, C. Assessment of tropical grasses and legumes for phytoremediation of Petroleum-contaminated soil [J]. Water, Air and Soil Pollution,2005, 165(1-4):195-209.
[87]土壤理化分析实验指导书[M].北京林业大学,2002.
[88]宁雯.石油污染盐碱土壤的微生物修复技术研究[D].中国石油大学,2009,26-28.
[89]Casida L C, Klem D A, Santoro T. Soil dehydrogenase activity [J]. Soil Science,1964,98: 371-376.
[90]关松荫.土壤酶及其研究法[M].北京：农业出版社,1986.
[91]张丽莉,陈利军,刘桂芬,等.污染土壤的酶学修复研究进展[J].应用生态学报,2003,14(12)：2342-2346.
[92]Spychalla J P, Desborough S L. Superoxide dismutase, catalase and alpha tocopherol content of stored potato tubers [J]. Plant Physiology,1990,94(3):1214-1218.
[93]Wu Y X, Von Tiedemann A. Impact of fungicides on active oxygen species and antioxidant enzymes in spring barley (Hordeum vulgare L.). Exposed to ozone [J]. Environmental Pollution,2002,116(1):37-47.
[94]Nakano Y, Asada K. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts [J]. Plant Cell Physiology,1981,22:867-880.
[95]Turkan I, Bor M, Ozdemir F, Koca H. Differential responses of lipid peroxidation and antioxidants in the leaves of drought-tolerant P. acutifolius Gray and drought-sensitive P. vulgaris L. subjected to polyethylene glycol mediated water stress[J]. Plant Science,2005, 168(1):223-231.
[96]赵世杰,许长成,邹琦.植物组织中丙二醛测定方法的改进[J].植物生理学通讯,1994,30(3)：207-210.
[97]Omura T, Sato R. The carbon monoxide-binding pigment of liver microsomes [J]. Journal of Biological Chemistry,1964,239(7):2370-2378.
[98]张志良,瞿伟菁.植物生理学试验指导[M].(第三版).北京：高等教育出版社,2003.
[99]王景升.种子科学与技术[M].沈阳：辽宁科技出版社,1991.367-369.
[100]勾影波,苏永春.东北高寒地区麦田土壤酶活性的季节变化与环境因素关系的研究[J].生态学杂志,1996,15(4)：63-65.
[101]Dhruva K J,Sharma G D, Mishra R R. Soil microbial population numbers and enzyme activities in relation to altitude and forest degradation[J]. Soil Biol. Biochem,1992, 8:761-767.
[102]Vance E. D., Brookes P. C., Jenkinson D.S.An extraction method formeasuring soil microbial biomass C [J]. SoilBio.l Biochem.,1987,19(6):703-707.
[2]Gogoi BK, Dutta NN, Go swami P, Krishna Mohani TR.A case study of bioremediation of petroleum-hydrocarbon contaminated soil at a crude oil spill site [J]. Advances in Environmental Research 2003,7(4):767-782.
[3]刘五星,骆永明,滕应.石油污染土壤的生态风险评价和生物修复Ⅰ.一株具有乳化石油能力的细菌分离鉴定[J].土壤学报,2006,43(3)：461-466.
[4]张乃明,段永蒽,毛昆明.土壤环境保护[M].北京：中国农业科学技术出版社,2002.
[5]卜淑君.石油化学工业固体废物治理[M].北京：中国环境科学出版社,1992,12.
[6]任华峰,单德臣,李淑芹.石油污染土壤微生物修复技术的研究进展[J].东北农业大学学报,2004,35(3)：373-376.
[7]陈鹤建.原油在土壤中的渗透及降解规律[J].油气田环境保护,2000,10(4)：14-15.
[8]吕志萍,程龙飞.石油污染土壤中石油含量对玉米的影响[J].油气田环境保护,2001,11(1)：36-37.
[9]孟紫强.环境毒理学[M].北京：中国环境科学出版社,2003,163.
[10]袁红莉,杨金水,王占生.降解石油微生物菌种的筛选及降解特性[J].中国环境科学,2003,23(2)：157-161.
[11]孙铁布,李培军,周启星.土壤污染形成机理与修复技术[M].北京：科学出版社,2005,86-87.
[12]S Hayat, Iqbal Ahmad, ZM Azam. Effect of long term application of oil refinery waste water on soil health with special reference to microbiological characteristics [J]. Bioresearch Technology,2002,84:159-163.
[13]李永涛,吴启堂.土壤污染治理方法研究[J].农业环境保护,1997,16(3)：118-122.
[14]任华峰,单德臣,李淑芹.石油污染土壤微生物修复技术的研究进展[J].东北农业大学学报,2004,35(3)：373-376.
[15]钱署强,刘铮.污染土壤修复技术介绍[J].化工进展,2000(4)：10-12.
[16]陆秀君,郭书海,孙清.石油污染土壤的修复技术研究现状及展望[J].沈阳农业大学学报,2003,34(1)：63-67.
[17]L Zhang, P Somasundaran, V Ososkov etal. Flotation of hydrophobic contaminants from soil [J]. Colloids and Surfaces,2001,117:235-246.
[18]于晓丽.落地原油对土壤污染及治理技术[J].农业环境与发展,2000,17(3)：28-29.
[19]吕荣湖,王嘉麟,陈琴仪.热碱水溶液清洗-气浮分离对原油污染土壤的处理[J].油气田环境保护,1994,4(3)：3-8.
[20]谢飞,吴芳云,刘建刚.洗涤法处理含油土壤的研究[J].油气田环境保护,1997,7(1)：5-9.
[21]童蕾,魏昌华,赵中一.表面预处理+热洗涤法处理含油污泥的方法研究[J].安徽化 工,2005,137(5)：45-47.
[22]K西莫查.净化污染土壤的新的有效技术[J].国外金属矿选矿,2001(2)：43-45.
[23]Maria S,Irena B,Sergey O,et al. Effect of bio-surfactants on crude oil description and mobilization in a soil system [J]. Environment International,2005,31:155-161.
[24]N Vasudevan, P Rajaram. Bioremediation of oil sludge-contaminated soil [J]. Environment International,2001,26:409-411.
[25]罗孝俊,杨卫东,党志.有机物污染的土壤治理方法及研究进展[J].矿物岩石地球化学通报,2000,19(2)：130-135.
[26]段云霞,隋红,韩振为.生物通风修复石油污染土壤的研究进展[J].环境保护科学,2003,29(12)：25-27.
[27]刘其友,赵东风,张云波.石油污染土壤生物修复技术的研究进展[J].油气田环境保护,2005,15(4)：14-16.
[28]徐向阳,冯孝善.五氯酚(PCP)污染土壤厌氧生物修复技术的初步研究[J].应用生态学报,2001,12(3)：439-442.
[29]VENOSA AD, SUIDANMT, WRENNBA, et al. Bioremediation of experimental oil spill on the shoreline of Delaware Bay [J]. Environmental Science and Technology,1996,30(5): 1764-1775.
[30]罗孝俊,杨卫东,党志.有机物污染的土壤治理方法及研究进展[J].矿物岩石地球化学通报,2000,19(2)：130-135.
[31]Cunningham S D, ANDERSON T A, Schwab A P, etal. Phytoremediation of soils contaminated with organic pollutants [J]. Advances in Agronomy,1996,56 (1):55-114.
[32]Jordahl J L, FOSTER L, Schnoor J L, etal. Effect of hybrid poplar trees on microbial populations important to hazardous waste bioremediation [J]. Environmental Toxicology and Chemistry,1997,16 (6):1318-1321.
[33]赵玉霞,杨珂.石油污染土壤修复技术研究综述[J].环境科技,2009,22(1)：60-63.
[34]沈铁孟,黄国强,李凌.石油污染土壤生物通风修复及其强化技术[J].环境污染治理技术与设备,2002,3(7)：67-69.
[35]丁克强,郑昭佩,孙铁珩.石油污染土壤的生物降解研究[J].生态学杂志,2001,20(4)：16-18.
[36]Ayotamuno M J, Kogbara R B, Ogaji S O T, etal.Bioremediation of a crude-oil polluted agricultural-soil at Port Harcourt, Nigeria [J]. App 1 Energy,2006,83 (11):1249-1257.
[37]叶淑红,丁鸣,马达.微生物修复辽东湾油污染湿地研究[J].环境科学,2005,26(5)：143-146.
[38]高彦征,凌婉婷,朱利中.黑麦草对多环芳烃污染土壤的修复作用及机制[J].农业环境科学学报,2005,24(3)：498-502.
[39]宋玉芳,许华夏,任丽萍.两种植物条件下土壤中矿物油和多环芳烃PAHs)的生物修复研究[J].应用生态学报,2001,12(1)：108-112.
[40]Muratova A Y, Turkovskaya O V, Hubner T, et al. Studies of the efficacy of alfalfa and reed in the phytoremediation of hydrocarbon polluted soil [J]. App 1 Bioremediation,2003, 39(6):599-605.
[41]MerklN, Schultze-kraf t R, Infante C. Assessment of tropical grasses and legumes for phytoremediation of petroleum contaminated soils [J]. Water Air Soil Pollution,2005, 165(1-4):195-209.
[42]张福锁,曹一平.根际动态过程与植物营养[J].土壤学报,1992,29(3)：239-250.
[43]Anderson T A, Guthrie E A, Walton BT. Bioremediation in the rhizophere, plant roots and associated microbes clean contaminated soil [J]. Environ. Sci. Technol.,1993,27:2630-2636.
[44]Anderson T A. Bioremediation through Rhizosphere Technology [M]. Washington:DC Amer Chemi Soci,1993,71-157.
[45]丁克强,骆永明,刘世亮.黑麦草对土壤中苯并[a]芘动态变化的影响[J].土壤学报,2004,41(3)：348-353.
[46]Zhang X X, Cheng S P, Zhu C J et al. Microbial PAH-degradation in soil:Degradation pathways and contributing factors[J]. Pedo-sphere,2006,12(2):151-156.
[47]欧阳威,刘红,于勇勇.高羊茅对微生物强化修复石油污染土壤影响的研究[J].环境污染治理技术与设备,2006,7(1)：94-971.
[48]李方敏,姚金龙,王琼山.修复石油污染土壤的植物筛选[J].中国农学通报,2006,22(9)：429-4311.
[49]孙东平,胡凌燕,周伶俐.高效石油降解微生物堆制法处理油污土壤[J].环境科学与技术,2007,30(6)：86-891.
[50]蔺昕,李培军,台培东.石油污染土壤植物-微生物修复研究进展[J].生态学杂志,2006,25(1)：93-1001.
[51]易筱筠,党志,石林.有机污染物污染土壤的植物修复[J].农业环境保护,2002,21(5)：477-479.
[52]Alkorta I, Garbisu C. Phytore mediation of organic contaminants in soils [J]. Bioresearch Techno,2001,79(3):273-276.
[53]唐世荣,黄昌勇,朱祖祥.利用植物修复污染土壤研究进展[J].环境科学进展,1996,4(6)：10-16.
[54]凌婉婷,朱利中,高彦征.植物根对土壤中PAHs的吸收及预测[J].生态学报,2005,25(9)：2320-2325.
[55]焦杏春,陈素华,沈伟然.水稻根系对多环芳烃的吸着与吸收[J].环境科学,2006,27(4)：760-764.
[56]Parrish Z D, White J C, Isleyen M, et al. Accumulation of weathered polycyclic aromatic hydrocarbons (PAHs) by plant and earthworm species [J]. Chemosphere,2006,64(4): 609-618.
[57]郜红建,蒋新,常江.根分泌物在污染土壤生物修复中的作用[J].生态学杂志,2004,23(4)：135-139.
[58]Liste H, Felgentreu D. Crop growth, cultivable bacteria, and degradation of petrol hydrocarbons (PHCs) in a long-term contaminated field soil [J]. App l Soil Ecol,2006, 31(1-2):43-52.
[59]陈嫣,李广贺,张旭.石油污染土壤植物根际微生态环境与降解效应[J].清华大学学报(自然科学版),2005,45(6)：784-787.
[60]Kirk J L, Klironomos J N, Lee H, et al. The effects of perennial ryegrass and alfalfa on microbial abundance and diversity in petroleum contaminated soil. Environ Pollute, 2005,133 (3):455-465.
[61]Parrish Z D,Banks M K,Schwab A P. Effect of root death and decay on dissipation of polycyclic aromatic hydrocarbons in the chemosphere of yellow sweet clover and tall fescue[J]. Environ Qua 1,2005,34 (1):207-216.
[62]陈静,王学军,胡俊栋.腐殖酸对土壤持留多环芳烃的影响[J].农业环境科学学报,2005,24(5)：849-853.
[63]Del Arco J P, Franca F P. Influence of oil contamination levels on hydrocarbon biodegradation in sandy sediment [J]. Environ Pollute,2001,112(3):515-519.
[64]Trinidad P V O, Sacral L G, Rizzo A C L, etal. Biro mediation of a weathered and a recently oil-contaminated soil from Brazil:a comparison study [J]. Chemosphere,2005,58 (4): 515-522.
[65]Lin Qianxin, Mendelssohn I A.The combined effects of phytoremediation and biodegradation in enhancing habitat restoration and oil degradation of petroleum contaminated wetlands [J]. Ecol Engineering,1998,10 (3):263-274.
[66]宋玉芳,孙铁珩,许华夏.表面活性剂TW-80对土壤中多环芳烃生物降解的影响[J].应用生态学报,1999,10(2)：230-232.
[67]Kechavarzi C, Pettersson K, Leeds Harrison P, etal. Root establishment of perennial ryegrass (L. perenne) in diesel contaminated subsurface soil layers [J]. Environ Pollute,2007,145 (1): 68-74.
[68]Merkl N, Schultze kraft R, Infante C. Phytore mediation in the tropics-influence of heavy crude oil on root morphological characteristics of grained [J]. Environ Pollute,2005,138 (1): 86-91.
[69]Singh O V, Jain R K. Phytore mediation of toxic aromatic pollutants from soil [J]. App l Micro b Bio techno,2003,63(2):128-135.
[70]欧阳威,刘红,于勇勇.高羊茅对微生物强化修复石油污染土壤影响的研究[J].环境污染治理技术与设备,2006,7(1)：94-97.
[71]Huang Xiaodong, El AlawiY, Gurska J, et al. A m ulti2 process phytore mediation system for decontamination of persistent total petroleum hydrocarbons (TPHs) from soils [J]. J Microchemist,2005,81 (1):139-147.
[72]Roy S, Labelle S, Mehta P, etal. Phytore mediation of heavy metal and PAHs contaminated brown field sites [J].Plant Soil,2005,272 (1-2):277-290.
[73]丁克强,骆永明.苜蓿修复重金属铜和有机物苯并[a]芘复合污染土壤的研究[J].农业环境科学学报,2005,24(4)：766-770.
[74]沈铁孟,黄国强,李凌.石油污染土壤生物通风修复及其强化技术[J].环境污染治理技术与设备,2002,3(7)：67-69.
[75]LIX, Feng Y, Sawatsky N. Importance of soil water relations in assessing the endpoint of bioremediation soils:1.Plant growth [J]. Plant and soil,1997,192:219-226.
[76]孙铁布,李培军,周启星.土壤污染形成机理与修复技术[M].北京：科学出版社,2005,8-87.
[77]丁克强,郑昭佩,孙铁晰.石油污染土壤的生物降解研究明[J].生态学杂志,2001,20(4)：16-18.
[78]Ayotamuno, MJ, Kogbara, RB, O gaji, SOT, et al. Bioremediation of a crude-oil polluted agricultural-soil at Port Harcourt, Nigeria [J]. Applied Energy,2006,83(11):1249-1257.
[79]叶淑红,丁鸣,马达.微生物修复辽东湾油污染湿地研究[J].环境科学,2005,26(5)：143-146.
[80]何良菊,魏德洲,张维庆.土壤微生物处理石油污染的研究[J].环境科学进展,1999,7(3)：110-115.
[81]任磊,黄廷林.石油污染土壤的生物修复技术[J].安全与环境学报,2001,1(1-2)：50-54.
[82]丁克强,骆永明.生物修复石油污染土壤[J].土壤,2001,4：179-184.
[83]高彦征,凌婉婷,朱利中.黑麦草对多环芳烃污染土壤的修复作用及机制[J].农业环境科学学报,2005,24(3)：498-502.
[84]宋玉芳,许华夏,任丽萍.两种植物条件下土壤中矿物油和多环芳烃PAHs)的生物修复研究[J].应用生态学报,2001,12(1)：105-112.
[85]Muratova, A Y, Turkovskaya, O V, Hubner, T, etal. Studies of the efficacy of alfalfa and reed In the Phytoremediation of hydrocarbon-Polluted soil [J].Applied Biochemistry and Microbiology,2003,39(6):599-605.
[86]Merkl, Schultze-kraft, R, Infante, C. Assessment of tropical grasses and legumes for phytoremediation of Petroleum-contaminated soil [J]. Water, Air and Soil Pollution,2005, 165(1-4):195-209.
[87]土壤理化分析实验指导书[M].北京林业大学,2002.
[88]宁雯.石油污染盐碱土壤的微生物修复技术研究[D].中国石油大学,2009,26-28.
[89]Casida L C, Klem D A, Santoro T. Soil dehydrogenase activity [J]. Soil Science,1964,98: 371-376.
[90]关松荫.土壤酶及其研究法[M].北京：农业出版社,1986.
[91]张丽莉,陈利军,刘桂芬,等.污染土壤的酶学修复研究进展[J].应用生态学报,2003,14(12)：2342-2346.
[92]Spychalla J P, Desborough S L. Superoxide dismutase, catalase and alpha tocopherol content of stored potato tubers [J]. Plant Physiology,1990,94(3):1214-1218.
[93]Wu Y X, Von Tiedemann A. Impact of fungicides on active oxygen species and antioxidant enzymes in spring barley (Hordeum vulgare L.). Exposed to ozone [J]. Environmental Pollution,2002,116(1):37-47.
[94]Nakano Y, Asada K. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts [J]. Plant Cell Physiology,1981,22:867-880.
[95]Turkan I, Bor M, Ozdemir F, Koca H. Differential responses of lipid peroxidation and antioxidants in the leaves of drought-tolerant P. acutifolius Gray and drought-sensitive P. vulgaris L. subjected to polyethylene glycol mediated water stress[J]. Plant Science,2005, 168(1):223-231.
[96]赵世杰,许长成,邹琦.植物组织中丙二醛测定方法的改进[J].植物生理学通讯,1994,30(3)：207-210.
[97]Omura T, Sato R. The carbon monoxide-binding pigment of liver microsomes [J]. Journal of Biological Chemistry,1964,239(7):2370-2378.
[98]张志良,瞿伟菁.植物生理学试验指导[M].(第三版).北京：高等教育出版社,2003.
[99]王景升.种子科学与技术[M].沈阳：辽宁科技出版社,1991.367-369.
[100]勾影波,苏永春.东北高寒地区麦田土壤酶活性的季节变化与环境因素关系的研究[J].生态学杂志,1996,15(4)：63-65.
[101]Dhruva K J,Sharma G D, Mishra R R. Soil microbial population numbers and enzyme activities in relation to altitude and forest degradation[J]. Soil Biol. Biochem,1992, 8:761-767.
[102]Vance E. D., Brookes P. C., Jenkinson D.S.An extraction method formeasuring soil microbial biomass C [J]. SoilBio.l Biochem.,1987,19(6):703-707.