污染土壤中铜、镉的植物有效性及其调控研究
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摘要
土壤重金属污染严重影响了土壤环境质量,并引发了诸多食品安全问题。植物是与土壤相互作用最为密切的生物,因此关于土壤重金属的植物有效性评价、重金属污染土壤的修复和安全利用等问题一直是生态环境研究领域的热点。本文在建立与检验土壤Cu、Cd的植物有效性评估模型的基础上,系统研究了土壤Cu、Cd复合污染对植物生长、生理指标的影响和污染土壤中重金属有效性的调控措施,旨在为污染土壤的科学管理与安全利用提供依据。
(1)污染土壤中Cu、Cd的植物有效性评估模型的建立与检验
本文首先以某污染企业周边的黄棕壤-水稻籽粒系统(n=7)为研究对象,采用一元线性回归的统计方法,分析0.lmol·L-1 HCl、0.5mol·L-1 HCl、2.5% HOAc、5%HOAc. DTPA (pH7.3)、0.05 mol·L-1 EDTA、0.01 mol·L-1 CaCl2、0.05mol·L-1 CaCl2、0.5 mol·L-1NH4OAc、1.0 mol·L-1 NH4OAc、0.1 mol·L-1 NaNO3、1.0 mol·L-1 NH4 NO3、0.5 mol·L-1 NH4 NO3+0.02 mol·L-1 EDTA、0.1 mol·L-1 C6H8O7(柠檬酸)和0.1 mol·L-1C2H2O4(草酸)等11种(共15个处理)化学提取剂提取的土壤Cu、Cd化学有效态含量与作物体内Cu、Cd含量的相关性,确定适宜的提取方法。结果表明,0.05mol·L-1 EDTA、DTPA(pH7.3)、0.1 mol·L-1 HCl、0.1mol·L-1 C6H8O7(柠檬酸)和2.5% HoAc等5种提取剂能有效地反映黄棕壤Cu、Cd的植物有效性。在此基础上,利用不同污染企业周边的红壤-白菜系统(n=28)检验了上述5种提取剂的适用性。
由于土壤Cu、Cd的植物有效性受到土壤、重金属和植物三方面因素的综合影响,故已筛选出的5种提取剂均无法同时表征不同土壤-植物系统中Cu、Cd的植物有效性。采用总权重离子冲量(ωI=(?)ω(Ci1/n))校正土壤Cu和Cd的毒性差异,其中Cu和Cd的权重值ω分别为1和6i=1；该指标不考虑中性和酸性土壤性质差异对Cu、Cd植物有效性的影响,碱性土壤中Cu、Cd的植物有效性较低,可对提取量减半校正；利用植物含水率校正植物对重金属的吸收差异。校正后,0.1mol·L-1HCl提取的土壤Cu、Cd含量可同时表征上述两个系统中土壤Cu、Cd的植物有效性。其中,土壤Cu、Cd的总权重离子冲量(ωI)与植物体内的总权重离子冲量极显著相关,相关系数达0.901(n=35,P<0.01)。
为了建立土壤Cu、Cd的植物有效性评估模型,选择代表性土壤中的黑土、暗棕壤、棕壤、黄棕壤、砖红壤、赤红壤、红壤、黄壤、紫色土、灰钙土、潮土和水稻土等12个土类共57个不同土样为供试对象,分析测定了土壤的基本理化性质(pH、有机质、速效磷、阳离子代换量、粘粒含量和土壤全Cu、全Cd),以0.lmol·L-1 HCl提取的Cu、Cd含量表征土壤重金属的植物有效性,利用多元线性回归的方法建立土壤Cu、Cd的植物有效性与土壤基本理化性质关系的评估模型。本试验结果表明,土壤中Cd的植物有效性与土壤中全Cd、有机质和速效磷含量呈显著的正相关关系,而与土壤pH呈显著的负相关关系。土壤Cu的植物有效性与土壤全Cu、全Cd和速效磷含量显著正相关,与土壤pH显著负相关。由此可见,在考虑Cu的植物有效性时,必须考虑Cd的影响。采用总权重离子冲量的形式可同时表征复合污染土壤中Cu、Cd的植物有效性：lgωI植物=0.995 lgωI土+0.1531gP速效+0.0841gSOM-0.058pH-0.365 (r=0.875, F=42.570, n=57, P<0.001).
在南昌地区化工、钢铁、电镀和塑料4种类型6个企业周边采集土壤-蔬菜样品(n=75)以检验评估模型的适用性。一元线性回归的分析结果表明,土壤Cu、Cd的植物有效性预测值与实测值之间呈极显著正相关关系(r=0.88,n=75,P<0.001),说明复合污染土壤中Cu、Cd的植物有效性主要受到土壤Cu和Cd的全量、pH、有机质和速效磷含量的影响,且采用总权重离子冲量建立的评估方程能较好地预测土壤Cu、Cd的植物有效性。
(2)土壤Cu、Cd复合污染对植物生长、生理指标的影响
评估和预测土壤中Cu、Cd的植物有效性是为了针对不同污染水平的土壤如何选择不同的利用方式,以实现科学合理的管理目标。因此,本文进一步探讨了土壤Cu、Cd复合污染对黑麦草和海州香薷两种植物种子萌发、幼苗生长和生理指标的影响。
Cu、Cd胁迫对黑麦草和海州香薷种子的萌发实验结果表明,种子萌发过程主要依靠自养,因北Cu、Cd胁迫对种子萌发带来的影响不大。水培实验结果表明,当溶液中Cu和Cd的浓度分别为10 mg·L-1和30 mg·L-1时,黑麦草的苗长受到极显著的胁迫作用(P<0.01)；当Cu和Cd的浓度分别为30 mg·L-1和5 mg·L-1时,海州香薷的苗长受到极显著的胁迫作用(P<0.01)。在本实验条件下,溶液中低浓度的Cu和Cd分别使海州香薷和黑麦草的苗长表现出毒性兴奋现象,说明海州香薷是Cu的耐性植物而黑麦草为Cd的耐性植物。
生物盆栽实验结果表明,两种植物地下部对Cu、Cd的吸收系数>地上部的吸收系数。红壤pH对黑麦草影响较大,土壤pH<5.0时黑麦草就无法正常生长。对于弱酸性红壤(pH6.0左右),全Cu<80 mg·kg-1时,黑麦草的生物量、叶绿素含量、抗氧化酶活性、根系活力和MDA含量与对照相比差异不大。当土壤中Cu含量达到230 mg·kg-1时,尽管黑麦草地上部分减产不明显,但黑麦草的生化指标已受到了明显的抑制,且重金属的吸收系数较大。由于黑麦草是Cd的耐性植物,Cd含量的增加未对黑麦草的生长和生理指标表现出显著差异。因此,为保证牧草的品质安全,黑麦草不适宜种在Cu、Cd重污染土壤上。海州香薷对土壤Cu含量和土壤酸性均有较高的耐性,在酸性红壤上(pH5.0左右),与对照相比,土壤中Cu含量为80 mg·kg-1时还能促进海州香薷的生长；当Cu含量达230 mg·kg-1时,海州香薷的生长则受到显著抑制。与黑麦草不同,当土壤Cd含量由0.05 mg·kg-1增加到15.97 mg·kg-1时,海州香薷的生长和生理指标受到显著的抑制作用。
叶绿素含量、POD酶活性、SOD酶活性,根系活力和MDA含量等生理指标均能反映Cu、Cd对海州香薷和黑麦草生长的胁迫作用。土壤蛋白酶、脲酶、过氧化物酶和酸性磷酸酶可以指示土壤Cu、Cd复合污染的程度。Cu和Cd之间表现为协同作用,并且这种协同作用在高Cu含量时更大。
(3)复合污染土壤中Cu、Cd植物有效性的调控措施
最后,本研究进一步探索了调控复合污染土壤中Cu、Cd有效性的具体措施。由于壳聚糖可提高土壤中Cu、Cd的活性,而二氧化钛可通过羟基基团的吸附作用钝化土壤中的Cu、Cd活性,所以采用“海州香薷联合壳聚糖活化诱导”和“海州香薷联合二氧化钛钝化”两种“植物+改良剂”联合的方法,比较两者对复合污染土壤中Cu、Cd有效性的调控效果的差异。结果表明,施用两种改良剂后仍可用植物的叶绿素含量、抗氧化酶系统、MDA含量和根活力等指标反映Cu、Cd对植物生长的胁迫作用。两种调控措施均可提高海州香薷地上部对Cu、Cd的积累量,因此优于单一植物修复的效果。“海州香薷联合壳聚糖活化诱导”的调控措施适用于含Cu量较低的复合污染红壤；而“海州香薷联合二氧化钛钝化”的调控措施更适用于含Cu量较高的复合污染红壤。
Heavy metals (HMs) pollution in soils has seriously affected soil quality and led to lots of food safety problems. Since plants have great interrelationship with soil, in the ecological environment research field there are hot discussion about the assessment of the phytoavailability of HMs in soil, remediation of soil contaminated with HMs and its safety utilization. With the purpose of improving the polluted soil management and its safety utilization, this study built and tested the statistical model of Cu, Cd phytoavailablity in contaminated soils, and focused on the effect of contaminated soil on plant growth and the control measures.
To built and test the model of Cu, Cd phytoavailability in contaminated soils
The first part of this study was to set up the appropriate extracting methods about phytoavailability of Cu, Cd in soils. Yellow brown earth-rice(oryza sativa) grain system near one polluted enterprises was studied. Chemical extractants, including 0.1mol·L-1 HC1, 0.5mol·L-1 HC1,2.5% HOAc,5% HOAc, DTPA(pH7.3),0.05 mol·L-1 EDTA,0.01 mol·L CaCl2,0.05ml·L-1 CaCl2,0.5 mol·L-1NH4OAc,1.0 mol·L-1 NH4OAc,0.1 mol·L-1 NaNO3, 1.0 mol·L-1NH4 NO3,0.5 mol·L-1 NH4NO3+0.02 mol·L-1 EDTA,0.1mol·L-1 C6H8O7(citric acid) and 0.1mol·L-1 C2H2O4(oxalic acid), were used to extract Cu and Cd from the soil samples. The correlation of the amount of Cu, Cd extracted from soil and the amount of Cu, Cd in plant was analyzed using the single linear regression models. The results showed that, among the eleven extractants, 0.05mol·L-1 EDTA, DTPA (pH7.3), 0.1mol·L-1 HC1, 0.1mol·L-1 C6H8O7 (citric acid) and 2.5% HoAc could reflect the phytoavailability of Cu, Cd in the test soils. And the 5 extractants were proved valid in the red soil-pakcoi (Brassica chinensis) system adjacent to different polluted enterprises.
It was difficult for any extractant to characterize the phytoavailablity of Cu and Cd in different soil-plant system simultaneously, since the phytoavailability was determined by interaction of soil, HMs and plant. Weight ion impulse (ωI=(?)ω(Ci1/n)) was used to correct toxic difference of Cu and Cd, and the weight index of Cu and Cd was 1 and 6 respectively. The extracted content of alkaline soil should be cut half for soil correction. In addition, the water content was used to correct the absorption difference among different plants. After the correction from these three factors, total WI of Cu-Cd extracted by 0.1mol·L-1 HCl could well reflect the phytoavailablity of the test soils. The total WI in the soil has significant correlationship with the total WI in the plant(r=0.901, n=35, P<0.01).
Then, the study was to build a statistical model of Cu,Cd phytoavailablity in different kinds of soils. The parameters included a series of physicochemical properties (including pH, soil organic matter(SOM), available phosphorous content(Pavailable), CEC and clay content, total-Cu and total-Cd) of 57 soil samples that covered 12 soil classes, i.e., black soil, dark brown soil, brown soil, yellow brown soil, latosol, latosolic red soil, red soil, zheltozem, purple soil, light sierozem, alluvial soil and paddy soil. The Cu, Cd phytoavailable contents were extracted by 0.lmol·L-1 HC1. The statistical model of Cd phytoavailablity and Cu phytoavailability against soil physicochemical properties were analyzed with SPSS 13.0. Under the experimental condition, there was significant positive correlation between Cd phytoavailabilty and total-Cd, SOM and Pavailable and negative correlation with pH. And there was positive correlation between Cu phytoavailability and total-Cu, total-Cd and Pavailable, and negative correlation with pH. However, the phytoavailability of Cu was affected by total-Cd significantly. The statistical model expressed in terms of total WI could characterize the relationship between the phytoavailability of Cu, Cd and physicochemical properties in different soils. The model could be mathematically described by lgWIpian=0.995 lgWIsoil+0.1531gPavailable+0.841gSOM-0.058pH-0.365(r=0.875, F=42.570, n=57, P<0.001).
The statistical model was tested and applied by the red soil-vegetable system in the adjacent area of Nanchang. Soil and vegetable samples were taken from the sites around six enterprises including iron and steel, chemical industry, electroplating and plastic manufactory in this area. Predicted phytoavailability was significant correlation with the actual content, while the main related factors were total Cu-Cd, SOM, pH and Pavailable (r=0.88,n=75, P<0.001).
Effects of Cu, Cd contamination on plant growth and physiological index
The purpose of predicting phytoavailability of Cu and Cd in soils is to provide the basis for utilization and management of HMs contaminated soil and to ensure the sustainable development. The second part of the study discussed the germination, seedling growth and physiological quality of ryegrass(Lolium perenne L.) and Elsholtzia Splendens under Cu-Cd combined contamination.
The germination test results showed the influence of Cu and Cd on plant germination was little, because plant germination mainly depended on self-nutrition. The height of the seedling could reflect the stress of Cu and Cd. The seedling height of ryegrass treated with Cu or Cd at 10mg·L-1 or 30 mg·L-1 respectively was significantly restrained. The seedling height of Elsholtzia Splendens was significantly restrained when Cu or Cd at 30mg·L-1 or 5mg·L-1 respectively. The height of ryegrass was significant enlonged when the concentration of Cd was low. Therefore it could be concluded that ryegrass was tolerant species of Cd and the same for Elsholtzia Splendens to Cu.
Ryegrass and Elsholtzia's pot experiment results showed that absorption coefficient of Cu and Cd in root was greater than that in shoot. Ryegrass stopped growing when soil pH was lower than 5.0. When total Cu in soil was less than 80 mg·kg-1 with soil pH about 6.0, the biomass, chlorophyll content, antioxidant enzymes, root vigor and MDA content of ryegrass had little difference with the CK treatment. When total Cu was more than 230mg·kg-1, physiological index of the plant had been stressed significantly, and the absorption coefficient of Cu and Cd increased significantly. The physiological index of ryegrass, being a Cd tolerant plant, had little difference with the other treatment under the same copper concentration. In order to ensure the forage safety, ryegrass could not be planted in heavy polluted soil. When total Cu in soil was less than 80 mg·kg-1, Elsholtzia biomass was higher than CK treatment. The biomass was stressed significantly while total Cu was more than 230 mg·kg-1.
All of the physiological index in this study, including chlorophyll content, antioxidant enzymes, root vigor and MDA content of the plant, had been stressed significantly by Cu and Cd. Soil proteinase, urease, peroxidase and phosphatase could indicate Cu, Cd contamination. Interaction between Cu and Cd was synergic under Cu, Cd combined contamination, and the higher Cu concentration, synergy stronger. The results were also demonstrated in the solution culture experiments.
Control on phytoavailability of Cu-Cd in contaminated soil
Finally, the control on phytoavailability of Cu-Cd in contaminated soil were studied. Chitosan could improve phytoavailability of Cu-Cd in red soil whereas TiO2 had the counter-productive reduction ability of heavy metal in that TiO2 can absorb Cu and Cd by hydroxyl group. So the combination control measures, such as "Elsholtzia and chitosan" and "Elsholtzia and TiO2" were adapted to comparing control effects with sole phytoremediation. The results showed that physiological index, such as chlorophyll content, antioxidant enzyme activities, root vigor and MDA content, could reflect the stress of Cu-Cd combined contamination on the growth of plant, no matter what chemical amendment was used. The total content of Cu-Cd in the plant with the combination measure was greater than that with sole phytoremediation, so the effect of former was better than that of the latter. Furthermore, "Elsholtzia and chitosan" measure was suited for lighter contaminated soil, while the other combination measure was suited for heavy contaminated soil.
(1)污染土壤中Cu、Cd的植物有效性评估模型的建立与检验
本文首先以某污染企业周边的黄棕壤-水稻籽粒系统(n=7)为研究对象,采用一元线性回归的统计方法,分析0.lmol·L-1 HCl、0.5mol·L-1 HCl、2.5% HOAc、5%HOAc. DTPA (pH7.3)、0.05 mol·L-1 EDTA、0.01 mol·L-1 CaCl2、0.05mol·L-1 CaCl2、0.5 mol·L-1NH4OAc、1.0 mol·L-1 NH4OAc、0.1 mol·L-1 NaNO3、1.0 mol·L-1 NH4 NO3、0.5 mol·L-1 NH4 NO3+0.02 mol·L-1 EDTA、0.1 mol·L-1 C6H8O7(柠檬酸)和0.1 mol·L-1C2H2O4(草酸)等11种(共15个处理)化学提取剂提取的土壤Cu、Cd化学有效态含量与作物体内Cu、Cd含量的相关性,确定适宜的提取方法。结果表明,0.05mol·L-1 EDTA、DTPA(pH7.3)、0.1 mol·L-1 HCl、0.1mol·L-1 C6H8O7(柠檬酸)和2.5% HoAc等5种提取剂能有效地反映黄棕壤Cu、Cd的植物有效性。在此基础上,利用不同污染企业周边的红壤-白菜系统(n=28)检验了上述5种提取剂的适用性。
由于土壤Cu、Cd的植物有效性受到土壤、重金属和植物三方面因素的综合影响,故已筛选出的5种提取剂均无法同时表征不同土壤-植物系统中Cu、Cd的植物有效性。采用总权重离子冲量(ωI=(?)ω(Ci1/n))校正土壤Cu和Cd的毒性差异,其中Cu和Cd的权重值ω分别为1和6i=1；该指标不考虑中性和酸性土壤性质差异对Cu、Cd植物有效性的影响,碱性土壤中Cu、Cd的植物有效性较低,可对提取量减半校正；利用植物含水率校正植物对重金属的吸收差异。校正后,0.1mol·L-1HCl提取的土壤Cu、Cd含量可同时表征上述两个系统中土壤Cu、Cd的植物有效性。其中,土壤Cu、Cd的总权重离子冲量(ωI)与植物体内的总权重离子冲量极显著相关,相关系数达0.901(n=35,P<0.01)。
为了建立土壤Cu、Cd的植物有效性评估模型,选择代表性土壤中的黑土、暗棕壤、棕壤、黄棕壤、砖红壤、赤红壤、红壤、黄壤、紫色土、灰钙土、潮土和水稻土等12个土类共57个不同土样为供试对象,分析测定了土壤的基本理化性质(pH、有机质、速效磷、阳离子代换量、粘粒含量和土壤全Cu、全Cd),以0.lmol·L-1 HCl提取的Cu、Cd含量表征土壤重金属的植物有效性,利用多元线性回归的方法建立土壤Cu、Cd的植物有效性与土壤基本理化性质关系的评估模型。本试验结果表明,土壤中Cd的植物有效性与土壤中全Cd、有机质和速效磷含量呈显著的正相关关系,而与土壤pH呈显著的负相关关系。土壤Cu的植物有效性与土壤全Cu、全Cd和速效磷含量显著正相关,与土壤pH显著负相关。由此可见,在考虑Cu的植物有效性时,必须考虑Cd的影响。采用总权重离子冲量的形式可同时表征复合污染土壤中Cu、Cd的植物有效性：lgωI植物=0.995 lgωI土+0.1531gP速效+0.0841gSOM-0.058pH-0.365 (r=0.875, F=42.570, n=57, P<0.001).
在南昌地区化工、钢铁、电镀和塑料4种类型6个企业周边采集土壤-蔬菜样品(n=75)以检验评估模型的适用性。一元线性回归的分析结果表明,土壤Cu、Cd的植物有效性预测值与实测值之间呈极显著正相关关系(r=0.88,n=75,P<0.001),说明复合污染土壤中Cu、Cd的植物有效性主要受到土壤Cu和Cd的全量、pH、有机质和速效磷含量的影响,且采用总权重离子冲量建立的评估方程能较好地预测土壤Cu、Cd的植物有效性。
(2)土壤Cu、Cd复合污染对植物生长、生理指标的影响
评估和预测土壤中Cu、Cd的植物有效性是为了针对不同污染水平的土壤如何选择不同的利用方式,以实现科学合理的管理目标。因此,本文进一步探讨了土壤Cu、Cd复合污染对黑麦草和海州香薷两种植物种子萌发、幼苗生长和生理指标的影响。
Cu、Cd胁迫对黑麦草和海州香薷种子的萌发实验结果表明,种子萌发过程主要依靠自养,因北Cu、Cd胁迫对种子萌发带来的影响不大。水培实验结果表明,当溶液中Cu和Cd的浓度分别为10 mg·L-1和30 mg·L-1时,黑麦草的苗长受到极显著的胁迫作用(P<0.01)；当Cu和Cd的浓度分别为30 mg·L-1和5 mg·L-1时,海州香薷的苗长受到极显著的胁迫作用(P<0.01)。在本实验条件下,溶液中低浓度的Cu和Cd分别使海州香薷和黑麦草的苗长表现出毒性兴奋现象,说明海州香薷是Cu的耐性植物而黑麦草为Cd的耐性植物。
生物盆栽实验结果表明,两种植物地下部对Cu、Cd的吸收系数>地上部的吸收系数。红壤pH对黑麦草影响较大,土壤pH<5.0时黑麦草就无法正常生长。对于弱酸性红壤(pH6.0左右),全Cu<80 mg·kg-1时,黑麦草的生物量、叶绿素含量、抗氧化酶活性、根系活力和MDA含量与对照相比差异不大。当土壤中Cu含量达到230 mg·kg-1时,尽管黑麦草地上部分减产不明显,但黑麦草的生化指标已受到了明显的抑制,且重金属的吸收系数较大。由于黑麦草是Cd的耐性植物,Cd含量的增加未对黑麦草的生长和生理指标表现出显著差异。因此,为保证牧草的品质安全,黑麦草不适宜种在Cu、Cd重污染土壤上。海州香薷对土壤Cu含量和土壤酸性均有较高的耐性,在酸性红壤上(pH5.0左右),与对照相比,土壤中Cu含量为80 mg·kg-1时还能促进海州香薷的生长；当Cu含量达230 mg·kg-1时,海州香薷的生长则受到显著抑制。与黑麦草不同,当土壤Cd含量由0.05 mg·kg-1增加到15.97 mg·kg-1时,海州香薷的生长和生理指标受到显著的抑制作用。
叶绿素含量、POD酶活性、SOD酶活性,根系活力和MDA含量等生理指标均能反映Cu、Cd对海州香薷和黑麦草生长的胁迫作用。土壤蛋白酶、脲酶、过氧化物酶和酸性磷酸酶可以指示土壤Cu、Cd复合污染的程度。Cu和Cd之间表现为协同作用,并且这种协同作用在高Cu含量时更大。
(3)复合污染土壤中Cu、Cd植物有效性的调控措施
最后,本研究进一步探索了调控复合污染土壤中Cu、Cd有效性的具体措施。由于壳聚糖可提高土壤中Cu、Cd的活性,而二氧化钛可通过羟基基团的吸附作用钝化土壤中的Cu、Cd活性,所以采用“海州香薷联合壳聚糖活化诱导”和“海州香薷联合二氧化钛钝化”两种“植物+改良剂”联合的方法,比较两者对复合污染土壤中Cu、Cd有效性的调控效果的差异。结果表明,施用两种改良剂后仍可用植物的叶绿素含量、抗氧化酶系统、MDA含量和根活力等指标反映Cu、Cd对植物生长的胁迫作用。两种调控措施均可提高海州香薷地上部对Cu、Cd的积累量,因此优于单一植物修复的效果。“海州香薷联合壳聚糖活化诱导”的调控措施适用于含Cu量较低的复合污染红壤；而“海州香薷联合二氧化钛钝化”的调控措施更适用于含Cu量较高的复合污染红壤。
Heavy metals (HMs) pollution in soils has seriously affected soil quality and led to lots of food safety problems. Since plants have great interrelationship with soil, in the ecological environment research field there are hot discussion about the assessment of the phytoavailability of HMs in soil, remediation of soil contaminated with HMs and its safety utilization. With the purpose of improving the polluted soil management and its safety utilization, this study built and tested the statistical model of Cu, Cd phytoavailablity in contaminated soils, and focused on the effect of contaminated soil on plant growth and the control measures.
To built and test the model of Cu, Cd phytoavailability in contaminated soils
The first part of this study was to set up the appropriate extracting methods about phytoavailability of Cu, Cd in soils. Yellow brown earth-rice(oryza sativa) grain system near one polluted enterprises was studied. Chemical extractants, including 0.1mol·L-1 HC1, 0.5mol·L-1 HC1,2.5% HOAc,5% HOAc, DTPA(pH7.3),0.05 mol·L-1 EDTA,0.01 mol·L CaCl2,0.05ml·L-1 CaCl2,0.5 mol·L-1NH4OAc,1.0 mol·L-1 NH4OAc,0.1 mol·L-1 NaNO3, 1.0 mol·L-1NH4 NO3,0.5 mol·L-1 NH4NO3+0.02 mol·L-1 EDTA,0.1mol·L-1 C6H8O7(citric acid) and 0.1mol·L-1 C2H2O4(oxalic acid), were used to extract Cu and Cd from the soil samples. The correlation of the amount of Cu, Cd extracted from soil and the amount of Cu, Cd in plant was analyzed using the single linear regression models. The results showed that, among the eleven extractants, 0.05mol·L-1 EDTA, DTPA (pH7.3), 0.1mol·L-1 HC1, 0.1mol·L-1 C6H8O7 (citric acid) and 2.5% HoAc could reflect the phytoavailability of Cu, Cd in the test soils. And the 5 extractants were proved valid in the red soil-pakcoi (Brassica chinensis) system adjacent to different polluted enterprises.
It was difficult for any extractant to characterize the phytoavailablity of Cu and Cd in different soil-plant system simultaneously, since the phytoavailability was determined by interaction of soil, HMs and plant. Weight ion impulse (ωI=(?)ω(Ci1/n)) was used to correct toxic difference of Cu and Cd, and the weight index of Cu and Cd was 1 and 6 respectively. The extracted content of alkaline soil should be cut half for soil correction. In addition, the water content was used to correct the absorption difference among different plants. After the correction from these three factors, total WI of Cu-Cd extracted by 0.1mol·L-1 HCl could well reflect the phytoavailablity of the test soils. The total WI in the soil has significant correlationship with the total WI in the plant(r=0.901, n=35, P<0.01).
Then, the study was to build a statistical model of Cu,Cd phytoavailablity in different kinds of soils. The parameters included a series of physicochemical properties (including pH, soil organic matter(SOM), available phosphorous content(Pavailable), CEC and clay content, total-Cu and total-Cd) of 57 soil samples that covered 12 soil classes, i.e., black soil, dark brown soil, brown soil, yellow brown soil, latosol, latosolic red soil, red soil, zheltozem, purple soil, light sierozem, alluvial soil and paddy soil. The Cu, Cd phytoavailable contents were extracted by 0.lmol·L-1 HC1. The statistical model of Cd phytoavailablity and Cu phytoavailability against soil physicochemical properties were analyzed with SPSS 13.0. Under the experimental condition, there was significant positive correlation between Cd phytoavailabilty and total-Cd, SOM and Pavailable and negative correlation with pH. And there was positive correlation between Cu phytoavailability and total-Cu, total-Cd and Pavailable, and negative correlation with pH. However, the phytoavailability of Cu was affected by total-Cd significantly. The statistical model expressed in terms of total WI could characterize the relationship between the phytoavailability of Cu, Cd and physicochemical properties in different soils. The model could be mathematically described by lgWIpian=0.995 lgWIsoil+0.1531gPavailable+0.841gSOM-0.058pH-0.365(r=0.875, F=42.570, n=57, P<0.001).
The statistical model was tested and applied by the red soil-vegetable system in the adjacent area of Nanchang. Soil and vegetable samples were taken from the sites around six enterprises including iron and steel, chemical industry, electroplating and plastic manufactory in this area. Predicted phytoavailability was significant correlation with the actual content, while the main related factors were total Cu-Cd, SOM, pH and Pavailable (r=0.88,n=75, P<0.001).
Effects of Cu, Cd contamination on plant growth and physiological index
The purpose of predicting phytoavailability of Cu and Cd in soils is to provide the basis for utilization and management of HMs contaminated soil and to ensure the sustainable development. The second part of the study discussed the germination, seedling growth and physiological quality of ryegrass(Lolium perenne L.) and Elsholtzia Splendens under Cu-Cd combined contamination.
The germination test results showed the influence of Cu and Cd on plant germination was little, because plant germination mainly depended on self-nutrition. The height of the seedling could reflect the stress of Cu and Cd. The seedling height of ryegrass treated with Cu or Cd at 10mg·L-1 or 30 mg·L-1 respectively was significantly restrained. The seedling height of Elsholtzia Splendens was significantly restrained when Cu or Cd at 30mg·L-1 or 5mg·L-1 respectively. The height of ryegrass was significant enlonged when the concentration of Cd was low. Therefore it could be concluded that ryegrass was tolerant species of Cd and the same for Elsholtzia Splendens to Cu.
Ryegrass and Elsholtzia's pot experiment results showed that absorption coefficient of Cu and Cd in root was greater than that in shoot. Ryegrass stopped growing when soil pH was lower than 5.0. When total Cu in soil was less than 80 mg·kg-1 with soil pH about 6.0, the biomass, chlorophyll content, antioxidant enzymes, root vigor and MDA content of ryegrass had little difference with the CK treatment. When total Cu was more than 230mg·kg-1, physiological index of the plant had been stressed significantly, and the absorption coefficient of Cu and Cd increased significantly. The physiological index of ryegrass, being a Cd tolerant plant, had little difference with the other treatment under the same copper concentration. In order to ensure the forage safety, ryegrass could not be planted in heavy polluted soil. When total Cu in soil was less than 80 mg·kg-1, Elsholtzia biomass was higher than CK treatment. The biomass was stressed significantly while total Cu was more than 230 mg·kg-1.
All of the physiological index in this study, including chlorophyll content, antioxidant enzymes, root vigor and MDA content of the plant, had been stressed significantly by Cu and Cd. Soil proteinase, urease, peroxidase and phosphatase could indicate Cu, Cd contamination. Interaction between Cu and Cd was synergic under Cu, Cd combined contamination, and the higher Cu concentration, synergy stronger. The results were also demonstrated in the solution culture experiments.
Control on phytoavailability of Cu-Cd in contaminated soil
Finally, the control on phytoavailability of Cu-Cd in contaminated soil were studied. Chitosan could improve phytoavailability of Cu-Cd in red soil whereas TiO2 had the counter-productive reduction ability of heavy metal in that TiO2 can absorb Cu and Cd by hydroxyl group. So the combination control measures, such as "Elsholtzia and chitosan" and "Elsholtzia and TiO2" were adapted to comparing control effects with sole phytoremediation. The results showed that physiological index, such as chlorophyll content, antioxidant enzyme activities, root vigor and MDA content, could reflect the stress of Cu-Cd combined contamination on the growth of plant, no matter what chemical amendment was used. The total content of Cu-Cd in the plant with the combination measure was greater than that with sole phytoremediation, so the effect of former was better than that of the latter. Furthermore, "Elsholtzia and chitosan" measure was suited for lighter contaminated soil, while the other combination measure was suited for heavy contaminated soil.
引文
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