摘要
有色金属矿产资源的开采和冶炼产生了大量的尾矿。尾矿堆存形成大片尾矿废弃地,不仅占用大量土地资源,而且容易对周围环境造成二次污染。植物修复技术能够实现多种目的,如稳定尾矿、控制污染、美化环境；而在尾矿废弃地上种植具有多种用途、经济价值较高的植物正逐渐成为一种新兴的尾矿复垦趋势。近年来,螯合剂在重金属污染土壤植物修复中的应用得到了普遍的关注。大量研究表明,螯合剂不仅能够提高污染土壤中重金属的生物有效性,而且能提高植物对重金属的积累。因此,基于以上研究背景设计了一系列试验,主要研究：(1)乙二胺二琥珀酸(EDDS)和乙二胺四乙酸(EDTA)对不同年代尾矿和矿区附近污染土壤中重金属和矿质元素的浸提；(2)以尾矿与土壤按0：100、25：75、50：50、75：25、100：0比例混合作为栽培基质,研究添加EDTA对铅锌尾矿废弃地玉米生长及Pb、Zn积累的影响；(3)在上述试验的基础上,以尾矿与土壤按1：1比例混合作为培养基质,研究单独添加NTA、EDTA以及二者复合添加对蓖麻幼苗生长及Pb、Zn积累的影响；(4)以尾矿与土壤按1：1比例混合作为培养基质,研究单独添加NTA、EDTA以及二者复合添加对Pb、Zn在蓖麻根、茎、叶亚细胞中分布的影响；(5)对覆土铅锌尾矿库中生长的植物重金属含量进行了研究。试图揭示螯合剂提高土壤重金属生物有效性,促进植物对重金属吸收及向地上部运输的作用机理,为螯合剂在植物修复铅锌尾矿废弃地中的应用提供理论依据。主要结果如下：
(1)浸提实验结果表明,两种螯合剂对两尾矿样品中Pb的浸提率为EDDS大于EDTA;二者对Cu和Zn的浸提率都比较低。两种螯合剂对土壤样品中Cu和Zn的提取效率高于Pb。对矿质元素来说,螯合剂的存在明显增加Mn的提取率,Fe的提取率也有所增加；
(2)玉米盆栽实验结果表明：抑制玉米生长的主要因素是尾矿极端贫瘠,Pb、Zn含量过高。添加EDTA前,玉米在所有处理中都能正常生长,植株没有出现明显的毒害症状。添加EDTA后玉米生物量显著下降,并且在改良基质中尾矿所占比例越高,下降幅度越大。EDTA可以显著提高玉米对Pb、Zn的积累能力,其中对Pb的积累能力大于Zn。基质比例为50：50时,添加EDTA后玉米对Pb、Zn的积累量虽然比基质100：0低,但是玉米的生物量、株高和根系各指标都比基质100：0下降幅度小。故尾矿和土壤比例为50：50的基质改良方式比较合理,可用于铅锌尾矿废弃地的复垦；
(3)蓖麻盆栽实验结果表明：添加螯合剂后蓖麻生物量和根系形态参数均显著下降,并且EDTA添加比例越高,下降幅度越大。其中,根系形态参数中根尖数下降最明显,与对照相比差异极显著(P<0.01)。各螯合处理蓖麻叶绿素a、b和叶绿素a+b与对照相比均显著下降(P<0.05),且叶绿素b比叶绿素a下降幅度略大。蓖麻叶片中SOD. CAT和POD三种酶活性均随螯合处理中EDTA添加量的增加先上升后下降。添加螯合剂后蓖麻各部位Pb. Zn含量明显增加,且Pb的增幅比Zn大。添加螯合剂显著提高了蓖麻对Pb. Zn (?)(?)积累能力,其中蓖麻对Pb的积累能力大于Zn。添加螯合剂明显促进了Pb. Zn从根部向地上部的转移,转移系数在添加3mmol/kg EDTA和3mmol/kg NTA处理时达最大值。蓖麻对Pb和Zn有较强的耐性和积累能力,可用于铅锌尾矿废弃地植物修复的进一步研究；
(4)蓖麻中铅锌的亚细胞分布实验结果表明：Pb. Zn在蓖麻细胞内的分布特征与其吸收和积累Pb. Zn的能力密切相关,施加NTA和EDTA能显著促进Pb. Zn在蓖麻体内的积累并且影响其在各亚细胞组分中的分布。在蓖麻根部,细胞壁是Pb. Zn的主要结合位点,其次为细胞质部分(除单独添加NTA处理外)；在茎、叶中,细胞质部分是Pb、 Zn的主要结合位点,其次为细胞壁部分。三种螯合处理下,蓖麻根中Pb、Zn在细胞壁的分布呈加强趋势,有从细胞器向细胞壁转移的趋势；蓖麻茎和叶中Pb、Zn在细胞质的分布呈加强趋势,添加螯合剂促进了Pb、Zn在茎、叶细胞中的区室化;
(5)对覆土铅锌尾矿库上随表土引入并成功定居的乡土植物研究表明,10种优势植物对Zn、Pb和Cu的富集系数均小于1,其地上部(茎、叶)Zn、Pb和Cu含量均未达到超富集植物的临界含量标准。山油麻、黄毛楤木、商陆、山莓等植物从根部向地上部运输Zn、Cu的能力较强,但积累量较低,对三种金属的吸收均表现为叶>根>茎的规律,是良好的Zn、Cu耐性植物。五节芒对Zn、Pb和Cu表现出一定的富集潜力,且适应性强、生物量大,可用于铅锌尾矿废弃地修复的进一步研究。
以上试验结果表明：(1)两尾矿样品中,EDDS对Pb的浸提率均大于EDTA,二者对Cu、Zn的浸提率均较低；(2)尾矿和土壤混合比例为1：1时,添加EDTA处理中玉米对Pb、Zn的提取效果最好；(3)添加3mmol/kg EDTA和3mmol/kg NTA的处理中,蓖麻生长所受抑制较小,Pb、Zn积累量较高,螯合提取Pb、Zn的效果最好；(4)螯合剂作用下,在蓖麻根部,细胞壁是Pb、Zn的主要结合位点；在茎、叶中,细胞质部分是Pb、Zn的主要结合位点；(5)覆土铅锌尾矿库定居植物中存在对其植被恢复具有一定修复潜力的乡土植物。
Tailings resulted from mining and smelting activities of non-ferrous metal mineral resources. Tailings storging form great tailings waste lands, which not only occupy land but may also cause secondary pollution of the surrounding environment.Revegetation of tailing waste lands is a general practice to stabilize tailings, prevent secondary pollution and beautify the environment; planting the crops that can be used for multiple application ways, bring high economic value will become a rising current about tailings reclamation. Recently, many researches have been focused on the effects of chelating agents on the phytoremediation of heavy metal contaminated soil. Some researches showed that chelating agents not only were able to increase heavy metal bioavailability in contaminated soils, but also enhanced heavy metals accumulation of plants.These studies were mainly focused on:(1) Extraction of heavy metals and mineral elements in tailing of different times and agricultural contaminated soil around mine area using biodegradable (EDDS) and Non-biodegradable (EDTA) Chelators;(2) Pot experiments were conducted to investigate the growth and lead-zinc accumulation of maize seedlings in amendment substrates mixed with lead-zinc tailings and soil with the addition of EDTA. The ratios of lead-zinc tailings to soil in pot substrates were0:100,25:75,50:50,75:25, and100:0, marked as TA00, TA25, TA50, TA75, and TA100, respectively;(3) Pot experiments were conducted to study the effects of single and combined addition of EDTA and NTA on growth and heavy metal accumulation in castor seedlings, in which the ratio of Pb-Zn tailings to soil in the substrates was1:1;(4) Pot experiments were conducted to study the effects of single and combined addition of EDTA and NTA on the subcellular distribution of Pb and Zn in the roots, stems and leaves of castor seedlings.(5) A Survey on heavy-metal content in plants in lead-zinc tailings dam covering soil. The aim of the present studies is to elucidate the mechanisms of increasing heavy metal bioavailability in soils and increasing heavy metal absorption and translocation from soils to plant shoots by chelating agents, which provides a theoretical basis for the phytoremediation on the lead-zinc tailing waste lands. The main results were shown as following:
(1) The results of chelating agents extraction showed that the extraction efficiency for Pb by EDDS was higher than EDTA from two mine tailings simples; the extraction efficiency of Cu and Zn by EDDS and EDTA were similar, and all lower. The extraction efficiency of Cu and Zn by EDDS and EDTA was higher than Pb. The extraction rates of Mn increased evidently at the presence of the two chelators, and the solution of Fe were also slightly enhanced.
(2) The results of pot experiments by maize seedlings showed that the main factors restricting the maize growth were the poor nutrition and rich Pb and Zn with high concentrations in substrates. Maize grew normally in test substrates without EDTA, and no apparent phyotoxity was observed. However, biomass of maize clearly decreased with the addition of EDTA, and the higher decrease degree of the maize biomass was found in the amendment substrates with larger proportion of lead-zinc tailings. The addition of EDTA significantly enhanced the accumulation of Pb and Zn by maize, and the accumulation amount of Pb by maize was higher than that of Zn. The accumulation amount of Pb and Zn in maize was smaller in substrates with50%tailing and50%soil than that in100%tailings, however, the weaker decrease degree of biomass, average of plant height, and root indexes of maize was observed for the former than the latter. We prospose that the amendment of50%soil in tailings seems reasonable for reclamation of lead-zinc tailing waste lands.
(3) The results of pot experiments by castor seedlings showed that the biomass and root morphological parameters of castor clearly decreased with the addition of chelating agents, and greater decreases in them were observed in castor grown in chelating agents containing higher proportions of EDTA. The decrease of root tips was more evident than the rest of root morphological parameters, and which reached extremely significant level compared with the control at the same time (P<0.01). Compared with the CK treatment, chlorophyll a, b, and a+b decreased significantly(P<0.0.5), the decrease extent of chlorophyll b is more than chlorophyll a. The activity of SOD, CAT and POD increased first and then decreased with the increase of EDTA in chelating agents. The contents of Pb and Zn by castor generally increased with addition of chelating agents, the increment of Pb contents was higher than Zn. The addition of chelating agents significantly enhanced the accumulation of Pb and Zn in castor, and more Pb accumulated than Zn. The addition of chelating agents advanced obviously the translocation of Pb and Zn from roots to shoots in castor, the maximum value of translocation factor of Pb and Zn were got at the addition of3mmol/kg EDTA and3mmol/kg NTA. Castor have strong endurance and the ability of heavy metal accumulation, and it is reasonable for reclamation of Pb-Zn tailing waste lands.
(4) The results showed that the subcellular distributions of Pb and Zn had significant relationship with the ability of heavy metal uptake and accumulation in castor. The addition of EDTA and NTA could evidently advance the absorption and accumulation of Pb and Zn in castor and affect on their distribution in cell wall and organelle fractions. Most of Pb and Zn was bound to the cell wall fraction and secondly cytoplasm fraction (except the addition of NTA singly) in root of castor. Most of Pb and Zn was bound to cytoplasm fraction and secondly the cell wall fraction in stems and leaves of castor. Under the inducement of EDTA and NTA, there were increasing Pb and Zn bound to cell wall, which mostly came from organelle fraction in roots of castor, there were increasing Pb and Zn bound to cytoplasm fraction in stems and leaves of castor, the addition of EDTA and NTA enhanced the compartmentalizing of Pb and Zn in stems and leaves of castor.
(5) The study about native plants which followed by topsoil and colonized successfully on lead-zinc tailings dam covering soil showed the accumulation factors of Zn, Pb and Cu about ten species dominant plants were all less than1; Zn, Pb and Cu contents of their shoots and leaves did not reach the standard of hyperaccumulator. Abelmoschus moschatus Medic, Aralia decaisneana Hance, Phytolacca acinosa roxb and Rubus corchorifolius L.f. exhibited higher transportation abilities for Zn and Cu from roots to shoots. But accumulation abilities were low, the accumulation of Zn, Pb and Cu were all listed in the order of leaves> roots> shoots, and three kinds of plants were well metal-tolerant plant. Miscanthus Jloridulus exhibited higher accumulation potentiality for Zn, Pb and Cu, it has strong adaptability and great biomass, can be used for metal immobilization of Pb/Zn tailings waste lands.
In conclusion, these results indicated:(1)The extraction efficiency for Pb by EDDS was higher than EDTA from two mine tailings, the extraction efficiency of Cu and Zn by EDDS and EDTA were similar, and all lower;(2) When the ratio of lead-zinc tailings to soil was1:1, the extraction effect of Pb and Zn by maize was the most significant in all of amendment substrates;(3) The inhibition that castor growth beared was smaller than rest chelating treatments, and accumulation amount of Pb and Zn by castor was higher than that of rest, the extraction effect of Pb and Zn by castor was the most significant in all of that under the addition of3mmol/kg EDTA and3mmol/kg NTA;(4) Under the inducement of EDTA and NTA, most of Pb and Zn was bound to the cell wall fraction in root of castor, most of Pb and Zn was bound to cytoplasm fraction in stems and leaves of castor;(5) There was some native plants those can be used for revegetation in plants colonization on lead-zinc tailings dam covering soil.
引文
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