CN114235727A - 一种重金属污染土壤修复效果的生态评估方法 - Google Patents
一种重金属污染土壤修复效果的生态评估方法 Download PDFInfo
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Abstract
本发明属于土壤修复评估技术领域,公开了一种重金属污染土壤修复效果的生态评估方法,包括:采集样品,且所述样品为经调理剂修复后目标区域内的土壤样品和农作物样品;获取所述土壤样品的理化性质评级A、微生物活性评级B及Cd含量评级C;获取所述农作物样品中的Cd含量评级D;计算所述目标区域内的土壤修复效果P,且P=iA+jB+kC+sD;式中,i为理化性质评级A的权重,j为微生物活性评级B的权重,k为Cd含量评级C的权重,s为Cd含量评级D的权重;综上,在本发明所提供的评估方法中,不仅考量了土壤与农作物中重金属的含量,还设计了修复土壤的理化性质及微生物活性的相关指标,相比于现有的评价方法更加全面、准确和科学。
Description
技术领域
本发明属于土壤修复评估技术领域,具体涉及一种重金属污染土壤修复效果的生态评估方法。
背景技术
近年来,由于工农业的急速发展,以及产品生产、消费、遗弃过程的加快,导致越来越大面积的土壤受到重金属及有机污染物的污染。
目前,重金属污染土壤修复方法主要分为物理化学修复及生物修复两类。其中:物理化学修复技术主要是通过添加化学调理剂改变Cd在土壤中的赋存形态、降低其生物有效性,从而使农产品达到安全生产。生物修复则是利用微生物群落和其多样性的变化来实现对重金属污染土壤的生态调节作用。
在现有技术中,一些发达国家已经建立起各类土壤修复标准用于评价土壤修复效果,然而在国内对于重金属污染土壤修复效果的评价,仍采用传统的环境质量风险评价模型,存在修复效果评价结果不够全面可信的问题,从而也不利于进行土地利用及土地规划决策。
发明内容
鉴于此,为解决上述背景技术中所提出的问题,本发明的目的在于提供一种重金属污染土壤修复效果的生态评估方法。
为实现上述目的,本发明提供如下技术方案:一种重金属污染土壤修复效果的生态评估方法,包括:
采集样品,且所述样品为经调理剂修复后目标区域内的土壤样品和农作物样品;
获取所述土壤样品的理化性质评级A、微生物活性评级B及Cd含量评级C;获取所述农作物样品中的Cd含量评级D;
计算所述目标区域内的土壤修复效果P,且P=iA+jB+kC+sD;式中,i为理化性质评级A的权重,j为微生物活性评级B的权重,k为Cd含量评级C的权重,s为Cd含量评级D的权重。
优选的,所述调理剂采用矿物型调理剂、有机型调理剂或微生物型调理剂。
优选的,在所述目标区域内,所述调理剂的修复用量为3t·hm-1。
优选的,所述矿物型调理剂至少包括硅酸钙、熟石灰、硫酸钾、无水硫酸镁和九水合硝酸铁。
优选的,获取所述土壤样品的理化性质评级A时,至少获取所述土壤样品的:pH值、有机碳含量、碱解氮含量、有效磷含量和速效钾含量。
优选的,获取所述土壤样品的的Cd含量评级C时,采用DTPA浸提法测定所述土壤样品的的Cd含量。
优选的,获取所述农作物样品中的Cd含量评级D时,采用石墨炉原子吸收光谱仪测定所述农作物样品中Cd的含量。
优选的,采用石墨炉原子吸收光谱仪测定所述农作物样品中Cd的含量时包括:
将所述农作物样品晾晒至恒重;
依次脱粒、脱壳、研磨和消解处理晾晒后的所述农作物样品,获得消解液;
采用石墨炉原子吸收光谱仪测定所述消解液中Cd的含量。
优选的,获取所述土壤样品的微生物活性评级B时,至少获取微生物活力和微生物量。
优选的,所述微生物活力采用土壤酶活性作为评价指标,且所述土壤酶至少包括蔗糖酶、脲酶和酸性磷酸酶。
本发明与现有技术相比,具有以下有益效果:
在本发明所提供的评估方法中,不仅考量了土壤与农作物中重金属的含量,还设计了修复土壤的理化性质及微生物活性的相关指标,由此覆盖了化学、微生物及植物方面的评估内容,综合考虑农田功能及污染修复实际情况,适用于全面评价污染修复工作及大范围的修复工作评价,相比于现有的评价方法更加全面、准确和科学。
附图说明
图1为本发明的四个实验组中土壤样品Cd和水稻样品Cd含量的示意图;
图2为本发明的四个实验组中土壤酶活性的示意图;
图3为本发明的四个实验组的土壤样品中细菌和真菌的群落分布图;
图4为本发明的四个实验的土壤样品中细菌和真菌在OTU水平上的聚类分析图;
图5为本发明的四个实验的土壤样品中细菌和真菌的群落结构的NMDS排序图;
图6为本发明的四个实验的土壤样品中细菌和真菌的群落结构的RDA分析图;
具体实施方式
下面将结合本发明实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
在本发明中提供了一种重金属污染土壤修复效果的生态评估方法,包括:
采集样品,且所述样品为经调理剂修复后目标区域内的土壤样品和农作物样品;
获取所述土壤样品的理化性质评级A、微生物活性评级B及Cd含量评级C;获取所述农作物样品中的Cd含量评级D;
计算所述目标区域内的土壤修复效果P,且P=iA+jB+kC+sD;式中,i为理化性质评级A的权重,j为微生物活性评级B的权重,k为Cd含量评级C的权重,s为Cd含量评级D的权重。
基于上述方法,在本发明中还提供如下试验:
于2020年3月至2020年7月,在广东省兴宁市寺岗村水稻田(24°34′N,115°69′E)开展田间试验。
一、实验组构建
设农作物为水稻,并且设置四组实验组,具体
实验组一为无修复处理的对照组(CK)。
实验组二为采用矿物型调理剂修复处理的试验组(T1),具体矿物型调理剂至少包括硅酸钙、熟石灰、硫酸钾、无水硫酸镁和九水合硝酸铁等成分。
实验组三为采用有机型调理剂修复处理试验组(T2),具体有机型调理剂的主要成分为蚕沙和烟梗(有机质含量≥47%),N+P2O5+K2O≥35%。
实验组四为采用微生物型调理剂修复处理试验组(T3),具体微生物型调理剂为复合微生物肥料,其有效活性菌数≥0.2亿·g-1(枯草芽孢杆菌、地衣芽孢杆菌、米曲霉),N+P2O5+K2O≥8%。
上述四组实验组均采用相同的处理方式对目标区域内土壤进行修复处理:每个目标区域为5亩,处理时实验组二至实验组四中的三种理剂修的修复用量均为3t·hm-1,且对相应的目标区域内土壤重复处理3次。
二、样品采集与分析
在水稻成熟期进行样品采集,具体采用5点取样法采集上述四个实验组所对应的目标区域内的土壤样品和水稻样品。
(1)土壤样品的理化性质分析
采用电位法测定pH值,采用重铬酸钾-油浴法测定有机碳含量,采用碱解扩散法测定碱解氮含量,采用碳酸氢钠提取-钼锑抗比色法测定有效磷含量,采用乙酸铵提取-原子吸收法测定速效钾含量。获得图下表的测定结果:
pH | 有机碳g·kg<sup>-1</sup> | 碱解氮mg·kg<sup>-1</sup> | 有效磷mg·kg<sup>-1</sup> | 速效钾mg·kg<sup>-1</sup> | |
CK | 5.20±0.03c | 28.47±0.10d | 204.98±0.99d | 5.53±0.00d | 51.20±0.33c |
T1 | 5.44±0.02a | 37.54±0.14b | 235.31±1.97b | 14.04±0.00b | 65.41±0.07b |
T2 | 5.31±0.01b | 41.72±0.04a | 257.27±3.45a | 15.91±0.00a | 83.35±1.06a |
T3 | 5.39±0.01a | 34.49±0.14c | 229.20±0.25c | 12.05±0.19c | 83.49±0.21a |
(2)关于土壤样品的微生物活性分析
2a)土壤全DNA提取及土壤微生物量测定
土壤全DNA提取采用试剂盒提取方法,主要步骤为细胞破碎-DNA溶出-吸附-纯化-洗脱,完成DNA提取后进行测序分析。具体,依照如下方式对测序得到的原始数据进行质控检测、过滤:
去除,i.平均质量分数小于20分以及长度小于50bp的序列;ii.Barcode错配1个碱基及以上的序列;iii.引物中错配大于2个碱基的序列;
使用FLASH,并基于重叠序列长度大于10bp、重叠区域不允许有模糊碱基的标准对序列进行拼接;
用mothur去除嵌合体。
由上,得到高质量序列,并按照97%的相似性阈值划分OTU,同时去除只含有1条序列的OTU。为了保证不同样品测序深度一致,我们对所有样品的序列按相同的序列数进行抽平,抽平后的数据用于后续的注释及统计分析。
选取每个OTU中最长的序列作为该OTU的代表序列,使用BLASTn在NCBI非冗余数据库中对代表序列进行检索和比对,以1×e-20做为E值的最小阈值,去除E>1×e-20的参考序列,然后从剩下的序列中选取得分最高的参考序列对该OTU进行注释。如果所有检索到的参考序列所对应的E值都大于1×e-20,则标记该代表序列尚无同源序列。
根据检索结果,在T1至T3样品中,细菌仅有4.1%~5.5%的序列未检索到相似序列或相对丰度低于1%,剩下的序列由图3A所示的放线菌门、变形菌门、氯曲菌门、酸杆菌门、厚壁菌门、硝基螺菌门、脱硫菌门、粘球菌门、类杆菌门、金霉素菌门、扁平菌门、MBNT15、Sva0485、疣微菌门组成。真菌一共注释到6个门,占检索序列的87%~90%,由图3B所示的子囊菌门、担子菌门、被子菌门、罗兹菌门、壶菌门、球囊菌门组成。
2b)土壤微生物活力测定
以土壤酶活性作为评价指标,且土壤酶至少包括蔗糖酶、脲酶和酸性磷酸酶。具体,土壤蔗糖酶、脲酶和酸性磷酸酶活性测定参见吴金水等(2006)的方法,并获得图2所示的测定结果。
(3)关于土壤样品和水稻样品的Cd含量分析
采用DTPA浸提法测定土壤样品的Cd含量,测定结果如图1上图所示;
采用石墨炉原子吸收光谱仪测定水稻样品中Cd的含量,具体包括:将水稻样品晾晒至恒重;依次脱粒、脱壳、研磨和消解处理晾晒后的水稻样品,获得消解液;采用石墨炉原子吸收光谱仪测定所述消解液中Cd的含量,测定结果如图1下图所示。
三、结果分析
(1)关于理化性质的测定结果分析
根据上述表1可知,不同实验组通过改变土壤理化性质来有效改善肥力和土壤质量,从而提高作物产量。从表1中可以看出调理剂的添加提高了土壤pH值,缓解了土壤酸化的情况;不同调理剂处理中土壤有机碳的含量分别增加了32%、47%和21%,且较大幅度的提高了土壤碱解氮、有效磷和有效钾等速效养分的含量,改善了土壤营养状况,土壤碱解氮、有效磷和速效钾分别增加了12%~15%、118%~188%和28%~63%。
(2)关于Cd含量的测定结果分析
结合图1可知,与对照相比,三种调理剂均显著降低了土壤样品DTPA-Cd含量,具体降幅分别为13%、48%和28%。对水稻样品Cd含量而言,三种调理剂均显著降低了其含量,最大降幅达42%。
由上可见,调理剂施用后土壤样品Cd和水稻样品Cd含量均有明显下降,其中实验组三(T2)中土壤样品Cd含量下降最为显著,但对水稻样品Cd含量的降低效果不佳。
(3)关于土壤样品的微生物活性的测定结果分析
结合图2可知,三种调理剂均显著增加了土壤蔗糖酶和脲酶的活性,其中实验组三(T2)对蔗糖酶活性的提升作用最大,增幅为151%;实验组四(T3)使脲酶活性增加了80%。对土壤酸性磷酸酶而言,实验组二(T1)降低了其活性,而实验组三(T2)增加了其活性。
作为土壤的活跃组成成分,微生物区系组成及微生物数量与土壤的理化性质变化密切相关。具体,结合图3可知,其中放线菌门和变形菌门的相对丰度在所有调理剂处理中均较高。方差分析表明,调理剂处理(T1、T2和T3)显著降低了脱硫菌门和类杆菌门的相对丰度;其中实验组二(T1)和实验组三(T2)的调理剂显著提高了土壤中放线菌门和金霉素菌门的相对丰度,实验组四(T3)的调理剂显著提高了酸杆菌门的相对丰度。
另外,通过对所获得的OTU进行热图聚类分析获得图4所示的分析结果,图中A为细菌分析结果,B图为真菌分析结果,且由图4可知,不同调理剂处理之间其OTU的相对丰度明显不一样。然而,细菌的聚类分析和真菌的有相似规律,即均有分化成矿物型调理剂和其他调理剂两个类别,然后再进行有机型和微生物型调理剂处理的分类。这也暗示出施加矿物型调理剂对细菌和真菌的物种组成的影响与其他两种调理剂处理方式显著不同。
不同调理剂处理下细菌和真菌的群落结构的NMDS排序图如图5所示,在图5中显示调理剂类型对细菌和真菌这两类微生物的群落结构的变化都有着显著影响,样品均能按照调理剂类型聚类在一起,而在不同的调理剂处理之间形成分开。相似性分析是一种非参数检验分析方法,常被用来检验两组或多组间差异是否显著大于组内差异,从而判断分组类型是否有意义。我们的实验结果显示,细菌的ANOSIM分析R=0.929(P<0.001),而真菌的ANOSIM分析R=0.772(P<0.001),由此,均可认为这四种处理之间的细菌和真菌的群落结构存在明显差异。
在生态学领域中,冗余分析(RDA)是常用的进行约束排序分析的方法,其目的是分析“解释变量(一般为环境因子矩阵)”对“响应变量(一般为物种矩阵)”的影响情况。通过RDA分析获得图6所示的分析图,且由图6发现所测得的理化因子对细菌和真菌微生物群落的解释度分别为73.49%和47.08%,其中轴1分别能解释65.43%和28.52%。对细菌的群落结构而言,施用实验组二(T1)能够在轴1上与对照显著分开,在轴2上与另外两种调理剂处理分开;另外,实验组三(T2)及实验组四(T3)均能够在轴2上与对照显著分开。而对真菌的群落结构来说,实验组三(T2)及实验组四(T3)均能在轴2上与对照显著分开。999次蒙特卡洛检验显示全磷是影响细菌和真菌微生物群落结构共同的主要环境因子。另外土壤有机碳、全氮、碱解氮、有效磷和全钾分别是影响细菌和真菌微生物群落结构的其他环境因子。土壤全磷与实验组二(T1)在细菌群落呈现出明显的正比关系,而与实验组四(T3)处理呈反相关;土壤中的有机碳、全氮、碱解氮、有效磷含量则明显与实验组三(T2)成正相关关系。
尽管已经示出和描述了本发明的实施例,对于本领域的普通技术人员而言,可以理解在不脱离本发明的原理和精神的情况下可以对这些实施例进行多种变化、修改、替换和变型,本发明的范围由所附权利要求及其等同物限定。
Claims (10)
1.一种重金属污染土壤修复效果的生态评估方法,其特征在于,包括:
采集样品,且所述样品为经调理剂修复后目标区域内的土壤样品和农作物样品;
获取所述土壤样品的理化性质评级A、微生物活性评级B及Cd含量评级C;获取所述农作物样品中的Cd含量评级D;
计算所述目标区域内的土壤修复效果P,且P=iA+jB+kC+sD;式中,i为理化性质评级A的权重,j为微生物活性评级B的权重,k为Cd含量评级C的权重,s为Cd含量评级D的权重。
2.根据权利要求1所述的一种重金属污染土壤修复效果的生态评估方法,其特征在于:所述调理剂采用矿物型调理剂、有机型调理剂或微生物型调理剂。
3.根据权利要求2所述的一种重金属污染土壤修复效果的生态评估方法,其特征在于:在所述目标区域内,所述调理剂的修复用量为3t·hm-1。
4.根据权利要求2或3所述的一种重金属污染土壤修复效果的生态评估方法,其特征在于:所述矿物型调理剂至少包括硅酸钙、熟石灰、硫酸钾、无水硫酸镁和九水合硝酸铁。
5.根据权利要求1所述的一种重金属污染土壤修复效果的生态评估方法,其特征在于:获取所述土壤样品的理化性质评级A时,至少获取所述土壤样品的:pH值、有机碳含量、碱解氮含量、有效磷含量和速效钾含量。
6.根据权利要求1所述的一种重金属污染土壤修复效果的生态评估方法,其特征在于:获取所述土壤样品的的Cd含量评级C时,采用DTPA浸提法测定所述土壤样品的的Cd含量。
7.根据权利要求1或6所述的一种重金属污染土壤修复效果的生态评估方法,其特征在于:获取所述农作物样品中的Cd含量评级D时,采用石墨炉原子吸收光谱仪测定所述农作物样品中Cd的含量。
8.根据权利要求7所述的一种重金属污染土壤修复效果的生态评估方法,其特征在于,采用石墨炉原子吸收光谱仪测定所述农作物样品中Cd的含量时包括:
将所述农作物样品晾晒至恒重;
依次脱粒、脱壳、研磨和消解处理晾晒后的所述农作物样品,获得消解液;
采用石墨炉原子吸收光谱仪测定所述消解液中Cd的含量。
9.根据权利要求1所述的一种重金属污染土壤修复效果的生态评估方法,其特征在于:获取所述土壤样品的微生物活性评级B时,至少获取微生物活力和微生物量。
10.根据权利要求9所述的一种重金属污染土壤修复效果的生态评估方法,其特征在于:所述微生物活力采用土壤酶活性作为评价指标,且所述土壤酶至少包括蔗糖酶、脲酶和酸性磷酸酶。
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