紫茉莉对铅胁迫的响应及耐性研究
摘要
矿业废弃地的治理和植被恢复是我国面临的紧迫任务之一,也是可持续发展战略优先关注的问题之一,而尾矿废弃地更是金属矿山最常见、最难恢复的矿业废弃弃地,还存在潜在的环境风险。植物稳定技术利用耐性植物的机械固定作用减轻风蚀和水蚀以及根系吸附和根际沉淀作用实现尾矿的长期稳定,降低重金属的生物有效性,是一种比较有前景的环境友好技术。在湖南湘潭和吉首等地矿区调查时发现紫茉莉具有生长迅速、生物量大、适应性强等特点,是一种较为理想修复重金属污染土壤的耐性植物。目前国内外关于紫茉莉在铅胁迫下生理生态响应的研究较少,因此,开展紫茉莉铅胁迫下生理生态响应和耐性机理的研究,将为其应用于重金属污染环境修复技术提供科学依据,进而推动金属矿业废弃地植物稳定技术的商业化应用。
在大量野外调查的基础上,通过营养液培养实验,综合运用电感耦合等离子体原子发射光谱仪、显微成像系统、透射电子显微镜技术、傅里叶变换红外光谱分析技术等分析手段比较系统地研究不同浓度铅胁迫条件下紫茉莉生物量和器官组织的变化、矿质元素的吸收和累积、不同器官组织化学组分的变化规律,以期为铅污染场地植物稳定提供理论依据。通过研究,取得以下主要结果:
1.紫茉莉不同器官组织对铅的吸收存在差异性,根系对铅的积累最强,其次是叶、茎。随着生长介质中铅浓度水平的升高,根、茎和叶组织铅含量呈增加态势。紫茉莉吸收的铅绝大部分都积累在植株的根系部位,向地上部分转移量非常少,不同铅处理条件下茎叶的转移系数均小于0.04。这既可降低铅向植株地上部分(茎、叶)的迁移、减轻铅对地上部分营养器官和生殖器官的毒害作用,又能将植株吸收铅固定在根系部位,降低Pb的生物有效性。
2.不同铅处理条件下,紫茉莉根、茎和叶的大量营养元素含量变化不大,并未随生长介质中铅浓度水平的升高呈明显降低,甚至部分处理条件下出现一定程度的增加。Cu、Mn、Zn等微量元素在根、茎和叶等器官的含量均呈现出在较低铅处理浓度条件下逐渐升高,而在较高浓度铅处理水平下呈下降趋势。在紫茉莉的整个生命周期内,没有发现Ca、Mg、Fe等元素缺乏的现象,大量营养元素的稳态维持可能是紫茉莉对铅胁迫具有较强耐性的一种反馈机制。
3.不同铅处理条件下,紫茉莉均能正常生长,即使在生长介质铅含量高达到1000μmol·L~(-1)时,紫茉莉仍然能够完成整个生命周期,生物量与对照植株相比没有显著差异。在200μmol·L~(-1)铅胁迫条件下紫茉莉的根部组织结构与对照相比无显著差异;当铅含量达到1000μmol·L~(-1)时植株的根部疏导组织虽然出现了一定程度的减少,但是表皮、皮层、中柱维管组织未发生明显变化,而叶片组织结构基本没有受到影响。这表明紫茉莉有较强的铅耐性,使其有应用植物稳定技术修复铅污染环境的潜力。
4.傅立叶变换红外光谱分析研究发现,紫茉莉根部某些羧酸、糖类、蛋白质、氨基酸等物质含量出现波动,在中低浓度铅处理下升高,在高浓度下下降,如在2920cm~(-1)处羧酸的峰高在200μmol·L~(-1)铅处理前上升,在500μmol·L~(-1)开始下降。这些物质能够螯合和络合重金属,降低重金属在植物体内活性,可以明显增强植物对重金属的耐性。在不同铅处理水平条件下,紫茉莉茎和叶中羧酸、糖类、蛋白质、氨基酸等物质含量差异不显著,这可能是由于紫茉莉吸收的铅主要集中在根系部位,向植株地上部分(茎、叶)转移少的缘故。
Environmental management and rehabilitation of mine wasteland is one of the pressing needs to be addressed for social and economic development to be healthy and sustainable in China. Tailing wasteland from metalliferous mines often poses the extreme stressful conditions and is tougher for ecological restoration. As a newly emerged environment -friendly technology, phytostabilization is defined as the use of metal -tolerant plant species to reduce the mobility and bioavailability of heavy metals in the substrates either by immobilization or by prevention of migration. Compared with traditional methods, phytostabilization is a cost-effective and environment-friendly technology. Mirabilis jalapa Linn is a plant found in Xiangtan and Jishou tailing wasteland. There are less researches on the toxicity and the tolerance machanisms of M. jalapa to lead up to now. And this would restrain its application on phytostabilization.
To elucidate systematically the changing of biomass and organs tissue structure, the uptake and accumulation of mineral elements and the alteration of chemistry components in M. jalapa organs to different lead stress, hydroponic experiment were applied in this study based on a lot of field survey by means of a series of instrumentations, such as Inductively -coupled plasma optical emission spectroscopy (ICP-OES), Transmission Electron Microscope (TEM), paraffin- embedded tissue slice and Fourier transform infrared(FTIR). The major results obtained are as follows:
1. The distribution of lead among roots, stems and leaves in M. jalapa is different. The ICP-OES analysis revealed that the levels of Pb in roots,stems and leaves of the plants increased while the concentration of Pb~(2+) increasing in the culture solution. The results showed that Pb distribution sequence is: roots > leaves > stems.And most of Pb was accumulated in roots, few Pb was transferred in shoots, meanwhile the translocation factor<0.04. This could be one of Pb defense mechanisms employed by M. jalapa. The results indicated that M. jalapa could alleviate toxic effect on vegetative organs and reproductive organs by low transportation ability of Pb from roots to shoots and also reduce the bioavailability of Pb by the Pb immobilization in roots.
2. In the present work, there existed no significant effects of Pb stress on the concentration of Macronutrients such as Ca、Mg、Fe in different organs of M.jalapa. These elements were not decreasing evidently while the concentration of Pb~(2+) increasing in the culture solution,but increasing in some treatment. The uptake and accumulation of trace elements Cu、Mn、Zn in M. jalapa was researched. All the result showed that the concentrations of Cu、Mn、Zn was rising under the low Pb stress and decreasing under high Pb stress. Macronutrients such as Ca、Mg、Fe was enough in M. jalapa during the whole growth cycle. The homeostasis of macronutrients maintained may revealed that M. jalapa had the strong resistance to Pb stress.
3. In our study, the biomass of M. jalapa had no significant differences between all rates. The results showed that plant grew normally and did not show any symptoms of lead toxicity during the period of treatment even at 1000μmol·L~(-1) Pb in the growth medium. There were no significant differences between 200μmol·L~(-1) Pb treatment and control plants in M. jalapa roots anatomize tissue structure. Meanwhile, the trachea of root was significant decreasing under 1000μmol·L~(-1) Pb stress, but the structure of epidermis cells, cortical cells and fibrovascular tissue maintained complete. And the tissue structure of leaf was not effected even the Pb treatment reached 1000μmol·L~(-1) basically. Therefore, there is great potential for using M.jalapa in the application of phytostabilization for lead contaminated mining wasteland because of its strong Pb tolerance.
4. The FTIR of M. jalapa showed the chemical composition such as organic acids, carbohydrate, protein and amino acids was effected by the Pb stress in roots. The absorbance of the dominating bands near 3420, 2920, 1610, 1060 cm~(-1) which correspond to organic acids, carbohydrate, protein and amino acids rose firstly and then decreased in root tissues. These material could reduce the availability and toxicity of heavy metal by chelation and strengthen the tolerance to heavy metal. But M. jalapa did not show obvious changes in leaves and stems because less Pb was transported into shoot tissues in all treatments. This could be a kind of feedback mechanism of Mirabilis jalapa Linn under Pb stress.
在大量野外调查的基础上,通过营养液培养实验,综合运用电感耦合等离子体原子发射光谱仪、显微成像系统、透射电子显微镜技术、傅里叶变换红外光谱分析技术等分析手段比较系统地研究不同浓度铅胁迫条件下紫茉莉生物量和器官组织的变化、矿质元素的吸收和累积、不同器官组织化学组分的变化规律,以期为铅污染场地植物稳定提供理论依据。通过研究,取得以下主要结果:
1.紫茉莉不同器官组织对铅的吸收存在差异性,根系对铅的积累最强,其次是叶、茎。随着生长介质中铅浓度水平的升高,根、茎和叶组织铅含量呈增加态势。紫茉莉吸收的铅绝大部分都积累在植株的根系部位,向地上部分转移量非常少,不同铅处理条件下茎叶的转移系数均小于0.04。这既可降低铅向植株地上部分(茎、叶)的迁移、减轻铅对地上部分营养器官和生殖器官的毒害作用,又能将植株吸收铅固定在根系部位,降低Pb的生物有效性。
2.不同铅处理条件下,紫茉莉根、茎和叶的大量营养元素含量变化不大,并未随生长介质中铅浓度水平的升高呈明显降低,甚至部分处理条件下出现一定程度的增加。Cu、Mn、Zn等微量元素在根、茎和叶等器官的含量均呈现出在较低铅处理浓度条件下逐渐升高,而在较高浓度铅处理水平下呈下降趋势。在紫茉莉的整个生命周期内,没有发现Ca、Mg、Fe等元素缺乏的现象,大量营养元素的稳态维持可能是紫茉莉对铅胁迫具有较强耐性的一种反馈机制。
3.不同铅处理条件下,紫茉莉均能正常生长,即使在生长介质铅含量高达到1000μmol·L~(-1)时,紫茉莉仍然能够完成整个生命周期,生物量与对照植株相比没有显著差异。在200μmol·L~(-1)铅胁迫条件下紫茉莉的根部组织结构与对照相比无显著差异;当铅含量达到1000μmol·L~(-1)时植株的根部疏导组织虽然出现了一定程度的减少,但是表皮、皮层、中柱维管组织未发生明显变化,而叶片组织结构基本没有受到影响。这表明紫茉莉有较强的铅耐性,使其有应用植物稳定技术修复铅污染环境的潜力。
4.傅立叶变换红外光谱分析研究发现,紫茉莉根部某些羧酸、糖类、蛋白质、氨基酸等物质含量出现波动,在中低浓度铅处理下升高,在高浓度下下降,如在2920cm~(-1)处羧酸的峰高在200μmol·L~(-1)铅处理前上升,在500μmol·L~(-1)开始下降。这些物质能够螯合和络合重金属,降低重金属在植物体内活性,可以明显增强植物对重金属的耐性。在不同铅处理水平条件下,紫茉莉茎和叶中羧酸、糖类、蛋白质、氨基酸等物质含量差异不显著,这可能是由于紫茉莉吸收的铅主要集中在根系部位,向植株地上部分(茎、叶)转移少的缘故。
Environmental management and rehabilitation of mine wasteland is one of the pressing needs to be addressed for social and economic development to be healthy and sustainable in China. Tailing wasteland from metalliferous mines often poses the extreme stressful conditions and is tougher for ecological restoration. As a newly emerged environment -friendly technology, phytostabilization is defined as the use of metal -tolerant plant species to reduce the mobility and bioavailability of heavy metals in the substrates either by immobilization or by prevention of migration. Compared with traditional methods, phytostabilization is a cost-effective and environment-friendly technology. Mirabilis jalapa Linn is a plant found in Xiangtan and Jishou tailing wasteland. There are less researches on the toxicity and the tolerance machanisms of M. jalapa to lead up to now. And this would restrain its application on phytostabilization.
To elucidate systematically the changing of biomass and organs tissue structure, the uptake and accumulation of mineral elements and the alteration of chemistry components in M. jalapa organs to different lead stress, hydroponic experiment were applied in this study based on a lot of field survey by means of a series of instrumentations, such as Inductively -coupled plasma optical emission spectroscopy (ICP-OES), Transmission Electron Microscope (TEM), paraffin- embedded tissue slice and Fourier transform infrared(FTIR). The major results obtained are as follows:
1. The distribution of lead among roots, stems and leaves in M. jalapa is different. The ICP-OES analysis revealed that the levels of Pb in roots,stems and leaves of the plants increased while the concentration of Pb~(2+) increasing in the culture solution. The results showed that Pb distribution sequence is: roots > leaves > stems.And most of Pb was accumulated in roots, few Pb was transferred in shoots, meanwhile the translocation factor<0.04. This could be one of Pb defense mechanisms employed by M. jalapa. The results indicated that M. jalapa could alleviate toxic effect on vegetative organs and reproductive organs by low transportation ability of Pb from roots to shoots and also reduce the bioavailability of Pb by the Pb immobilization in roots.
2. In the present work, there existed no significant effects of Pb stress on the concentration of Macronutrients such as Ca、Mg、Fe in different organs of M.jalapa. These elements were not decreasing evidently while the concentration of Pb~(2+) increasing in the culture solution,but increasing in some treatment. The uptake and accumulation of trace elements Cu、Mn、Zn in M. jalapa was researched. All the result showed that the concentrations of Cu、Mn、Zn was rising under the low Pb stress and decreasing under high Pb stress. Macronutrients such as Ca、Mg、Fe was enough in M. jalapa during the whole growth cycle. The homeostasis of macronutrients maintained may revealed that M. jalapa had the strong resistance to Pb stress.
3. In our study, the biomass of M. jalapa had no significant differences between all rates. The results showed that plant grew normally and did not show any symptoms of lead toxicity during the period of treatment even at 1000μmol·L~(-1) Pb in the growth medium. There were no significant differences between 200μmol·L~(-1) Pb treatment and control plants in M. jalapa roots anatomize tissue structure. Meanwhile, the trachea of root was significant decreasing under 1000μmol·L~(-1) Pb stress, but the structure of epidermis cells, cortical cells and fibrovascular tissue maintained complete. And the tissue structure of leaf was not effected even the Pb treatment reached 1000μmol·L~(-1) basically. Therefore, there is great potential for using M.jalapa in the application of phytostabilization for lead contaminated mining wasteland because of its strong Pb tolerance.
4. The FTIR of M. jalapa showed the chemical composition such as organic acids, carbohydrate, protein and amino acids was effected by the Pb stress in roots. The absorbance of the dominating bands near 3420, 2920, 1610, 1060 cm~(-1) which correspond to organic acids, carbohydrate, protein and amino acids rose firstly and then decreased in root tissues. These material could reduce the availability and toxicity of heavy metal by chelation and strengthen the tolerance to heavy metal. But M. jalapa did not show obvious changes in leaves and stems because less Pb was transported into shoot tissues in all treatments. This could be a kind of feedback mechanism of Mirabilis jalapa Linn under Pb stress.
引文
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