镉超积累植物及植物镉积累特性转基因改良研究进展

植物提取修复技术是一项既经济又环保的土壤镉(Cd)污染修复技术,该技术的关键是筛选Cd超积累植物或利用基因工程手段改良植物以提高其Cd积累能力。人们已发现遏兰菜等7种Cd超积累植物及美人蕉等潜在的Cd超积累植物。还发现了许多与Cd耐受和积累能力有关的基因:(1)编码与Cd积累、耐受有关酶的基因,如细菌中的ACC(1-aminocyclopropane-1-carboxylic acid),植物中的PCS(Phytochelatin Synthase)基因;(2)编码金属结合蛋白的基因:MT(Metallothionein)、转运蛋白(P-type ATPase、ABC型转运器)基因;(3)其它相关基因:Hvhsp17、PvSR2(Phaseolus vulgaris stress-related gene number 2)等。并将其中的一些基因转入到其它生物中,提高了其对Cd的耐受性和积累量,为实现Cd污染土壤修复的目标奠定基础。     

小球藻生物富集锌、镉机制的研究

藻类对许多重金属具有较强的生物富集能力,小球藻（Chlorella spp.)分别在不同系列Zn^2 ,Cd^2 浓度下培养8d间隔2d,测定小球藻的生物量及Zn和Cd的含量。结果表明,小球藻随水体中Zn^2 和Cd^2 的浓度不同,在重重金属水体中暴露时间不同,具有不同的富集速率和能力。     

超积累植物伴矿景天镉耐受基因SpMT2的分离及功能鉴定

超积累植物由于其对重金属具有地上部超积累以及超耐受等特性,不仅是研究植物离子转运及毒性耐受的理想模式,而且在植物修复的发展和应用中具有不可替代的作用。伴矿景天是近年在我国境内发现的一种景天科镉(Cd)/锌(Zn)超积累植物。为鉴定其富集和耐受Cd的关键基因,笔者构建了其酵母表达cDNA文库,利用酵母的遗传互补系统筛选到一个极大提高了酵母抗Cd能力的基因SpMT2。SpMT2属于富含半胱氨酸(Cys)的金属硫蛋白(Metallothionein)家族。亚细胞定位表明SpMT2表达于酵母细胞质中,并特异地提高酵母对Cd的抗性。进一步研究发现SpMT2的表达显著降低了酵母液泡中Cd含量,但酵母吸收的总Cd含量无显著变化。推测抗性增加是由于SpMT2在酵母细胞质中通过螯合Cd从而降低Cd对酵母的毒害。qRT-PCR分析表明SpMT2在伴矿景天的根和地上部都高丰度表达,且不受Cd诱导变化。鉴于SpMT2也定位于植物细胞质中,结合上述结果,推测SpMT2可能在伴矿景天细胞质中螯合Cd,在降低Cd毒害的同时可能还保持Cd在细胞质中的流动性,从而在Cd长途转运过程中也发挥重要作用。     

镉和砷在植物中的积累及其分子调控机制研究进展

植物源食物是人类摄入有毒金属元素镉(Cd)和砷(As)的主要途径。深化植物对Cd和As积累途径分子机制的认识,有助于培育可食部分中低有毒金属元素含量的作物新种质。该文基于近年来有关植物Cd和As积累在主要模式植物中所取得的实质性研究进展,对植物介导Cd和As吸收的转运蛋白、As形态和生物转化机制以及控制Cd和As根-地上部转运效率和分配的关键因子等方面的研究进展进行综述,并对未来的研究前景进行了展望。     

石油烃对沙蚕镉生物富集特性及金属硫蛋白诱导的影响

多毛类沙蚕作为海陆交错带的关键性物种,常被作为环境监测的指示生物.我国海陆交错带沙蚕常见种——双齿围沙蚕经常遭受各种污染物\[包括重金属镉(Cd)和石油烃\]的胁迫.本研究采用慢性微宇宙试验,以暴露于单一和复合Cd和石油烃条件下的沙蚕为研究对象,重点考察石油烃对沙蚕Cd生物富集特性及诱导金属硫蛋白的影响.结果表明: 单一Cd胁迫条件下,沙蚕对重金属Cd的生物累积随暴露浓度的升高而显著上升,生物富集系数随暴露时间的延长呈上升趋势;复合胁迫条件下,与对照组相比,石油烃的添加可以显著增加沙蚕对重金属Cd的生物累积.Cd可以诱导沙蚕金属硫蛋白(MT)的表达,但当沙蚕体内Cd含量达到180 mg·kg^-1DM时,MT基本饱和;石油烃不能显著诱导沙蚕MT合成,与单一Cd处理相比,石油烃的添加能够显著影响Cd诱导的MT含量.这说明尽管石油烃本身不能直接影响MT合成,但在与Cd共存时可以调节MT的表达,因此在利用野生沙蚕MT作为监测指标时要具体区分蛋白表达的潜在诱导因素.     

脱金属硫蛋白与镉离子的络合作用及构象研究

用圆二色（ＣＤ）谱地研究兔肝脱金属硫属蛋白的两个亚型与Ｃｄ＾２＋的络合作用及对重组ＭＴ构象的影响。观测了ａｐｏ－ＭＴ垢巯基在空气和室温下的稳定性。在ＰＨ４．７１,镉重组ＭＴ１的ＣＤ谱特征峰在２５７ｎｍ（＋）,２３８ｎｍ（－）,２２６ｎｍ（＋）与镉诱导的天然ＭＴ１相同。在空气存在和ＰＨ７．９０的ＣＤ谱只有２４３ｎｍ（＋）一个峰。向两亚型分别加入７ｅｑＣｄ＾２＋测定ＣＤ谱随ＰＨ值的变化,发现在ＰＨ２．     

镉诱导家兔肝脏含锌金属硫蛋白的纯化及鉴定

预先给家兔皮下注射CdCl_2四次,取肝脏经匀浆、离心、乙醇沉淀后,通过Sephadex G-50凝胶过滤层析,DEAE Sepharose Fast Flow离子交换层析和Sephadex G-25层析柱脱盐,得到MT的两个“亚型”—MT-1和MT-2,对其进行鉴定,其紫外扫描图谱示在280和250nm处无吸收峰,而最大吸收在220nm处。MT-1和MT-2均不含镉和铜,而每分子结合6个锌。用简化巯基试剂测定,MT-1含18.8个巯基,MT-2含19.1个巯基;HPLC分析MT主要以单体存在,也有少量二聚体。氨基酸组成与Cd-MT一致。整个纯化过程采用示波极谱法和火焰原子吸收光谱法测锌含量。结果表明前者可代替后者。     

镉不同积累型作物品种对镉吸收、转运和积累特性的研究进展

农田土壤镉污染问题日趋严重, 利用可食部位低积累镉的农作物品种成为保障食品安全的有效策略之一。镉在作物可食部位的累积与镉的吸收转运有关。分析了根系对镉的吸收、镉向地上部的运输和分配及镉在细胞内的分布和结合形态与作物积累镉的关系, 指出作物品种间的镉积累差异受以上多种因素的调控, 但不同作物之间影响镉积累的主要因素存在差异。同时提出在对低镉积累作物品种的筛选和机制研究方面还有很多工作需要开展。     

RNA干涉P-Hsp70基因表达对不同温度下方斑瓢虫成虫寿命及繁殖力的影响

【目的】本研究旨在明确热激蛋白P-Hsp70基因表达对不同温度下方斑瓢虫Propylaea quatuordecimpunctata成虫寿命和繁殖力的影响。【方法】采用显微注射递送dsRNA的方法对方斑瓢虫成虫的P-Hsp70基因进行干涉,利用qRT-PCR检测方法计算P-Hsp70基因的干扰效率,同时观察P-Hsp70基因被干扰后方斑瓢虫成虫暴露在不同温度（32、35和38℃）下的存活和繁殖情况。【结果】显微注射递送dsRNA的方法可有效抑制方斑瓢虫成虫P-Hsp70基因的表达。显微注射ds-P-Hsp70处理组方斑瓢虫成虫在各个温度下的存活率、寿命和繁殖力均显著低于未注射dsRNA的对照组。在32、35和38℃下,显微注射dsRNA 24 h后方斑瓢虫成虫存活率较对照组分别降低了15.5%、16.7%和56.8%,成虫平均寿命分别缩短40.6%、51.2%和65.8%。在32和35℃下,显微注射dsRNA成虫繁殖力分别降低了37.5%和100.0%;在38℃时,无论注射dsRNA与否,成虫均未能产卵。【结论】热激蛋白P-Hsp70基因在方斑瓢虫成虫抵御高温胁迫过程中具有非常重要的作...     

镉中毒大鼠睾丸与肝脏金属硫蛋白表达的时相研究

啮齿目动物睾丸对镉毒性较肝脏更敏感.为阐明睾丸的镉毒性分子机制,比较了肝脏与睾丸金属硫蛋白（MT）表达的时相变化.mRNA采用RT-PCR技术分析并用光密度扫描定量;蛋白质定量用ELISA方法.结果显示,睾丸中存在MT,镉中毒后MT1与MT2 mRNA明显升高,但MT没有相应增加;肝脏镉中毒后MTmRNA与MT均明显升高.结果提示：镉虽然能诱导睾丸MTmRNA的转录,但没有促进其MT的合成,这可能是睾丸对镉毒性与致癌作用较肝脏更敏感的重要原因.     

Isolation and characterization of microsatellite loci in Babylonia areolata and cross-species amplification in Babylonia formosae habei

Nine novel microsatellite primer pairs were presented for Babylonia areolata, representing the first microsatellite markers available for this genus. Levels of polymorphism were variable with 2 to 11 alleles per locus and expected heterozygosities ranging from 0.073 to 0.907 in 27 individuals of the population from which the loci were isolated. We found significant heterozygote deficit at one locus that might be attributable to null alleles. We were successful at cross-amplifying six loci in the congeneric B. formosae habei. These markers are therefore potentially useful for conservation studies, population structure assessment, ecological analyses and linkage map construction.     

Cadmium uptake and bioaccumulation in selected cultivars of durum wheat and flax as affected by soil type

Accumulation of cadmium (Cd) in crop plants is of great concern due to the potential for food chain contamination through the soil-root interface. Although Cd uptake varies considerably with plant species, the processes which determine the accumulation of Cd in plant tissues are affected by soil factors. The influence of soil type on Cd uptake by durum wheat (Triticum turgidum var. durum L.) and flax (Linum usitatissimum L.) was studied in a pot experiment under environmentally controlled growth chamber conditions. Four cultivars/lines of durum wheat (Kyle, Sceptre, DT 627, and DT 637) and three cultivars/lines of flax (Flanders, AC Emerson, and YSED 2) were grown in two Saskatchewan soils: an Orthic Gray Luvisol (low background Cd concentration; total/ABDTPA extractable Cd: 0.12/0.03 mg kg^-1, respectively) and a Dark Brown Chernozem (relatively high background Cd concentration; total/ABDTPA Cd: 0.34/0.17 mg kg^-1 respectively). Plant roots, stems, newly developed heads, and grain/seeds were analyzed for Cd concentration at three stages of plant growth: two and seven weeks after germination, and at plant maturity. The results showed that Cd bioaccumulation and distribution within the plants were strongly affected by both soil type and plant cultivar/line. The Cd concentration in roots leaves and stems varied at different stages of plant growth. However, all cultivars of both plant species grown in the Chernozemic soil accumulated more Cd in grain/seeds than plants grown in the Orthic Gray Luvisol soil. The different Cd accumulation pattern also corresponded to the levels of ABDTPA extractable and metal-organic complex bound soil Cd found in both soils. Large differences were found in grain Cd among the durum wheat cultivars grown in the same soil type, suggesting the importance of rhizosphere processes in Cd bioaccumulation and/or Cd transport processes within the plant. Distribution of Cd in parts of mature plants showed that durum grain contained up to 21 and 36% of the total amount of Cd taken up by the plants for the Orthic Gray Luvisol and Chernozemic soils, respectively. These results indicate the importance of studying Cd speciation, bioaccumulation and cycling in the environment for the management of agricultural soils and crops.     

Low-molecular-weight organic acids in rhizosphere soils of durum wheat and their effect on cadmium bioaccumulation

Cadmium (Cd) accumulation has been found to vary between cultivars of durum wheat (Triticum turgidum var. durum), and it is hypothesized that low-molecular-weight organic acids (LMWOAs) produced at the soil-root interface (rhizosphere) may play an important role in the availability and uptake of Cd by these plants. The objective of this study, therefore, was to (1) investigate the nature and quantity of LMWOAs present in the rhizosphere of durum wheat cultivars Arcola (low Cd accumulator) and Kyle (high Cd accumulator) grown in three different soils: Yorkton, Sutherland and Waitville, and (2) determine the relationship between Cd accumulation in these plants and LMWOAs present in the rhizosphere. Plants were grown for two weeks in pot-cultures under growth chamber conditions. Oxalic, fumaric, succinic, L-malic, tartaric, citric, acetic, propionic and butyric acids were found and quantified in the water extracts of rhizosphere soil, with acetic and succinic acids being predominant. No water extractable LMWOAs were identified in the bulk soil. Total amount of LMWOAs in the rhizosphere soil of the high Cd accumulator (Kyle) was significantly higher than that for the low Cd accumulator (Arcola) in all three soils. Furthermore, large differences in amounts of LMWOAs were found in the rhizosphere soil for the same cultivars grown in different soils and followed the pattern: Sutherland > Waitville > Yorkton. Extractable soil Cd (M NH4Cl) and Cd accumulation in the plants also followed the same soil sequence as LMWOA production. Cadmium accumulation by the high and low Cd accumulating cultivars was proportional to the levels of LMWOAs found in the rhizosphere soil of each cultivar. These results suggest that the differing levels of LMWOAs present in the rhizosphere soil played an important role in the solubilization of particulate-bound Cd into soil solution and its subsequent phytoaccumulation by the high and low Cd accumulating cultivars.     

Stereoselective bioaccumulation,transformation, and toxicity of triadimefon in Scenedesmus obliquus

In this study the stereoselective bioaccumulation and transformation of triadimefon and the toxicity of triadimefon and its metabolite triadimenol to the green algae Scenedesmus obliquus were studied. In growth inhibition experiments, triadimenol was more toxic than triadimefon, and (1S,2R)‐triadimenol, which has the largest fungicidal activity, presented the highest toxicity to the algae. In bioaccumulation experiments, triadimefon was rapidly taken up by algae cells, and the decrease in the concentration of triadimefon was accompanied by an increase in triadimenol. The transformation of S‐(+)‐ triadimefon was faster than that of the R‐(−)‐enantiomer, resulting in four triadimenol stereoisomers at different forming rates: B2 (1S, 2S) > B1 (1R, 2R) > A2 (1S, 2R) > A1 (1R, 2S). Thus, it is necessary to explore the enantioselective toxicology and ecological fate of these chiral pesticides in an environmental risk assessment. Also, their metabolites should be paid specific attention to since they may pose higher ecological risks.     

Melatonin increases tissue accumulation and toxicity of cadmium in the bank vole (<Emphasis Type="Italic">Clethrionomys glareolus</Emphasis>)

Recent study has shown that a short photoperiod increases the accumulation and toxicity of cadmium (Cd) in the bank vole as compared to a long photoperiod. Since many of the effects of photoperiod on physiological processes in small mammals are transduced by the pineal gland and its hormone melatonin, in this study the effect of subchronic melatonin injection (7 mol/kg/day for 6 weeks) on the hepatic, renal and intestinal Cd accumulation in the bank voles raised under a long photoperiod and exposed to dietary Cd (0.9 mol/g) was examined. Simultaneously, histological examinations of the liver and kidneys, and analyses of metallothionein (MT) and lipid peroxidation were carried out. Melatonin co-treatment brought about a significant increase in the hepatic (61%), renal (79%) and intestinal (77%) Cd concentrations as compared to those in the Cd alone group. However, the concentrations of MT in the liver and kidneys of the Cd + melatonin co-treated bank voles did not differ from those in the Cd alone group. Also, histopathological changes in the liver (infiltration of leukocytes) and kidneys (glomerular swelling and a focal tubular cell degeneration) as well as an increase (2-fold) in the renal lipid peroxidation occurred only in animals from the Cd + melatonin group. These data indicate that (1) subchronic melatonin injection has similar effect on the tissue accumulation and toxicity of Cd to that produced by a short photoperiod and (2) the Cd-induced toxicity in the liver and kidneys of melatonin co-treated bank voles is probably due to increased Cd accumulation and decreased synthesis of MT.     

Cadmium status of soils and plants from a long-term fertility experiment in southeast Norway

To determine whether the use of phosphate fertilizer resulted in measurable cadmium accumulation in soils and crops harvested, soil and plant samples were collected from some selected treatments of a seventy year-old fertilizer experiment at Møystad, southeast Norway. Soil samples after extraction with Aqua Regia or 1M NH₄NO₃ and plant samples after digestion were analyzed for Cd. Cadmium balance based on fertilizer and atmospheric inputs and crop removal and leaching losses was worked out. Neither the total nor the available (NH₄NO₃-extractable) Cd in the soil was significantly affected by Cd added through fertilizer, though a tendency of higher Cd in soils from the plots receiving higher amounts of fertilizer was seen. The same trend was also observed for the Cd concentration in plants. Annual Cd input rates (fertilizer and atmosphere) varied from 1.20 to 2.57 g Cd ha^-1 y^-1 and the Cd removal (crops and leaching) rates varied from 1.16 to 1.79 g Cd ha^-1 y^-1. The balance calculations based on the seventy years data indicated Cd accumulation in the soil was <1 g Cd ha^-1 y^-1, but that increasing the doses of either commercial fertilizer or farm yard manure would likely result in increased accumulation of the element. This may have a negative impact because the available soil Cd content would be increased at a faster rate, resulting in increased plant uptake. Although Cd tended to accumulate as a result of P fertilization, the rate of increase was slow. The annual increase in the total Cd content of fertilized plots varied from 0.04 to 0.12% indicating that it may take from 800 to 2000 years, depending upon the fertilizer input, to accumulate Cd equivalent to that currently present in the soil.     

镉胁迫下萝卜基因组DNA甲基化敏感扩增多态性分析

应用甲基化敏感扩增多态性(MSAP)技术分析了重金属镉(cd)胁迫处理后萝卜基因组DNA甲基化程度的变化。结果表明,经50、250和500mg/L CdCl_2处理后,MSAP比率分别为37%、43%和51%,均高于对照(34%);全甲基化率(双链C~mCGG)分别为23%、25%和27%,而其对照为22%,表明重金属CdCl_2胁迫后,某些位点发生了重新甲基化。萝卜叶片DNA中总甲基化水平的增加与CdCl_2处理浓度呈显著正相关。甲基化变异可分为重新甲基化、去甲基化、不定类型以及与对照相同的甲基化模式等类型,Cd胁迫处理引起的植株基因组DNA甲基化程度的提高主要是重新甲基化。     

Cadmium toxicity in crop plants and its alleviation by arbuscular mycorrhizal (AM) fungi: An overview

Cadmium (Cd), a toxic metal released into agricultural settings induces numerous changes in plant growth and physiology. The main known mechanisms of Cd toxicity include its affinity for sulfhydryl groups in proteins and its ability to replace some essential metals in active sites of enzymes, thus causing inhibition of enzyme activities and protein denaturation. This article reviews detrimental effects of Cd toxicity on the functional biology of plants and summarizes the mechanisms that are activated by plants to prevent the absorption or to detoxify Cd ions such as synthesis of antioxidants, osmolytes, phytochelatins, metallothioneins, etc. Arbuscular mycorrhizal (AM) fungi are reported to be present on the roots of plants growing in metal-contaminated soils and play an important role in metal tolerance. Through mycorrhizal symbiosis, heavy metals are immobilized in the rhizosphere through precipitation in the soil matrix, adsorption onto the root surface or accumulation within roots, and compartmentalized in aboveground parts of the plant. This article unfolds the potential role of AM fungi in enhancing Cd tolerance of plants.