Levels of Betaine and Betaine Aldehyde Dehydrogenase Activity in the Green Leaves, and Etiolated Leaves and Roots of Barley

The accumulation of betaine and the induction of betaine aldehydedehydrogenase, which catalyzes the last step in the synthesisof betaine, were analyzed in salt-stressed barley leaves. Whenhydroponically grown barley plants were transferred to a mediumthat contained 200 mM NaCl, the levels of both betaine and thetotal extractable betaine aldehyde dehydrogenase activity inthe leaves increased approximately 7-fold and 3-fold when calculatedon the basis of total leaf protein, respectively, over the courseof 7 days. Betaine aldehyde dehydrogenase activity was alsodetected in either etiolated leaves or roots of barley plantsgrown under aseptic conditions. Betaine was detected in bothetiolated leaves and roots at levels that were about 20% ofthat in green leaves when calculated on a fresh weight basis. ¹ This research was supported financially by a research grantfrom the Ministry of Education, Science and Culture (63560080) (Received March 9, 1990; Accepted May 29, 1990)     

Employment of associative bacteria for the inoculation of barley plants cultivated in soil contaminated with lead and cadmium

In laboratory experiments, the rhizobacteria Azospirillum lipoferum 137, Arthrobacter mysorens 7, Agrobacterium radiobacter 10, and Flavobacterium sp. L30 were found to have a relatively high resistance to the toxic heavy metals lead and cadmium (except that strain L30 was found to be sensitive to Cd). When introduced by means of seed bacterization, the heavy metal-resistant strains actively colonized the rhizosphere of barley plants cultivated in uncontaminated and contaminated soils. In both pot and field experiments, seed bacterization improved the growth of barley plants and the uptake of nutrient elements from soil contaminated with Pb and Cd. The bacterization also prevented the accumulation of Pb and Cd in barley plants, thereby mitigating the toxic effect of these heavy metals on the plants.     

Enzymatic characterization of peroxisomal and cytosolic betaine aldehyde dehydrogenases in barley

Betaine aldehyde dehydrogenase (BADH; EC 1.2.1.8) is an important enzyme that catalyzes the last step in the synthesis of glycine betaine, a compatible solute accumulated by many plants under various abiotic stresses. In barley ( Hordeum vulgare L.), we reported previously the existence of two BADH genes ( BBD1 and BBD2 ) and their corresponding proteins, peroxisomal BADH (BBD1) and cytosolic BADH (BBD2). To investigate their enzymatic properties, we expressed them in Escherichia coli and purified both proteins. Enzymatic analysis indicated that the affinity of BBD2 for betaine aldehyde was reasonable as other plant BADHs, but BBD1 showed extremely low affinity for betaine aldehyde with apparent K_m of 18.9 μ M and 19.9 m M , respectively. In addition, V_max/K_m with betaine aldehyde of BBD2 was about 2000-fold higher than that of BBD1, suggesting that BBD2 plays a main role in glycine betaine synthesis in barley plants. However, BBD1 catalyzed the oxidation of ω-aminoaldehydes such as 4-aminobutyraldehyde and 3-aminopropionaldehyde as efficiently as BBD2. We also found that both BBDs oxidized 4- N -trimethylaminobutyraldehyde and 3- N -trimethylaminopropionaldehyde.     

Employment of Rhizobacteria for the Inoculation of Barley Plants Cultivated in Soil Contaminated with Lead and Cadmium

In laboratory experiments, the rhizobacteria Azospirillum lipoferum 137, Arthrobacter mysorens7, Agrobacterium radiobacter 10, and Flavobacterium sp. L30 were found to have a relatively high resistance to the toxic heavy metals lead and cadmium (except that strain L30 was found to be sensitive to Cd). When introduced by means of seed bacterization, the heavy metal–resistant strains actively colonized the rhizosphere of barley plants cultivated in uncontaminated and contaminated soils. In both pot and field experiments, seed bacterization improved the growth of barley plants and the uptake of nutrient elements from soil contaminated with Pb and Cd. The bacterization also prevented the accumulation of Pb and Cd in barley plants, thereby mitigating the toxic effect of these heavy metals on the plants.     

Capability of selected crop plants for shoot mercury accumulation from polluted soils: phytoremediation perspectives

High-biomass crops can be considered as an alternative to hyperaccumulator plants to phytoremediate soils contaminated by heavy metals. In order to assess their practical capability for the absorption and accumulation of Hg in shoots, barley, white lupine, lentil, and chickpea were tested in pot experiments using several growth substrates. In the first experimental series, plants were grown in a mixture of vermiculite and perlite spiked with 8.35 microg g(-1) d.w. of soluble Hg. The mercury concentration of the plants' aerial tissues ranged from 1.51 to 5.13 microg g(-1) d.w. with lentil and lupine showing the highest values. In a second experiment carried out using a Hg-polluted soil (32.16 microg g(-1) d.w.) collected from a historical mining area (Almadén, Spain), the crop plants tested only reached shoot Hg concentration up to 1.13 microg g(-1) d.w. In the third experimental series, the Almadén soil was spiked with 1 microg g(-1) d.w. of soluble Hg; as a result, mercury concentrations in the plant shoots increased approximately 6 times for lupine, 5 times for chickpea, and 3.5 times for barley and lentil, with respect to those obtained with the original soil without Hg added. This marked difference was attributed to the low availability of Hg in the original Almadin soil and its subsequent increase in the Hg-spiked soil. The low mercury accumulation yields obtained for all plants do not make a successful decontamination of the Almadén soils possible byphytoremediation using crop plants. However, since the crops tested can effectively decrease the plant-available Hg level in this soil, their use could, to some extent, reduce the environmental risk of Hg pollution in the area.     

Betaine accumulation in salt-stressed sorghum

Two analytical methods for measuring betaine were compared in a study of betaine accumulation in salt-stressed sorghum. Spectrophotometric determination of betaine as the p -bromophenacyl ester is highly sensitive and specific. However, a periodide assay was found to be more convenient for screening numerous plant samples without undue sacrifice in accuracy.
The accumulation of betaine in grain sorghum [ Sorghum bicolor (L.) Moench] cvs NK 265 and Double TX was measured in salt-stressed plants grown hydroponically and in the field and in drought-stressed potted plants. Neither drought nor mild salinity (-0.2 MPa) stress was effective in stimulating betaine accumulation. However, when the osmotic potential of the culture solution was lowered to -0.8 MPa, betaine levels in the shoots rose rapidly for 12 days after initiation of salination, and then declined, apparently because of dilution by plant growth. In young leaf blades, betaine was strongly accumulated up to 70–75 μmol (g dry weight)⁻¹; the concentration in leaf sheaths was less than 6 μmol (g dry weight)⁻¹.
In the field, betaine levels in salt-stressed sorghum increased 6- to 7-fold over the basal level of the control plants. In a comparable study of two wheat species ( Triticum aestivum L. cv. Probred and T. durum Desf. cv. 1000-D), betaine increased only 3- to 4-fold over unstressed plants.     

金毛狗对重金属的富集特性的研究

为探讨金毛狗[Cibotium barometz(L.) J. Sm.]对重金属的富集能力,在广东省选取6个样点(南岭、南昆山、白云山、大岭山、梧桐山、西樵山)采集金毛狗的叶片、根状茎和根际土壤,采用ICP-MS测定9种重金属元素(Cr、Mn、Ni、Cu、Zn、As、Cd、Hg、Pb)的含量。结果表明,样地土壤已受到不同程度的重金属污染,土壤中Cd和Hg含量均高于广东省土壤背景值,分别为背景值的1.61~4.82倍和4.74~11.79倍。西樵山土壤中Cd含量最大,南岭土壤中Hg含量最大。在9种元素中,金毛狗对Hg的转运系数最高,达4.8,对Cd的富集系数最高,达2.2,Cu和Cd元素的转运系数和富集系数均大于1。这说明金毛狗对重金属元素的富集能力较弱而转运能力较强。     

Optimum utilization of sewage sludge of low and high metal content for grain production on arid lands

Summary Two municipal sludges, one from a highly industrialized city, Chicago, Il, and another from a little industrialized, highly agricultural area, Tucson, AZ are compared for winter barley production on Pima c 1 (Typic torrifluvent). Both sludges were responsible for highly significant additions of Zn, Cu, Ni, Cd and P to the soil each year when applied at the rates of 100 mt/ha singly and 20 mt/ha each year for 4 years. Nitrogen responses for barley straw and grain were observed from both sludges. Tucson sludge appears to be attractive as a potential fertilizer, not only as an NPK source, but also for its organic matter and minimal amounts of heavy metals. The Chicago sludge with relatively high levels of heavy metals, particularly Cd, appears poorly suited as a fertilizer, if used for an extended period of time, because of the plant's tendency to take up elevated levels of certain heavy metals. Some parts of barley plants proved to be a better indicator of heavy metal uptake and concentration than others. The diagnostic-tissue test promises to be a useful tool to warn against undesirable accumulation of heavy metals. Fortunately, when compared with other plant parts, the heavy metal in grain was the least altered as a result of continued sewage sludge use on arid land. The soil's neutral to slightly alkaline pH and the presence of lime throughout the soil profile appeared to be critical factors in keeping plant uptake of heavy metals relatively low as compared with soils of other climates.     

金属矿区芒草种群对重金属的积累及其与土壤特性的关系

通过分析大型综合金属矿区中经历不同污染强度与污染时间胁迫的芒草（Miscanthus sinensis）种群对4种主要重金属的积累状况,初步揭示芒草对这些重金属的积累特性与土壤重金属含量的关系。结果表明,1芒草根茎叶对4种重金属的的积累顺序为：根〉叶〉茎;2芒草对Cd、Pb的积累量与土壤中这两种重金属含量之间存在显著（P〈0．05）正相关关系;对Cu、Zn的积累量与土壤含量之间无显著相关,主要是因为土壤最高Cu与Zn含量已超过芒草对这两种元素积累所需的最大量,成为对芒草构成胁迫的主要因子。在该矿区的酸性条件下,芒草对Pb、Zn、Cu3种重金属的吸收率随pH值升高而升高,pH接近的样地,芒草的吸收率主要受土壤重金属含量的影响。结合各种群对四种重金属的积累状况判断,强度胁迫下的种群可能已发生耐性分化,从而产生较其它种群更强的耐重金属特性。总体上芒草是一种多重金属耐性植物,对这四种重金属的耐性顺序是：Cd〈Cu〈Zn-Pb。     

Enhanced toxic metal accumulation in engineered bacterial cells expressing Arabidopsis thaliana phytochelatin synthase

Phytochelatins (PCs) are metal-binding cysteine-rich peptides, enzymatically synthesized in plants and yeasts from glutathione in response to heavy metal stress by PC synthase (EC 2.3.2.15). In an attempt to increase the ability of bacterial cells to accumulate heavy metals, the Arabidopsis thaliana gene encoding PC synthase (AtPCS) was expressed in Escherichia coli. A marked accumulation of PCs was observed in vivo together with a decrease in the glutathione cellular content. When bacterial cells expressing AtPCS were placed in the presence of heavy metals such as cadmium or the metalloid arsenic, cellular metal contents were increased 20- and 50-fold, respectively. We discuss the possibility of using genes of the PC biosynthetic pathway to design bacterial strains or higher plants with increased abilities to accumulate toxic metals, and also arsenic, for use in bioremediation and/or phytoremediation processes.     

Heavy metals accumulation and distribution in durum wheat and barley grown in contaminated soils under Mediterranean field conditions

Abstract

In the framework of a phytoremediation project in the Apulia region (Italy) a field experiment was carried out in multi-metal contaminated soils. The accumulation and distribution of metals in different plant parts of durum wheat and barley were studied. Further, the application of Bacillus licheniformis strain BLMB1 to soil was evaluated as a means to enhance metal accumulation in plants. The translocation and the bioconcentration factors indicated that wheat and barley do not act as metal accumulators in the field conditions tested, thus phytoextraction by these species would not be recommended as a soil remediation alternative. Application of B. licheniformis improved the accumulation of all metals in roots of wheat and barley, and increased Cd, Cr, and Pb contents in the shoots of barley. Low health risk for humans and animals was evaluated to exist if straw and grain from both cereal crops grown in these contaminated sites are consumed.     

Heavy metal accumulation in wheat and barley: The effects of soil presence and liquid manure amendment

An experiment was undertaken to evaluate the effect of liquid manure amendment on heavy metal accumulation in wheat and barley. For this purpose, both kinds of seedlings were grown simultaneously in a Petri dish, while wheat seedlings were also grown in pots containing unpolluted agricultural soil. All of the seedlings were irrigated with one of the three prepared solutions: artificial rainwater solution, heavy metal solution and liquid manure solution containing NH₄NO₃, H₃PO₄ and KOH along with equal amounts of heavy metals as in the second solution. Twenty days later, 1 g of plant tissue was digested with the mixture of HNO₃ and H₂O₂ for ICP-OES/HG-ICP-OES analysis. The results showed that the uptake of arsenic and mercury was highest for both plants grown in a Petri dish. Furthermore, the wheat grown in a Petri dish also had a high content of nickel, cadmium and copper, while the pot-grown wheat contained high amounts of iron and manganese, probably due to the adsorption of nickel, cadmium, copper and mercury on soil phases. The lower uptake of all heavy metals was observed after the amendment of liquid manure, with the exception of manganese in wheat and mercury in all plants.     

芜湖钢铁厂周边土壤及油菜籽中镉、铜、锌、铅含量和形态分布研究

研究了污染土壤、油菜籽中Cd、Cu、Zn、Pb含量、形态分布特征和重金属富集状况及可能存在的生物毒性.结果表明,土壤中Cd、Zn、Pb以铁锰氧化物结合态、Cu以残留态占5种形态最高比例,分别为31.1%、39.3%、53.79%、46.24%;Cd、Pb交换态比例较高,为23.47%、16.32%,Cu、Zn的交换态比例较小,为3.14%、0.54%;土壤中不同重金属与各重金属形态相关关系有差别,5种重金属形态转化为有效态重金属难易程度不同;油菜籽和油菜籽壳中不同重金属累积趋势有差异,Cu易在油菜籽壳中累积,Cd、Zn、Pb易在油菜籽中累积;油菜籽中不同重金属累积率不同,Cd累积率最高,为0.56.油菜籽中重金属累积率与土壤中重金属总量呈显著负相关关系(P＜0.05),土壤中重金属的形态、转化差异是此种负相关关系的主要原因;油菜籽中Cd、Cu、Pb以氯化钠态为主,分别为32.50%、22.94%、34.69%,Zn以EDTA态为主,为45.97%.油菜籽中重金属形态可能影响其毒性,但其毒性的人类膳食风险还需进一步研究证实.油菜籽中重金属形态与油菜中重金属总量相关性不好.     

Distribution of nutrients and phytotoxic metal ions in the soil and in two forest floor plant species of a beech (Fagus sylvatica L.) stand

In a beech (Fagus sylvatica L.) stand in north-west Germany vegetation of two transects (25m:1m and 20m:1m) was mapped and contents of macronutrients (Ca, Mg and K), micronutrients (Fe, Mn, Zn and Cu), and potentially phytotoxic metals (Pb, Cd, Ni and Al) were measured in different soil compartments and in roots, rhizomes, stems and leaves of two forest floor plant species (Mercurialis perennis L. and Polygonatum multiflorum L.). NH₄Cl extractable cation contents, pH and other soil variables were also determined.The highest macronutrient contents could be found in the leaves of M. perennis and P. multiflorum. Heavy metals and Al accumulated in the roots. Correlation analysis suggests a considerable translocation of Zn and Cd between below- and above-ground organs of both investigated forest floor plants. No significant correlation was found between the contents of the other elements in the below- and above-ground parts.Available data indicate a considerable uptake by the plants not only of nutrients, but also of heavy metals from the upper mineral soil. Amounts of heavy metals and Al solubilized in the presence of NH₄Cl increased with decreasing pH, whereas levels of soluble Ca and Mg were maximal at high pH-values of the extracts. It can be concluded that element uptake in the investigated plants is indirectly controlled by the pH of the upper mineral soil.     

Compatible solute accumulation and stress-mitigating effects in barley genotypes contrasting in their salt tolerance

The accumulation of compatible solutes is often regarded as a basic strategy for the protection and survival of plants under abiotic stress conditions, including both salinity and oxidative stress. In this work, a possible causal link between the ability of contrasting barley genotypes to accumulate/synthesize compatible solutes and their salinity stress tolerance was investigated. The impact of H(2)O(2) (one of the components of salt stress) on K(+) flux (a measure of stress 'severity') and the mitigating effects of glycine betaine and proline on NaCl-induced K(+) efflux were found to be significantly higher in salt-sensitive barley genotypes. At the same time, a 2-fold higher accumulation of leaf and root proline and leaf glycine betaine was found in salt-sensitive cultivars. The total amino acid content was also less affected by salinity in salt-tolerant cultivars. In these, potassium was found to be the main contributor to cytoplasmic osmolality, while in salt-sensitive genotypes, glycine betaine and proline contributed substantially to cell osmolality, compensating for reduced cytosolic K(+). Significant negative correlations (r= -0.89 and -0.94) were observed between Na(+)-induced K(+) efflux (an indicator of salt tolerance) and leaf glycine betaine and proline. These results indicate that hyperaccumulation of known major compatible solutes in barley does not appear to play a major role in salt-tolerance, but rather, may be a symptom of salt-susceptibility.     

锰胁迫和性别竞争交互处理下沙棘雌雄幼苗生理响应特征

沙棘(Hippophae rhamnoides)是重要的雌雄异株人工林防护树种,但对其环境胁迫的性别响应差异研究不足,性别竞争与胁迫因子的交互效应响应特征尚不清楚。为了探讨锰胁迫和性别竞争交互处理下沙棘雌雄植株的生理响应特征和耐受能力,旨在为沙棘修复土壤重金属污染提供实践指导,该文研究了锰胁迫(4 000 mg·kg^-1)和3种不同性别组合模式(雌雄、雌雌、雄雄)处理下沙棘的生理响应,分别测定雌雄沙棘叶片中叶绿素、过氧化物酶(peroxidase,POD)、超氧化物歧化酶(superoxide dismutase,SOD)、丙二醛(malondialdehyde,MDA)、总酚(total phenols, TP)、游离脯氨酸(free proline,Pro)、可溶性糖(soluble sugar,SS)、甜菜碱(glycine betaine, GB)以及锰含量。结果表明:(1)锰胁迫下,在所有竞争组合中,性间竞争的雄株(M/FM)SOD活性最高,而MDA含量与对照相比未有明显升高,表明雄株的抗氧化能力更好,膜氧化损伤程度更小。(2)锰胁迫时M/FM积累了更多的游离脯氨酸,表现出更好的渗透调节能力和耐受能力。(3)交互效应分析显示性别互作和锰胁迫交互处理显著影响了沙棘雌雄叶片的光合色素、抗氧化酶活性和渗透调节能力; 主成分分析显示SOD、POD、MDA、叶绿素b(chlorophyll b, Chlb)、SS、Pro可作为重要的生理响应指示参数。该研究结果对于利用沙棘修复土壤重金属污染可提供一定的参考。     

设施菜地重金属的剖面分布特征

以山东省寿光市种植1～12年的蔬菜大棚及相邻的小麦地为对象,研究了设施菜地中As、Cd、Cu、Zn、Cr、Ni 6种重金属在土壤剖面中的分布特征.结果表明：在0～150 cm土层内,随着土层深度的增加,设施菜地与小麦地中6种重金属含量均呈下降趋势;与小麦地相比,相同土层设施菜地中6种重金属的含量均明显较高,设施菜地0～20 cm土层中As、Cd、Cu、Zn、Cr、Ni的含量分别比小麦地相同土层高出35.0%、146.2%、65.6%、36.4%、21.5%和14.0%,120～150 cm土层分别高出10.6%、.5%、19.4%、20.2%、15.2%和9.3%,20～120 cm各土层中的含量亦高于小麦地.设施菜地0～20 cm土层中,Cd、Cu、Zn含量与种植年限呈显著正相关,随着种植年限的增加,其累积速率分别为0.027、1.153和2.830 mg·kg^-1·a^-1;设施菜地中6种重金属的含量与土壤有机碳、全氮含量呈正相关,除Ni和As外的4种重金属含量亦与土壤全磷含量呈极显著相关,有机肥等的施用可能在一定程度上导致设施菜地中重金属的累积.     

攀钢冶炼渣堆土壤与优势植物的重金属含量

采用原子吸收分光光度法测定攀钢西渣场冶炼渣堆土壤和6科12种优势植物中Mn、Pb、 Ni、 Cu、Cd等5种重金属含量,并计算优势植物对重金属的富积系数和转移系数.结果表明:渣堆土壤中重金属含量Mn最高(3869.14 mg/kg),次后顺序为Pb>Ni>Cu>Cd;植物与土壤的重金属分布基本一致;所测优势植物中,多数植物对重金属的富积系数较低,而转移系数却较高,如天名精对Cu的转移系数为5.1,羽芒菊对Pb转移系数为3.3,五月艾对Cd的转移系数为6.0,其中8种植物(天名精、羽芒菊等)对Mn的转移系数均大于1.该结果为重金属污染土壤的植物修复提供了参考物种,同时也为植物重金属耐受机制的研究提供了筛选对象.     

Enhanced Toxic Metal Accumulation in Engineered Bacterial Cells Expressing Arabidopsis thaliana Phytochelatin Synthase

Phytochelatins (PCs) are metal-binding cysteine-rich peptides, enzymatically synthesized in plants and yeasts from glutathione in response to heavy metal stress by PC synthase (EC 2.3.2.15). In an attempt to increase the ability of bacterial cells to accumulate heavy metals, the Arabidopsis thaliana gene encoding PC synthase (AtPCS) was expressed in Escherichia coli. A marked accumulation of PCs was observed in vivo together with a decrease in the glutathione cellular content. When bacterial cells expressing AtPCS were placed in the presence of heavy metals such as cadmium or the metalloid arsenic, cellular metal contents were increased 20- and 50-fold, respectively. We discuss the possibility of using genes of the PC biosynthetic pathway to design bacterial strains or higher plants with increased abilities to accumulate toxic metals, and also arsenic, for use in bioremediation and/or phytoremediation processes.