Lhcb2 gene expression analysis in two ecotypes of Sedum alfredii subjected to Zn/Cd treatments with functional analysis of SaLhcb2 isolated from a Zn/Cd hyperaccumulator

The Lhcb2 gene from hyperaccumulator Sedum alfredii was up-regulated more than three-fold while the non-hyperaccumulator accumulated one or two-fold higher amount of the mRNA than control plants under different concentrations of Cd²⁺ for 24 h. Lhcb2 expression was up-regulated more than five-fold in a non-hyperaccumulator S. alfredii when exposed to 2 μM Cd²⁺ or 50 μM Zn²⁺ for 8 d and the hyperaccumulator had over two-fold more mRNA abundance than the control plants. Over-expression of SaLhcb2 increased the shoot biomass by 14–41% and the root biomass by 21–57% without Cd²⁺ treatment. Four transgenic tobacco lines (L5, L7, L10 and L11) possessed higher shoot biomass than WT plants with Cd²⁺. Four transgenic lines (L7, L8, L10 and L11) accumulated 6–35% higher Cd²⁺ amounts in shoots than the wild type plants.     

Root adaptations to cadmium-induced oxidative stress contribute to Cd tolerance in the hyperaccumulator Sedum alfredii

Short-term responses of Sedum alfredii roots to Cd exposure was compared in Cd hyperaccumulator (HE) and nonhyperaccumulating ecotype (NHE). Cadmium exposure significantly inhibited root elongation and induced loss of plasma membrane integrity and lipid peroxidation of roots tips in the NHE, whereas these effects were much less pronounced in the HE plants. A strong accumulation of reactive oxygen species with increasing Cd concentration was noted in the NHE root tips, but not in HE. After Cd exposure, a dose-dependent decrease in oxidized glutathione and marked increase in reduced glutathione and non-protein thiols were observed in root tips of HE, but were not seen in the NHE plants. These results suggest that the HE tolerates high Cd in the environment through the differential adaptations against Cd-induced oxidative stress.     

Dynamics of zinc uptake and accumulation in the hyperaccumulating and non-hyperaccumulating ecotypes of Sedum alfredii Hance

Sedum alfredii Hance has been identified as a Zn-hyperaccumulating plant species native to China. The characteristics of Zn uptake and accumulation in the hyperaccumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) of S. alfredii were investigated under nutrient solution and soil culture conditions. The growth of HE was normal up to 1000 μM Zn in nutrient solution, and 1600 mg Zn kg⁻¹ soil in a Zn-amended soil. Growth of the NHE was inhibited at Zn levels ≥250 μM in nutrient solution. Zinc concentrations in the leaves and stems increased with increasing Zn supply levels, peaking at 500 and 250 μM Zn in nutrient solution for the HE and the NHE, respectively, and then gradually decreased or leveled off with further increase in solution Zn. Minimal increases in root Zn were noted at Zn levels up to 50 μM; root Zn sharply increased at higher Zn supply. The maximum Zn concentration in the shoots of the HE reached 20,000 and 29,000 mg kg⁻¹ in the nutrient solution and soil experiments, respectively, approximately 20 times greater than those of the NHE. Root Zn concentrations were higher in the NHE than in the HE when plants were grown at Zn levels ≥50 μM. The time-course of Zn uptake and accumulation exhibited a hyperbolic saturation curve: a rapid linear increase during the first 6 days in the long-term and 60 min in the short-term studies; followed by a slower increase or leveling off with time. More than 80% of Zn accumulated in the shoots of the HE at half time (day 16) of the long-term uptake in 500 μM Zn, and also at half time (120 min) of the short-term uptake in 10 μM ⁶⁵Zn²⁺. These results indicate that Zn uptake and accumulation in the shoots of S. alfredii exhibited a down-regulation by internal Zn accumulated in roots or leaves under both nutrient solution and soil conditions. An altered Zn transport system and increased metal sequestration capacity in the shoot tissues, especially in the stems, may be the factors that allow increased Zn accumulation in the hyperaccumulating ecotype of S. alfredii. Section Editor: F. J. Zhao     

外源钙离子对东南景天生长及锌积累的影响

采用水培试验,研究了外源添加不同浓度钙离子（Ca²⁺）对两种生态型东南景天生物量、根系形态及体内锌、钙、硫含量的影响.结果表明：随着外源Ca²⁺浓度的上升,两种生态型东南景天的干物质量均增加,且超积累生态型地上部增加显著(P<0.05);超积累生态型根长和根表面积增加,而非超积累生态型降低;超积累生态型根、茎、叶锌含量随着外源Ca²⁺浓度的增加而上升,但各处理间差异不显著(P>0.05),非超积累生态型地上部锌含量显著降低(P<0.05).非超积累生态型根、茎、叶钙含量与外源Ca²⁺浓度呈显著正相关(P<0.05),超积累生态型根系硫含量与外源Ca²⁺浓度呈极显著正相关(P<0.01).外源Ca²⁺对超积累生态型东南景天锌吸收及积累有促进作用,而Ca²⁺浓度的升高抑制了非超积累生态型东南景天对锌的吸收.适当增加外源Ca²⁺可促进超积累生态型东南景天生长,改善其锌积累能力.     

Lead induced changes in the growth and antioxidant metabolism of the lead accumulating and non-accumulating ecotypes of Sedum alfredii

The phytotoxicity and antioxidative adaptations of lead （Pb） accumulating ecotype （AE） and non-accumulating ecotype （NAE） of Sedum alfredii Hance were investigated under different Pb treatments involving 0, 0.02 mmol/L Pb, 0.1 mmol/L Pb and 0.1 mmol/L Pb/0.1 mmol/L ethylenediaminetetraacetic acid （EDTA） for 6days. With the increasing Pb level, the Pb concentration in the shoots of AE plants enhanced accordingly, and EDTA supply helped 51% of Pb translocation to shoots of AE compared with those treated with 0.1 mmol/L Pb alone. Moreover, the presence of EDTA alleviated Pb phytotoxicity through changes in plant biomass, root morphology and chlorophyll contents. Lead toxicity induced hydrogen peroxide （H2O2） accumulation and lipid peroxidation in both ecotypes of S. alfredii. The activities of superoxide dismutase （SOD）, guaiacol peroxidase （G-POD）, ascorbate peroxidase, and dehydroascorbate reductase elevated in both leaves and roots of AE as well as in leaves of NAE with the increasing Pb levels, but SOD and G-POD declined in roots of NAE. Enhancement in glutathione reductase activity was only detected in roots of NAE while a depression in catalase activity was recorded in the leaves of NAE. A significant enhancement in glutathione and ascorbic acid （AsA） levels occurred in both ecotypes exposed to Pb and Pb/EDTA treatment compared with the control, however, the differences between these two treatments were insignificant. The dehydroascorbate （DHA） contents in roots of both ecotypes were 1.41 to 11.22-fold higher than those in leaves, whereas the ratios of AsA to DHA （1.38 to 6.84） in leaves altering more to the reduced AsA form were much higher than those in roots. These results suggested that antioxidative enzymes and antioxidants play an important role in counteracting Pb stress in S. alfredii.     

Structural and functional variability in root-associated bacterial microbiomes of Cd/Zn hyperaccumulator Sedum alfredii

Applied Microbiology and Biotechnology - Interactions between roots and microbes affect plant’s resistance to abiotic stress. However, the structural and functional variation of...     

Root foraging for zinc and cadmium requirement in the Zn/Cd hyperaccumulator plant Sedum alfredii

Positive root response to metals may enhance metal accumulation for greater requirement in hyperaccumulators. The effects of spatially heterogeneous Zn/Cd addition on root allocation, metal accumulation, and growth of the Zn/Cd hyperaccumulator Sedum alfredii were assessed in a pot experiment. Young shoots of S. alfredii were grown with or without supplied Zn/Cd. Two concentrations were used of each metal, and each metal concentration had one homogeneous and two heterogeneous treatments. Growth increased by 1.6–3.2 times with the increasing overall dose of Zn/Cd addition, and shoot biomass was positively correlated with shoot Zn/Cd concentration (P?<?0.001). In all heterogeneous treatments, the plants consistently allocated approximately 90% of root biomass to the metal-enriched patches, and shoot Zn/Cd contents were greater than or similar to those in the homogeneous treatment at each metal concentration. Plants in the control treatment showed symptoms of Zn deficiency, although their shoots had Zn concentrations 100-fold higher than the critical deficiency value for most plants. We conclude that S. alfredii has evolved root foraging mechanisms associated with its greater requirements for Zn/Cd. These results could have important implications both for phytoremediation and for investigation of positive role of Cd in higher plants.     

东南景天对锌、镉复合污染的反应及其对锌、镉的吸收和积累特性

Sedum alfredii Hance has been identified as a new Zn-hyperaccumulator native to China. In this study, responses and metal accumulation of S. alfredii were examined under Zn/Cd complex polluted conditions. The results showed that optimal growth of S. alfredii in terms of the maximum dry matter yield was observed at Zn/Cd complex level of 500/100 祄ol/L. Plant cadmium (Cd) or zinc (Zn) concentrations increased with increasing Cd or Zn supply. During the 20 d treatment, the highest Cd concentration in the leaves reached 12.1 g/kg at Zn /Cd level of 50/400 祄ol/L and that of Zn in the stems was 23.2 g/kg at Zn/Cd level of 1 000/50 祄ol/L. The distribution of Cd in different plant parts decreased in the order: leaf ＞ stem≥ root, whereas that of Zn was: stem ＞ leaf ≥ root. The accumulation of Cd and Zn in the shoots and roots of S. afredii increased with the increasing of Zn/Cd supply levels, peaked at Zn/Cd levels of 250/400 and 500/100 祄ol/L, respectively. The highest Cd and Zn uptake by the shoots was approximately 5 and 11 mg/plant, and was over 20 and 10 times higher than those in the roots, respectively. Zn supply at levels ≤ 500 祄ol/L increased plant Cd concentrations, whereas high Zn supply decreased root Cd but did not affect leaf Cd concentrations in S. alfredii. Low Cd supply increased Zn concentration in the leaves, but Cd supply higher than 50 祄ol/L considerably reduced root Zn concentrations, especially at low Zn level. These results indicate that S. alfredii can tolerate high Zn/Cd complex levels and has an extraordinary ability to hyperaccumulate not only Zn but also Cd. It could provide a new valuable plant material for understanding the mechanisms responsible for co-hyperaccumulation of Zn and Cd as well as for phytoremediation of the Cd/Zn complex polluted soils.     

Cadmium tolerance and hyperaccumulation in a new Zn-hyperaccumulating plant species (Sedum alfredii Hance)

Plant and Soil - Sedum alfredii Hance has been identified as a new zinc (Zn) hyperaccumulating plant species. In this study, the effects of cadmium (Cd) supply levels (control, 12.5, 25, 50, 100,...     

东南景天对锌、镉复合污染的反应及其对锌、镉的吸收和积累特性

东南景天(Sedum alfredii Hance)已被鉴定为一种中国原生的新的锌超积累植物。 本文主要研究了锌(Zn)、镉(Cd)复合处理水平对东南景天的生长及其对锌、镉的吸收积累特性的影响。 结果表明,在Zn/Cd复合水平为500/100 祄ol/L时,植物生长最佳。Zn/Cd在东南景天叶片、茎部和根系中含量随着Zn/Cd处理水平的提高而增高。在Zn/Cd 复合水平为50/400 祄ol/L时茎叶中Cd含量达最高,其中叶片Cd含量达12.1 g/kg;在Zn/Cd 复合水平为1 000/50 祄ol/L时茎叶中Zn含量达最高, 其中茎中Zn含量达 23.2 g/kg。 植株各部位Cd含量的分布为:叶片>茎>>根系,而Zn在体内的分布为: 茎>叶片>>根系。 Zn、Cd在地上部和根部的积累量也随着处理水平的提高而增加,分别在Zn/Cd复合水平为250/400和500/100 祄ol/L下达高峰值。 东南景天地上部积累最高Zn和Cd的量分别达11和5 g/plant, 其比根系的积累量分别大10和25余倍。 Zn、Cd对东南景天的生长、吸收积累的相互作用依赖于Zn/Cd复合水平和植物部位。 在适宜Zn/Cd 处理水平范围内,Zn和Cd的吸收和积累具有相互促进作用。 高Zn或高Cd处理均抑制了植物对Zn和Cd的吸收和积累。 本研究结果表明,东南景天不仅具有忍耐高Zn/Cd复合污染,而且具有超量积累Zn和Cd的特异能力。 它为进一步研究植物Zn、Cd     

Effects of bacteria on enhanced metal uptake of the Cd/Zn-hyperaccumulating plant, Sedum alfredii

To investigate the effects of bacteria (Burkholderia cepacia) on metal uptake by the hyperaccumulating plant, Sedum alfredii, a hydroponic experiment with different concentrations of Cd and Zn was conducted. It was found that inoculation of bacteria on S. alfredii significantly enhanced plant growth (up to 110% with Zn treatment), P (up to 56.1% with Cd treatment), and metal uptake (up to 243% and 96.3% with Cd and Zn treatment, respectively) in shoots, tolerance index (up to 134% with Zn added treatment), and better translocation of metals (up to 296% and 135% with Cd and Zn treatment, respectively) from root to shoot. In the ampicillin added treatment with metal addition, stimulation of organic acid production (up to an increase of 133% of tartaric acid with Cd treatment) by roots of S. alfredii was observed. The secretion of organic acids appears to be a functional metal resistance mechanism that chelates the metal ions extracellularly, reducing their uptake and subsequent impacts on root physiological processes.     

Quantitative proteomics analysis of leaves from two Sedum alfredii (Crassulaceae) populations that differ in cadmium accumulation

Due to their extraordinary capacity to hypertolerate and hyperaccumulate heavy metals in above‐ground tissues, hyperaccumulator plant species have gained wide attention from researchers seeking biotechnologies to manage environmental heavy metal pollution. However, the molecular basis of hyperaccumulation is still far from being fully understood. Here, we used iTRAQ to perform a quantitative proteomics study of the leaves of Sedum alfredii (Crassulaceae) from hyperaccumulating population (HP) and non‐hyperaccumulating population (NHP). A total of 248 proteins had constitutively higher levels in HP leaves than in NHP leaves. Cadmium (Cd) treatment led to the induction of 13 proteins in HP leaves and 33 proteins in NHP leaves. Two proteins were induced by Cd in both HP leaves and NHP leaves. The annotations for many of the proteins that were higher in HP leaves and proteins that were induced by Cd treatments were associated with vacuolar sequestration, cell wall/membrane modification, and plant defense. In addition to establishing a global empirical foundation for the study of proteins in S. alfredii, our findings relating to the differential constitutive and inducible expression of proteins open potential new research avenues and bolster previously reported suppositions about Cd hyperaccumulation in hyperaccumulator plants.     

Enhanced Cd extraction of oilseed rape (Brassica napus) by plant growth-promoting bacteria isolated from Cd hyperaccumulator Sedum alfredii Hance

Four plant growth-promoting bacteria (PGPB) were used as study materials, among them two heavy metal-tolerant rhizosphere strains SrN1 (Arthrobacter sp.) and SrN9 (Bacillus altitudinis) were isolated from rhizosphere soil, while two endophytic strains SaN1 (Bacillus megaterium) and SaMR12 (Sphingomonas) were identified from roots of the cadmium (Cd)/zinc (Zn) hyperaccumulator Sedum alfredii Hance. A pot experiment was carried out to investigate the effects of these PGPB on plant growth and Cd accumulation of oilseed rape (Brassica napus) plants grown on aged Cd-spiked soil. The results showed that the four PGPB significantly boosted oilseed rape shoot biomass production, improved soil and plant analyzer development (SPAD) value, enhanced Cd uptake of plant and Cd translocation to the leaves. By fluorescent in situ hybridization (FISH) and green fluorescent protein (GFP), we demonstrated the studied S. alfredii endophytic bacterium SaMR12 were able to colonize successfully in the B. napus roots. However, all four PGPB could increase seed Cd accumulation. Due to its potential to enhance Cd uptake by the plant and to restrict Cd accumulation in the seeds, SaMR12 was selected as the most promising microbial partner of B. napus when setting up a plant–microbe fortified remediation system.     

镉在东南景天中的亚细胞分配

采用差速离心技术,比较研究了Cd在矿山生态型东南景天(Sedum alfredii Hance)根、茎、叶中的亚细胞分布。结果表明:矿山生态型东南景天对Cd具有很好的忍受耐和累积能力,非矿山生态型东南景天则不具有这种能力。Cd在矿山生态型东南景天根、茎、叶各部分的亚细胞分配满足F1(细胞壁部分)>F3(可溶部分)>F2(细胞器及膜)规律,且Cd在细胞壁部分的分配占绝对优势;同时矿山生态型东南景天地上部有很好的Cd累积能力。与众多超累积植物相似的Cd亚细胞分配规律和地上部的良好的Cd累积能力。对此植物的Cd亚细胞分布完全不同于非矿山生态型东南景天的情况作了比较分析,结果显示矿山生态型东南景天很可能是一种新的镉超累积种质资源。     

镉在东南景天中的亚细胞分配

采用差速离心技术,比较研究了Cd在矿山生态型东南景天(Sedum alfredii Hance)根、茎、叶中的亚细胞分布.结果表明:矿山生态型东南景天对Cd具有很好的忍受耐和累积能力,非矿山生态型东南景天则不具有这种能力.Cd在矿山生态型东南景天根、茎、叶各部分的亚细胞分配满足F1(细胞壁部分)>F3(可溶部分)>F2(细胞器及膜)规律,且Cd在细胞壁部分的分配占绝对优势;同时矿山生态型东南景天地上部有很好的Cd累积能力.与众多超累积植物相似的Cd亚细胞分配规律和地上部的良好的Cd累积能力.对此植物的Cd亚细胞分布完全不同于非矿山生态型东南景天的情况作了比较分析,结果显示矿山生态型东南景天很可能是一种新的镉超累积种质资源.