Feasibility of enhanced phytoextraction of Zn contaminated soil with Zn mobilizing and plant growth promoting endophytic bacteria

Three bacterial endophytes of Sedum alfredii, VI8L2, II8L4 and VI8R2, were examined for promoting soil Zn bioavailability and Zn accumulation in S. alfredii. Results showed that three strains were re-introduced into S. alfredii rhizosphere soils under Zn stress and resulted in better plant growth, as roots biomass increased from 80% to 525% and shoot biomass from 11% to 47% compared with the uninoculated ones. Strains IVsLz, II8L4 and IVsR2 significantly increased shoot and root Zn concentrations in the ZnCO3 contaminated soil. Inoculation with strain IVsL2 resulted in 44% and 39% higher shoot and root Zn concentrations, while strain IV8R2 significantly decreased shoot Zn concentration in the Zn3（PO4）2 contaminated soils. In the aged contaminated soil, isolates IVsL2, IIsL4 and IVsR2 significantly increased root Zn concentration, but decreased shoot Zn concentration of Sedum alfredii. It suggested that endophytes might be used for enhancing phytoextraction efficiency.

Growth Response and Metal Accumulation of Sedum alfredii to Cd/Zn Complex-Polluted Ion Levels

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 mumol/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 mumol/L and that of Zn in the stems was 23.2 g/kg at Zn/Cd level of 1000/50 mumol/L. The distribution of Cd in different plant parts decreased in the order: leaf > stem greater than or equal to root, whereas that of Zn was: stem > leaf greater than or equal to root. The accumulation of Cd and Zn in the shoots and roots of S. alfredii increased with the increasing of Zn/Cd supply levels, peaked at Zn/Cd levels of 250/400 and 500/100 mumol/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 less than or equal to 500 mumol/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 mumol/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.

Differences of Uptake and Accumulation of Zinc in Four Species of Sedum

Four species of Sedum L. were grown in nutrient solution for the comparison of their Zn uptake and accumulation. S. alfredii Hance showed much greater tolerance to Zn than the other three species. Shoot and root yields of S. sarmentosum Bunge, S. bulbiferum Makino, and S. emarginatum Migo decreased with increasing Zn concentration in the solution, while shoot and root yields of S. alfredii increased when Zn concentration was ≤80 mg·L -1. At 80 mg·L -1 Zn, Zn concentration in shoots of S. alfredii reached 19.09 mg·g -1. S. alfredii was also more efficient in Zn translocation from roots to shoots, while Zn concentration in shoots was much higher than in roots. However, this was not the case for the other three species. The results showed that S. alfredii is a Zn hyperaccumulator and could be useful for the phytoremediation of Zn-contaminated soils.

Sedum alfredii : A New Lead-Accumulating Ecotype

hi a survey of plant population, Sedum alfredii Hance, a new lead (Pb)-tolerant and lead (Pb)accumulating ecotype, was found in an old Pb/Zn mining area in Zhejiang Province of China. The growth and Pb content of plant ecotypes being able to and unable to accumulate Pb were studied by hydroponic culture with different concentrations of Pb(NO3)(2). Growth of shoots of accumulating ecotype was not affected by Ph treatments up to 320 mg/L, whereas that of non-accumulating ecotype was inhibited in all Ph treatments. The Ph concentrations in the roots and shoots of accumulating ecotype increased with increasing of Pb level in the nutrient solution. The maximum Ph concentrations in the shoots and roots of accumulating ecotype were 514 mg/kg and 13 922 mg/kg, 2.27 times and 2.62 times as much as that of non-accumulating ecotype, respectively. The highest rate of Pb accumulation of accumulating ecotype was 8.62 mug/plant/d, 7.16 times as much as that of non-accumulating ecotype. Due to its fast growth rate and high Pb-accumulating ability, from a phytoremediation perspective, accumulating ecotype of S. alfredii is a potential plant species for Pb removal from contaminated soils.

Subcellular Distribution of Cadmium in Mining Ecotype Sedum alfredii

The mining ecotype Sedum alfredii Hance could tolerate and grow normally in a nutritive solution containing cadmium (Cd) as high as 400 mumol/L. Under such a high Cd concentration, the subcellular accumulation of Cd in root, stem and leaf of this plant was found to be the highest in the cell wall, less in the soluble fraction and lowest in the cell organs. The mode of subcellular distribution of Cd in the mining ecotype S. alfredii was similar to other hyper accumulators of heavy metals, in which Cd was distributed more in the aerial part of plant. The results suggest that the mining ecotype S. alfredii is a new species of Cd hyperaccumulator.

Photooxidation in Leaves of Facultative CAM Plant Sedum spectabile at C_3 and CAM Mode

The switch from C-3 to CAM pathway was induced by water stress in a C-3/CAM intermediate plant Sedum spectabile Boreau. Typical CAM criteria were observed upon 15 d of withholding water. Leaf delta(13)C value (-%) and water content showed a linear correlation fashion. Chlorophyll fluorescence parameters and antioxidative capacity were altered by water stress. Phi(PSII) and q(P) were reduced by 50% and 34% of the control, respectively, while NPQ rose ca. 180%. SOD activity and ability to scavenge DPPH. free radical went down but membrane permeability changed slightly. However, when an additional photooxidation by MV with high PPFD was carried out with leaf discs from watered (C-3 mode) and drought plants (CAM mode), q(P) and Phi(PSII) in leaves at induced CAM mode stage continuously decreased to a very low level. High 1 - q(P) value (0.86) and 1 - q(P)/NPQ ratio (>1) indicated the presence of high reduction state and unbalance of light energy budget. Together with the marked loss of membrane integral, it was evidenced that photooxidative damage was more serious in the induced CAM mode than in the C-3 mode. No advantage of photooxidation tolerance was found at the induced CAM expression stage of the facultative CAM plant, as compared with its C-3 mode stage, and also with the constitutive CAM plants reported previously. The differences in photooxidation sensitivity between the inducible CAM expressing plant and the constitutive CAM plant were discussed.

铅胁迫对景天属和花生属植物生理特性和根系分泌物的影响

在水培条件下,用1000mg/L铅(Pb)处理景天属(Sedum)的野生种植物紫叶景天(Sedum telephium)和栽培种植物佛甲草(Sedum lineare),花生属(Arachis)的野生种植物蔓花生(Arachis duranensis)和栽培种植物花生(Arachis hypogaea)后发现,景天属植物佛甲草和花生属植物蔓花生、花生等叶片相对电导率、MDA含量都明显升高,叶绿素含量、根系活力不同程度降低,生物量积累明显下降;来自非洲草原的紫叶景天基本上不受Pb胁迫的影响。Pb处理下景天属和花生属植物根系分泌的总有机酸和可溶性糖含量均有所增加,且表现出明显的种间差异。在Pb胁迫下,花生属植物的有机酸分泌强度要明显高于景天属植物,野生种类的分泌强度要明显超过栽培种类。景天属植物根系分泌的有机酸主要种类为草酸、酒石酸、苹果酸和柠檬酸;花生属植物根系分泌的有机酸主要种类为柠檬酸、草酸和琥珀酸。

金叶景天扦插繁殖技术

近年来,彩叶地被植物被广泛应用于园林景观绿化中,金叶景天更是以其自身的优良特性,让园林工作者青睐有加。以金叶景天茎段为扦插材料,研究不同基质和激素浓度对金叶景天生根率、成活率等的影响。结果表明,以河沙∶蛭石∶珍珠岩=1∶1∶1混合的基质植株生长最旺盛,成活率最高;在此基础上,用50 mg/L IBA处理的茎段长势最好,用100 mg/L NAA处理的茎段长势次之。

松塔景天组织培养与快速繁殖的研究

以松塔景天幼嫩叶片作为外植体,利用含有不同浓度配比植物生长调节剂的培养基分别进行愈伤组织的诱导、不定芽的诱导、生根培养及驯化移栽,建立了松塔景天组织培养和快速繁殖体系。试验结果表明:诱导愈伤组织的最适培养基为MS+6-BA 0.5mg/L+NAA 0.05mg/L,愈伤组织诱导率达87.78%;诱导不定芽的最适培养基为MS+6-BA 1.0mg/L+NAA0.1mg/L,不定芽诱导率达88.89%;组培苗最佳生根培养基为1/2MS+IBA 0.3mg/L,生根率达91.33%;组培苗的移栽成活率达84%。

IBA处理及不同配方营养液对水培景天三七生长的影响

采用水培法,以景天三七(Sedum aizoon L.)为试验材料,研究不同浓度吲哚丁酸(IBA)处理及不同营养液配方和浓度对景天三七插穗生根、新叶数、植株增高的影响。结果表明,用IBA 150 mg/L浓度处理景天三七插穗后,再置于1/2山崎配方营养液中培养最有利于水培景天三七的生长,这为景天三七水培的发展和推广提供了理论依据。

景天三七根、茎、花、叶中总黄酮含量测定

景天三七(Sedum aizoon L.)根、茎、花、叶样品经粉碎后以甲醇为溶剂进行索氏提取,以芦丁为对照品,NaNO_(2)-A1(NO_(3))_(3)-NaOH作显色剂,在优选的最大吸收波长365 nm处检测,建立总黄酮测定的紫外分光光度法并对其含量进行测定。结果表明,在浓度8~40μg/g吸光度线性关系良好,回归方程为y=0.0175x+0.04(R^(2)=0.9999),景天三七不同部位低、中、高3个浓度芦丁添加回收率在92%~94%,RSD≤5%;景天三七根、茎、花、叶中总黄酮含量分别为(29.44±1.37)、(27.96±0.87)、(9.79±0.36)和(16.16±0.50)mg/g。研究建立了一种简单、准确的景天三七不同部位总黄酮的测定方法,为景天三七不同部位总黄酮检测及有针对性地开发利用提供科学依据。

Zn Accumulation and Subcellular Distribution in the Zn Hyperaccumulator Sedurn alfredii Hance

Zn accumulation and subcellular distribution in leaves of the hyperaccumulating ecotype （HE） and non-hyperaccumulating ecotype （NHE） of Sedum alfredii Hance were studied using radiotracer and gradient centrifugation techniques. Leaf Zn accumulation in the HE of S. alfredii was 18.5-26.7 times greater than that in the NHE when the plants were grown at 1-500μmol Zn L-1. Leaf section uptake of 65Zn was highly dependent on external Zn levels. Greater 65Zn uptake in HE was noted only at external Zn levels 〉 100μmol L-1. Zinc subcellular distribution in the leaves of the two ecotypes of S. alfredii was： cell wall 〉 soluble fraction 〉 cell organelle. However, more Zn was distributed to the leaf cell wall and soluble fractions for HE than for NHE. In the leaf of HE, 91%-94% of the Zn was found in the cell walls and the soluble fraction and only 6%-9% Zn was distributed in the cell organelle fraction. For NHE, about 20%-26% Zn was recovered in the cell organelle fraction. In stems, Zn distribution to the ceil wail fraction was approximately two fold greater in the HE than that in the NHE. For the hyperaccumulating ecotype of S. alfredii, the cell wall and the vacuole played a very important role in Zn tolerance and hyperaccumulation.

Selection of appropriate organic additives for enhancing Zn and Cd phytoextraction by hyperaccumulators

Chelant-enhanced phytoextraction is one of the most promising technologies to remove heavy metals from soil. The key of the technology is to choose suitable additives in combination with a suitable plant. In the present study, laboratory batch experiment of metal solubilization, cress seeds germination were undertaken to investigate the metal-mobilizing capability and the phytotoxicity of organic additives, including ethylene diamine triacetic acid （EDTA）, citric acid, acetic acid, oxalic acid, glutamine and monosodium glutamate waste liquid （MGWL） from food industry. Experiments in pots were carried out to study the effects of the additives on Zn and Cd phytoextraction. Furthermore, a leaching experiment with lysimeter was performed to evaluate the environmental risks of additive-induced leaching to underground water. The results showed that EDTA had a strong mobilizing ability for Zn and Cd, followed by mixed reagent （MR） and MGWL. MGWL and acetic acid at 5 mmol equivalent per liter resulted in seed germination index less than 2%. Experiments in pots verified the phytotoxicity of acetic acid and MGWL. Addition of the mixed reagent at 6--10 mmol/kg significantly increased Zn phytoextraction by Thlaspi caerulescens. The same for EDTA and the mixed reagent at 10 mmol/kg by Sedum dfredii. But only mixed reagents could significantly increase Cd phytoextraction by the studied hyperaccumulators. This suggested that the strong chelant was not always the good agent to enhance phytoextraction. S. alfredii combined with 2--10 mmol/kg soil MR was preferred for phytoremediation of Cd/Zn contaminated soils in southern China, this could result in high phytoextraction of Cd/Zn and reduce the leaching risk to underground water than EDTA assisted phytoextration.

Zinc adsorption and desorption characteristics in root cell wall involving zinc hyperaccumulation in Sedum alfredii Hance

Radiotracer techniques were employed to characterize 65Zn adsorption and desorption in root-cell-wall of hyperac-cumulating ecotype (HE) and non-hyperaccumulating ecotype (NHE) species of Sedum alfredii Hance. The results indicated that at the end of a 30 min short time radioisotope loading period, comparable amounts of 65Zn were accumulated in the roots of the two ecotypes Sedum alfredii, whereas 2.1-fold more 65Zn remains in NHE root after 45-min desorption. At the end of 60 min uptake period, no difference of 65Zn accumulation was observed in undesorbed root-cell-wall of Sedum alfredii. However, 3.0-fold more 65Zn accumulated in desorbed root-cell-wall of NHE. Zn2+ binding in root-cell-wall preparations of NHE was greater than that in HE under high Zn2+ concentration. All these results suggested that root-cell-wall of the two ecotypes Sedum alfredii had the same ability to adsorb Zn2+, whereas the desorption characteristics were different, and with most of 65Zn binding on root of HE being available for loading into the xylem, as a result, more 65Zn was translocated to the shoot.

Comparison between sequential and single extraction procedures for metal speciation in fresh and dried Sedum Plumbizincicola

Sequential and single extraction procedures were applied to both fresh and dried Sedum Plumbizincicola leaves and stems.The extractants, different from those of soil, sediment or sewage sludge metal fractions, were water, 80%(v/v) ethanol, 1 mol/L Na Cl,2% HAc and 0.6 mol/L HCl. Zn, Cd and Cu in the extracts and samples were measured by flame atomic adsorption spectrometry. In sequential extraction procedures, water soluble form and ethanol soluble form are the main fractions for Zn, while water soluble form and Na Cl soluble form for Cd, and comparatively uniform distribution for Cu with the residue form most and HCl soluble form second. Single extraction procedures are used to compare the extraction efficiencies of the five reagents to screen appropriate extractants and operating conditions for liquid extraction to deal with large amount of harvested metal-contained biomass, which will pose a threat to the environment if treated improperly. The sequences of extraction efficiencies are HCl>Na Cl≈HAc>Water≈Ethanol for Zn and HCl≈Na Cl≈HAc>Water>Ethanol for Cd. As for Cu, all the five extractants cannot effectively extract Cu, but HCl achieves a higher efficiency(>70% in fresh samples, and 45%-60% in dried samples). Besides, extraction efficiencies for most extractants in fresh samples are higher than those in dried samples, and extraction efficiencies of stems and leaves for the five extractants are close. The two extraction procedures can obtain high degree of accuracy with the relative standard deviation(RSD)lower than 10%, and metal recoveries are controlled between 80%-120% with most of 90%-110%.

Differential generation of hydrogen peroxide upon exposure to zinc and cadmium in the hyperaccumulating plant specie(Sedum alfredii Hance)

Sedum alfredii Hance has been identified as zinc(Zn) and cadmium(Cd) co-hyperaccumulator.In this paper the relationships of Zn or Cd hyperaccumulation to the generation and the role of H2O2 in Sedum alfredii H.were examined.The results show that Zn and Cd contents in the shoots of Sedum alfredii H.treated with 1000 μmol/L Zn2+ and/or 200 μmol/L Cd2+ increased linearly within 15 d.Contents of total S,glutathione(GSH) and H2O2 in shoots also increased within 15 d,and then decreased.Total S and GSH contents in shoots were higher under Cd2+ treatment than under Zn2+ treatment.However,reverse trends of H2O2 content in shoots were obtained,in which much higher H2O2 content was observed in Zn2+-treated shoots than in Cd2+-treated shoots.Similarly,the microscopic imaging of H2O2 accumulation in leaves using H2O2 probe technique showed that much higher H2O2 accumulation was observed in the Zn2+-treated leaf than in the Cd2+-treated one.These results suggest that there are different responses in the generation of H2O2 upon exposure to Zn2+ and Cd2+ for the hyperaccumulator Sedum alfredii H.And this is the first report that the generation of H2O2 may play an important role in Zn hyperaccumulation in the leaves.Our results also imply that GSH may play an important role in the detoxification of dissociated Zn/Cd and the generation of H2O2.

Screening of Anti-tumor Effective Extracts from Sedum bulbiferum Makino and HPLC Determination of Quercetin and Kaempferol in This Herbal Medicine

[Objectives] To investigate the anti-tumor activity of Sedum bulbiferum Makino in vitro,and establish a HPLC method for determination of quercetin and kaempferol in S. bulbiferum. [Methods] The inhibitory activities of different extracts and total flavonoids of S. bulbiferum on proliferation of three kinds of cancer cells( Hep G2,EC109,SW480) were tested by MTT assay. HPLC method for determination of quercetin and kaempferol in S. bulbiferum was established. [Results]The growth and proliferation of the three kinds of cancer cells were all significantly inhibited by ethyl acetate fraction and total flavonoids isolated from S. bulbiferum. With each extraction concentration increasing,stronger anti-tumor activity was found. The linear ranges of quercetin and kaempferol were 0. 03-0. 36 μg( R = 0. 999 9) and0. 08-0. 96 μg( R = 0. 999 9),and the average recoveries were 98. 90%( RSD = 1. 15%) and 98. 27%( RSD = 1. 70%),respectively.[Conclusions]S. bulbiferum has significant anti-tumor activity in vitro. The HPLC method established was accurate,reproducible,and could be used for quality control of this crude drug.

Integrated Fertilization Regimes Boost Heavy Metals Accumulation and Biomass of Sedum alfredii Hance

The hyperaccumulator Sedum alfredii Hance(S.alfredii)may be employed for zinc(Zn)and cadmium(Cd)-polluted soil remediation.However,the low phytoremediation efficiency,related to the low biomass production,limits its use with that purpose.In this experiment,nitrogen(N),phosphorus(P),and potassium(K)fertilizers,and organic manure were applied to investigate the phytoremediation ability of S.alfredii.Hydroponic and pot experiments were conducted using Zn-Cd polluted soil.The hydroponic experiment indicated that appropriate fertilizer application could increase(p<0.05)the amount of accumulated Zn and Cd in S.alfredii.When N supply ranged from 0.5 to 2.5 mmol L^(−1),it could improve growth and accumulation of Zn and Cd in whole plants of S.alfredii.The 1 mmol L^(−1) N was an optimal N dosage for shoot biomass production and Cd accumulation in shoots,while the 2.5 mmol L^(−1) was an optimal N dosage for Zn accumulation in shoots.Both low(<0.05 mmol L^(−1))and high(>0.8 mmol L^(−1))P supply decreased growth,and Zn/Cd accumulation in whole plants of the studied species.The 0.1 mmol L^(−1) P was an optimal dosage for S.alfredii biomass production and Zn/Cd accumulation in shoots.The supply levels within the range from 0.3 to 1 mmol L^(−1) K could significantly improve the biomass production of S.alfredii and its capability to accumulate Zn and Cd in the biomass.The 0.5 mmol L^(−1) K was an optimal dosage for the whole biomass production and Zn accumulation in shoots,while the 1 mmol L^(−1) was an optimal K dosage for Zn accumulation in shoots,which was 17.2%higher than the control.Moreover,the soil pot experiment showed that the combination of organic(fermented manure)and inorganic fertilizers made significant effects on the Zn and Cd-polluted soil remediation by S.alfredii.These effects varied,however,with the application of different proportions of N,P,K and organic matter.The Zn accumulation by S.alfredii reached the highest efficiency ability under the highest fertilizer mixing rate(N:50 mg kg^(−1),P:40 mg kg^(−1),K:100 mg kg^(−1),organic matter:1%).Even more,S.alfredii showed the strongest ability to accumulate Cd with a lower fertilizer mixing rate(N:25mg kg^(−1),P:20mg kg^(−1),K:50 mg kg^(−1),organic matter:0.5%).

Optimization of Extraction Conditions for SOD in Sedum aizoon by the Response Surface Method

This paper aimed to optimize extracting conditions of SOD in Sedum aizoon by response surface method. First,the single factor optimization tests were carried out. Three single factors were time,p H and liquid-solid ratio respectively. By Design expert 8. 0. 5 software,17 kinds of test scheme were designed. The protein yield of each scheme was obtained,and Design expert 8. 0. 5 software was used to conduct variance analysis and interactive analysis among factors,to obtain the corresponding mathematical model. The results showed that in single factor experiments,the optimal time was 1 h,p H was 7. 8,and liquid-solid ratio was 15:1. The interactive analysis indicated that the optimal scheme for extracting SOD in Sedum aizoon was time of 0. 98 h,p H of 7. 79,liquid-solid ratio of 13. 71:1. By the method of pyrogallol autoxidation,the extracted SOD enzyme activity was measured,namely 12 U/g. The results could provide a reference for further utilization and research of Sedum aizoon resource.