Phytoextraction of Heavy Metals from Highly Contaminated Soils Using Sauropus androgynus

It has been found that Sauropusandrogynus has a strong adaptive capacity in multiple heavy-metal-contaminated soils. Field trials in highly heavy-metal-contaminated soils were conducted to investigate the extraction efficiency of Cd, Cu, Pb, and Zn by S. androgynus. The yield of S. androgynus could reach 10.01 t ha^?1, and the amount of Cd, Cu, Pb, and Zn extracted by the plant was 57.36, 218.57, 2078.11, and 19.64 g ha^?1, respectively. S. androgynus removed 0.35%, 0.01%, 0.03%, and 1.37% of the total soil Cd, Cu, Pb, and Zn in one growing season, respectively. By comparing the extracting capability and yield with some hyperaccumulators reported in the literature, S. androgynus was considered to be a highly effective plant to extract heavy metals (Cd, Cu, Pb, and Zn). The plant should be useful for phytoextraction of soils contaminated by the heavy metals.     

新疆杨愈伤组织原生质体的游离与纯化

目的:以愈伤组织为材料,研究新疆杨原生质体的游离、纯化。方法:以新疆杨愈伤组织为材料,采用简单试验设计和方差分析方法,对新疆杨原生质体游离的影响因素进行研究,并利用二乙酸荧光素染色法观察原生质体活力。结果:适宜新疆杨愈伤组织原生质体游离的较适宜条件是:CPW+2.0%纤维素酶R-10+1.0%离析酶R-10+1.0%果胶酶Y-23+0.6 mol/L甘露醇,酶解温度27℃,酶解时间8 h。在此条件下,原生质体产量达8.5×106个/(g.FW),活力达83.6%。原生质体纯化可采用蔗糖等密度离心法,较适蔗糖浓度为30%。结论:研究筛选出的酶解因素组合与等密度离心条件较适宜新疆杨愈伤组织原生质体的游离和纯化。     

Phytoextraction of soil cadmium and zinc by microbes-inoculated Indian mustard (Brassica juncea)

Abstract

A pot experiment was carried out to evaluate the effect of Pseudomonas fluorescens and Trichoderma harzianum inoculation on the uptake of zinc (Zn) and cadmium (Cd) by Indian mustard (Brassica juncea) from the soil having three different concentrations of Zn (300, 600, 900 mg/kg) and Cd (5, 10, 15 mg/kg) separately. Microbial inoculation resulted in significantly better plant growth, available metal content and their uptake than control (without microbes). Available Zn was enhanced, ca.1.6- and 1.4-fold and Cd ca. 2.5- and 1.8-fold, by P. fluorescens and T. harzianum, respectively. P. fluorescens resulted in an increase in Zn uptake by 113.9, 51.9 and 58.4% and T. harzianum by 42.6, 32.1 and 33.9% over control from soils having 300, 600 and 900 mg Zn, respectively, while of the corresponding results for Cd were 110.2, 48.9 and 58.1% with P. fluorescens and 42.6, 30.9 and 33.4% with T. harzianum from soil having 5, 10 and 15 mg Cd, respectively, after 90 days of treatment. In general the rate of metal uptake was higher during the initial 30 days and declined later.     

Multiple shoot regeneration from root organ cultures of Populus alba x P. grandidentata

Excised roots of various ages from Crandon and Hansen clones of Populus alba x P. grandidentata were tested for their regeneration capacity. Sixty-day-old excised roots that contained root tips were found to be most suitable. The highest number of shoots (an average of 111 shoots/root segment with Crandon and 98 with Hansen) was obtained by adding 22M and 14M zeatin to the medium, respectively. The two clones of hybrid poplar responded similarly to growth regulator treatments; however, the number of shoots produced was greater from the root organs derived from Crandon clones. Regenerated shoots were rooted in basal Woody Plant Medium without any growth regulators. Successful transplantation into soil and growth was achieved with all plants.     

Phytoextraction and Accumulation of Boron and Selenium by Poplar (Populus) Hybrid Clones

There has been much interest recently in central California for reusing drainage water to grow trees. A sand-culture study was conducted to investigate the accumulation of boron (B) and selenium (Se) in eight hybrid poplar (Populus) clones irrigated with synthetic agricultural effluent containing increasing levels of chloride salt, B, and Se. Electrical conductivity (EC) ranged from 1.5 to 15 dS m^-1, B levels from 1 to 5 mg L^-1, and Se levels from 100 to 500 μg L^-1. Compared with all tree organs, the leaves accumulated the greatest concentrations of B and Se at the time of harvest. The results show that pooled leaf B concentrations were positively correlated with EC levels (r = 0.78, P < 0.001) and negatively correlated (r = -0.53, P < 0.001) with leaf dry matter for all clones at all tested B levels. Combined leaf and stem Se data show, respectively, a significant decrease (P < 0.05 level) in tissue accumulation of Se with increased salinity. Toxicity symptoms (e.g., burning leaf margins, shoot die back) occurred in most clones grown at 12 and 15 dS m^-1 treatments leading to leaf abscission. Based on the data, clone 49177 (Populus trichocarpa × P. deltoidus) best tolerated the tested parameters among the clones and accumulated the greatest amount of B and Se. The moderate ability of the Populus species to remove and accumulate B and Se from saline effluent is most effective at salinity levels less than 7 dS m^-1.     

Cadmium Removal Potential from Contaminated Soil by Tagetes erecta L. and Panicum maximum: Influence of Soil pH

The potential for cadmium (Cd) removal from contaminated soil by two species—marigold (Tagetes erecta L.) and Guinea grass (Panicum maximum)—was investigated in pot culture experiments in a greenhouse in triplicate. The concentration of Cd was varied from 50 to 200 mg kg^?1 and the pH was varied from 5.0 to 7.5 to investigate the effect of pH on Cd uptake. The results showed that total biomass of Guinea grass was around nine and seven times higher than that of marigold for Cd treatments of 50 and 100 mg kg^?1 at pH 5.0, respectively. Total cadmium uptake at Cd treatments of 50 and 100 mg kg^?1 at pH 5.0 by Guinea grass was 19.28 ± 3.14 and 36.06 ± 4.28 mg kg^?1, respectively, and for marigold was 15.66 ± 4.17 and 20.38 ± 3.24 mg kg^?1, respectively. The total Cd uptake by Guinea grass was 1.23 and 1.77 higher than that of marigold at Cd treatments of 50 and 100 mg kg^?1, respectively, at pH 5.0 due to higher biomass. The maximum Cd uptake by marigold and Guinea grass occurred at pH 5.0 at Cd treatment of 100 mg kg^?1. The results clearly show that the two species behave very differently for Cd uptake. Guinea grass is easy to grow, drought tolerant and, due to its higher biomass, it can be used for remediation of Cd-contaminated soil.     

Cryopreservation of white poplar (Populus alba L.) by vitrification of in vitro-grown shoot tips

 Shoot tips from in vitro-grown, cold-hardened stock plants of white poplar (Populus alba L.) were successfully cryopreserved at –196  °C by one-step vitrification. After preculturing at 5  °C for 2 days on hormone-free MS medium containing different sucrose concentrations, and loading for 20 min with 2 m glycerol and 0.4 m sucrose, shoot tips were treated with the PVS2 vitrification solution and plunged directly into liquid nitrogen. Best survival rate (90%) was obtained when shoot tips were precultured on 0.09 m sucrose, hormone-free MS medium, vitrified by exposure to PVS2 solution for 60 min at 0  °C and, following cryopreservation, rewarmed at 40  °C and washed in 1.2 m sucrose solution for 20 min. Regrowth was improved by plating shoot tips on a gelled MS medium containing 1.5 μm N⁶-benzyladenine plus 0.5 μm gibberellic acid, while shoot rooting was achieved on MS medium containing 3 μm indole-3-butyric acid. Following this procedure, almost 60% rooted shoots were obtained from cryopreserved shoot tips. Received: 1 February 1999 / Revision received: 3 May 1999 · Accepted: 21 May 1999     

Seasonal Variation of Photoinhibition of Photosynthesis in Bark from Populus Tremula L.

In the bark of Populus tremula L. photochemical efficiency of photosystem 2 (PS2) determined as Fv/Fm decreased during winter. The strongest reduction was found after cold periods. The degree of reduction depended on irradiance since the lowest levels of Fv/Fm were found on the sun-exposed side of the stem and below thin phellem. Therefore, photoinhibition was partly responsible for the reduction in Fv/Fm. The photochemical efficiency of PS2 recovered in late April about a month before the trees got leaves. In the laboratory, Fv/Fm recovered within about a week under low irradiance at 20 °C. Rapid recovery of photochemical efficiency of PS2 in the bark may be important to reduce respiratory loss of CO2 from the stem before the trees get leaves.     

Reduction of Cadmium Availability to Tobacco (Nicotiana tabacum) Plants using Soil Amendments in Low Cadmium-contaminated Agricultural Soils: A Pot Experiment

Cadmium (Cd) concentration in field-grown tobacco leaves usually ranges from < 0.5 to 5 mg Cd kg^–1 dry matter (DM). Reducing bioavailability of soil Cd by adding amendments to the soil could be suitable to mitigate Cd uptake by tobacco plants. However, little is known on the effect of inorganic amendments on agricultural soils with low Cd concentrations. Therefore, we performed a pot experiment with tobacco plants that were grown during 56 days in two neutral to alkaline agricultural soils with low total Cd concentrations (soil 1 = 0.4, soil 2 = 0.7 mg kg^–1). Both soils were amended or not with 1 or 5% of sepiolite, zeolite, hydroxyapatite and apatite II™. Major and trace elements were measured in mid-stalk position leaves. Soil metals were measured in a DTPA soil extraction to assess the effect of the amendments on metal bioavailability. Some amendments significantly reduced Cd concentration in tobacco leaves, but the effect differed between the two soils tested. In soil 1, the use of zeolite at the 1% dose was the most efficient, reducing the average Cd concentration from 0.6 to 0.4 mg kg^–1. In soil 2, the 5% hydroxyapatite treatment led to the maximal reduction in Cd concentration (50%), with an average final Cd concentration in leaves of 0.7 mg kg^–1 (control: 1.5 mg kg^–1). There was a dose effect for some amendments in soil 2 (containing more Cd), suggesting a reduced efficiency of the amendment at the lowest addition rate. DTPA extractable Cd and Zn measured at the end of the pot experiment were correlated to the metal concentrations in tobacco leaves suggesting that (1) the reduction in leaf Cd concentration was due to a reduction in metal availability to tobacco and (2) DTPA may be a suitable extractant to estimate Cd availability to tobacco plants in these two soils. In addition, a batch experiment was performed with the same soils to test a larger number of amendments, including the four tested in the pot experiment. Results were compared to those of the pot experiment to assess whether a batch experiment may predict the efficiency of an amendment on a given soil. It gave results compatible with those from the pot experiment except for the sepiolite and highlighted the broad range of potential amendments available for heavy metal remediation in crop plants.     

通过农杆菌介导法将兔防御素NP-1基因导入毛白杨(P.tomentosa)

采用叶盘法将兔防御素NP－1基因导入毛白杨后,经PCR分析、Southern杂交检测与筛选获得了转基因植株。并经体外抑菌实验表明,转NP－1基因毛白杨植株组织提取物对某些微生物的生长具有明显的抑制作用。     

Chemical Constituents from the Seeds of Ziziphus jujuba var. Spinosa (Bunge) Hu

To search for new and bioactive minor components from traditional Chinese medicines, a new compound, named jujuphenoside (1), was isolated from the seeds of Ziziphusjujuba var. spinosa (Bunge)Hu. The structure of jujuphenoside was elucidated by spectral and chemical methods, particularly twodimensional nuclear magnetic resonance analysis. Together with the new compound, 22 known compounds were also isolated and identified from the seeds of Z. jujuba var. spinosa, among which, epiceanothic acid (2) was first obtained from natural resources, whereas compounds 7-16 were first obtained from this plant.     

Phytoextraction Potential of Wild Type and 35S-GSHI Transgenic Poplar Trees (Populus × Canescens) for Environmental Pollutants Herbicide Paraquat,Salt Sodium,Zinc Sulfate and Nitric Oxide In Vitro

Phytoextraction potentials of two transgenic (TR) poplar (Populus × canescens) clones TRggs11 and TRlgl6 were compared with that of wild-type (WT) following exposure to paraquat, zinc sulfate, common salt and nitric oxide (NO), using a leaf-disc system incubated for 21 days on EDTA-containing nutritive WPM media in vitro. Glutathione (GSH) contents of leaf discs of TRlgl6 and TRggs11 showed increments to 296% and 190%, respectively, compared with WT. NO exposure led to a twofold GSH content in TRlgl6, which was coupled with a significantly increased sulfate uptake when exposed to 10^?3 M ZnSO₄. The highest mineral contents of Na, Zn, Mn, Cu, and Mo was observed in the TRggs11 clone. Salt-induced activity of catalase enzyme increased in both TR clones significantly compared with WT under NaCl (0.75% and 1.5%) exposure. The in silico sequence analyses of gsh1 genes revealed that P. × canadensis and Salix sachalinensis show the closest sequence similarity to that of P. × canescens, which predicted an active GSH production with high phytoextraction potentials of these species with indication for their use where P. × canescens can not be grown.     

Aerobic biodegradation of methyl tert-butyl ether (MTBE) by pure bacterialcultures isolated from contaminated soil

Summary Methyl tert-Butyl Ether (MTBE) has been used in gasoline as a substitute for lead-based additives, which have been demonstrated to be toxic. MTBE however, is persistent in soil and water, showing high affinity for water and low affinity for soil, and has become an important contaminant. Therefore, the aim of this work was to isolate and identify soil microorganisms capable of degrading MTBE. Two samples were taken from a gasoline-contaminated soil at a service station and 59 different bacterial strains were isolated by enrichment culture with three consecutive selective transfers. Biochemical and morphological characterization of the bacterial isolates classified them into the following groups: Bacillus, Rhodococcus, Micrococcus, Aureobacterium and Proteus. Twelve strains were selected for evaluation of MTBE biodegradation depending on visual growth and biomass production of the isolates in minimal salt broth. Six strains significantly reduced MTBE concentration (22–37%) compared to an abiotic control after 5 days of incubation. Although it has been considered that MTBE is degraded mainly by cometabolism, our results demonstrate that these microorganisms are able to reduce MTBE concentration when MTBE is the sole source of carbon.     

Application of genetic markers to the discrimination of European Black Poplar (Populus nigra) from American Black Poplar (P. deltoides) and Hybrid Poplars (P. x canadensis) in Switzerland

European Black Poplar (Populus nigra) is considered a rare and endangered tree species because of severe reduction of its natural riverine habitat and potential hybridisation with the related non-indigenous taxa P. deltoides and P. x canadensis. As it is difficult to distinguish these taxa solely based on their morphology, we applied a PCR-based assay with an easy-to-use and robust molecular marker set (cpDNA trnL-trnF/RsaI RFLP, nDNA win3 and nDNA POPX/MspI RFLP) in order to identify pure P. nigra. Different plant tissues could be used for fast and standardised DNA extraction. The application of the three marker types was tested on a number of different Populus taxa, and they were also used for the verification of pure P. nigra in a sample of 304 putative P. nigra individuals from Switzerland. Cross-checking of the DNA data with those using a traditional allozyme approach resulted in complete agreement. The availability of molecular identification methods is an important prerequisite for the conservation of European Black Poplar, because pure, non-introgressed plant material can then be used in restoration projects of European floodplains.     

Metallo-proteinase from the seedlings of kale (Brassica oleracea L. var. sabellica):

Metallo-proteinase from 8-d-old seedlings of kale was isolated. The enzyme was extracted with 1% NaCl, concentrated by ammonium sulfate and finally purified by high-performance liquid chromatography. The isolated enzyme had a molecular weight of 22.4 kDa and showed a maximum activity at pH 9.0 using casein as a substrate. Proteolytic activity of proteinase was inhibited by chelators. The inhibition by ethylenediaminetetraacetate (EDTA) was abolished by some divalent metals ions, especially by Zn²⁺. The enzyme showed activity against the synthetic peptides Suc-Ala-Ala-Pro-Leu-pNA and Suc-Ala-Ala-Pro-Phe-pNA, and hydrolized the following peptide bonds in the oxidized insulin B-chain: Leu6-Cya7, Leu15-Tyr16, Leu17—Val18 and Phe25-Tyr26.Abbreviations EDTA ethylenediaminotetraacetic acid - HPLC high-performance liquid chromatography - NEM N-ethylmaleimide - PCMB p-mecuribenzoic acid - PMSF phenylmethylsulfonyl fluoride This work was supported by the University Science Programme, Ministry of National Education, and Polish Academy of Science, Warsaw, Poland.     

中国黄耆属簇毛黄耆亚属的叶表皮特征及其系统学意义

在光学显微镜和扫描电镜下,观察了国产黄耆属簇毛黄耆亚属16种1变种的叶表皮特征。结果表明,气孔器在各种植物的上、下表皮均有分布,多为无规则型,也有不等细胞型;叶表皮细胞形状有不规则形与多边形,表皮细胞垂周壁有平直、浅波状、波状或深波状。叶表皮细胞形状与垂周壁的式样可以分为四种类型,这四种类型与亚属的分组有一定的对应关系。在扫描电镜下可见表皮细胞上有角质层,气孔下陷,气孔的外拱盖及其内缘特征在亚属内都比较一致。表皮细胞角质层的纹饰在个别类群中有一定的变异,对种类的鉴定有一定的意义。     

Regeneration of plants from hypocotyl protoplasts of rapessed (Brassica napus L. var. oleifera) cultivars

Protoplast cultures were prepared from hypocotyls of ten spring rapeseed cultivars. Protoplasts from all genotypes tested formed calli, and shoots were regenerated from calli of nine of the genotypes at frequencies varying from 15 to 76%. The regenerating cultivars fell into a high regenerating group (>60% and a low regenerating group <25%).     

Differential expression of two SOD (Superoxide dismytase) genes from small radish (Rhaphanus sativus L. var.sativus)

We have isolated two Superoxide dismutase cDNA clones (RsCu/ZnSod andRsFeSod) from small radish (Raphanus sativus L.) by cDNA library screening.RsCu/ZnSod is 563 bp long, with an open reading frame of 153 amino acids, and corresponds to a protein of predicted molecular mass 15.1 kDa and a pl of 5.44. The 823-bp RsFeSod has an ORF of 213 amino acids, corresponding to a protein of predicted molecular mass 25.4 kDa and a pl of 8.77. Their nucleotide and deduced amino acid sequences show the highest homology with those ofArabidopsis. Genomic Southern blot analysis, using each cDNA clone as probe, has revealed that the SOD genes are present as at least two copies in the small radish genome. Nondenaturing polyacrylamide gels for SOD activity has demonstrated the presence of several isozymes, depending on the organ type and developmental stage. TheseRsSod genes also have differential expression patterns in response to treatments with white light, xenobiotics, UV, osmoticums, plant hormones, and salicylic acid. Therefore, we suggest that they are involved in an antioxidative defense mechanism against stress induced by environmental change.