WO2001036963A2 - Procede pour deceler la presence de metaux lourds et/ou de polluants organiques au moyen d'une suspension cellulaire vegetale - Google Patents

Procede pour deceler la presence de metaux lourds et/ou de polluants organiques au moyen d'une suspension cellulaire vegetale Download PDF

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Publication number
WO2001036963A2
WO2001036963A2 PCT/EP2000/011384 EP0011384W WO0136963A2 WO 2001036963 A2 WO2001036963 A2 WO 2001036963A2 EP 0011384 W EP0011384 W EP 0011384W WO 0136963 A2 WO0136963 A2 WO 0136963A2
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Prior art keywords
gst
suspension culture
cell suspension
activity
detection
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PCT/EP2000/011384
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German (de)
English (en)
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WO2001036963A3 (fr
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Peter Schröder
Reinhard Debus
Andrea Wenzel
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Gsf-Forschungszentrum Fur Umwelt Und Gesundheit Gmbh
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Priority to AU21597/01A priority Critical patent/AU2159701A/en
Priority to EP00985047A priority patent/EP1230547A2/fr
Publication of WO2001036963A2 publication Critical patent/WO2001036963A2/fr
Publication of WO2001036963A3 publication Critical patent/WO2001036963A3/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/18—Water
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/24—Earth materials
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5097—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving plant cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/91—Transferases (2.)
    • G01N2333/9116—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5)
    • G01N2333/91165—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5) general (2.5.1)
    • G01N2333/91171—Transferases (2.) transferring alkyl or aryl groups other than methyl groups (2.5) general (2.5.1) with definite EC number (2.5.1.-)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2520/00—Use of whole organisms as detectors of pollution

Definitions

  • the present invention relates to a method for the detection of heavy metals and / or organic pollutants in a test solution, for example a soil solution or water sample, using plant cell suspension cultures, in particular using cell suspension cultures of gymnosperms, such as Picea and Pwws cultures, or other cell cultures capable of lignification.
  • the method according to the invention comprises the steps: a) adding a test solution to a plant cell suspension culture, b) incubating the cell suspension culture and c) determining the H 2 O content, the GSH (glutathione, reduced) / GSSG (glutathione, oxidized) - Content and the glutathione S-transferase activity.
  • Explosives such as 2,4,6-trinitrotoluene, nitrocellulose, 2,4-dinitrotoluene, 2,6-dinitrotoluene, 2-amino-4-nitrotoluene, hexogen and 1,3 are used primarily in contaminated soils of former armaments sites and military training areas - Dinitronaphthalene, igniters, such as lead trinitroresorcinate, lead azide, tetrazene and mercury fullminate, accompanying substances such as phthalates, diphenylamine and dioctyl phthalate, and heavy metals (ions) such as cadmium, lead, arsenic and mercury 2+ .
  • igniters such as lead trinitroresorcinate, lead azide, tetrazene and mercury fullminate
  • accompanying substances such as phthalates, diphenylamine and dioctyl phthalate
  • heavy metals (ions) such as cadmium, lead, arsenic
  • Soil decontamination mostly takes place through grain separation and
  • Soil washing This has the advantage that a large part of the soil can be brought back to its place of origin; all that remains for disposal is the highly contaminated fine grain and the process water.
  • the process water is cleaned by filtration and biological methods with high efficiency, and the largest amount of water can be returned to the water cycle after appropriate analysis and toxicity testing.
  • the biggest problem with this approach is the availability of the relevant toxicity tests and their practical handling in routine activities.
  • the object of the invention is to provide a toxicity test or a detection method with which both inorganic pollutants, for example heavy metals, and organic pollutants in soil solutions and water samples can be detected reliably, sensitively, quickly and in a standardized manner.
  • Plants are increasingly being used as test organisms, since they are characterized by the fact that they are already specific in a small number React quantities of pollutants. As sessile organisms, plants are highly exposed to the influences of pollutants. The plant metabolism reacts to external influences with great flexibility and is able to ward off pollutants of biogenic and anthropogenic origin. This is done either with the help of biogenic metabolic intermediates that are in the
  • Plants can be influenced by the influence of pollutants: they can be strongly induced or inhibited in response to some substances.
  • the first-mentioned publication shows the general inducibility of GST. References to the effect or diagnosis of individual substances or substance mixtures are not disclosed there.
  • the second publication mentioned above describes the inhibition of GST and shows that GST enzymes can be influenced by foreign substances that are substrates for GST.
  • the third publication mentioned above also describes xenobiotics as inducers for GST, which are substrates for GST. Xenobiotics without substrate character or heavy metals as inducers of the GST and in particular methods for the detection of xenobiotics based on the
  • the method according to the invention not only represents a reliable toxicity test for heavy metals and organic pollutants, it is also able, particularly by combining different detection parameters, to reliably differentiate between different pollutants and to determine them both qualitatively and quantitatively.
  • the object of the invention is therefore achieved by a method which comprises the steps: a) adding a test solution to a plant cell suspension culture, b) incubating the cell suspension culture and c) determining the H 2 O 2 content, the GSH (glutathione, reduced) / GSSG (glutathione, oxidized) content and the glutathione S-transferase activity.
  • the cell suspension culture is a suspension of gymnosperms, particularly preferably a suspension of a pine tree (Pinaceae) and most preferably a suspension of Picea abies.
  • any cell culture can be used whose cells are able to form lignin or whose cells can be stimulated to form lignin.
  • pinus cultures or cultures of Ginkgo biloba are particularly suitable for the detection method according to the invention.
  • Cell suspension culture suitably conditioned by targeted hormone and sugar addition for the detection method.
  • the person skilled in the art is familiar with suitable phytohormones and other substances for adjusting the cell culture.
  • the oxidative burst is an HO secretion of living cells induced by pathogens or stress as a defense reaction to attacks from outside.
  • the H 2 O 2 content can be determined in various ways; the
  • the oxidative burst is preferably determined based on the method of Messner & Boll (1994, Plant Cell, Tissue & Organ Cult. 39: 69-73). Other methods for determining the oxidative burst are disclosed, for example, in Messner and Schröder (1999, Journal of Applied Botany 73: 6-10).
  • GSH / GSSG content Numerous methods are also available to the person skilled in the art for determining the GSH / GSSG content.
  • the method of silage is preferred Cepeda et al. (1991, Plant Cell Physiol. 32: 1179-1185).
  • HPLC methods are particularly suitable, possibly with pre- and post-column derivatization.
  • GSH and GSSG should be measured discriminatively.
  • the GST activity is generally determined by extracting the enzyme from the cell suspension and then measuring the GST enzyme activity in the protein extract.
  • GST activity can also be determined using molecular biological techniques
  • Nucleic acid level are determined, with hybridization, DNA chip and PCR techniques being particularly suitable here.
  • Biosensors are also useful in the context of the invention for determining GST activity. With modern PCR methods such as the PCR-select kit from Clontech, for example, it is possible to infer the amount of enzyme and enzyme activity from the amount of transcript.
  • Cultivation of plant cell suspension cultures is based on the guidelines for testing plant protection products in the approval process, Part IV, 3-2 / 1.
  • Residue behavior test rapid test for metabolizability and Degradation of organic crop protection agents in plant cell cultures, the Federal Biological Research Center for Agriculture and Forestry, Braunschweig (1992).
  • the method according to the invention is suitable for determining
  • Heavy metals especially lead, cadmium, arsenic, and organic xenobiotics, especially dinitrotoluenes, in soil eluates and water samples.
  • xenobiotics is used here in the sense of environmental chemicals, i.e. for substances and compounds that are foreign to a certain ecosystem or organism.
  • the incubation period should if possible not be longer than 24 hours.
  • a cell suspension culture of spruce (Picea abies (L.) Karst.) was prepared as described by Messner and Boll (1994, Plant Cell, Tissue and Organ Culture 39: 69-78), Messner and Berndt (1990, Z. Naturforschung 45c: 614-620).
  • the heterotrophic cell suspension culture requires both light and sucrose to grow. For this purpose it is in a modified Murashige and Skoog medium (MS medium; 1962, Physiol. Plant
  • the nutrient medium additionally contains 3% (w / v) sucrose, the phytohormones benzylaminopurine (4.4 ⁇ M) and Naphtylacetic acid (16.1 ⁇ M).
  • the nutrient medium is grown in a shaking machine at 27 ° C., 56 rpm and continuous incidence of light (Fluora fluorescent lamps; Osram L40 W / 77, 1000 lux). After 3 days, the used medium is poured off and replaced with 300 ml of fresh medium.
  • the cultures are halved by 1) allowing the cells to sediment, 2) pouring off the supernatant medium, 3) reducing the cell volume by half and 4) adding 300 ml of fresh medium to half of the cells. This rhythm ensures the constant availability of standardized cell material.
  • the Picea abies stock culture described above is divided after 7 days of growth. Each 12 g wet cell weight is transferred under sterile conditions into 1000 ml baffled flasks containing 120 ml fresh MS
  • the determination of the oxidative burst is carried out based on the method of Messner & Boll (1994, supra).
  • the HO formed is measured by the decrease in the amount of phenol red in the batch. In this reaction catalyzed by peroxidase, phenol red is destroyed by H 2 O 2 . The more HO 2 is formed, the less phenol red is present in the medium. This decrease can be measured on the photometer and related to the HO concentration using calibration curves.
  • a batch with 0.6 ml of an elicitor preparation from Rhizosphaera kalkhoffi serves as a positive control.
  • An approach that contains only cells, peroxidase solution and phenol red serves as a negative control.
  • PC A perchloric acid
  • BPDS bathophenanthroline disulfonic acid
  • the sample is centrifuged off at 10,000 rpm for 10 min and the supernatant is filtered off using Minisart RC4 filters (0.45 ⁇ m pore size). 100 ⁇ l of the filtrate are then used for HPLC analysis on a 3-aminopropyl spherisorb Column (5 ⁇ m, 4.6 x 200 mm) was used. Detection is carried out using a UV detector at 365 nm. Elution of the thiols is carried out using a gradient of mobile solvent A (80% methanol) and mobile solvent B (64% methanol, 0.5 M sodium acetate, 10% acetic acid). The gradient increases after 5 min. of 20% solvent B within 20 min. to 99% »solvent B, remains 5 min. at
  • GSH GSH in the range of 5-250 ⁇ M and for GSSG in the range of 5-200 ⁇ M.
  • concentration the ratio of the GSH or GSSG peak to the internal standard ⁇ -Glu-Glu peak is formed and the respective concentrations per g fresh weight are calculated.
  • the processing of the plant material can be carried out according to standard methods for the isolation of plant GST, e.g. according to Schröder and Berkau
  • the GST is separated from other proteins of the crude extract by fractional ammonium sulfate precipitation in two steps, from 0-40% and from 40-80%> saturation.
  • the amounts of salt required for the individual precipitation steps are slowly (30 min.) Added to the crude extract cooled on ice with constant stirring. The mixture is then left for a further 30 min. continue stirring so that the solution balance can be established.
  • 20 min. Centrifuged at 20,000 rpm and discarded the pellet.
  • the precipitated protein is 30 min. Pelleted at 20,000 rpm by centrifugation. The pellet is resuspended in 2.5 ml of 20 mM Tris / HCl, pH 7.0.
  • the ammonium sulfate is removed from the extract again by gel filtration.
  • Pre-packed PD 10 gel filtration columns with Sephadex G-25 material from Pharmacia Biotech, Freiburg are used. As indicated by the manufacturer, the columns are equilibrated with 20 ml buffer (20 mM Tris / HCl, pH 7.0). 2.5 ml of the protein solution to be desalinated are applied and eluted from the column with 3.5 ml of buffer.
  • Extraction method for vegetable protein can be used. The procedure is as follows: After the end of the incubation, the cells are sucked off as described and frozen in liquid nitrogen. The cells are ground up and an aliquot of the work-up buffer is added. Instead of the first centrifugation, the extract is filtered through several layers of Miracloth. This extract represents the crude extract. This is followed by a fractionated ammonium sulfate
  • the enzyme activity of the GST is determined in the protein extract after PD-10 gel filtration. Protein determination in the cell culture extracts is carried out according to the method of Bradford (1976, Anal. Biochem. 72: 248-254).
  • the GST activity can be assigned to individual isoenzymes with different substrate specificity, in addition to the model substrate l-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), p-nitrobenzoyl chloride (pNBC) and 1,2-epoxy-3 (p-nitrophenoxy) propane (EPNP) was used to determine the enzyme activity.
  • the above-mentioned substrates are conjugated with reduced glutathione (GSH) in the course of the enzyme-catalyzed reaction.
  • GSH reduced glutathione
  • the linear increase in the product can vary with each substrate-specific wavelength can be measured in the photometer.
  • GSH and the individual substrates are bidest. Prepared water or in ethanol and stored on ice.
  • 0.1 M potassium phosphate buffer with different pH is stored at room temperature (20 ° C).
  • 540 ⁇ l KPP buffer and 20 ⁇ l GSH and substrate solution are pipetted into 1.5 ml cuvettes and mixed well.
  • the reaction is started by adding an aliquot of the enzyme sample.
  • An approach in which the enzyme sample is replaced by water is used as the blank value.
  • the photometric determination of the GST activity induced by CDNB is carried out according to the method of Habig et al. (1974, J. Biol. Chem. 249: 1730-1739), which is induced by DCNB and pNBC, is carried out based on the method of Götzberger (1994, diploma thesis, University of Ulm), see also Schröder et al. (1997, Appl. Botany 71: 31-37).
  • To measure GST (EPNP) activity the method of Fjellstedt et al. (1973, J. Biol. Chem. 248: 3702-3707).
  • Heavy metals and organic compounds can be distinguished.
  • the organic compounds 2,4-DNT and 2,6-DNT react. Apparently, they do not generate oxidative stress on the cells.
  • the GSH content increases by 83% and the GSSG content decreases by 34% in the treated cells.
  • the GSSG level increases by 226% compared to the control, the GSH level decreases contrast, by 20% compared to the control. You can see an opposite picture of the effect of the two metals.
  • 2,4-DNT and 2,6-DNT have a weak effect in the selected concentration range.
  • the activities change significantly at a concentration of 1 ppm. They move in an interval of 25% - 25%. It is striking that the activities at a concentration of 8 ppm do not differ much from those at 1 ppm.
  • 2,6-DNT you also get significant activity changes at 1 ppm. In contrast to 2,4-DNT, all values are above the control.
  • Concentration by lyophilization is a preferred embodiment of the method according to the invention.
  • test system it is also important in the context of the test system according to the invention that, in addition to pathogens, only heavy metals and not also the tested organic substances trigger an oxidative burst.
  • the heavy metals cause reactions here even in the low concentrations that are present in authentic soil eluates. This makes it possible to make qualitative statements as to whether the soil contamination found is heavy metals or organic compounds.
  • the test system presented is therefore useful for determining the potential effects of xenobiotics in soil and water samples.

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Abstract

L'invention concerne un procédé pour déceler la présence de métaux lourds et/ou de polluants organiques dans une solution pour essai, par exemple une solution du sol ou un prélèvement d'eau, au moyen de cultures de suspensions cellulaires végétales, notamment au moyen de cultures de suspensions cellulaires de gymnospermes, par ex. Picea abies. Ce procédé comprend les étapes suivantes : addition d'une solution pour essai à une culture de suspension cellulaire végétale, b) incubation de la culture de suspension cellulaire, et c) détermination de la teneur en H2O2, en GSH (glutathion, réduit)/GSSG (glutathion, oxydé) et de l'activité de la glutathion S-transférase.
PCT/EP2000/011384 1999-11-18 2000-11-16 Procede pour deceler la presence de metaux lourds et/ou de polluants organiques au moyen d'une suspension cellulaire vegetale WO2001036963A2 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
AU21597/01A AU2159701A (en) 1999-11-18 2000-11-16 Method for detecting heavy metals and/or organic noxious substances using plant cell suspensions
EP00985047A EP1230547A2 (fr) 1999-11-18 2000-11-16 Procede pour deceler la presence de polluants au moyen d'une suspension cellulaire vegetale

Applications Claiming Priority (2)

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DE19955604.0 1999-11-18
DE1999155604 DE19955604C2 (de) 1999-11-18 1999-11-18 Verfahren zum Nachweis von Schwermetallen und organischen Schadstoffen unter Verwendung pflanzlicher Zellsuspensionen

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Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102128915A (zh) * 2010-12-02 2011-07-20 南京大学 利用蚯蚓生物标志物诊断土壤污染状况的方法
CN103675233A (zh) * 2013-11-26 2014-03-26 南京信息工程大学 利用农作物标志物检测土壤污染的方法
CN103837632A (zh) * 2014-03-07 2014-06-04 山东省科学院海洋仪器仪表研究所 液相色谱流动注射化学发光法测量水体中镉的装置及方法
CN106442805A (zh) * 2016-11-23 2017-02-22 新疆农业科学院农产品贮藏加工研究所 一种库尔勒香梨果实石细胞中磨木木质素提取的方法

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Publication number Priority date Publication date Assignee Title
CN108479738B (zh) * 2018-05-09 2020-11-13 大连理工大学 一种疏水层析介质、制备方法及其应用
CN109115546A (zh) * 2018-08-22 2019-01-01 滑县恒琢信息科技有限公司 一种绿化园林重金属土壤污染物治理取样系统

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2144543A (en) * 1983-08-01 1985-03-06 Kernforschungsanlage Juelich Method of determining environmental pollutants
DE4433384A1 (de) * 1994-09-09 1996-03-14 Ufz Leipzighalle Gmbh Ökotoxikologische Stoffwechseluntersuchung mit stabilen Isotopen

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2144543A (en) * 1983-08-01 1985-03-06 Kernforschungsanlage Juelich Method of determining environmental pollutants
DE4433384A1 (de) * 1994-09-09 1996-03-14 Ufz Leipzighalle Gmbh Ökotoxikologische Stoffwechseluntersuchung mit stabilen Isotopen

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 10. Juni 1998 (1998-06-10) MASERTI B E ET AL: "Preliminary results on cadmium and mercury tolerance mechanisms in Nicotiana sp." Database accession no. PREV199800354752 XP002169820 & SCIENCE OF THE TOTAL ENVIRONMENT, Bd. 213, Nr. 1-3, 10. Juni 1998 (1998-06-10), Seiten 273-277, ISSN: 0048-9697 *
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1992 SCHROEDER PETER ET AL: "Biomarker for organic xenobiotics in spruce (Picea abies L.): Dynamics of the detoxification enzyme glutathione S-transferase." Database accession no. PREV199395101402 XP002169814 & ANGEWANDTE BOTANIK, Bd. 66, Nr. 5-6, 1992, Seiten 174-179, ISSN: 0066-1759 in der Anmeldung erwähnt *
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1992 SCHROEDER PETER ET AL: "Characterization of glutathione S-transferase from dwarf pine needles (Pinus mugo Turra)." Database accession no. PREV199395003832 XP002169815 & TREE PHYSIOLOGY, Bd. 11, Nr. 2, 1992, Seiten 151-160, ISSN: 0829-318X *
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1994 TAUSZ MICHAEL ET AL: "Stress-physiological Investigations and Chromosomal Analysis on Norway spruce (Picea abies (L.) Karst.)- A Field Study." Database accession no. PREV199598287005 XP002169819 & PHYTON (HORN), Bd. 34, Nr. 2, 1994, Seiten 291-308, ISSN: 0079-2047 *
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1996 RIJSTENBIL J W ET AL: "HPLC analysis of nonprotein thiols in planktonic diatoms: Pool size, redox state and response to copper and cadmium exposure." Database accession no. PREV199799340582 XP002169816 & MARINE BIOLOGY (BERLIN), Bd. 127, Nr. 1, 1996, Seiten 45-54, ISSN: 0025-3162 *
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1998 AMICUCCI E ET AL: "Oxidative burst and electrolyte leakage induced by sulfhydryl blockers and by membrane permeabilizing reagents in different organs of Egeria densa." Database accession no. PREV199900145951 XP002169818 & PROTOPLASMA, Bd. 205, Nr. 1-4, 1998, Seiten 93-100, ISSN: 0033-183X *
DATABASE BIOSIS [Online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; Dezember 1998 (1998-12) LANGE ANKE ET AL: "Purification and characterization of glutathione S-transferase from needles of air polluted Scots pine (Pinus sylvestris L.) trees." Database accession no. PREV199900111265 XP002169817 & JOURNAL OF APPLIED BOTANY, Bd. 72, Nr. 5-6, Dezember 1998 (1998-12), Seiten 207-211, ISSN: 0949-5460 *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102128915A (zh) * 2010-12-02 2011-07-20 南京大学 利用蚯蚓生物标志物诊断土壤污染状况的方法
CN103675233A (zh) * 2013-11-26 2014-03-26 南京信息工程大学 利用农作物标志物检测土壤污染的方法
CN103837632A (zh) * 2014-03-07 2014-06-04 山东省科学院海洋仪器仪表研究所 液相色谱流动注射化学发光法测量水体中镉的装置及方法
CN103837632B (zh) * 2014-03-07 2015-06-24 山东省科学院海洋仪器仪表研究所 液相色谱流动注射化学发光法测量水体中镉的装置及方法
CN106442805A (zh) * 2016-11-23 2017-02-22 新疆农业科学院农产品贮藏加工研究所 一种库尔勒香梨果实石细胞中磨木木质素提取的方法

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