1、BS ISO 18763:2016Soil quality Determinationof the toxic effects ofpollutants on germination andearly growth of higher plantsBSI Standards PublicationWB11885_BSI_StandardCovs_2013_AW.indd 1 15/05/2013 15:06BS ISO 18763:2016 BRITISH STANDARDNational forewordThis British Standard is the UK implementati
2、on of ISO 18763:2016.The UK participation in its preparation was entrusted to TechnicalCommittee EH/4, Soil quality.A list of organizations represented on this committee can beobtained on request to its secretary.This publication does not purport to include all the necessaryprovisions of a contract.
3、 Users are responsible for its correctapplication. The British Standards Institution 2016.Published by BSI Standards Limited 2016ISBN 978 0 580 86044 7ICS 13.080.30Compliance with a British Standard cannot confer immunity fromlegal obligations.This British Standard was published under the authority
4、of theStandards Policy and Strategy Committee on 31 July 2016.Amendments/corrigenda issued since publicationDate T e x t a f f e c t e dBS ISO 18763:2016 ISO 2016Soil quality Determination of the toxic effects of pollutants on germination and early growth of higher plantsQualit du sol Dtermination d
5、es effets toxiques des polluants sur la germination et la croissance primaire des plantes suprieuresINTERNATIONAL STANDARDISO18763First edition2016-07-01Reference numberISO 18763:2016(E)BS ISO 18763:2016ISO 18763:2016(E)ii ISO 2016 All rights reservedCOPYRIGHT PROTECTED DOCUMENT ISO 2016, Published
6、in SwitzerlandAll rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized otherwise in any form or by any means, electronic or mechanical, including photocopying, or posting on the internet or an intranet, without prior written permission. Permission ca
7、n be requested from either ISO at the address below or ISOs member body in the country of the requester.ISO copyright officeCh. de Blandonnet 8 CP 401CH-1214 Vernier, Geneva, SwitzerlandTel. +41 22 749 01 11Fax +41 22 749 09 47copyrightiso.orgwww.iso.orgBS ISO 18763:2016ISO 18763:2016(E)Foreword ivI
8、ntroduction v1 Scope . 12 Normative references 13 Terms and definitions . 14 Principle 35 Reagents, test organisms and media . 36 Apparatus and materials 47 Treatment and preparation of samples 68 Procedure. 69 Measurement . 810 Calculation of the percentage inhibition . 911 Reference chemical .1012
9、 Precision 1013 Validity criteria .1114 Test report 11Annex A (informative) Application of the phytotoxicity test in transparent test plates on natural and artificial soils and soil materials, with different plant species 12Annex B (informative) Assemblage of test plates for the phytotoxicity test 1
10、4Annex C (informative) International interlaboratory comparisons on the phytotoxicity test .15Bibliography .20 ISO 2016 All rights reserved iiiContents PageBS ISO 18763:2016ISO 18763:2016(E)ForewordISO (the International Organization for Standardization) is a worldwide federation of national standar
11、ds bodies (ISO member bodies). The work of preparing International Standards is normally carried out through ISO technical committees. Each member body interested in a subject for which a technical committee has been established has the right to be represented on that committee. International organi
12、zations, governmental and non-governmental, in liaison with ISO, also take part in the work. ISO collaborates closely with the International Electrotechnical Commission (IEC) on all matters of electrotechnical standardization.The procedures used to develop this document and those intended for its fu
13、rther maintenance are described in the ISO/IEC Directives, Part 1. In particular the different approval criteria needed for the different types of ISO documents should be noted. This document was drafted in accordance with the editorial rules of the ISO/IEC Directives, Part 2 (see www.iso.org/direct
14、ives).Attention is drawn to the possibility that some of the elements of this document may be the subject of patent rights. ISO shall not be held responsible for identifying any or all such patent rights. Details of any patent rights identified during the development of the document will be in the I
15、ntroduction and/or on the ISO list of patent declarations received (see www.iso.org/patents).Any trade name used in this document is information given for the convenience of users and does not constitute an endorsement.For an explanation on the meaning of ISO specific terms and expressions related t
16、o conformity assessment, as well as information about ISOs adherence to the WTO principles in the Technical Barriers to Trade (TBT) see the following URL: Foreword - Supplementary informationThe committee responsible for this document is ISO/TC 190, Soil quality, Subcommittee SC 4, Biological method
17、s.iv ISO 2016 All rights reservedBS ISO 18763:2016ISO 18763:2016(E)IntroductionEcotoxicological testing of test soils or waste materials to be disposed on soil are required to assess the potential environmental risk resulting from soil pollution or the disposal of wastes such as sewage sludge on far
18、mland. There is also a need to monitor the quality of soil after reclamation of industrial sites. Therefore, a very practical and rapid germination and growth test has been developed based on seed germination and seedling growth in controlled environmental conditions.The assay, which does not requir
19、e any pretreatment of the seeds, is performed in “transparent test plates”, incubated vertically, to allow the roots and the shoots of the germinated seeds to be seen. After 72 h exposure, a picture of the transparent test plates is taken and can be analysed “by image analysis” for multiple endpoint
20、s, such as percentage of seed germination and of length of roots and shoots. To account for the plant species variability in sensitivity, the assays are performed with the seeds of three plant species: one monocotyl (Sorghum saccharatum) and two dicotyls (Lepidium sativum and Sinapis alba).A major a
21、dvantage of this test is that after the shooting and storing of the pictures of the test plates, the measurements by image analysis can be postponed to any appropriate timing.Reference or standard soils can be used as negative controls, such as, for example, the ISO standard artificial soil accordin
22、g to ISO 11269-1 and ISO 11269-2.Commercially available seeds, with a shelf life longer than one year, allow the use of this test at any time of the year.Two International interlaboratory comparisons demonstrated that the test provides good results.A substantial number of studies report data on the
23、application of this test on various types of soils and soil materials with several types of plant species. ISO 2016 All rights reserved vBS ISO 18763:2016BS ISO 18763:2016Soil quality Determination of the toxic effects of pollutants on germination and early growth of higher plants1 ScopeThis Interna
24、tional Standard describes a technique for determining the effects of soil and soil-related materials on the seed germination and early growth of higher plants. These endpoints are useful indicators for the assessment of the quality of a soil as a habitat for organisms. This International Standard is
25、 applicable to all soils in which soil organisms are active and may be used to evaluate: the effects on plants due to toxicity of solid or liquid chemicals contaminating soil or materials (compost, sludge, waste) and chemicals added to soil; the changes in the soil effect on plants after restoration
26、 measures.2 Normative referencesThe following documents, in whole or in part, are normatively referenced in this document and are indispensable for its application. For dated references, only the edition cited applies. For undated references, the latest edition of the referenced document (including
27、any amendments) applies.ISO 11269-1, Soil quality Determination of the effects of pollutants on soil flora Part 1: Method for the measurement of inhibition of root growthISO 11269-2, Soil quality Determination of the effects of pollutants on soil flora Part 2: Effects of contaminated soil on the eme
28、rgence and early growth of higher plantsISO/TS 20281, Water quality Guidance on statistical interpretation of ecotoxicity data3 Terms and definitionsFor the purposes of this document, the following terms and definitions apply.3.1artificial soilmixture of sand, kaolinite, peat and calcium carbonate p
29、repared according to ISO 11269-1 and ISO 11269-23.2control soilreference or standard soil used as a control and as a medium for preparing dilution series with test soils or a reference substance3.3reference soiluncontaminated site-specific soil (e.g. collected in the vicinity of a contaminated site)
30、 with similar properties (nutrient concentrations, pH, organic carbon content and texture) as the test soil3.4standard soilfield-collected soil or artificial soil whose main properties (e.g. pH, texture, organic matter content) are within a known rangeNote 1 to entry: The properties of standard soil
31、s can differ from the test soil.INTERNATIONAL STANDARD ISO 18763:2016(E) ISO 2016 All rights reserved 1BS ISO 18763:2016ISO 18763:2016(E)EXAMPLE Euro-soils1, artificial soil2, LUFA soil.1)3.5test soileither a natural or an artificial clean soil that is spiked with the test substance or a contaminate
32、d natural soil (a site soil)53.6seeding emergenceappearance of a visible seedling above the surface of the cover materialSOURCE: ISO 17126:2005, 3.1, modified3.7germinationappearance of a root of at least 1 mm of length3.8pure watergrade of water, produced, for example, by single distillation, by de
33、-ionization, by ultra-filtration or by reverse osmosis53.9root lengthlength of the root from seed to root tip3.10shoot lengthlength of the part that grows upward, from seed to tip3.11water saturationmaximum water content that a soil can retain against gravity under undisturbed soil conditions, conve
34、ntionally stated as water content two days to three days after full saturation with waterSOURCE: ISO 11074:2015, 2.1.5 field capacity, modified3.12water saturated soilsoil which has reached its maximum water content3.13water-holding capacitymass of water that evaporates from soil saturated with wate
35、r when the soil is dried to constant mass at 105 C, divided by the dry mass of the soil73.14negative controlany well-characterized material or substance that, when tested by a specific procedure, demonstrates the suitability of the procedure to yield a reproducible, appropriately negative, non-react
36、ive or minimal response in the test systemSOURCE: ISO 10993-10:2010, 3.12, modified3.15effect percentagepercentage decrease of the seed germination and the growth of the plant roots and/or shoots in the test soil in comparison to the control soil1) Euro-soils, artificial soil and LUFA soil are examp
37、les of suitable products available commercially. This information is given for the convenience of users of this document and does not constitute an endorsement by ISO of these products.2 ISO 2016 All rights reservedBS ISO 18763:2016ISO 18763:2016(E)4 PrincipleThis method compares the seed germinatio
38、n and early growth of monocotyledonous and dicotyledonous plants in a test soil and/or a series of mixtures with a control soil. This method may also be used for the testing of compost, sludge or waste.Seeds of one monocotyledonous plant, such as Sorghum saccharatum (L.) Moench, and two dicotyledono
39、us plants, such as Lepidium sativum L. and Sinapis alba L., are exposed to the test material under controlled conditions. After (72 1) h, the number of germinated seeds is recorded and the length of the roots of the test plants is measured in the test soil and in the control soil.If different seed s
40、pecies are used, the length of the incubation period may be adjusted, depending on the time of germination of the seeds and the growth speed of the roots.The test makes use of unique flat and shallow transparent test plates (6.3) composed of two compartments, the lower one of which contains hydrated
41、 soil.Seeds of the selected test plants are positioned at equal distance near the middle ridge of the test plate (6.3) on a black filter paper (6.5) placed on top of the hydrated soil.After closing the test plates (6.3) with their transparent cover, the test plates are placed vertically in a holder
42、(6.4) and incubated at (25 1) C for (72 1) h.At the end of the incubation period, the length of each root (and shoot, if wished) can be measured directly with a ruler and recorded.Alternatively, a “digital” picture is taken of the test plates (6.3) with the germinated plants (either with a digital c
43、amera, a webcam camera or a flatbed paper scanner) for storage in a computer file. The subsequent root length measurements are performed by image analysis. The analyses on germination and root growth can then be made immediately or postponed to any appropriate timing.NOTE The same procedure can be a
44、pplied to also measure the shoot height, if wished. Calculation of the shoot/root length ratio is a possible additional effect parameter.5 Reagents, test organisms and media5.1 Water.Pure water having a conductivity below 10 S/cm.5.2 Test organisms.The test organisms are seeds of one monocotyledonou
45、s plant, such as Sorghum saccharatum (L.) Moench, and two dicotyledonous plants, such as Lepidium sativum L. and Sinapis alba L.Investigations have been performed not only with the three plant species indicated in 5.2 but also with other monocotyl and dicotyl plant species. A synthesis on these publ
46、ished studies is given in Annex A.Seeds coated with insecticides and/or fungicides should be avoided.5.3 Control soil.Either reference or standard soils can be used as the control soil, if unhindered growth of the test plants in these soils can be expected.When comparing the root elongation in soils
47、 of known and unknown quality, the control soil and soil under test should be of the same textural class, and be as similar as practicable in all respects other than the presence of the chemical or contaminant being investigated. Indeed, significant differences in soil characteristics other than the
48、 presence of contaminant may lead to differences in root lengths and may induce false positive test results. ISO 2016 All rights reserved 3BS ISO 18763:2016ISO 18763:2016(E)Alternatively, artificial soil according to ISO 11269-1 and ISO 11269-2 may be used. The substrate called “artificial soil” has
49、 the following composition:Percentage expressed on dry-mass basisSphagnum peat finely ground and with no visible plant remains 10 %Kaolinite clay containing not less than 30 % kaolinite 20 %Industrial quartz sand (dominant fine sand with more than 50 % of particle size 0,05 mm to 0,2 mm)69 %NOTE As indicated in ISO 11269-1, 5 % peat have proven to be sufficient for maintaining the desired structure of the artificial soil (with a corresponding increase of the sand percentage to 75 %). A lower percentage of kaolini
