SIST-TS CEN ISO/TS 29843-2:2014
(Main)Soil quality - Determination of soil microbial diversity - Part 2: Method by phospholipid fatty acid analysis (PLFA) using the simple PLFA extraction method (ISO/TS 29843-2:2011)
Soil quality - Determination of soil microbial diversity - Part 2: Method by phospholipid fatty acid analysis (PLFA) using the simple PLFA extraction method (ISO/TS 29843-2:2011)
This part of ISO/TS 29843 specifies a simple method for the extraction of only phospholipid fatty acids (PLFA) from soils.
ISO/TS 29843-1 specifies an extended method for the extraction and determination of both PLFA and PLEL from soils.
Bodenbeschaffenheit - Bestimmung der Diversität von Bodenmikroorganismen - Teil 2: Verfahren mittels Phospholipidfettsäure(PLFA)-Analyse unter Verwendung des einfachen PLFA-Extraktionsverfahrens (ISO/TS 29843-2:2011)
Dieser Teil von ISO/TS 29843 legt ein einfaches Verfahren ausschließlich zur Extraktion von Phospholipid-fettsäuren (PLFA) aus Böden fest.
ISO/TS 29843 1 legt ein erweitertes Verfahren zur Extraktion und Bestimmung von Phospholipidfettsäuren (PLFA) und Phospholipidetherlipiden (PLEL) aus Böden fest.
Qualité du sol - Détermination de la diversité microbienne du sol - Partie 2 : Méthode par analyse des acides gras phospholipidiques (PLFA) en utilisant la méthode simple d'extraction des PLFA (ISO/TS 29843-2:2011)
L'ISO/TS 29843-2:2011 spécifie une méthode simple permettant d'extraire uniquement les acides gras phospholipidiques (PLFA) du sol.
Kakovost tal - Določanje mikrobne raznolikosti tal - 2. del: Analiza fosfolipidnih maščobnih kislin (PLFA) z enostavno ekstrakcijsko metodo PLFA (ISO/TS 29843-2:2011)
Ta del standarda ISO/TS 29843 določa enostavno metodo za ekstrakcijo samo fosfolipidnih maščobnih kislin (PLFA) iz tal.
ISO/TS 29843-1 določa razširjeno metodo za ekstrakcijo določanje PLFA in PLEL iz tal.
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
SIST-TS CEN ISO/TS 29843-2:2014
01-december-2014
.DNRYRVWWDO'RORþDQMHPLNUREQHUD]QROLNRVWLWDOGHO $QDOL]DIRVIROLSLGQLK
PDãþREQLKNLVOLQ 3/)$ ]HQRVWDYQRHNVWUDNFLMVNRPHWRGR3/)$ ,6276
Soil quality - Determination of soil microbial diversity - Part 2: Method by phospholipid
fatty acid analysis (PLFA) using the simple PLFA extraction method (ISO/TS 29843-
2:2011)
Qualité du sol - Détermination de la diversité microbienne du sol - Partie 2 : Méthode par
analyse des acides gras phospholipidiques (PLFA) en utilisant la méthode simple
d'extraction des PLFA (ISO/TS 29843-2:2011)
Ta slovenski standard je istoveten z: CEN ISO/TS 29843-2:2014
ICS:
13.080.30 Biološke lastnosti tal Biological properties of soils
SIST-TS CEN ISO/TS 29843-2:2014 en
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
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SIST-TS CEN ISO/TS 29843-2:2014
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SIST-TS CEN ISO/TS 29843-2:2014
TECHNICAL SPECIFICATION
CEN ISO/TS 29843-2
SPÉCIFICATION TECHNIQUE
TECHNISCHE SPEZIFIKATION
October 2014
ICS 13.080.30
English Version
Soil quality - Determination of soil microbial diversity - Part 2:
Method by phospholipid fatty acid analysis (PLFA) using the
simple PLFA extraction method (ISO/TS 29843-2:2011,
Corrected version 2012-02-01)
Qualité du sol - Détermination de la diversité microbienne Bodenbeschaffenheit - Bestimmung der Diversität von
du sol - Partie 2 : Méthode par analyse des acides gras Bodenmikroorganismen - Teil 2: Verfahren mittels
phospholipidiques (PLFA) en utilisant la méthode simple Phospholipidfettsäure(PLFA)-Analyse unter Verwendung
d'extraction des PLFA (ISO/TS 29843-2:2011) des einfachen PLFA-Extraktionsverfahrens (ISO/TS 29843-
2:2011)
This Technical Specification (CEN/TS) was approved by CEN on 11 August 2014 for provisional application.
The period of validity of this CEN/TS is limited initially to three years. After two years the members of CEN will be requested to submit their
comments, particularly on the question whether the CEN/TS can be converted into a European Standard.
CEN members are required to announce the existence of this CEN/TS in the same way as for an EN and to make the CEN/TS available
promptly at national level in an appropriate form. It is permissible to keep conflicting national standards in force (in parallel to the CEN/TS)
until the final decision about the possible conversion of the CEN/TS into an EN is reached.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, Former Yugoslav Republic of Macedonia, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania,
Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and United
Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Avenue Marnix 17, B-1000 Brussels
© 2014 CEN All rights of exploitation in any form and by any means reserved Ref. No. CEN ISO/TS 29843-2:2014 E
worldwide for CEN national Members.
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SIST-TS CEN ISO/TS 29843-2:2014
CEN ISO/TS 29843-2:2014 (E)
Contents Page
Foreword .3
2
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SIST-TS CEN ISO/TS 29843-2:2014
CEN ISO/TS 29843-2:2014 (E)
Foreword
The text of ISO/TS 29843-2:2011, Corrected version 2012-02-01 has been prepared by Technical Committee
ISO/TC 190 “Soil quality” of the International Organization for Standardization (ISO) and has been taken over
as CEN ISO/TS 29843-2:2014 by Technical Committee CEN/TC 345 “Characterization of soils” the secretariat
of which is held by NEN.
Attention is drawn to the possibility that some of the elements of this document may be the subject of patent
rights. CEN [and/or CENELEC] shall not be held responsible for identifying any or all such patent rights.
According to the CEN-CENELEC Internal Regulations, the national standards organizations of the following
countries are bound to announce this Technical Specification: Austria, Belgium, Bulgaria, Croatia, Cyprus,
Czech Republic, Denmark, Estonia, Finland, Former Yugoslav Republic of Macedonia, France, Germany,
Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland,
Portugal, Romania, Slovakia, Slovenia, Spain, Sweden, Switzerland, Turkey and the United Kingdom.
Endorsement notice
The text of ISO/TS 29843-2:2011, Corrected version 2012-02-01 has been approved by CEN as
CEN ISO/TS 29843-2:2014 without any modification.
3
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SIST-TS CEN ISO/TS 29843-2:2014
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SIST-TS CEN ISO/TS 29843-2:2014
TECHNICAL ISO/TS
SPECIFICATION 29843-2
First edition
2011-09-15
Corrected version
2012-0-
Soil quality — Determination of soil
microbial diversity —
Part 2:
Method by phospholipid fatty acid
analysis (PLFA) using the simple PLFA
extraction method
Qualité du sol — Détermination de la diversité microbienne du sol —
Partie 2: Méthode par analyse des acides gras phospholipidiques
(PLFA) en utilisant la méthode simple d’extraction des PLFA
Reference number
ISO/TS 29843-2:2011(E)
©
ISO 2011
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SIST-TS CEN ISO/TS 29843-2:2014
ISO/TS 29843-2:2011(E)
COPYRIGHT PROTECTED DOCUMENT
© ISO 2011
All rights reserved. Unless otherwise specified, no part of this publication may be reproduced or utilized in any form or by any means,
electronic or mechanical, including photocopying and microfilm, without permission in writing from either ISO at the address below or ISO’s
member body in the country of the requester.
ISO copyright office
Case postale 56 • CH-1211 Geneva 20
Tel. + 41 22 749 01 11
Fax + 41 22 749 09 47
E-mail copyright@iso.org
Web www.iso.org
Published in Switzerland
ii © ISO 2011 – All rights reserved
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SIST-TS CEN ISO/TS 29843-2:2014
ISO/TS 29843-2:2011(E)
Contents Page
Foreword .iv
Introduction . v
1 Scope . 1
2 Normative references . 1
3 Symbols and abbreviated terms (except chemical products and reagents) . 1
4 Principle . 1
5 Test materials . 2
5.1 Soil . 2
5.2 Reagents . 2
5.3 Apparatus . 4
6 Procedures . 5
6.1 Lipid extraction (Bligh-Dyer extraction) . 5
6.2 Separation of lipids by SI column . 5
6.3 Derivatization — Transmethylation — Clean-up . 5
6.4 PLFA analysis . 6
Bibliography . 7
© ISO 2011 – All rights reserved iii
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SIST-TS CEN ISO/TS 29843-2:2014
ISO/TS 29843-2:2011(E)
Foreword
ISO (the International Organization for Standardization) is a worldwide federation of national standards 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 organizations, 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.
International Standards are drafted in accordance with the rules given in the ISO/IEC Directives, Part 2.
The main task of technical committees is to prepare International Standards. Draft International Standards
adopted by the technical committees are circulated to the member bodies for voting. Publication as an
International Standard requires approval by at least 75 % of the member bodies casting a vote.
In other circumstances, particularly when there is an urgent market requirement for such documents, a technical
committee may decide to publish other types of document:
— an ISO Publicly Available Specification (ISO/PAS) represents an agreement between technical experts in
an ISO working group and is accepted for publication if it is approved by more than 50 % of the members
of the parent committee casting a vote;
— an ISO Technical Specification (ISO/TS) represents an agreement between the members of a technical
committee and is accepted for publication if it is approved by 2/3 of the members of the committee
casting a vote.
An ISO/PAS or ISO/TS is reviewed after three years in order to decide whether it will be confirmed for a further
three years, revised to become an International Standard, or withdrawn. If the ISO/PAS or ISO/TS is confirmed,
it is reviewed again after a further three years, at which time it must either be transformed into an International
Standard or be withdrawn.
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.
ISO/TS 29843-2 was prepared by Technical Committee ISO/TC 190, Soil quality, Subcommittee SC 4,
Biological methods.
ISO/TS 29843 consists of the following parts, under the general title Soil quality — Determination of soil
microbial diversity:
— Part 1: Method by phospholipid fatty acid analysis (PLFA) and phospholipid ether lipids (PLEL) analysis
— Part 2: Method by phospholipid fatty acid analysis (PLFA) using the simple PLFA extraction method
In this corrected version, the missing Equation (1) in 6.1 has been added.
iv © ISO 2011 – All rights reserved
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SIST-TS CEN ISO/TS 29843-2:2014
ISO/TS 29843-2:2011(E)
Introduction
Phospholipids are essential components of membranes of all living cells. Extracted from soil samples in fatty
acid form (PLFA: phospholipid fatty acids) or ether-linked isoprenoid side chains (PLEL: phospholipid ether
lipid), they provide quantitative and qualitative insights into the soil’s viable/active microbial biomass. Cellular
enzymes hydrolyse and release the phosphate group within minutes to hours following cell death (Reference [1]).
The determination of total PLFA and PLEL provides a quantitative measure of the viable biomass of soil, i.e.
microorganisms from all three domains of the biosphere (bacteria, fungi and archaebacteria). PLFA and PLEL
can also allow for taxonomic differentiation within complex microbial communities (References [2] and [3]).
This approach is now well established in soil ecology and serves as a phenotypic, and thus complementary,
tool to genotypic (molecular genetic) approaches for determining microbial diversity. Apart from taxonomic
descriptions, the PLFA technique enables the determination of physiological changes within microbial consortia.
For instance, the monoenic PLFA 16:1ω7c and 18:1ω7c are increasingly converted to the cyclopropyl fatty acids
cy17:0 and cy19:0 in Gram-negative bacteria in response to environmental stress (Reference [4]).
Different methodologies are available for the determination of soil fatty acids. These methodologies present
different levels of complexity when applied and provide different levels of resolution in the description of soil
microbial communities. ISO/TS 29843-1 deals with the generally called “extended PLFA extraction method” while
this part of ISO/TS 29843 deals with the generally called “simple PLFA extraction method” (References [5] and[6]).
This part of ISO/TS 29843, which deals with the simple PLFA extraction method, is accessible to most research
and analytical laboratories involved in soil sciences. This methodology can be used for a wide range of soils. It
provides a broad diversity measurement of a soil microbial community at the phenotypic level. It can be applied
to biomass estimation and can be used to differentiate microbial communities among different soil samples
(with the aid of an adapted statistical method). This method is especially adapted for detecting rapid changes
in the soil microbial community str
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