SIST EN 17722:2025
(Main)Plant biostimulants - Determination of mycorrhizal fungi
Plant biostimulants - Determination of mycorrhizal fungi
This European Standard establishes the methodology for the determination of Mycorrhizal fungi in plant biostimulant products in accordance to the future Regulation of EU fertilising products.
Pflanzen-Biostimulanzien - Bestimmung von Mykorrhizapilzen
Diese Norm wurde entwickelt, um ein horizontales Verfahren für die Zählung und die Bestimmung der Gattung/Spezies [1] [2] [3] von Mykorrhizapilzen in mikrobiellen Pflanzen-Biostimulanzien in Übereinstimmung mit der Verordnung (EU) 2019/1009 des Europäischen Parlaments und des Rates bereitzustellen.
Biostimulants des végétaux - Détermination des champignons mycorhiziens
La présente norme a été élaborée pour fournir une méthode horizontale pour le dénombrement et la détermination du genre/de l’espèce [1], [2], [3] des champignons mycorhiziens dans un biostimulant microbien des végétaux conformément au Règlement UE 2019/1009 du Parlement Européen et du Conseil.
Rastlinski biostimulanti - Določanje mikoriznih gliv
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
01-februar-2025
Nadomešča:
SIST-TS CEN/TS 17722:2023
Rastlinski biostimulanti - Določanje mikoriznih gliv
Plant biostimulants - Determination of mycorrhizal fungi
Pflanzen-Biostimulanzien - Bestimmung von Mykorrhizapilzen
Biostimulants des végétaux - Détermination des champignons mycorhiziens
Ta slovenski standard je istoveten z: EN 17722:2024
ICS:
65.080 Gnojila Fertilizers
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
EN 17722
EUROPEAN STANDARD
NORME EUROPÉENNE
November 2024
EUROPÄISCHE NORM
ICS 65.080 Supersedes CEN/TS 17722:2022
English Version
Plant biostimulants - Determination of mycorrhizal fungi
Biostimulants des végétaux - Détermination des Pflanzen-Biostimulanzien - Bestimmung von
champignons mycorhiziens Mykorrhizapilzen
This European Standard was approved by CEN on 26 August 2024.
CEN members are bound to comply with the CEN/CENELEC Internal Regulations which stipulate the conditions for giving this
European Standard the status of a national standard without any alteration. Up-to-date lists and bibliographical references
concerning such national standards may be obtained on application to the CEN-CENELEC Management Centre or to any CEN
member.
This European Standard exists in three official versions (English, French, German). A version in any other language made by
translation under the responsibility of a CEN member into its own language and notified to the CEN-CENELEC Management
Centre has the same status as the official versions.
CEN members are the national standards bodies of Austria, Belgium, Bulgaria, Croatia, Cyprus, Czech Republic, Denmark, Estonia,
Finland, France, Germany, Greece, Hungary, Iceland, Ireland, Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway,
Poland, Portugal, Republic of North Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and
United Kingdom.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2024 CEN All rights of exploitation in any form and by any means reserved Ref. No. EN 17722:2024 E
worldwide for CEN national Members.
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 6
2 Normative references . 6
3 Terms and definitions . 6
4 Methods for the quantification of mycorrhizae . 9
4.1 General . 9
4.2 How to prepare the initial sample .10
4.2.1 General .10
4.2.2 Liquid (water-based) formulations .10
4.2.3 Liquid (oil-based) emulsifiable concentrate (EC) formulations .10
4.2.4 Solid wettable powder (WP) formulations .10
4.2.5 Solid water dispersible granules (WDG) formulations .10
4.2.6 Solid pellets, granules, microgranules (slow release) formulations .11
4.2.7 Solid substrate .11
4.2.8 Optional procedure for homogenization for solid .11
4.3 Enumeration methods .11
4.3.1 General .11
4.3.2 Method N°1: Spore isolation and counting with MTT .11
4.3.3 Method N°2: Procedure for the clearing and the staining of the root specimens and for
the enumeration of spores/vesicles in the stained root samples .15
4.3.4 Enumeration of UPM [9] in the product using Method N°1 + Method N°2 .18
4.3.5 Method N°3: Endomycorrhizae bioassay .18
4.3.6 Method N°4: Ectomycorrhizae and ericoid mycorrhizae count on plates .26
5 Molecular characterization and identification of mycorrhizal isolates .30
5.1 General .30
5.2 Materials and equipment .30
5.3 Method for the molecular characterization and identification of mycorrhiza isolates
....................................................................................................................................................................30
5.3.1 Spores cleaning .30
5.3.2 DNA extraction .31
5.3.3 Preparation for polymerase chain reaction (PCR) .32
5.3.4 Preparation for gel-electrophoresis .34
5.3.5 Direct sequencing (outsourced sequencing lab) .35
6 Method of molecular characterization and identification for ectomycorrhizae and
ericoid mycorrhizae .36
6.1 General .36
6.2 Materials .36
6.2.1 Fungal material .36
6.2.2 Molecular biology kits/chemicals .36
6.2.3 Equipment .37
6.3 Detailed description of the method .37
6.3.1 Material preparation .37
6.3.2 DNA extraction and quality check .38
6.3.3 PCR amplification of ITS sequences . 38
6.3.4 Gel electrophoresis and PCR product visualization . 39
Annex A (informative) Repeatability and reproducibility of the method . 40
A.1 Materials used in the interlaboratory comparison study . 40
A.2 Interlaboratory comparison results . 41
Annex ZA (informative) Relationship of this European Standard and the essential
requirements of Regulation (EU) 2019/1009 making available on the market of EU
fertilising products aimed to be covered . 43
Bibliography . 44
European foreword
This document (EN 17722:2024) has been prepared by Technical Committee CEN/TC 455 “Plant
biostimulants”, the secretariat of which is held by AFNOR.
This European Standard shall be given the status of a national standard, either by publication of an
identical text or by endorsement, at the latest by May 2025, and conflicting national standards shall be
withdrawn at the latest by May 2025.
Attention is drawn to the possibility that some of the elements of this document may be the subject of
patent rights. CEN shall not be held responsible for identifying any or all such patent rights.
This document supersedes CEN/TS 17722:2022.
— the European foreword has been updated;
— the Introduction has been updated;
— the Bibliography has been re-numbered;
— Clause 4 has been reworked;
— Clause 5 has been reworked;
— Clause 6 has been reworked;
— Annex A on repeatability and reproducibility of the method has been added;
— Annex ZA has been added.
This document has been prepared under a standardization request addressed to CEN by the European
Commission. The Standing Committee of the EFTA States subsequently approves these requests for its
Member States.
For the relationship with EU Legislation, see informative Annex ZA, which is an integral part of this
document.
Any feedback and questions on this document should be directed to the users’ national standards body.
A complete listing of these bodies can be found on the CEN website.
According to the CEN-CENELEC Internal Regulations, the national standards organisations of the
following countries are bound to implement this European Standard: Austria, Belgium, Bulgaria, Croatia,
Cyprus, Czech Republic, Denmark, Estonia, Finland, France, Germany, Greece, Hungary, Iceland, Ireland,
Italy, Latvia, Lithuania, Luxembourg, Malta, Netherlands, Norway, Poland, Portugal, Republic of North
Macedonia, Romania, Serbia, Slovakia, Slovenia, Spain, Sweden, Switzerland, Türkiye and the United
Kingdom.
Introduction
The European Committee for Standardization (CEN) was requested by the European Commission (EC) to
draft European Standards or European Standardization deliverables to support the implementation of
Regulation (EU) 2019/1009 of 5 June 2019 [1] laying down rules on the making available on the market
of EU fertilising products (“FPR” or “Fertilising Products Regulation”).
This standardization request, presented as SR M/564 and relevant amendments, also contributes to the
Communication on “Innovating for Sustainable Growth: A Bio economy for Europe”. The interest in plant
biostimulants has increased significantly in Europe as a valuable tool to use in agriculture.
Standardization was identified as having an important role in order to promote the use of biostimulants.
The work of CEN/TC 455 seeks to improve the reliability of the supply chain, thereby improving the
confidence of farmers, industry, and consumers in biostimulants, and will promote and support
commercialisation of the European biostimulant industry.
WARNING — Persons using this document should be familiar with normal laboratory practice. This
document does not purport to address all of the safety problems, if any, associated with its use. It is the
responsibility of the user to establish appropriate safety and health practices and to ensure compliance
with any national regulatory conditions.
IMPORTANT — It is absolutely essential that tests conducted in accordance with this document be
carried out by suitably trained staff.
1 Scope
This document specifies a horizontal method for the enumeration and genus/species determination [2],
[3], [4] of mycorrhizal fungi in microbial plant biostimulants.
This document is applicable to the blends of fertilizing products where a blend is a mix of at least two of
the following component EU fertilising products categories: Fertilizers, Liming Materials, Soil Improvers,
Growing Media, Plant Biostimulants and where the following category Plant Biostimulants is the highest
percentage in the blend by mass or volume, or in the case of liquid form by dry mass. If Plant
Biostimulants is not the highest percentage in the blend, the European Standard for the highest
percentage of the blend applies. In case a blend of fertilizing products is composed of components in equal
quantity or in case the component EU fertilising products used for the blend have identical formulations ,
the user decides which standard to apply.
2 Normative references
There are no normative references in this document.
3 Terms and definitions
For the purposes of this document, the following terms and definitions apply.
ISO and IEC maintain terminology databases
...
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