FprEN 17722
(Main)Plant biostimulants - Determination of mycorrhizal fungi
Plant biostimulants - Determination of mycorrhizal fungi
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 fertilizing 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.
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
Le présent document spécifie une méthode horizontale pour le dénombrement et la détermination du genre/de l’espèce [2], [3], [4] des champignons mycorhiziens dans les biostimulants microbiens des végétaux.
Le présent document est applicable aux mélanges de fertilisants constitués d’au moins deux composants appartenant aux catégories de fertilisants UE suivantes : engrais, amendements minéraux basiques, amendements du sol, supports de culture, biostimulants des végétaux et, dans lesquels la catégorie Biostimulants des végétaux représente le pourcentage le plus élevé du mélange en masse ou en volume, ou dans le cas d’un liquide, en masse sèche. Si les biostimulants des végétaux ne représentent pas le pourcentage le plus élevé du mélange, la Norme européenne relative au composant représentant le pourcentage le plus élevé du mélange s’applique. Dans le cas où un mélange de fertilisants contient des composants en quantité égale ou dans le cas où les composants des fertilisants UE utilisés pour le mélange ont des formulations identiques, l’utilisateur décide quelle norme appliquer.
Rastlinski biostimulanti - Določanje mikoriznih gliv
General Information
Relations
Standards Content (Sample)
SLOVENSKI STANDARD
oSIST prEN 17722:2023
01-maj-2023
Rastlinski biostimulanti - Ugotavljanje prisotnosti in števila 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: prEN 17722
ICS:
65.080 Gnojila Fertilizers
oSIST prEN 17722:2023 en,fr,de
2003-01.Slovenski inštitut za standardizacijo. Razmnoževanje celote ali delov tega standarda ni dovoljeno.
oSIST prEN 17722:2023
oSIST prEN 17722:2023
DRAFT
EUROPEAN STANDARD
prEN 17722
NORME EUROPÉENNE
EUROPÄISCHE NORM
April 2023
ICS 65.080 Will supersede 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 draft European Standard is submitted to CEN members for enquiry. It has been drawn up by the Technical Committee
CEN/TC 455.
If this draft becomes a European Standard, 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.
This draft European Standard was established by CEN 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.
Recipients of this draft are invited to submit, with their comments, notification of any relevant patent rights of which they are
aware and to provide supporting documentation.
Warning : This document is not a European Standard. It is distributed for review and comments. It is subject to change without
notice and shall not be referred to as a European Standard.
EUROPEAN COMMITTEE FOR STANDARDIZATION
COMITÉ EUROPÉEN DE NORMALISATION
EUROPÄISCHES KOMITEE FÜR NORMUNG
CEN-CENELEC Management Centre: Rue de la Science 23, B-1040 Brussels
© 2023 CEN All rights of exploitation in any form and by any means reserved Ref. No. prEN 17722:2023 E
worldwide for CEN national Members.
oSIST prEN 17722:2023
prEN 17722:2023 (E)
Contents Page
European foreword . 4
Introduction . 5
1 Scope . 7
2 Normative references . 7
3 Terms and definitions . 7
4 Methods for the quantification of mycorrhiza .10
4.1 General .10
4.2 How to prepare the initial sample .12
4.2.1 General .12
4.2.2 Liquid – water-based formulations .12
4.2.3 Liquid – oil-based (emulsifiable concentrate EC) formulations .12
4.2.4 Solid – wettable powder (WP) formulations .12
4.2.5 Solid – water dispersible granules (WDG) formulations .12
4.2.6 Solid – pellets, granules, microgranules (slow release) formulations.13
4.2.7 Solid – substrate .13
4.3 Enumeration methods .13
4.3.1 General .13
4.3.2 Method N° 1: Spore isolation and counting MTT .13
4.3.3 Method N° 2: Procedure for clearing and staining root specimens and enumeration of
vesicles in the stained root samples .15
4.3.4 Enumeration of the total number of UPM in the product using Method N°1 + Method
N°2 .18
4.3.5 Method N°3: Endomycorrhiza Bioassay .18
4.3.6 Method N°4: Ectomycorrhiza and Ericoid count .26
5 Molecular characterization and identification of mycorrhiza isolates .31
5.1 General .31
5.2 Materials and equipment .31
5.3 Method for the molecular characterization and identification of mycorrhiza isolates
....................................................................................................................................................................31
5.3.1 Spores cleaning .31
5.3.2 DNA extraction .32
5.3.3 Preparation for PCR .33
5.3.4 Preparation for gel-electrophoresis .35
5.3.5 Direct sequencing (outsourced sequencing lab) .36
6 Method of molecular characterization and identification for ectomycorrhiza and
ericoid .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 method .38
6.3.1 Material preparation .38
6.3.2 DNA extraction and quality check .38
oSIST prEN 17722:2023
prEN 17722:2023 (E)
6.3.3 PCR amplification of ITS sequences . 39
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
oSIST prEN 17722:2023
prEN 17722:2023 (E)
European foreword
This document (prEN 17722:2023) has been prepared by Technical Committee CEN/TC 455 “Plant
Biostimulants”, the secretariat of which is held by AFNOR.
This document is currently submitted to the CEN enquiry.
This document will supersede CEN/TS 17722:2022.
This document has been prepared under a Standardization Request given to CEN by the European
Commission and the European Free Trade Association, and supports essential requirements of EU
Directive(s) / Regulation(s).
For relationship with EU Directive(s) / Regulation(s), see informative Annex ZA, which is an integral part
of this document.
oSIST prEN 17722:2023
prEN 17722:2023 (E)
Introduction
This document was prepared by the experts of CEN/TC 455 “Plant Biostimulants”. 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 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 M/564 Amd1, also contributes to the Communication on “Innovating for
Sustainable Growth: A Bio economy for Europe”. Working Group 5 “Labelling and denominations”, was
created to develop a work program as part of this standardization request.
Technical committee CEN/TC 455 “Plant Biostimulants” was established to carry out the work program
that will prepare a series of standards. 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 plant 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
plant biostimulants, and will promote and support commercialisation of the European plant biostimulant
industry.
The plant biostimulants used in agriculture can be applied in multiple ways: on soil, on plants, as seed
treatment, etc. A microbial plant biostimulant consists of a microorganism or a consortium of
microorganisms, as referred to in Component Material Category 7 of Annex II of the EU Fertilising
Products Regulation.
This document is applicable to all plant biostimulants in agriculture based on live microorganisms
belonging to the mycorrhiza.
Table 1 summarizes many of the agro-ecological principles and the role played by plant biostimulants.
Table 1 — Agro-ecological principles and the role played by plant biostimulants
Increase biodiversity
By improving soil microorganism quality/quantity
Reinforce biological regulation and interactions
By reinforcing plant-microorganism interactions
— symbiotic exchanges i.e. mycorrhiza
— symbiotic exchanges i.e. rhizobiaciae/fava
— secretions mimicking plant hormones (i.e. trichoderma)
By regulating plant physiological processes
— e.g. growth, metabolism, plant development
Improve biogeochemical cycles
— improve absorption of nutritional elements
— improve bioavailability of nutritional elements in the soil
— stimulate degradation of organic matter
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.
oSIST prEN 17722:2023
prEN 17722:2023 (E)
IMPORTANT — It is absolutely essential that tests conducted in accordance with this document be
carried out by suitably trained staff.
oSIST prEN 17722:2023
prEN 17722:2023 (E)
1 Scope
This standard was developed to provide a horizontal method for enumeration and genera/species
determination [1], [2], [3] of mycorrhizal fungi in microbial plant biostimulant in accordance with the
Regulation (EU) 2019/1009 of the European Parliament and of the Council.
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 d
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