BSI PD CEN/TS 17722:2022
$167.15
Plant biostimulants. Determination of mycorrhizal fungi
Published By | Publication Date | Number of Pages |
BSI | 2022 | 44 |
This document was developed to provide a horizontal method for enumeration and genera/species determination [1], [2], [3] of mycorrhizal fungi in plant biostimulants products in accordance with the EU Fertilising Products Regulation.
PDF Catalog
PDF Pages | PDF Title |
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2 | undefined |
9 | 1 Scope 2 Normative references 3 Terms and definitions |
11 | 4 Methods for the quantification of mycorrhiza 4.1 General |
13 | 4.2 How to prepare the initial sample 4.2.1 General 4.2.2 Liquid – water-based formulations 4.2.3 Liquid – oil-based (emulsifiable concentrate EC) formulations 4.2.4 Solid – wettable powder (WP) formulations 4.2.5 Solid – water dispersible granules (WDG) formulations |
14 | 4.2.6 Solid – pellets, granules, microgranules (slow release) formulations 4.2.7 Solid – substrate 4.3 Enumeration methods 4.3.1 General 4.3.2 Method N° 1: Spore isolation and counting MTT 4.3.2.1 Procedure for enumeration of spores |
15 | 4.3.2.2 Viability of spores |
16 | 4.3.3 Method N° 2: Procedure for clearing and staining root samples and enumeration of vesicles in the stained root samples 4.3.3.1 Materials and equipment |
17 | 4.3.3.2 Procedure |
19 | 4.3.4 Enumeration of the total number of UPM in the product using Method N°1 + Method N°2 4.3.5 Method N°3: Endomycorrhiza bioassay 4.3.5.1 Principle of the MPN (Most Probable Number of mycorrhizal propagule) |
24 | 4.3.5.2 Protocols and examples |
27 | 4.3.6 Method N°4: Ectomycorrhiza and ericoid count 4.3.6.1 General 4.3.6.2 Sample preparation 4.3.6.3 Serial dilution 4.3.6.4 Plate counts of ectomycorrhiza in sterile diluent |
28 | 4.3.6.5 Spread-plate counting with modified Melin Norkrans Medium (MNM) agar 4.3.6.6 Calculation |
31 | 5 Molecular characterization and identification of mycorrhiza isolates 5.1 General 5.2 Materials and equipment 5.3 Method for the molecular characterization and identification of mycorrhiza isolates 5.3.1 Spores cleaning |
32 | 5.3.2 DNA extraction |
33 | 5.3.3 Preparation for PCR 5.3.3.1 Primer sequences 5.3.3.2 Master mix preparation |
34 | 5.3.3.3 Procedure for running PCR 5.3.3.4 PCR program for each primer set |
35 | 5.3.4 Preparation for gel-electrophoresis 5.3.4.1 Materials and equipment 5.3.4.2 Procedure |
36 | 5.3.5 Direct sequencing (outsourced sequencing lab) 6 Method of molecular characterization and identification for ectomycorrhiza and ericoid 6.1 General |
37 | 6.2 Materials 6.2.1 Fungal material 6.2.2 Molecular biology kits/chemicals |
38 | 6.2.3 Equipment 6.3 Detailed description of method 6.3.1 Material preparation |
39 | 6.3.2 DNA extraction and quality check 6.3.3 PCR amplification of ITS sequences |
40 | 6.3.4 Gel electrophoresis and PCR product visualization |