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BS EN 17718:2024 Plant biostimulants. Determination of Rhizobium spp., 2024
- undefined
- European foreword
- Introduction
- 1 Scope
- 2 Normative references
- 3 Terms and definitions
- 4 Enumeration of Rhizobium spp., Mesorhizobium spp., Ensifer spp., Bradyrhizobium spp. [Go to Page]
- 4.1 General
- 4.2 Sample preparation [Go to Page]
- 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
- 4.2.6 Solid pellets, granules, microgranules - slow release - formulations
- 4.2.7 Solid substrate
- 4.3 Serial dilution
- 4.4 Preparation of the culture media
- Figure 1 — Scheme of serial dilutions [Go to Page]
- 4.5 Spread plate counting
- 4.6 Plate counts of Rhizobiaceae in sterile diluent
- 4.7 Growing media
- Table 2 —Growing media for the growth of the different microorganisms [Go to Page]
- 4.8 Calculation
- 5 Species determination of Rhizobium spp., Mesorhizobium spp., Ensifer spp., Bradyrhizobium spp. via genetic analysis [Go to Page]
- 5.1 General
- 5.2 Preparation of the sample for genomic DNA extraction [Go to Page]
- 5.2.1 Isolation and preparation of the microorganism
- 5.2.2 Sample concentration
- 5.2.3 DNA extraction and storage
- 5.2.4 Partial PCR amplification of the 16S rRNA genes [Go to Page]
- 5.2.4.1 PCR amplification
- 5.2.4.2 Visualization of PCR products
- 5.2.4.3 Sequencing of partial 16S rRNA genes
- 5.2.4.4 Whole-genome sequencing
- Annex A (normative) Formula of culture media7F
- A.1 YT (Tryptone-Yeast Medium) for Rhizobium spp., Mesorhizobium spp. and Ensifer spp.
- A.1.1 Ingredients of YT medium
- Table A.1 — Ingredients of YT medium
- A.1.2 Method to prepare YT media
- A.2 Modified Yeast extract mannitol agar medium (YEMA)alternative medium for Rhizobium spp.
- A.2.1 Ingredients of YEMA medium (and suggested suppliers)
- Table A.2 — Ingredients of YEMA medium
- A.2.2 Method to prepare YEMA medium
- A.3 Reasoner's 2A Agar Medium (R2A) for Bradyrhizobium spp.
- A.3.1 Ingredients of R2A medium (and suggested suppliers)
- Table A.3 — Ingredients of R2A media
- A.3.2 Method to prepare Reasoner's 2A Agar (R2A) medium
- A.4 Congo Red stock solution for YEMA medium
- A.4.1 Congo Red stock solution medium preparation
- Table A.4 — Ingredients of Congo red stock solution medium
- A.5 0,1 M Phosphate Buffered Saline (PBS)
- A.6 Nutrient broth
- A.7 Buffered Peptone Water
- A.7.1 Composition
- A.7.2 Preparation
- Annex B (informative) Repeatability and reproducibility of the method
- B.1 Materials used in the interlaboratory comparison study
- Table B.1 — Materials tested in the interlaboratory study for the enumeration and determination of Rhizobium spp., Mesorhizobium spp., Ensifer spp., or Bradyrhizobium spp. in plant biostimulant
- B.2 Interlaboratory comparison results
- Table B.2 — Results of the interlaboratory comparison study of the enumeration of Rhizobium spp., Mesorhizobium spp., Ensifer spp., or Bradyrhizobium spp. in biostimulants
- Table B.3 — Samples distributed for the interlaboratory comparison study of the determination of Rhizobium spp., Mesorhizobium spp., Ensifer spp., or Bradyrhizobium spp. in biostimulants
- 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
- Table ZA.1 — Correspondence between this European Standard and Regulation (EU) 2019/1009
- Bibliography [Go to Page]