Methode01 20x20 crushed

Initial concept

We randomly select a 20cm x 20cm square in the plot, if possible avoiding edge effects. We cut everything in this square above ground level. The cultivated plant as well as any weeds growing in between. All this plants are store in a ziplock with an unique code. They will then be extracted as one entities to rate the chimiodiversity of the parcelle.

Methodology

Collecte

The 20cm X 20cm square is draw on the ground using a measuring tape. The location is randomly chose in on of the side row of the plot (the middle row is reserved for measuring yield and cannot be touched). All the crops, weeds and others wich are found in this square are cut at their base and put in zip-lock prelabbeled with an unique code. The entire zip-lock is then immersed in liquid nitrogen. The sample are store at -80°C waiting for the extraction process.

Each type of agricuture is replicated four times on the DOK experiment. That is allowing us to have four replications of our experiment. That means we have four samples of above-ground-biodynamic specimens and four samples of above-ground-conventional specimens.

Extraction

The entire contents of the zip-lock are transfered to a paper bag and put in the dry-freezer for 72 hours.

After lyophilization all the sample are weighted. They are then placed in a blender with a proportional amount of 200ml/40g of the dbgi extraction mix. The plants are blended until a homogeneous fluid is obtained. Six grams of this fluid are set appart in a falcon tube and 34 milliter of the dbgi mix are added to reconstituate the ratio of dbgi extraction. A second tube is completely filled with plant extract and stored at -80°C for future extractions.

Alternately the sample can be dry crushed in the blender and store as a dry powder. The dbgi mix can be add to the powder in the right proportion.

The tube are then centrifuged and the extraction process is performed in the same way as the dbgi extraction process.

Necessary change in method

During the collecte the zip-lock partially destroyed in liquid nitrogen. It is necessary to find another means of storage. Paper bag can be use for the liquide nitrogen and the dry-freezer. But zip lock are indispensable after the lyophilisation to keep the sample dry.