Morphologic and agronomic trait characterisation method

In order to rationalise observations and benefit from the experience embodied in some statutory evaluations, UPOV (International Union for the Protection of New Varieties of Plants) reference documents were accessed. On the UPOV website, countries in charge of the “DUS” testing for O. viciifolia are recorded. These services are responsible for testing whether the variety is distinct (D) from any other variety and that it is sufficiently uniform (U) and stable (S) in terms of maintaining its characters across years. The examination generates a description of the variety, using its relevant characteristics (e.g. plant height, leaf shape, time of flowering), by which it can be defined as a variety. The countries mentioned are Czech Republic, Germany, Spain, France, Hungary, Poland and Ukraine. Since cultivation of O viciifolia has declined in recent years, only one reference document from the German federal authority Bundessortenamt (BSA) was available. These German national guidelines (Table 5) were used in DUS (Distinctness Uniformity and Stability) testing of O. viciifolia in the 1990’s.

for O. viciifolia

Trait Mark Explanation

Plant height in year of sowing 3, 5, 7 Small, medium, tall Leaf colour in year of sowing 3, 5, 7 Light green, medium green, dark

green

Plant height in spring 3, 5, 7 Small, medium, tall Leaf colour in spring 3, 5, 7 Light green, medium green, dark

green

Time of flowering 3, 5, 7 Early, medium, late

Leaf length 3, 5, 7 Small, medium, long

Leaflet length 3, 5, 7 Small, medium, long

Stem length 3, 5, 7 Small, medium, long

Inflorescence length 3, 5, 7 Small, medium, long

Stem thickness 3, 5, 7 Thin, medium, thick

Plant height at green seed stage

3, 5, 7 Small, medium, tall

Plant colour at green seed stage

3, 5, 7 Light green, medium green, dark green

This DUS protocol was used as an initial basis for development of an O viciifolia evaluation protocol within this project. Normally, new varieties are assessed against designated reference varieties, however as no reference varieties were known, the BSA notation based on the 3, 5, 7 numbers had to be adapted.

2.2.4.1. Agronomical characterisation

Agronomical characterisation was performed on 162 accessions from 2007 to 2010. Eleven traits were measured or characterised, sometimes several times in 2008, 2009 and or 2010. They are listed below.

The ‘survival’ of the accession was measured in October 2008 (notation from 1 to 9) and in April 2009 (number of alive plants left). It corresponds to the plant persistency measured either by scoring method (1 for no persistency to 9

with 100% persistency) or by number of alive plants.

The ‘score’ of the accession was measured in April 2008, June 2008, April 2009 and June 2010. It corresponds to the overall state of each plot. Plant vigour, infection by diseases and foliage density were all taken in account to give a representative score (1 for empty plot to 9 for ideal plot with healthy and vigorous plants).

The ‘flowering date’ was measured in 2008 and 2009. It is the date (day of the year) when a plot reached on average the full flowering stage (when 50% of the stems bear inflorescence with opened flowers in the lower half).

The ‘fusarium’ notation was measured in October 2008 and July 2009. It corresponds respectively to the presence (1) or absence (0) of Fusarium sp. infection symptoms and to the number of plants showing Fusarium sp. infection symptoms per plot.

The ‘mildew’ notation was taken in October 2008. It corresponds to the presence (1) or absence (0) of Erysiphe trifolii infection symptoms.

The accession ‘height’ was measured in 2008. It corresponds to the height (in cm) reached on average by full flowering stage plants measured from the soil surface.

The accession ‘weight’ was measured in 2008 and 2009. It corresponds to the dry matter production per plot (in g) in the first cut at full flowering stage estimated by fresh weight converted with dried sample.

The ‘soil cover’ accession abilities were measured in October 2008. It was assessed by a notation from 1 to 9 corresponding to the presence of plant covering the soil (1 for empty plot to 9 for no visible soil).

The ‘flower presence’ was measured in October 2008 and July 2009. It corresponds to the abundance of flowers that regrow after cutting (0 for no flower, 1 for few flowers and 2 for numerous flowers).

The accession ‘regrowth’ abilities were measured in July 2009. It was represented by a 1 to 9 notation and corresponds to the ability of a plant to regrow (1 for no regrowth after cut to 9 for vigourous regrowth).

2.2.4.2. Morphological characterisation

Preliminary morphological characterisation was performed on all accessions in 2008. Then, more complete evaluations were performed on the selected accessions in the second year (2009). Twelve traits were measured or characterised for 3 randomly selected plants in each selected plot, they are listed below.

The ‘leaf colour’ was measured in 2008. It was a 3 to 7 notation (3 for light green, 5 for medium and 7 for dark green).

The ‘flower colour’ was measured in 2008 and 2009. It was a 1 to 9 notation (1 for white flowers and 9 for red flowers).

The ‘stem colour’ was measured in 2008 and 2009. It was a 1 to 9 notation and corresponds to the largest stem colour from totally green (1) to totally red (9).

The ‘number of leaflets’ was measured in 2008 and 2009. It corresponds to the number of leaflets on the reference leaf (first mature leaf at the top of the largest stem).

The ‘leaflet width’ and ‘leaflet length’ were measured in 2008 and 2009. These correspond to the width and length (in cm) of the bottom leaflet on the reference leaf.

The ratio ‘leaflet length/leaflet width’ was calculated in 2008 and 2009.

The ‘inflorescence length’ and the ‘leaf length’ were measured in 2009. They correspond respectively to the measurement (in cm) of the reference inflorescence length (first inflorescence with 1/3 open flowers at the top of the largest stem) and of the reference leaf length.

The ‘habit’ was measured in 2008 and 2009. The notation from 1 to 4 corresponds to the plant growth habit (1: prostrate plant, 2: semi prostrate plant, 3: semi erect plant, 4: erect plant).

The ‘number of leaves per stem’ and the ‘number of stems per plant’ were measured in 2009. They correspond respectively to the number of leaves on the largest stem and to the number of stems on a randomly selected plant.

The ‘stem length’ was measured in 2009. It corresponds to the largest stem length in cm.

The ‘stem thickness’ was measured in 2008 and 2009. It corresponds to the largest stem thickness in mm.

The ‘number of inflorescences per stem’ was measured in 2009. It corresponds to the number of inflorescences on the largest stem.

The accession ‘homogeneity’ was measured in 2008 and 2009. This notation from 1 to 9 realised in full flowering stage (May to June) represented the plot homogeneity in terms of: inflorescence colour, stem colour, foliage colour, growth habit, height, phenological stage, inflorescence length, leaf shape (1 for a perfectly homogeneous plot to 9 for a completely non-homogeneous plot).

2.2.4.3. Selection of accessions of particular interest

A preliminary characterisation was attempted on all accessions in April 2008. As that many accessions represented too much material to analyse for the other partners of the project, a selection of accessions, representative of the diversity (geographic origin, Giant/Common, cultivar/wild) was further characterised (Appendix 1).