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3. MORPHOLOGICAL VARIATION...
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3.2. MATERIAL AND METHODS
In total 11 L. multiflorum (8 cultivars and 3 ecotypes) and 12 L. perenne (8 cultivars
and 4 ecotypes) populations were used in morphological analyses. The origin of all populations
and seed sources are listed in Annex 13.1. All experiments were conducted from 2002
to 2003. The overall experimental design is presented in Figure 13.1.1 (Annex 13.1). The
fluorescence test was used to confirm the species identity (Annex 13.2). Seedling roots of
L. multiflorum fluoresce when placed under UV light, and those of L. perenne do not fluoresce.
From 20 to 30 genotypes were analysed per a population.
3.2.1. Analysed characters and sampling methods
The characters were selected on the basis of Terrell (1968), Loos (1993a) and Bennett
(1997), plus following own observations. Where possible each plant was scored for 17
morphological characters. Seedling characters (leaf and root length) were scored together
with seedling root fluorescence determination i.e., on the 14 th day after germination. Adult
vegetative characters were scored during two crops i.e., in July and September 2002 and
then averaged. For each plant, three average-size basal leaves were removed from a plant
and protected with a paper envelope. The leaf length and width were measured next day and
the data were averaged for each plant. Flowering characters were studied during the second
crop in July. Whenever it was possible the analysis was done both in 2002 and 2003 and
then averaged. The plant height was determined before cut. The three well developed spikes
together with flag leaves were collected from each plant, placed in a paper bag and used for
spike, spikelet and floret character measurements. The areas of vegetative and flag leaves
were counted by multiplication of the leaf length by leaf width. The green and dry weight
were measured twice i.e., in July and September. Recovery after winter was analysed as
a number of plants survived for which at least 50% of a tuft started to grow and developed
into mature plants.
3.2.2. Statistical methods
Data were analysed using STATISTICA 7.1 software. Range of variation was plotted
on the basis of maximum and minimum values and standard deviation. Analysis of variance
was used to check for population, species, form (cultivar or ecotype) and interaction species
x form effects. The LSD test (Least Significant Difference) was used to examine differences
between all possible pairs of means (multiple comparisons). Each population was further
characterized by main effects i.e., deviation of its mean from the total mean per a species.
The F statistics was used for estimation of the significance of main effects.
The analysis of discriminant value was employed to estimate the participation of each
character in total variation of analysed populations. The discriminant value of each character
was estimated by partial lambda Wilks statistics. It has range from 0 to 1 and the lower lambda
Wilks statistics the higher discriminant value of a character. Thus, the value 1.00 means
that a character has no discriminant value. The discriminant analysis was made separately