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5. ORIGIN OF SEEDLING...
5.2.2. Level of seedling root fluorescence
To compare the level of seedling root fluorescence in annual and perennial ryegrass
in total 17 L. multiflorum and 26 L. perenne populations were used including cultivars
and wild ecotypes. Additionally, samples of the other Lolium species were analysed i.e.,
L. loliaceum, L. persicum, L. remotum, L. temulentum and L. rigidum. All populations are
described in Annex 13.1. About 300 seeds of each L. multiflorum and L. perenne population
were germinated at 24 o C for six days in order to obtain 240 equally developed
seedlings. In total 20 seedlings were transferred onto a single glass plate. In case of
L. loliaceum, L. persicum, L. remotum, L. temulentum and L. rigidum, due to low number of
seeds, only 100 seeds were used and about 80-90 seedlings were analysed. After 14 days
seedling roots were scored for the presence (+) and absence (-) of fluorescence under UV
light (Figure 5.2). All plants with fluorescent roots (+) and all plants with nonfluorescent roots
(-) were rescued for further analyses. Each population was analysed in four runs with three
randomly arranged replications in each run. The level of fluorescence was calculated in
relation to a total number of normal seedlings. Data were pooled for the four runs and they
were analysed using one-factor ANOVA with LSD test in STATISTICA 7.1. The squared
Mahalanobis distance was estimated between all species from the genus Lolium based on
fluorescence level. The Euclidean distance and Ward’s method were applied for clustering.
5.2.3. Development of populations for morphological analyses and analysed characters
Only these populations were analysed, in which both fluorescent and nonfluorescent
seedlings were found in sufficient amount to set up a field experiment. In total nine
L. multiflorum including seven cultivars and two ecotypes and nine L. perenne populations
including five cultivars and four ecotypes were used in morphological analyses. After estimation
of fluorescence of seminal roots the plants were transferred into pots and as soon as
they developed into multitillered plants, all plants were divided into 9-10 ramets and transplanted
to the experimental field in April and early May 2002. A randomized complete block
design with three replications was used. Each block consisted of three ramets as replicates