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mm depth were sinked into the soil between the rows to prevented spreading of stolons
to the neighbouring rows.
Morphometrical analysis: In May 2006, shoots were collected from the labelled shoot
clusters of the original habitats (controls) and others were collected from the
experimental plot. The following characteristics of the vegetative organs were
measured: (1) height at flowering, (2) lengths of the longest leaf-blade of lateral tillers,
(3) maximum widths of these leaves, (4) length of the flag leaf-blade, (5) and the flag
leaf sheath, (6) width of the flag leaf-blade, (7) length of the ligule of the flag leaves.
The following characters of the reproductive organs were measured: (8) length of the
panicle, (9) length of the longest panicle branch, (10) length of the spikelets from the
end of the lowest panicle branches, (11) length of the lower and (12) upper glume, (13)
length of the palea, (14) length of the lemma. We measured 15 individuals from each
population. The average of 10 data per individual was used for the following features: 2,
11, 12, 13 and 14. A slide-gauge was used for measuring.
AFLP method: For AFLP, each sample consisted of about one leaf per plant for ten
plants per population. ZenoGene kit (ZenonBio Ltd, Szeged, Hungary) was used for
DNA extraction following manufacturer’s instructions. Extracted DNA samples were
stored at –20 °C until used. After the AFLP procedure (K. Szabó et al., 2006) the final
amplified and selected PCR fragments were appeared by capillary electrophoresis with
ABI Prism 3100 Genetic Analyzer in fragment analysis mode at the Agricultural
Biotechnology Centre (Gödöllő, Hungary) using 360 mm length capillaries and POP-4
gel. GeneScan HD-400 Rox Size was applied as standard marker. AFLP fragments were
identified by GeneScan 3.7 computer program.
Data analysis: Morphological data were analysed by principal component analysis
using the R statistical program (R Development Core Team, 2005). For cluster analysis
of AFLP data each detected band above 80 threshold limit was scored as present (1) or
absent (0) in all samples. The unweighted pair group method using arithmetic means
(UPGMA) was used and a dendrogram was constructed based on Jaccard’s similarity
coefficient (Jaccard, 1908) by the statistical software package SPSS 11.0 for Windows
(SPSS Inc., USA). Genetic diversity and significant differences in the populations were
estimated by the proportion of polymorphic loci. The allele frequencies and
heterozygosity were estimated based on square root of recessive genotypes. These
estimates and Nei-genetical distances (1978) were analysed by TFPGA (Miller and
Mark, 1997).
RESULTS
Morphometrical analysis: We did not observe a sharp separation on the PCA biplot
diagram based on all features of the two species (Fig. 1). Clusters of the individuals of
the two species show complete overlap. However, the variables show unambiguously
two different tendencies, depending on whether they are vegetative (v1-v8, v14), or
reproductive (v9-v13) characteristics.
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