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Section 2. Biotechnology
Shehu Matilda, Department of Biologyy Faculty of Technical Sciences University of Vlora, Albania Plant Biotechnology PhD., candidate E-mail: [email protected] Zekaj (Trojani) Zhaneta, Department of Biotechnology, University of Tirana
CYTOLOGICAL DIVERSITY BETWEEN THREE POPULATIONS OF SCILLA AUTUMNALIS L. IN SOUTH ALBANIA
Abstract: In this study, three populations of S. autumnalis L. were investigated. The aim of the work is to analyse the cytological variation of this species, examined for the first time in Albania. The technique used for cytological study is the squash technique. Three cytotypes resulted for this species in Vlora area. For population of Portopalermo was realised chromosome formula, karyogram and idiogram.
Keywords: Scilla autumnalis L., cytological variation, cytotype, squash technique.
I. Introduction: Scilla autumnalis L. is a rather II. Materials and methods:
widely distributed and common species in the Medi- For the cytological analysis of S. autumnalis spe-
terranean region and Western Europe. In our coun- cies in the three habitats, the were used root tips.
try is wide distributed in all territory, but in other The slides were prepared using the squash technique.
countries (Romania) is critically endangered [1]. Is The technique of processing the herbal material for
one of the most studied species by many authors. Till the preparation of the chromosome slides passes
now it has been classified as a a single species, mainly through several stages: the prefixation of the roots
based on morphological traits. It is a species with a tips in icy water at 2 °C throughout the night [8].
large variation in the number, type and morphology Then were moved to Carnua (absolute ethanol: gla-
of the chromosomes. So far, 10 cytologically distinct cial acetic acid/3:l) for 3-12 hours at 4 °C. After fix-
populations have been detected at the species level ation in Karnua root tips were stored in alcohol 70%
[2; 3; 4; 5]. For the populations analyzed in Tunisia at 4 °C for several weeks. Then were transferred into
and Aegean regions, 8 cytotope was reported [6]. In acetic orcein 45% [9] for several hours at ambient
a study of 31 Scilla autumnalis L. populations from temperature and then was made double enzymatic
Spain and Portugal 4 cytotypes were found [7]. The treatment with pectinases and cellulase which was
aim of this study has been to add more information left in 4 °C overnight. The next day has been prepared
on the cytotypes, realised for the first time for these the slides with the squash technique. Observation
species in Albania. has been realized with microscope Optika at 100
x magnification. Measurement of chromosomes, chromosomal formula, karyogram and Idiogram was realized with Cromo II software [10]. Photos were realized with Apple iphone at ocular tube. III. Results and discusión Scilla autumnalis L. in three considerated habitats shows variation of chromosome number. For S. autumnalis L. Portopalermo we were able to de-
fine the chromosome number and chromosome morphology. For other two population we defined only chromosome number. Three cytotypes were found during investigation. The population of Du-kat results tetraploid with chromosome number 2n = 4 x = 28, while the population of Llogara appears hexaploid with chromosome number 2n = 6 x = 42 (Figure 1).
a) b) c)
Figure 1. S. autumnalis L. chromosomes: a) Portopalermo; b) Llogara; c) Dukat
Figure 2. Chromosomes of S. autumnalis L. Portopalermo (arrows indicate the presence of satellites)
Chromosome number defined for Portopaler- In this cytotype we encountered a pair of satellites mo population was 2n = 2x = 14 and chromosom- present in the pair of metacentric chromosomes. al formula: 2n = 2x = 5m + 2m0 + 3sm + 4st = 14. In the figure 2 appears the chromosome sketch
for population of Portopalermo where arrows topalermo. They represent the typology, number indicates the satellites. The (figure 3) shows the and position of satellites in the chromosomal hap-karyogram and idiogram of S. autumnalis L. Por- loid and diploid set.
DATIINSERITI E DATI ELABORATI Nun: 63 Taxon: Scilla autumnalis
Scala:100 Locus: Portopalermo Formula di Leuan: 5m + 2m0+3sm + 4st
Id Lu Co Tot r S SI T1 S2 T2
3 170 120 290 1.42
13 150 140 290 1.07
11 150 90 240 1.67
4 110 80 190 1.38
10 125 40 165 3.13
2 125 30 155 4.17
7 115 35 150 3.29
9 110 30 140 3.67
12 65 40 105 1.63 30TC
14 60 40 100 1.50 30TC
5 95 40 135 2.38
6 90 40 130 2.25
8 80 40 120 2.00
1 70 45 115 1.56
A
B
C D
Figure 3. Scilla autumnalis L. A. Table with data for chromosomes (long arm, short arm, arm ratio, satellite), B. Numerical Karyogram, C. Karyogram, D. Idiogram
During the study, it is noticed that the population otic stress factors (drought and cold) [13], therefore of Llogara, which is at higher altitude, has a higher polyploid individuals extend at higher altitudes than
level of ploidy. Many authors reported that elevation increases also the level of ploidy [11; 12]. The phenomenon of chromosome number variability is also related to the resistance ofpolyploid genotypes to abi-
diploid individuals. This is in accordance with our results, where individuals of Portopalermo exhibit diploid karyotype, while the populations of Dukat and Llogara located at higher altitudes exhibit poliploid.
Figure 4. Mitotic index of S. autumnalis L. in three habitats
According to the data of other authors, poly- features but in some cases polyploid plants may
ploid species exhibit different morphological and have slower rates of growth [17] due to slower
physiological traits that allow them to spread in cell division causing smaller mitotic index. This is
different geographic areas [14; 15; 16]. Usually in in accordance with our results. The population of
many plant species, polyploid individuals are su- Llogara with a higher level of ploidy results with a
perior to diploid ones in terms of morphological lower mitotic index (Figure 4).
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