Characterization of nuclear DNA content and chromosome numbers of Tulipa luanica Millaku, T. kosovarica Kit Tan, Shuka & Krasniqi and T. albanica Kit Tan & Shuka

The Balkan Peninsula is considered an important centre of native tulip species. Tulipa kosovarica and Tulipa luanica are new species recently discovered in Kosovo, and Tulipa albanica in Albania. The current study aims at the investigating the nuclear DNA content and chromosome number of these three tulipa species in order to provide for the first time data on their genome size and differences among these three Tulipa species. Analysis of nuclear DNA content was performed by flow cytometer (Partec CyFlow Space) in mature fresh leaves for each Tulipa species. Samples for chromosome analysis were taken from the root tip meristem of the bulbs. Results showed significantly higher amounts of nuclear DNA (2C) in T. luanica compared to T. kosovarica and T. albanica. The chromosome number for these three species was 2n = 2x = 24, while the chromosome sizes of T. luanica resulted larger, compared to that of T. kosovarica and T. albanica. A correlation between the nuclear DNA content and chromosome size was found among these tulipa species. Moreover, nuclear DNA content and chromosome sizes of T. luanica, T. kosovarica and T. albanica showed clear differences among these species

Effects of cold-stratification, gibberellic acid and potassium nitrate on seed germination of yellow gentian (Gentiana lutea L.)

The purpose of this study was to show the effect of cold-stratification, gibberellic acid and potassium nitrate on seed germination of yellow gentian (Gentiana lutea L.). The seeds of natural yellow gentian populations of the Albanian Alps (Kosovo) were collected in September 2010. Seeds placed in filter paper (plastic bag) and in mixture of sand-soil were cold-stratified (2±1°C) for 72 days. The effect of different concentrations of gibberellic acid (250, 500 and 1000 ppm GA3) and potassium nitrate (0.1, 0.2 and 0.3% KNO3) in final germination percentage (FGP), mean germination time (MGT) and germination index (GI) in non-stratified and stratified seeds were examined. Seeds stratified in filter paper and treated with 1000 ppm GA3 and 0.1% KNO3 showed significantly higher percentage of germination (FGP and GI), while seeds stratified in mixture of sand-soil showed the highest values of FGP and GI in treatments with 500 ppm GA3 and different concentrations of KNO3. The cold-stratification and GA3 treatments of yellow gentian seeds are suitable for the higher percentage of germinations.Key words: Gentiana lutea L., cold-stratification, seeds, germination, gibberellic acid (GA3), potassium nitrate (KNO3)

Determination of Nuclear DNA Content of Tulipa Taxons from Kosovo by Flow Cytometer and Usage in Ploidy Analysis and Classification

Tulipa cinsi farklı ploidy düzeylerine sahip yaklaşık olarak 100' ü tür olarak kabul görmekte olan 500 civarında taksonu içermektedir. Cinsin üyeleri bahar aylarında açan şahane çiçeklere sahiptir ve bu özelliklerinden dolayıda çok eski tarihlerden beri büyük bir ekonomik değere sahiptirler. Sunulan bu araştırma projesinin amacı flow sitometri ile Kosova' nın bazı bölgelerinde doğal olarak yayılış gösteren Tulipa taksonlarının çekirdek DNA içeriklerini ilk defa belirlemek ve elde edilen bilgiyi taksonların ploidy düzeylerinin belirlenmesi ve taksonomik teşhislerinde kullanmaktır. Çalışmada flow sitometri ile Kosova' nın iki farklı lokasyonundan toplanmış ve iki farklı türe (T. luanica ve T. kosovorica) ait 45 Tulipa genotipinin 2C çekirdek DNA içerikleri belirlenmiştir. Analizlerde florasan boya olarak propidium iodide ve internal standard olarak çavdar bitkisi kullanılmıştır. Elde edilen sonuçlara göre analiz edilen genotiplerin 2C çekirdek DNA içeriklerinin 42.58 pg ile 48.36 pg arasında değiştiği belirlenmiştir. T. luanica ve T. kosovorica türlerinin ortalama 2C çekirdek DNA içerikleri ise sırasıyla 46.80 ve 44.41 pg olarak belirlenmiştir. İki tür arasındaki çekirdek DNA içeriği bakımından farklılık küçük olmakla beraber istatistiki açıdan önemli bulunmuştur. Bu nedenle Tulipa türlerinin teşhisinde yararlı olabileceği belirlenmiştir. Türlerin çekirdek DNA içeriği ile ploidy düzeylerini ilişkilendirmek için klasik yöntemle her türden bir bitkinin kromozomları sayılmış ve her iki bitkininde Tulipa cinsi içerinde en yaygın ploidy düzeyi olan diploid (2x = 2n = 24) kromozom sayısına sahip oldukları gözlenmiştir. Benzer DNA içeriklerine sahip oldukları için diğer 43 bitkininde ploidy düzeylerinin diploid olduğu kabul edilmiştir. Sonuç olarak flow sitometri ile belirlenen çekirdek DNA içeriği bilgisi Tulipa türlerinin ploidi düzeylerinin belirlenmesi, ve taksonomik teşhisleri ile sınıflandırılmalarında son derece yararlı olacağı belirlenmiştir.Genus Tulipa includes approximatelly 500 taxons and about 100 of them already accepted as species. Members of the genus have spectacular flowers in spring and therefore, they have a big economic importance since very old times. The objectives of this research project were to determine nuclear DNA content of Tulipa taxons from Kosovo for the first time by using flow cytometer and use it in ploidy analysis and taxonomic identification. In the study, 2C nuclear DNA content of 45 Tulipa plants, representing two different Tulipa species (T. luanica ve T. kosovorica), collected from two different locations of Kosovo were determined. Propidium iodide was used as florescent dye in preparation of samples while rye was the internal standard. Based on the results of the study, 2C nuclear DNA content of Tulipa plants analysed in the study varied from 42.58 pg to 48.36 pg. However, the mean 2C nuclear DNA content of T. luanica ve T. kosovorica species were determined as 46.80 pg and 44.41 pg respectively. The mean 2C nuclear DNA content differences between two species species was small but, statistically significant. Therefore, the nuclear DNA content information can be usefull in taxonomic identification and classification of the species in the genus Tulipa. Nuclear DNA content of the taxons were correlated with their ploidy levels by counting mitotic chromosomes of two plants, one for each taxons with classical methods. The rest was assumed to have same ploidy levels since they have very similar nuclear DNA content. Based on the results of chromosome analysis, ploidy of all of the genotypes were diploid with 2x = 2n = 24 chromosome number. In conclusion, nuclear DNA content information determined by flow cytometer can be usefull in analysing ploidy of Tulipa taxons, and in their taxonomic identification and classification

Concentration of some metals in soil and plant organs and their biochemical profiles in Tulipa luanica, T. kosovarica and T. albanica native plant species

The purpose of this study was to determine the concentration of some metals (Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Zn, Ca and Mg) in soil of serpentine and limestone sites, their bioaccumulation and impact on some biochemical parameters in T. luanica, T. kosovarica and T. albanica plants. T. kosovarica and T. albanica grows in serpentine soil, while T. luanica grow in limestone soil. The research showed that concentrations of Cd, Co, Cr, Fe, Mn and Ni were significantly higher at serpentine soil sites in comparison with limestone sites, while concentrations of Pb, Cd, Co and Cr in bulbs, leaves and seeds were under the limit of detection. The concentration of Ni in plant samples of T. kosovarica was significantly higher in comparison with its concentration in T. albanica, but it was under the limit of detection in T. luanica. Moreover, concentrations of Al and Fe in leaves of T. kosovarica and T. albanica were higher in comparison with T. luanica. The concentration of Mg was significantly higher in T. kosovarica and T. albanica than in T. luanica. The delta-aminolevulinic acid dehydratase activity, malondialdehyde and glutathione contents in leaves of T. luanica were higher in comparison with T. kosovarica and T. albanica. In addition, the amounts of total chlorophyll and delta-aminolevulinic acid (ALA) in leaves of T. albanica were higher in comparison with T. kosovarica and T. luanica. Our findings show that target organs of metal accumulation in three Tulip species appears to be leaves[seeds[bulbs, while the biochemical parameters show that limestone sites represent a less stressful habitat for growing these plant species in comparison with serpentine sites

Activity of δ-Aminolevulinic Acid Dehydratase at Ramonda nathaliae and Ramonda serbica Plants During Dehydration and Rehydration

Introduction: Ramonda nathaliae and Ramonda serbica are resurrection plants belonging to homoiochlorophyllous desiccation-tolerant angiosperms. Chlorophyll biosynthesis is one of the most important metabolic pathways to tolerate desiccation in these plant species. Materials and methods: To better understand the early pathway steps of chlorophyll biosynthesis, we have analyzed the enzyme δ-aminolevulinic acid dehydratase (ALA-D) and contents of δ-aminolevulinic acid (ALA) and total chlorophyll as a final product during dehydration and rehydration stages for these plant species. Results: Our results showed that the activity of ALA-D in R. nathaliae and R. serbica plants rapidly decreased during dehydration and in the final stage of desiccation the activity of this enzyme was decreased by 79% and 86%, respectively. After rehydration of plants, the ALA-D activity was fully restored. In contrast, the ALA content of both plant species significantly increased during desiccation and decreased after 48 hr of rewatering. In each stage of dehydration or rehydration, a significant negative correlation was established between ALA-D activity and ALA content in both plant species. Conclusions: Total chlorophyll content was preserved more in R. nathaliae than in R. serbica during desiccation. Moreover, ALA-D activity was decreased to a minimal level but preserved its function during desiccation, and this suggests one possible mechanism of desiccation tolerance to retain the chlorophyll of these plant species

Concentration of some metals in soil and plant organs and their biochemical profiles in Tulipa luanica, T. kosovarica and T. albanica native plant species

The purpose of this study was to determine the concentration of some metals (Al, Cd, Co, Cr, Cu, Fe, Mn, Ni, Pb, Zn, Ca and Mg) in soil of serpentine and limestone sites, their bioaccumulation and impact on some biochemical parameters in T. luanica, T. kosovarica and T. albanica plants. T. kosovarica and T. albanica grows in serpentine soil, while T. luanica grow in limestone soil. The research showed that concentrations of Cd, Co, Cr, Fe, Mn and Ni were significantly higher at serpentine soil sites in comparison with limestone sites, while concentrations of Pb, Cd, Co and Cr in bulbs, leaves and seeds were under the limit of detection. The concentration of Ni in plant samples of T. kosovarica was significantly higher in comparison with its concentration in T. albanica, but it was under the limit of detection in T. luanica. Moreover, concentrations of Al and Fe in leaves of T. kosovarica and T. albanica were higher in comparison with T. luanica. The concentration of Mg was significantly higher in T. kosovarica and T. albanica than in T. luanica. The delta-aminolevulinic acid dehydratase activity, malondialdehyde and glutathione contents in leaves of T. luanica were higher in comparison with T. kosovarica and T. albanica. In addition, the amounts of total chlorophyll and delta-aminolevulinic acid (ALA) in leaves of T. albanica were higher in comparison with T. kosovarica and T. luanica. Our findings show that target organs of metal accumulation in three Tulip species appears to be leaves[seeds[bulbs, while the biochemical parameters show that limestone sites represent a less stressful habitat for growing these plant species in comparison with serpentine sites