ABSTRACT
Colchicum autumnale is a perennial, toxic plant that originated in Europe and North Africa. Although inedible, it is occasionally consumed accidentally because it resembles the edible Allium victorialis and other related species. This misidentification has led to episodes of food poisoning in Japan. However, determining the causative agent of a food poisoning outbreak by observing the sample visually or analyzing the chemical composition is challenging when dealing with small samples. Therefore, we developed a novel set of PCR primers that anneal to the internal transcribed spacer (ITS) region of C. autumnale ribosomal DNA, designed to detect the presence of C. autumnale in small samples. These primers successfully detected C. autumnale in all samples in which it was present, and did not give a positive PCR band in the 48 other distinct crop species tested, in which it was not present. Further, our method could amplify DNA from samples of C. autumnale that had been heat-treated and digested using artificial gastric fluids. Thus, this PCR strategy is highly specific and can be used to distinguish C. autumnale simply and rapidly from various other crops.
Subject(s)
Colchicum/classification , DNA, Plant/isolation & purification , Foodborne Diseases/diagnosis , DNA Primers , DNA, Ribosomal Spacer/genetics , Humans , Japan , Polymerase Chain ReactionABSTRACT
PURPOSE: Cancer is a long process that leads the organism to death and is associated with the normal cells acquiring the ability to divide permanently. Nowadays, the use of natural products in cancer therapy has a great importance. In addition, working with plants that are endemic to Turkey and determining the biological activities of these plant extracts, is extremely important due to the potential for new drug development. There is no comparative study available in the literature on the antitumor effects of Colchicum sanguicolle, a new found species of the genus Colchicum in Turkey, Crateagus microphylla, of the genus Crateagus and Centaurea antiochia of the genus Centaurea. In this study, we tried to demonstrate the antitumor effect of these plant extracts on HeLa and C 4-1 cells. METHODS: Five different doses (0.001, 0.01, 0.05, 0.25 and 0.5 mg/ml) of the three plant types were prepared and applied for 24, 48 and 72 hrs on the cervical cancer derived cell lines. Subsequently, the growth rate was evaluated with the mitochondrial dehydrogenase enzyme method. RESULTS: Colchicum sanguicolle extracts showed the most effective antitumor activity. For the Colchicum sanguicolle extract, the IC50 dose for HeLa cells was 0.01 mg/ml at 48 hrs, while for the C-4 I cells it was 0.001 mg/ml at 48 hrs. These results showed that C-4 I cells were more sensitive to the Colchicum sanguicolle extracts. Conclus?on: The results of from this study regarding the antitumor effect of plant extracts of endemic varieties of Turkey may have an important place in design and development of anticancer drugs and would make contributions to other studies to be conducted in this area.
Subject(s)
Antineoplastic Agents, Phytogenic/pharmacology , Colchicum/chemistry , Drug Screening Assays, Antitumor , Plant Extracts/pharmacology , Uterine Cervical Neoplasms/drug therapy , Antineoplastic Agents, Phytogenic/isolation & purification , Biomarkers, Tumor/metabolism , Cell Proliferation/drug effects , Cell Shape/drug effects , Cell Survival/drug effects , Colchicum/classification , Dose-Response Relationship, Drug , Female , HeLa Cells , Humans , Inhibitory Concentration 50 , Mitochondrial Proteins/metabolism , Oxidoreductases/metabolism , Phytotherapy , Plant Extracts/isolation & purification , Plants, Medicinal , Time Factors , Turkey , Uterine Cervical Neoplasms/enzymology , Uterine Cervical Neoplasms/pathologyABSTRACT
The study of phylogenetic relationships between 14 Colchicum taxa spread throughout Turkey was performed using a fluorescent-based amplified fragment length polymorphism (AFLP) technique. Five primer pair combinations were used in AFLP reactions. The data set was analyzed statistically using the NTSYS 2.1 software, and the neighbor-joining and maximum parsimony methods were implemented to generate phylogenetic trees. These analyses clustered the samples into 3 main clades. Both the neighbor-joining and maximum parsimony analyses resulted in similar topologies. Furthermore, supporting the phylogenetic trees, a similar grouping of 14 taxa was generated by principal component analysis. The AFLP analysis with 5 primer combinations was carried out to assess 14 taxa. Fragment sizes ranged from 54 to 462 bp in length for each primer combination. The average was 166 fragments per primer pair, primer B2 generated the highest number of bands (200), and primer B3 produced the lowest number of bands (112). A total of 834 polymorphic bands were scored. The cophenetic correlation coefficient between the data matrix and the cophenetic matrix for AFLP data was 0.72. Based on this molecular data, we concluded that the genetic diversity among these Turkish accessions is relatively high.
