Overcoming obstacles: Analysis of blood and semen stains washed with different chemicals with ATR-FTIR.

INTRODUCTION: Blood and semen stains are the most common biological stains encountered at crime scenes. The washing of biological stains is a common application that perpetrators use to spoil the crime scene. With a structured experiment approach, this study aims to investigate the effects of washing with various chemicals on the ATR-FTIR detection of blood and semen stains on cotton. MATERIALS AND METHODS: On cotton pieces, a total of 78 blood and 78 semen stains were applied, and each group of six stains was immersed or mechanically cleaned in water, 40% methanol, 5% sodium hypochlorite solution, 5% hypochlorous acid solution, 5 g/L soap dissolved pure water, and 5 g/L dishwashing detergent dissolved water. ATR-FTIR spectra gathered from all stains and analyzed with chemometric tools. RESULTS AND DISCUSSION: According to performance parameters of developed models, PLS-DA is a powerful tool for discrimination of washing chemical for both washed blood and semen stains. Results from this study show that FTIR is promising for use in detecting blood and semen stains that have become invisible to the naked eye due to washing of the findings. CONCLUSION: Our approach allows blood and semen to be detected on cotton pieces using FTIR combined with chemometrics, even though it is not visible to the naked eye. Washing chemicals also can be distinguished via FTIR spectra of stains.

The molecular mechanisms of vulpinic acid induced programmed cell death in melanoma.

BACKGROUNDS: Malignant melanoma is an aggressive skin tumor with a rapidly increasing incidence and there is not yet a successful treatment strategy. Vulpinic acid (VA) is derived from secondary metabolites from lichen species. In the current study, we, for the first time, investigated the anti-cancer effects of VA and the underlying mechanism VA induced programmed cell death in melanoma. METHODS: The anti-cancer effects of VA on melanoma cells were evaluated by the xCELLigence system, flow cytometry, caspase-3 activity and RT-PCR analysis. RESULTS: Our results showed that VA had a strong anti-proliferative effect on A-375 melanoma cells without damaging human epidermal melanocyte cells. Additionally, VA promoted apoptotic cell death through G2/M arrest and the activation of both intrinsic and extrinsic apoptosis pathways according to the analysis of 88 genes associated with apoptosis by qRT-PCR. CONCLUSIONS: Our findings suggest that VA could become an alternative topical and transdermal treatment strategy in the treatment of maligned melanoma cancer. However, further investigations are needed to assess the underlying molecular mechanism of VA mediated apoptotic cell death in the treatment of melanoma.

Identification and Characterization of SRS Genes in Phaseolus vulgaris Genome and Their Responses Under Salt Stress.

SHI-Related Sequence (SRS) transcription factors comprise a protein family with important roles in growth and development. However, the genome-wide study of the SRS protein family has not yet been carried out in the common bean. For this reason, the SRS family has been characterized in depth at both gene and protein levels and several bioinformatics methods have been used. As a result, 10 SRS genes have been identified and their proteins have been phylogenetically categorized into three major groups within the common bean. By investigating duplications that play a major role in the development of gene families, 19 duplication events have been identified in the SRS family (18 segmental and 1 tandem). In addition, using available RNAseq data, comparative expression analysis of Pvul-SRS genes was performed and expression changes in Pvul-SRS-1, 2, 4, 6, 7, and 10 genes were observed under both salt and drought stress. Five Pvul-SRS genes were selected based on RNAseq data (Pvul-SRS-1, 2, 4, 6, and 10) and screened with RT-qPCR in two common bean cultivars (Yakutiye 'salt-resistant' and Zulbiye 'salt-susceptible' cv.). These genes also showed different levels of expression between two common bean cultivars under salt stress conditions and this may explain the responses of Pvul-SRS genes against abiotic stress. In summary, this work is the first study in which in silico identification and characterization of Pvul-SRS genes have been examined at gene expression level. The results could therefore provide the basis for future studies of functional characterization of Pvul-SRS genes.

Genome-wide identification of CAMTA gene family members in Phaseolus vulgaris L. and their expression profiling during salt stress.

The calmodulin-binding transcriptional activator (CAMTA) family was first observed in tobacco (NtER1) during a screening for the CaM-binding proteins, which are known to be one of the fast response stress proteins. Due to the increased importance of plant transcription factors in recent years; genome-wide identification of CAMTA genes has been performed in several plant species, except for Phaseolus vulgaris. Therefore, our aim was to identify and characterize CAMTA genes in P. vulgaris via in silico genome-wide analysis approach. Our results showed a total of eight CAMTA genes that were identified and observed on five out of 11 chromosomes of P. vulgaris. Four gene couples were found to be segmentally-duplicated and these segmental duplication events were shown to occur from 29.97 to 92.06 MYA. The phylogenetic tree of CAMTA homologs from P. vulgaris, A. thaliana, and G. max. revealed three groups based on their homology and the intron numbers of Pvul-CAMTA genes, ranged from 11 to 12. According to the syteny analysis; CAMTA genes of P. vulgaris and G. max revealed higher similarity, because they have highly similar genomes compared to A. thaliana. All Pvul-CAMTA genes were targeted by miRNAs, which play a role in response mechanism of salt stress. To detect expression levels in different plant tissues, mRNA analysis of Pvul-CAMTA genes were performed using publicly available expression data in Phytozome v12.1. In addition, responses of Pvul-CAMTA genes to salt stress, were also examined via both RNAseq and qRT-PCR analysis. To identify and to obtain insight into biological functions of CAMTA genes in the genome of P. vulgaris, several analyses were conducted using many online and offline bioinformatic tools, genome databases and qRT-PCR analyses. Due to this study being the first in the identification of CAMTA genes in P. vulgaris, this study could be considered as an useful source for future CAMTA genes studies in either P. vulgaris or comparative different plant species.

Determination of Vulpinic Acid Effect on Apoptosis and mRNA Expression Levels in Breast Cancer Cell Lines.

OBJECTIVE: Breast cancer is one of the most common diseases among women worldwide and it is characterized by a high ratio of malignancy and metastasis and low rate of survival of patients. Due to limited treatment options, the discovery of alternative therapeutic agents and clarifying the molecular mechanism of breast cancer development may offer new hope for its treatment. Lichen secondary metabolites may be one of these therapeutic agents. METHODS: In this study, the effects of Vulpinic Acid (VA) lichen secondary metabolite on the cell viability and apoptosis of breast cancer cells and non-cancerous cell line were investigated. Quantitative polymerase chain reaction was also performed to determine changes in the expression of apoptosis-related genes at a molecular level. RESULTS: The results demonstrated that VA significantly inhibited the cell viability and induced apoptosis of human breast cancer cells. The highest rates of decreased growth were determined using the IC50 value of VA for 48h on MCF-7 breast cancer cell. Interestingly, VA treatment significantly reduced cell viability in all examined breast cancer cell lines compared to their non-cancerous human breast epithelial cell line. This is the first study on the investigation of the effects of VA on the molecular mechanisms associated with the expression of apoptosis-related genes in breast cancer cell lines. Results demonstrated that the gene expression of P53 genes was altered up to fourteen-fold levels in SK-BR-3 cell lines whereas it reached 2.5-fold in the MCF-12A cell line after treatment with VA. These observations support that VA induces apoptosis on the breast cancer cells compared with the non-cancerous human breast epithelial cell line. CONCLUSION: It is implicated that VA may be a promising novel molecule for the induction of apoptosis on breast cancer cells.

Genome-wide analysis of Phaseolus vulgaris C2C2-YABBY transcription factors under salt stress conditions.

The aim of this study was to identify and characterize the C2C2-YABBY family of genes by a genome-wide scale in common bean. Various in silico approaches were used for the study and the results were confirmed through common molecular biology techniques. Quantitative real-time PCR (qPCR) analysis was performed for identified putative PvulYABBY genes in leaf and root tissues of two common bean cultivars, namely Yakutiye and Zulbiye under salt stress condition. Eight candidate PvulYABBY proteins were discovered and the length of these proteins ranged from 173 to 256 amino acids. The isoelectric points (pIs) of YABBY proteins were between 5.18 and 9.34 and ranged from acidic to alkaline, and the molecular weight of PvulYABBYs were between 18978.4 and 28916.8 Da. Three segmentally duplicated gene couples among the identified eight PvulYABBY genes were detected. These segmentally duplicated gene couples were PvulYABBY-1/PvulYABBY-3, PvulYABBY-5/PvulYABBY-7 and PvulYABBY-6/PvulYABBY-8. The predicted number of exons among the PvulYABBY genes varied from 6 to 8 exons. Additionally, all genes found included introns within ORFs. PvulYABBY-2, -4, -5 and -7 genes were targeted by miRNAs of five plant species and a total of five miRNA families (miR5660, miR1157, miR5769, miR5286 and miR8120) were detected. According to RNA-seq analysis, all genes were up- or down-regulated except for PvulYABBY-1 and PvulYABBY-6 after salt stress treatment in leaf and root tissues of common bean. According to the qPCR analysis, six out of eight genes were expressed in the leaves but only four out of eight genes were expressed in the roots and these genes exhibited tissue- and cultivar-specific expression patterns.

Genome-wide identification of salinity responsive HSP70s in common bean.

The present study is aimed to identify and characterize HSP70 (PvHSP70) genes in two different common bean cultivars under salt stress. For this purpose various in silico methods such as RNAseq data and qRT-PCR analysis were used. A total of 24 candidate PvHSP70 gene were identified. Except for chromosome 4 and 7, these candidate PvHSP70 genes were distributed on the remaining chromosomes. While the lowest number of PvHSP70 genes was determined on chromosomes 1, 3, 5, 7, 9, 10 and 11 (one HSP70 gene), the highest number of PvHSP70s was on chromosomes 6 and 8 (seven HSP70 genes each). Three genes; PvHSP70-5, -9, and -10 were found to have no-introns. In addition, four tandemly and six segmentally duplicated gene couples were detected. A total of 13 PvHSP70 genes were targeted by miRNAs of 44 plant species and the most targeted genes were PvHSP70-5 and -23. The expression profile of PvHSP70 genes based on publicly available RNA-seq data was identified and salt treated leaf tissue was found to have more gene expression levels compared to the root. qRT-PCR analysis showed that the transcript concentrations of upregulated PvHSP70 genes in leaves of Zulbiye (sensitive) were mostly higher than those of Yakutiye (resistant). The present study revealed that PvHSP70 genes might play an important role in salt stress response for common bean cultivars and variability between cultivars also suggests that these genes could be used as functional markers for salt tolerance in common bean.

Roles of catalase (CAT) and ascorbate peroxidase (APX) genes in stress response of eggplant (Solanum melongena L.) against Cu(+2) and Zn(+2) heavy metal stresses.

Eggplant (Solanum melongena L.) is a good source of minerals and vitamins and this feature makes its value comparable with tomato which is economically the most important vegetable worldwide. Due to its common usage as food and in medicines, eggplant cultivation has a growing reputation worldwide. But genetic yield potential of an eggplant variety is not always attained, and it is limited by some factors such as heavy metal contaminated soils in today's world. Today, one of the main objectives of plant stress biology and agricultural biotechnology areas is to find the genes involved in antioxidant stress response and engineering the key genes to improve the plant resistance mechanisms. In this regard, the current study was conducted to gain an idea on the roles of catalase (CAT) and ascorbate peroxidase (APX) genes in defense mechanism of eggplant (S. melongena L., Pala-49 (Turkish cultivar)) treated with different concentrations of Cu(+2) and Zn(+2). For this aim, the steady-state messenger RNA (mRNA) levels of CAT and APX genes were determined by quantitative real-time PCR (qRT-PCR) in stressed eggplants. The results of the current study showed that different concentrations of Cu(+2) and Zn(+2) stresses altered the mRNA levels of CAT and APX genes in eggplants compared to the untreated control samples. When the mRNA levels of both genes were compared, it was observed that CAT gene was more active than APX gene in eggplant samples subjected to Cu(+2) contamination. The current study highlights the importance of CAT and APX genes in response to Cu(+2) and Zn(+2) heavy metal stresses in eggplant and gives an important knowledge about this complex interaction.

The determination of physiological and DNA changes in seedlings of maize (Zea mays L.) seeds exposed to the waters of the Gediz River and copper heavy metal stress.

In this study, the effects of the heavy metal-polluted waters of the Gediz River, which flow into the Aegean Sea, and different concentrations of copper (Cu) solutions on maize (Zea mays L.) seedlings are investigated with physiological parameters and random amplified polymorphic DNA (RAPD) assay. Results displayed physiologically a significant difference in root and stem length between the control seedlings and the seedlings grown with the waters of the Gediz River. Also, the certain ascending concentrations of copper solution (80, 160, 320, 640, and 1280 ppm) caused a significant decrease in root and stem length of seedlings compared to the control seedlings. As a result of the waters of the Gediz River and copper solution treatment, the changes occurred in RAPD profiles of seedlings observed as variations like increment and/or loss of bands compared with the control seedlings. And these changes were reflected as a decrease in genomic template stability (GTS, changes in RAPD profile) derived by genotoxicity. RAPD band profiles and GTS values showed consistent results with physiological parameter. In conclusion, the study revealed the environmental risk and negative effect of waters of the Gediz River on maize seedlings and the suitability of RAPD assay for the detection of environmental toxicology.

Evaluation of different isotherm models, kinetic, thermodynamic, and copper biosorption efficiency of Lobaria pulmonaria (L.) Hoffm.

UNLABELLED: The biosorption characteristics of Cu(II) ions from aqueous solution using Lobaria pulmonaria (L.) Hoffm. biomass were investigated. The biosorption efficiency of Cu(II) onto biomass was significantly influenced by the operating parameters. The maximum biosorption efficiency of L. pulmonaria was 65.3% at 10 mg/L initial metal concentration for 5 g/L lichen biomass dosage. The biosorption of Cu(II) ions onto biomass fits the Langmuir isotherm model and the pseudo-second-order kinetic model well. The thermodynamic parameters indicate the feasibility and exothermic and spontaneous nature of the biosorption. The effective desorption achieved with HCl was 96%. Information on the nature of possible interactions between the functional groups of the L. pulmonaria biomass and Cu(II) ions was obtained via Fourier transform infrared (FTIR) spectroscopy. The results indicated that the carboxyl (-COOH) and hydroxyl (-OH) groups of the biomass were mainly involved in the biosorption of Cu(II) onto L. pulmonaria biomass. The L. pulmonaria is a promising biosorbent for Cu(lI) ions because of its availability, low cost, and high metal biosorption and desorption capacities. IMPLICATIONS: Lobaria pulmonaria is a promising biosorbent for Cu(II) ions because of its availability, low cost, and high metal biosorption and desorption capacities. To the best of our knowledge, this is the first paper on the biosorption Cu by L. pulmonaria.

Air-quality biomonitoring: assessment of genotoxicity of air pollution in the Province of Kayseri (Central Anatolia) by use of the lichen Pseudevernia furfuracea (L.) Zopf and amplified fragment-length polymorphism markers.

Mixed air pollutants are considered a major cause of DNA damage in living organisms. In this study, samples of the lichen Pseudevernia furfuracea (L.) Zopf were used as bioindicators to assess the genotoxicity of air pollutants in the province of Central Anatolia, Kayseri. The study area is characterized by the presence of numerous industrial activities, such as steel works, glassworks, and ship-building, metallurgical, mechanical and chemical industries. In the study, two biomonitoring experiments were performed during the dry and wet seasons of 2005. P. furfuracea lichen samples were exposed to various pollutants at 12 monitoring sites, distributed throughout the different parts of the province, and each experiment lasted for a period of four weeks. Genotoxic effects of environmental pollutants were evaluated with amplified fragment-length polymorphism (AFLP) molecular markers. The results indicate that the mixture of pollutants might have contributed to the changes in the band patterns obtained by AFLP analysis, reflecting the presence of DNA damage. The average value of polymorphism obtained from the amplification of the primers used was 45.0% for the wet period and 64.6% for the dry period. Genomic template stability (GTS) ratios revealed that the highest values belong to the P. furfuracea samples from Rural Site I and Rural Site II (97.9%, 99.3% respectively for the dry season), the lowest values were from Shanty II and Urban Road Site I (85.8%, 85.2%, respectively for the wet season). The present results indicate that the licxhen species P. furfuracea, which is known for its bioindicator-biomonitor capacity, also has a high capacity as indicator of genotoxicity. AFLP markers are cheap, reliable and, therefore, an important tool for studying genotoxicity in lichen species.

Detection of superoxide radicals in tomato plants exposed to salinity, drought, cold and heavy metal stress using CMC-G-SOD biosensor.

A novel highly sensitive electrochemical carboxymethylcellulose-gelatin-superoxide dismutase biosensor was used for the determination of superoxide radicals enhancement in tomato plants exposed to salinity, drought, cold and heavy metal stress. The variations in superoxide radicals depending on abiotic stress was determined using biosensor. The superoxide radical production with regard to control rapidly was increased in tomato plants exposed to salinity, drought, cold and heavy metal stress. The superoxide radical enhancement in tomato plants exposed to salinity, drought, cold and heavy metal stress was successfully determined using carboxymethylcellulose-gelatin-superoxide dismutase biosensor.

Genotoxic effect of cadmium in okra seedlings: comparative investigation with population parameters and molecular markers.

Plants are considered as good bioindicators because of their significant role in food chain transfer. They are also easy to grow, adaptable to environmental stresses and can be used for assaying a range of environmental conditions in different habitats. Thus, many plant species have been used as bioindicators. In order to evaluate the genotoxic effect of cadmium, okra (Abelmoschus esculontus L.) seedlings were treated with different concentrations (30, 60, 120 mg I(-1)) of cadmium and investigated for their population parameters such as inhibition of root growth; total soluble protein content, dry weight and also the impact of metal on the genetic material by RAPD analysis. Root growth and total soluble protein content in okra seedlings were reduced with increased Cd concentrations. RAPD analysis indicated formation of new bands mostly at 60 and 120 mg I(-1) Cd treatments. Altered DNA band patterns and population parameters after Cd treatments suggest that okra could be used as an indicator to reveal the effects of genotoxic agents.

Characterization of stress induced by copper and zinc on cucumber (Cucumis sativus L.) seedlings by means of molecular and population parameters.

Contamination of plants with heavy metals could result in damage in DNA, such as mutations and cross-links with proteins. These altered DNA profiles may become visible in changes such as the appearance of a new band, or loss of an existing band, in the random amplified polymorphic DNA (RAPD) assay. In this study, various concentrations of copper and zinc salts were applied to cucumber seedlings during germination. Results displayed abnormalities in germination and also changes in root elongation, dry weight and total soluble protein level. All treatment concentrations (40, 80, 160, 240, 320, and 640mg/L) used in the study caused a decrease/delay in germination of the cucumbers to different extents. Inhibition or activation of root elongation was considered to be the first effect of metal toxicity in the tested plants. Application of the metal salts and the combined solutions on cucumber (Cucumis sativus L.) seedlings revealed similar consequences for total soluble protein level, dry weight and ultimately in inhibitory rates as well. The data obtained from RAPD band-profiles and genomic template stability (GTS) showed results that were consistent with the population parameters. In this regard, we conclude that molecular marker assays can be applied in combination with population parameters to measure genotoxic effects of heavy metals on plants.

Carboxymethylcellulose-gelatin-superoxidase dismutase electrode for amperometric superoxide radical sensing.

A novel, highly sensitive superoxide dismutase biosensor for the direct and simultaneous determination of superoxide radicals was developed by immobilization of superoxide dismutase within carboxymethylcellulose-gelatin on a Pt electrode surface. The parameters affecting the performance of the biosensor were investigated. The response of the CMC-G-SOD biosensor was proportional to O (2) (·-) concentration and the detection limit was 1.25 × 10(-3) mM with a correlation coefficient of 0.9994. The developed biosensor exhibited high analytical performance with wider linear range, high sensitivity and low response time. The biosensor retained 89.8% of its sensitivity after use for 80 days. The support system enhanced the immobilization of superoxide dismutase and promoted the electron transfer of superoxide dismutase minimizing its fouling effect. The biosensor was quite effective not only in detecting O (2) (·-) , but also in determining the antioxidant properties of acetylsalicylic acid-based drugs and the anti-radical activity of healthy and cancerous human brain tissues.

Assessment of air pollution genotoxicity by RAPD in Evernia prunastri L. Ach. from around iron-steel factory in Karabük, Turkey.

Lichens are widely used in biomonitoring studies of air pollution, either as bioindicators of air quality or as bioaccumulators of atmospheric deposition. Over the past decade, several molecular techniques have been developed to provide information on diversity, genotoxicology, genetic relationships, etc. The heavy metal contents of Evernia prunastri samples were determined by atomic absorption spectrometry. The Random Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR) method was used to describe the pattern of DNA band variation in the samples influenced by the environmental pollution. The study was designed to describe the level of pollution in an area contaminated with smoke and waste from an iron-steel factory, and to reveal the level of potential genotoxic agents around this source of pollution. The study also examined the suitability of the lichen samples for the detection of genotoxicity.

Evaluation of copper-induced stress on eggplant (Solanum melongena L.) seedlings at the molecular and population levels by use of various biomarkers.

Heavy-metal contamination is an important environmental problem in the world. It is known that high concentrations of heavy metals cause toxic damage to cells and tissues. In this study the effects of copper (Cu(2+)) contamination were determined at the molecular and population levels in eggplant (Solanum melongena L.) seedlings exposed to various concentrations of the metal ion. Inhibition of root growth, reduction in dry weight and total soluble protein content in the roots of eggplant seedlings were observed with increasing Cu(2+) concentrations. In ecotoxicology, analysis by random amplification of polymorphic DNA (RAPD) has been applied as a suitable biomarker assay for plants. For the RAPD analyses, nine RAPD primers were found to produce unique polymorphic band patterns and were subsequently used to produce a total of 80 and 168 bands in the roots of untreated and treated eggplant seedlings, respectively. The changes in RAPD profiles after Cu(2+) contamination were considered as variations, i.e. as gain and/or loss of bands compared with control seedlings. These results suggest that changes in genomic template stability could be detected with RAPD profiles and this result could be compared with the growth, dry weight and total soluble protein content of the seedlings grown at various Cu(2+) concentrations. The measurements of parameters at the molecular and population levels are fundamental to accumulate valuable information and to understand clearly the effect of a contaminant on an organism in ecotoxicology.

Assessment of air pollution genotoxicity by molecular markers in the exposed samples of Pseudevernia furfuracea (L.) Zopf in the province of Kayseri (Central Anatolia).

In living organisms heterogenous air pollutants are considered as a major source of DNA damage. In this study, we aimed to describe the DNA changes in Pseudevernia furfuracea (L.) Zopf samples exposed to pollution at various sites in Kayseri by RAPD (random amplified polymorphic DNA) analysis in order to reveal the pattern of genetic variation influenced by the environmental pollution. The study area is characterized by the presence of numerous industrial activities, such as steel works, glassworks, shipbuilding, and metallurgical, mechanical, chemical and food industries. Control samples were collected from Cat forest (Sivas) and exposed to pollution in dry and wet seasons in the province of Kayseri located in Central Anatolia. Results indicated that heterogeneous pollutants might have contributed to the changes in the band patterns obtained by RAPD analysis, reflecting the occurrence of DNA damage in the control samples.

Heavy metal accumulation in Pseudevernia furfuracea (L.) Zopf from the Karabük iron-steel factory in Karabük, Turkey.

Pseudevernia furfuracea (L.) Zopf lichen specimens were collected every 5 km starting from around an iron-steel factory located in the central area of Karabük province, up to Yenice Forest. Zn, Cu, Mn, Fe, Pb, Ni, Cd, Cr contents were analyzed in the samples collected from polluted and unpolluted areas. A Pseudevernia furfuracea (L.) Zopf sample from Yenice Forest was used as a control. The reason for this choise was the abundance of species diversity, and therefore sample collection might cause a very low impact on natural population density. The forest is among the 100 forested areas that must be urgently taken under protection according to WWF (World Wildlife Fund) researches. Results of the current study manifested significant variations among the contents of these elements between stations. As expected, the pollution sources, such as iron-steel factory, roads and railroads, industry, heavy traffic, and waste treatment plants, have major impact on the heavy metal accumulation in P. furfuracea (L.) Zopf, and, in accordance to their location, samples 8 and 10 displayed high element accumulation. Surprisingly, although Yenice Forest is under protection, results of our study showed that the region is becoming polluted by the influence of many pollution sources in the area. The present study also confirms the efficient metal accumulation capacity of lichens.

Phylogenetic relations of Rhizoplaca Zopf. from Anatolia inferred from ITS sequence data.

Like many lichen-forming fungi, species of the genus Rhizoplaca have wide geographical distributions, but studies of their genetic variability are limited. The information about the ITS rDNA sequences of three species of Rhizoplaca from Anatolia was generated and aligned with other species from other countries and also with the data belonging to Lecanora species. The examined species were collected from the volcanic rocks of Mount Erciyes which is located in the middle of Anatolia (Turkey). The sequence data aligned with eight other samples of Rhizoplaca and six different species of Lecanora were obtained from GenBank. The results support the concept maintained by Arup and Grube (2000) that Rhizoplaca may not be a genus separate from Lecanora. According to the phylogenetic tree, Rhizoplaca melanopthalma from Turkey with two different samples of R. melanopthalma from Arizona (AF159929, AF159934) and a sample from Austria formed a group under the same branch. R. peltata and R. chrysoleuca samples from Anatolia located in two other branches of the tree formed sister groups with the samples of the same species from different countries. Although R. peltata remained on the same branch with other samples of the same species from other countries it was placed in a different branch within the group. When the three species from Anatolia were considered alone, it was noticed that Rhizoplaca melanopthalma and Rhizoplaca peltata are phylogenetically closer to each other than Rhizoplaca chrysoleuca; the morphological characteristics also support this result.