orchid species have been recorded from this region and 85% of them are tuberous, most of them belonging to the genera Orchis, Serapias, Ophrys, Anacamptis, Dactylorhiza, Cephalanthera, Epipactis, and some other genera (Sezik, 2002). The tubers of terrestrial orchid species are very valuable due to the presence of glucomannose and are used to produce a special drink and ice cream having particular aroma and rheological properties (Kaya and Tekin, 2001; Dalar and Konczak, 2012, 2013). Although consumption of this drink is high, these plants are not cultivated and hence are collected from nature, leading to even disappearance of these groups of species from some areas (Kasparek and Grimm, 1999; Şekercioğlu et al., 2011). Although a significant number of research studies focused on the pharmaceutical properties, biology, propagation method, and genetic and phenotypic structure of these common groups of terrestrial species have been carried out, morphological and physiological characteristics of these species are still ambiguous in Abstract: Terrestrial orchid species are natural sources of salep and a closely related group of plant species widely distributed throughout Turkey. The phylogenetic relationship among fourteen different tuber-producing orchid species was investigated after analyzing phenotypic and genetic variation within and among the natural population through fifteen morphometric traits and ten random amplified polymorphic DNA (RAPD) primer combinations. Statistical analyses (principal component analysis (PCA), principal coordinate analysis (PCoA), and cluster analysis) using the generated data identified taxonomic and genetic distance within the studied plant samples. The results of PCA from morphological traits show that there are no major groupings within and among different species instead somehow overlapping with few distinctly characterized species. In addition, the UPGMA-based phenogram with Euclidean distance (0-1) produces five major clusters among the studied orchid species according to their taxonomic status with few exceptions. On the other hand, PCoA and the phylogenetic dendrogram with the coefficient (0.56-0.79) from RAPD band profiles determine the true genetic diversity of those species. Although both combinations of genetic and phenotypic characteristics reveal the phylogenetic relationship of some those studied species very effectively, they are not clear for others. These results suggest that in the natural population of terrestrial orchid species significant amounts of gene flow are ongoing at intra/interspecies level. Therefore, it is recommended that conservation studies of these groups of orchid species should be done as a geographical unit rather than according to taxonomic status.

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The objective of this study was to develop inexpensive and facile nanocomposites based on chitosan and organo-clay with the antimicrobial affectivity that provide the serious challenges caused by bacterial infections in various products such as food packaging materials. The chitosan with octadecylamine montmorillonite (ODA-MMT) nanocomposites by supplementing 1, 2.5, and 5 w% Nigella arvensis seed (black cumin) extract (CMBC-1, CMBC-2.5, and CMBC-5) were prepared chitosan from ionic liquid solutions in the presence of ODA-MMT and black cumin extract suspension. The effect of black cumin with different content on the structure and antimicrobial activity of the nanocomposite have been investigated. The interactions between the chitosan matrix, ODA-MMT, and black cumin extract at different conditions were characterized both physicochemically (FT-IR, SEM, and XRD) and biologically (antimicrobial). The results indicated that the formation of exfoliated nanostructure of nano composites was provided by loading of nanodispersed clay in matrix. Antimicrobial activity of CMBC nanocomposite film was evaluated using disc diffusion method against Gram-negative bacteria Escherichia coli ATCC 25922 and Salmonella enterica serotype Typhimurium SL 1344 and Gram-positives Staphylococcus aureus ATCC 25923 and Streptococcus mutans ATCC 25175. The antimicrobial activity studies of the CMBCs illustrated that the nanocomposites could more strongly inhibit the growth of the tested Gram-negative bacteria than Gram-positive bacteria within increased content of black cumin from 1 to 5 w%. To our knowledge, this is the first report on the antimicrobial effect of CMBC nanocomposite film. Such biomaterials within nontoxic and inexpensive properties will thus have great potential applications in the development of new packing materials that can effectively prevent the antimicrobial formation.