DNA Isolation from Desiccated Leaf Material for Plum Tree (Prunus domestica L.) Molecular Analysis

Conservation of biodiversity is very important, especially in species such as plum tree. One of the key aspects is the establishment of appropriate genetic markers that could be used in identifying provenances and also for breeding purposes. In this context, molecular characterization using SSR (short sequence repeats) markers is very useful and was chosen for our study. This article describes the assessment of a CTAB-based DNA extraction protocol for isolating and purifying DNA from the plum cultivars selected for molecular characterisation. The quality of DNA extracts was assessed by spectrophotometric measurement and their suitability for molecular analysis was evaluated using a SSR marker system. We concluded that the DNA extraction protocol is suitable for obtaining plum DNA that can be used for molecular analysis using SSR markers

Ecological analyses on benthic diatom and invertebrate communities from the Someșul Mic catchment area (Transylvania, Romania)

The present study focused on benthic diatom and invertebrate communities from the Someșul Mic catchment area, between Someșul Rece and Apahida localities. Five sites were chosen, and they were sampled in spring, summer and autumn 2014. The area experienced major human impacts on aquatic communities, all caused by the existence of numerous urban and rural centers: discharges of waste waters into the natural streams, hydro-technical works for the production of energy or for water storage, hydropeaking, pollution coming from point or diffuse sources. The presence and the indicator value of the diatom species identified in the study area, together with the relative abundance of benthic invertebrate taxa were used to characterize the ecological status of the five sampling sites. Both diatoms and invertebrates showed the highest ecological status at the sampling site located on the Someșul Rece River, while the sampling sites located in or downstream Cluj-Napoca displayed the most impacted conditions. However, eutrophic conditions were characteristic to all sampling sites, showing affected biotic communities even in habitats with low human impacts

Selection of DNA Isolation Method and PCR Protocol for the Study of Endemic Astragalus exscapus L. ssp. Transsilvanicus (Schur) Nyár

Astragalus exscapus (Fabaceae) is a melliferous, perennial plant, which fits within a rarity type characterized by having small populations of relatively high habitat specificity (Rabinowitz, 1981). Astragalus exscapus L. ssp. transsilvanicus (Schur) Nyár. is an endemic subspecies, which has been identified in the Transylvanian basin in 24 populations. In order to establish conservation strategies and to characterize polymorphism of the threatened subspecies, genetic and morphologic analysis has to be implemented. Our study describes the application and optimalization of SRAP procedures to A. exscapus, including sampling, DNA extraction, PCR amplification and electrophoresis. Sequence Related Amplified Polymorphism (SRAP) molecular marker system is going to be used on eight different populations of A. exscapus ssp. transsilvanicus to identify the degree of polymorphism which is essential for the evolution of species. The methodology was tested and optimized successfully in two repetitive experiments. The results confirmed the suitability of the DNA extraction method to be applied and the optimization of the SRAP technique for the large-scale studies we are performing on this subspecies

Preliminary testing of SRAP primers in order to establish genetic diversity of Astragalus exscapus L. subsp. transsilvanicus (Schur) Nyár.

The Carpathian List of Endangered Species contains eight Astragalus species mentioned to be vulnerable or endangered. One of them is the endemic Astragalus. exscapus L. subsp. transsilvanicus (Schur) Nyár., a rare perennial plant with only 24 populations remaining in the Transylvanian basin. Analysis of genetic diversity and population structure of this threatened subspecies is an important prerequisite for its conservation. The present study investigates eight different populations of A. exscapus using sequence related amplified polymorphism (SRAP) molecular markers. This technique is a relatively new PCR-based system, more reliable than random amplified polymorphic DNA (RAPD), and suitable for population structure analysis. This essay reports the screening of 64 SRAP primer combinations on 3 or 4 samples from every population with the intent of selecting the most reliable ones for subsequent analysis. PCR amplification products were assessed on 2% agarose gel and a total number of 1473 DNA fragments were visualized by EtBr staining. We selected fourteen primer combinations for further research based on clarity, reproducibility of amplified bands and high rates of polymorphism. The most efficient primer combination was Me8/Em3 which generated 42 visible DNA fragments. Â

Interaction-driven relaxation of two-level systems in glasses

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