A1 Geo-referenced inventory and characterization of current populations (distribution, demography, genetic diversity)

The global knowledge of a species range and features is essential for any conservation planning. Accordingly, it is necessary to collect information about the target species and its current habitat in all known populations. For each population of Calendula maritima we will estimate the numerical size and perimeter of each single unit, through sub-metric precision GPS device. In the aim to investigate the associated plant species, also phytosociological relevés will be carried out in order to analyze the ecological role and dynamism of the plant communities the target species belongs to. Moreover, since in the same range of the target species, the congener species Calendula fulgida is often present, hybridization and genetic pollution are a concrete threat. Accordingly, data on some morphological characters will be collected and a qualitative-quantitative estimate of the percentage of distribution of the different morphotypes will be provided. All the data will be incorporated in a georeferenced database, which is the starting point for all the monitoring activities related to the evolutionary trends at species and population level, planned within and beyond the project duration.

With regard to the problems of genetic pollution, a genetic characterization of C. maritima and related morphotypes will be carried out in order to discriminate the pure lineages, on which to concentrate conservation actions. Since no specific molecular markers for C. maritima exists, a de novo assembly of the transcriptome (the transcribed part of DNA) of the two species – C. maritima and C. fulgida – will be performed through new sequencing techniques (NGS – Next Generation Sequencing). The goal is to draw species-specific molecular markers that will allow to unequivocally discriminating the target species from hybrids. The activities involves the extraction of the total RNA and the following sequencing of the entire transcriptome of C. maritima and C. fulgida, using RNA sequencing techniques (RNASeq). Later on, the selected markers will be employed for genotyping all the populations of the target species in order to identify sure pure individuals to be propagated, reintroduced and disseminated in subsequent actions (Actions C1, C2, C3 and C7).

 

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