Commenced in January 2007
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2 Utilization of Developed Single Sequence Repeats Markers for Dalmatian Pyrethrum (Tanacetum cinerariifolium) in Preliminary Genetic Diversity Study on Natural Populations
Authors: F. Varga, Z. Liber, J. Jakše, A. Turudić, Z. Šatović, I. Radosavljević, N. Jeran, M. Grdiša
Abstract:
Dalmatian pyrethrum (Tanacetum cinerariifolium (Trevir.) Sch. Bip.; Asteraceae), a source of the commercially dominant plant insecticide pyrethrin, is a species endemic to the eastern Adriatic. Genetic diversity of T. cinerariifolium was previously studied using amplified fragment length polymorphism (AFLP) markers. However, microsatellite markers (single sequence repeats - SSRs) are more informative because they are codominant, highly polymorphic, locus-specific, and more reproducible, and thus are most often used to assess the genetic diversity of plant species. Dalmatian pyrethrum is an outcrossing diploid (2n = 18) whose large genome size and high repeatability have prevented the success of the traditional approach to SSR markers development. The advent of next-generation sequencing combined with the specifically developed method recently enabled the development of, to the author's best knowledge, the first set of SSRs for genomic characterization of Dalmatian pyrethrum, which is essential from the perspective of plant genetic resources conservation. To evaluate the effectiveness of the developed SSR markers in genetic differentiation of Dalmatian pyrethrum populations, a preliminary genetic diversity study was conducted on 30 individuals from three geographically distinct natural populations in Croatia (northern Adriatic island of Mali Lošinj, southern Adriatic island of Čiovo, and Mount Biokovo) based on 12 SSR loci. Analysis of molecular variance (AMOVA) by randomization test with 10,000 permutations was performed in Arlequin 3.5. The average number of alleles per locus, observed and expected heterozygosity, probability of deviations from Hardy-Weinberg equilibrium, and inbreeding coefficient was calculated using GENEPOP 4.4. Genetic distance based on the proportion of common alleles (DPSA) was calculated using MICROSAT. Cluster analysis using the neighbor-joining method with 1,000 bootstraps was performed with PHYLIP to generate a dendrogram. The results of the AMOVA analysis showed that the total SSR diversity was 23% within and 77% between the three populations. A slight deviation from Hardy-Weinberg equilibrium was observed in the Mali Lošinj population. Allele richness ranged from 2.92 to 3.92, with the highest number of private alleles observed in the Mali Lošinj population (17). The average observed DPSA between 30 individuals was 0.557. The highest DPSA (0.875) was observed between several pairs of Dalmatian pyrethrum individuals from the Mali Lošinj and Mt. Biokovo populations, and the lowest between two individuals from the Čiovo population. Neighbor-joining trees, based on DPSA, grouped individuals into clusters according to their population affiliation. The separation of Mt. Biokovo clade was supported (bootstrap value 58%), which is consistent with the previous study on AFLP markers, where isolated populations from Mt. Biokovo differed from the rest of the populations. The developed SSR markers are an effective tool for assessing the genetic diversity and structure of natural Dalmatian pyrethrum populations. These preliminary results are encouraging for a future comprehensive study with a larger sample size across the species' range. Combined with the biochemical data, these highly informative markers could help identify potential genotypes of interest for future development of breeding lines and cultivars that are both resistant to environmental stress and high in pyrethrins. Acknowledgment: This work has been supported by the Croatian Science Foundation under the project ‘Genetic background of Dalmatian pyrethrum (Tanacetum cinerariifolium /Trevir./ Sch. Bip.) insecticidal potential’- (PyrDiv) (IP-06-2016-9034) and by project KK.01.1.1.01.0005, Biodiversity and Molecular Plant Breeding, at the Centre of Excellence for Biodiversity and Molecular Plant Breeding (CoE CroP-BioDiv), Zagreb, Croatia.Keywords: Asteraceae, genetic diversity, genomic SSRs, NGS, pyrethrum, Tanacetum cinerariifolium
Procedia PDF Downloads 1141 Understanding Patterns of Hard Coral Demographics in Kenyan Reefs to Inform Restoration
Authors: Swaleh Aboud, Mishal Gudka, David Obura
Abstract:
Background: Coral reefs are becoming increasingly vulnerable due to several threats ranging from climate change to overfishing. This has resulted in increased management and conservation efforts to protect reefs from degradation and facilitate recovery. Recruitmentof new individuals are isimportant in the recovery process and critical for the persistence of coral reef ecosystems. Local coral community structure can be influenced by successful recruit settlement, survival, and growth Understanding coral recruitment patterns can help quantify reef resilience and connectivity, establish baselines and track changes and evaluate the effectiveness of reef restoration and conservation efforts. This study will examine the abundance and spatial pattern of coral recruits and how this relates to adult community structure, including the distribution of thermal resistance and sensitive genera and their distribution in different management regimes. Methods: Coral recruit and demography surveys were conducted from 2020 to 2022, covering 35 sites in 19coral reef locations along the Kenyan coast. These included marine parks, reserves, community conservation areas (CMAs), and open access areas from the north (Marereni) to the south (Kisite) coast of Kenya and across different reef habitats. The data was collected through the underwater visual census (UVC) technique. We counted adult corals (>10 cm diameter)of23 selected genera using belt transects (25 by 1 m) and sampling of 1 m2 quadrat (at an interval of 5m) for all coloniesless than 10 cm diameter. The benthic cover was collected using photo quadrats. The surveys were only done during the northeast monsoon season. The data wereanalyzed using the R program to see the distribution patterns and the Kruskal Wallis test to see whether there was a significant difference. Spearman correlation was also applied to assess the relationship between the distribution of coral genera in recruits and adults. Results: A total of 44 different coral genera were recorded for recruits, ranging from 3at Marereni to 30at Watamu Marine Reserve. Recruit densities ranged from 1.2±1.5recruit m-2 (mean±SD) at Likoni to 10.3± 8.4 recruit m-2 at Kisite Marine Park. The overall densityof recruitssignificantly differed between reef locations, with Kisite Marine Park and Reserve and Likonihaving significantly large differences from all the other locations, while Vuma, Watamu, Malindi, and Kilifi had significantly lower differences from all the other locations. The recruit generadensity along the Kenya coastwas divided into two clusters, one of which only included sites inKisite Marine Park. Adult colonies were dominated by Porites massive, Acropora, Platygyra, and Favites, whereas recruits were dominated by Porites branching, Porites massive, Galaxea, and Acropora. However, correlation analysis revealed a statistically significant positive correlation (r=0.81, p<0.05) between recruit and adult coral densities across the 23 coral genera. Marereni, which had the lowest densityof recruits, has only thermallyresistant coral genera, while Kisite Marine Park, with the highest recruit densities, has over 90% thermal sensitive coral genera. A weak positive correlation was found between recruit density and coralline algae, dead standing corals, and turf algae, whereas a weak negative correlation was found between recruit density and bare substrate and macroalgae. Between management regimes, marine reserves were found to have more recruits than no-take zones (marine parks and CMAs) and open access areas, although the difference was not significant. Conclusion: There was a statistically significant difference in the density of recruits between different reef locations along the Kenyan coast. Although the dominating genera of adults and recruits were different, there was a strong positive correlation between their coral communities, which could indicate self-recruitment processes or consistent distance seedings (of the same recruit genera). Sites such as Kisite Marine Park, with high recruit densities but dominated by thermally sensitive genera, will, on the other hand, be adversely affected by future thermal stress. This could imply that reducing the threats to coral reefs such as overfishingcould allow for their natural regeneration and recovery.Keywords: coral recruits, coral adult size-class, cora demography, resilience
Procedia PDF Downloads 124