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Unravelling wild carrot differentiation in Europe: preliminary data on a candidate gene approach

Publication . Nobre, T.; Ragonezi, C.; Arnholdt-Schmitt, B.

Carrot is an outcrossing species and levels of gene flow between populations, and even between wild and domesticated relatives, are expected to be high. Cases of natural hybridization and introgression of crops and wild relatives have been reported. Have these events diluted any putative habitat-adapted genotypes? In other words, can we still find a correlation between wild carrot genotypes and regional/local environment? We have chosen to start addressing this question using a member of the alternative oxidase (AOX) gene family. AOX genes seem to be linked to all kinds of abiotic and biotic stress reactions. Wild carrots were sampled in an environmental gradient across Western Europe. This gradient included sampling points with more deviating conditions, such as Sierra de Guadarrama or the central Pyrenees and the French Massif Central. Phylogenetic reconstruction on this molecular marker is to be combined with geographic, climatic, and ecological evidence. So far, the preliminary results suggest the existence of a biogeographical barrier at the Pyrenees, and higher gene diversity than initially expected. From an applied point of view, diversity of functional traits is much more relevant than species diversity. Gene transfer from wild to cultivated plants has contributed to the evolution of crop species. Providing that deterioration of genetic resources and biodiversity loss have not been drastic, gene transfer from wild plants has the potential to further contribute to a (targeted) improvement of cultivars.

2017Journal article

Open access

Reference genes selection and normalization of oxidative stress responsive genes upon different temperature stress conditions in Hypericum perforatum L

Publication . Velada, Isabel; Ragonezi, Carla; Arnholdt-Schmitt, Birgit; Cardoso, Hélia

Reverse transcription-quantitative real-time PCR (RT-qPCR) is a widely used technique for gene expression analysis. The reliability of this method depends largely on the suitable selection of stable reference genes for accurate data normalization. Hypericum perforatum L. (St. John’s wort) is a field growing plant that is frequently exposed to a variety of adverse environmental stresses that can negatively affect its productivity. This widely known medicinal plant with broad pharmacological properties (anti-depressant, anti-tumor, anti-inflammatory, antiviral, antioxidant, anti-cancer, and antibacterial) has been overlooked with respect to the identification of reference genes suitable for RT-qPCR data normalization. In this study, 11 candidate reference genes were analyzed in H. perforatum plants subjected to cold and heat stresses. The expression stability of these genes was assessed using GeNorm, NormFinder and BestKeeper algorithms. The results revealed that the ranking of stability among the three algorithms showed only minor differences within each treatment. The best-ranked reference genes differed between cold- and heat-treated samples; nevertheless, TUB was the most stable gene in both experimental conditions. GSA and GAPDH were found to be reliable reference genes in cold-treated samples, while GAPDH showed low expression stability in heat-treated samples. 26SrRNA and H2A had the highest stabilities in the heat assay, whereas H2A was less stable in the cold assay. Finally, AOX1, AOX2, CAT1 and CHS genes, associated with plant stress responses and oxidative stress, were used as target genes to validate the reliability of identified reference genes. These target genes showed differential expression profiles over time in treated samples. This study not only is the first systematic analysis for the selection of suitable reference genes for RT-qPCR studies in H. perforatum subjected to temperature stress conditions, but may also provide valuable information about the roles of genes associated with temperature stress responses.

2014Journal article

Open access