Vaccinium is a large genus of shrubs that includes a handful of economically important berry crops. Given the numerous hybridizations and polyploidization events, the taxonomy of this genus has remained the subject of long debate. In addition, berries and berry-based products are liable to adulteration, either fraudulent or unintentional due to misidentification of species. The availability of more genomic information could help achieve higher phylogenetic resolution for the genus, provide molecular markers for berry crops identification, and a framework for efficient genetic engineering of chloroplasts. Therefore, in this study we assembled five Vaccinium chloroplast sequences representing the economically relevant berry types: northern highbush blueberry (V. corymbosum), southern highbush blueberry (V. corymbosum hybrids), rabbiteye blueberry (V. virgatum), lowbush blueberry (V. angustifolium), and bilberry (V. myrtillus). Comparative analyses showed that the Vaccinium chloroplast genomes exhibited an overall highly conserved synteny and sequence identity among them. Polymorphic regions included the expansion/contraction of inverted repeats, gene copy number variation, simple sequence repeats, indels, and single nucleotide polymorphisms. Based on their in silico discrimination power, we suggested variants that could be developed into molecular markers for berry crops identification. Phylogenetic analysis revealed multiple origins of highbush blueberry plastomes, likely due to the hybridization events that occurred during northern and southern highbush blueberry domestication.
Vaccinium is an economically important genus of berry crops in the family Ericaceae. Given the numerous hybridizations and polyploidization events among Vaccinium species, the taxonomy of this genus has remained uncertain and the subject of long debate. Therefore, the availability of more genomic resources for Vaccinium can provide useful tools for phylogenetic resolution, species identification, authentication of berry food products, and a framework for genetic engineering. In this study, we assembled five Vaccinium chloroplast sequences representing the following berry types: northern highbush blueberry (V. corymbosum), southern highbush blueberry (V. corymbosum hybrids), rabbiteye blueberry (V. virgatum), lowbush blueberry (V. angustifolium), and bilberry (V. myrtillus). Two complete plastid genomes were achieved using long-read PacBio sequencing, while three draft sequences were obtained using short-read Illumina sequencing. Comparative analyses also included other previously available Vaccinium chloroplast sequences, especially the commercially important species V. macrocarpon (cranberry). The Vaccinium chloroplast genomes exhibited a circular quadripartite structure, with an overall highly conserved synteny and sequence identity among them. Despite their high similarity, we identified some polymorphic regions in terms of expansion/contraction of inverted repeats, gene copy number variation, simple sequence repeats, and single nucleotide polymorphisms. Phylogenetic analysis revealed multiple origins of highbush blueberry plastomes, likely due to the hybridization events during northern and southern highbush blueberry domestication. Additional whole plastome analyses including more samples and wild species will be useful to obtain a refined knowledge of the maternal breeding history of blueberries and increase phylogenetic resolution at low taxonomic levels.
The pathogen Xylella fastidiosa is a xylem-restricted, gram-negative bacterium that is known to cause diseases of many cultivated plant species. Recent outbreaks of X. fastidiosa diseases in Europe have brought attention to the impact of this pathogen, especially to perennial crops. Among the Prunus genus, X. fastidiosa is known to have a wide range of hosts, including plum, almond, peach, cherry, and apricot. Infected trees have reduced fruit quality, possibly resulting in unmarketable fruits, as well as reduced lifespan. There are no curative management options for X. fastidiosa diseases in Prunus; therefore, development of resistant or tolerant cultivars through breeding represents an efficient option to reduce the impact of this pathogen. In this context, the main objective of this study was to determine the occurrence of X. fastidiosa in germplasm of the Stone Fruit Breeding Program at the University of Florida located in Gainesville, FL, USA, under natural infection conditions. A total of 43 individuals representing 10 different genotypic groups within the Prunus genus were tested for the presence of X. fastidiosa. Additionally, we report a novel and easy sampling method using sawdust collected from tree trunks for the detection of this pathogen in Prunus and the development of an endogenous control for improving the diagnosis of this pathogen using real-time polymerase chain reactions. Our results showed a high incidence of X. fastidiosa in the germplasm tested, with more than 65% of the samples positive for the presence of the bacterial pathogen. However, X. fastidiosa was not detected in most of the P. mume samples tested, whereas almost all the P. mume × P. armeniaca hybrids were positive. Negative individuals were also identified in P. avium, P. campanulata, P. umbellata, and P. salicina × P. ceracifera. These trees have been planted in the field, exposed to natural infection for 4 to 11 years, and are considered to show field resistance. Finally, primers and probes based on the Prunus COX gene developed in this study can be used as an internal amplification control to enhance the interpretation of results of X. fastidiosa detection assays using sawdust samples.
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