Increase in Tomato Locule Number Is Controlled by Two Single-Nucleotide Polymorphisms Located

In tomato (Solanum lycopersicum) fruit, the number of locules (cavities containing seeds that are derived from carpels) varies from two to up to 10 or more. Locule number affects fruit shape and size and is controlled by several quantitative trait loci(QTLs). The large majority of the phenotypic variation is explained by two of these QTLs, fasciated (fas) and locule number (lc), that interact epistatically with one another. FAS has been cloned, and mutations in the gene are described as key factors leading to the increase in fruit size in modern varieties. Here, we report the map-based cloning of lc. The lc QTL includes a 1,600-bp region that is located 1,080 bp from the 3# end of WUSCHEL, which encodes a homeodomain protein that regulates stem cell fate in plants. The molecular evolution of lc showed a reduction of diversity in cultivated accessions with the exception of two single-nucleotide polymorphisms. These two single-nucleotide polymorphisms were shown to be responsible for the increase in locule number. An evolutionary model of locule number is proposed herein, suggesting that the fas mutation appeared after the mutation in the lc locus to confer the extreme high-locule-number phenotype

Relative quantification in seed GMO analysis: state of art and bottlenecks

International audienceReliable quantitative methods are needed to comply with current EU regulations on the mandatory labeling of genetically modified organisms (GMOs) and GMO-derived food and feed products with a minimum GMO content of 0.9 %. The implementation of EU Commission Recommendation 2004/787/EC on technical guidance for sampling and detection which meant as a helpful tool for the practical implementation of EC Regulation 1830/2003, which states that "the results of quantitative analysis should be expressed as the number of target DNA sequences per target taxon specific sequences calculated in terms of haploid genomes". This has led to an intense debate on the type of calibrator best suitable for GMO quantification. The main question addressed in this review is whether reference materials and calibrators should be matrix based or whether pure DNA analytes should be used for relative quantification in GMO analysis. The state of the art, including the advantages and drawbacks, of using DNA plasmid (compared to genomic DNA reference materials) as calibrators, is widely described. In addition, the influence of the genetic structure of seeds on real-time PCR quantitative results obtained for seed lots is discussed. The specific composition of a seed kernel, the mode of inheritance, and the ploidy level ensure that there is discordance between a GMO % expressed as a haploid genome equivalent and a GMO % based on numbers of seeds. This means that a threshold fixed as a percentage of seeds cannot be used as such for RT-PCR. All critical points that affect the expression of the GMO content in seeds are discussed in this paper

Multigenerational versus single generation studies to estimate herbicide resistance fitness cost in Arabidopsis thaliana

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Investigating a flower-insect forager network in a mountain grassland community using pollen DNA barcoding

International audienceFaced with the decline of pollinators, it is relevant to strengthen our understanding of the whole plant-pollinator web in semi-natural grasslands that serve as refuges for pollinator populations. The aim of this study was to explore the diversity of flower-foraging insects involved in pollen transfer in mountain semi-natural grasslands. Insects actively collecting pollen and/or nectar were caught in spring in six mountain semi-natural grasslands displaying a floristic richness gradient. Individual determinations of insects were made at the finest possible taxonomic scale and pollen loads were removed from the insect body. Using next-generation DNA sequencing, pollens were identified through the ribosomal DNA cistron using the ITS2 database and the ITS plant rDNA cistron sequences from Genbank. A total of 236 flower-foraging insects were collected. Diptera represented 82% of the total catches distantly followed by Hymenoptera (15%) and Apoidea (bees) (11%). Visual observations revealed that Diptera foraged on 16 of the 21 flower species visited by insects. DNA metabarcoding showed that 82% (191) of all of the collected insects were carrying pollen and 44% (104) were carrying two genera of plants or more. Our results demonstrate that Diptera are potential key-pollinators in mountain semi-natural grasslands that cannot be overlooked by the scientific community. However difficulties of taxonomic determination due to severe shortage of experts for Diptera have to be urgently overcome. Further studies on the link between pollen transfer and actual pollination in a global change context are also required. Moreover, our results support the idea that DNA metabarcoding provides accurate information about the plants-insects networks but it also pointed out sensitive issues, especially the necessity to build reliable national barcode databases

Development of a Potato Cultivar (Solanum tuberosum L.) Core Collection, a Valuable Tool to Prospect Genetic Variation for Novel Traits

International audienceThis study presents the development of a core collection capturing the genetic diversity of a collection of 350 tetraploid cultivated potato varieties (Solanum tuberosum L.). The core collection was established by using simple sequence repeats (SSR) data and the M strategy, which aims at maximizing the allelic diversity. A 48-core collection was defined which captured 99.5% of the SSR alleles used to establish it, and 96.9% of the SSR alleles which belonged to an independent set of markers. The defined core collection was further validated by analysing 35 agro-morphological traits. The class coverage value and the estimates of the Shannon-Weaver diversity index indicated a good representation of the phenotypic diversity in the core collection. Furthermore, the core set included accessions having the most desirable status for several agronomic traits. A linkage disequilibrium (LD) analysis, using data obtained with the SolCAP SNP array on the defined core collection, was performed. The population structure analysis showed that the core collection did not present a clear genetic structure. The linkage disequilibrium analysis carried out between markers located on the same pseudomolecule within 10,000 bp concluded that 41.3% of these pairs of SNP markers have a significant LD. We conclude that this core collection, representative of the genetic diversity of cultivated potato varieties, is a relevant tool for a first screening for genetic variation regarding novel traits of interest

Long Reads for Long-Term Goals : A Genome Resource for Lavander Genetic Improvement

International audienceLavender (Lavandula angustifolia L.) is both a patrimonial and economically important species, used for its essential oils in cosmetics and perfumery products. Naturally adapted to poor soil and dry environment, lavender represents a promising culture in the context of climate change. However, increasing pathogenic pressure related to climate change challenges present varieties yields and quality of essential oils. The cultivar 'Maillette', that is most-valued for the quality of its essential oil, is widely used in genetic improvement programs in France. Useful genomic resources available for these programs remain nevertheless scarce, despite a short-reads assembled genome available. To support the emerging programs for genetic improvement in lavender, we generated genomic resources based on long-reads sequences, in complement with Illumina short-reads sequences

Deciphering the genetic diversity of landraces with high-throughput SNP genotyping of DNA bulks: methodology and application to the maize 50k array Mariangela Arca

Genebanks harbor original landraces carrying many original favorable alleles for mitigating biotic and abiotic stresses. Their genetic diversity remains however poorly characterized due to their large within genetic diversity. We developed a high-throughput, cheap and labor saving DNA bulk approach based on SNP Illumina Infinium HD array to genotype landraces. Samples were gathered for each landrace by mixing equal weights from young leaves, from which DNA was extracted. We then estimated allelic frequencies in each DNA bulk based on fluorescent intensity ratio (FIR) between two alleles at each SNP using a two step-approach. We first tested either whether the DNA bulk was monomorphic or polymorphic according to the two FIR distributions of individuals homozygous for allele A or B, respectively. If the DNA bulk was polymorphic, we estimated its allelic frequency by using a predictive equation calibrated on FIR from DNA bulks with known allelic frequencies. Our approach: (i) gives accurate allelic frequency estimations that are highly reproducible across laboratories, (ii) protects against false detection of allele fixation within landraces. We estimated allelic frequencies of 23,412 SNPs in 156 landraces representing American and European maize diversity. Modified Roger’s genetic Distance between 156 landraces estimated from 23,412 SNPs and 17 SSRs using the same DNA bulks were highly correlated, suggesting that the ascertainment bias is low. Our approach is affordable, easy to implement and does not require specific bioinformatics support and laboratory equipment, and therefore should be highly relevant for large-scale characterization of genebanks for a wide range of species