Accuracy of r_GP as a function of the number of randomly selected SNPs used to compute the GRM.

<p>Results for the three traits under analysis are reported: a) Yield; b) Lodging; c) Starch content. Accuracies obtained with the A+I, G+I and A+G+I, models are represented in blue, green and red, respectively. The color of the dots show if each r_GP was significantly lower than the highest observed r_GP obtained with each model.</p

Optimizing Training Population Size and Genotyping Strategy for Genomic Prediction Using Association Study Results and Pedigree Information. A Case of Study in Advanced Wheat Breeding Lines

<div><p>Wheat breeding programs generate a large amount of variation which cannot be completely explored because of limited phenotyping throughput. Genomic prediction (GP) has been proposed as a new tool which provides breeding values estimations without the need of phenotyping all the material produced but only a subset of it named training population (TP). However, genotyping of all the accessions under analysis is needed and, therefore, optimizing TP dimension and genotyping strategy is pivotal to implement GP in commercial breeding schemes. Here, we explored the optimum TP size and we integrated pedigree records and genome wide association studies (GWAS) results to optimize the genotyping strategy. A total of 988 advanced wheat breeding lines were genotyped with the Illumina 15K SNPs wheat chip and phenotyped across several years and locations for yield, lodging, and starch content. Cross-validation using the largest possible TP size and all the SNPs available after editing (~11k), yielded predictive abilities (r_GP) ranging between 0.5–0.6. In order to explore the Training population size, r_GP were computed using progressively smaller TP. These exercises showed that TP of around 700 lines were enough to yield the highest observed r_GP. Moreover, r_GP were calculated by randomly reducing the SNPs number. This showed that around 1K markers were enough to reach the highest observed r_GP. GWAS was used to identify markers associated with the traits analyzed. A GWAS-based selection of SNPs resulted in increased r_GP when compared with random selection and few hundreds SNPs were sufficient to obtain the highest observed r_GP. For each of these scenarios, advantages of adding the pedigree information were shown. Our results indicate that moderate TP sizes were enough to yield high r_GP and that pedigree information and GWAS results can be used to greatly optimize the genotyping strategy.</p></div

Manhattan plots.

<p>GWAS results for the three traits under analysis are displayed: a) Yield; b) Lodging; c) Starch content.</p

Prediction accuracy as a function of the training set size.

<p>Results are displayed for the three traits under analysis: a) Yield; b) Lodging; c) Starch content. Three models were considered: a) A+I in blue; b) G+I in green; c) A+G+I in red. The color of the dots show if each r_GP was significantly lower than the highest observed r_GP obtained with each model.</p

Biometrical analysis of parental species and interspecific hybrids.

<p>a: Number of open flowers per plant; b: Quotient of flower diameter and flower length; c: Chlorophyll Content Index. Parental species and interspecific hybrids codes are provided in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137537#pone.0137537.t001" target="_blank">Table 1</a>.</p

Descriptive statistics for the traits considered.

<p>Descriptive statistics for the traits considered.</p

Traits estimated variance components and predictive abilities.

<p>Traits estimated variance components and predictive abilities.</p

Origin, ploidy and chromosome number from selected <i>Campanula</i> species.

<p>[<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137537#pone.0137537.ref025" target="_blank">25</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0137537#pone.0137537.ref030" target="_blank">30</a>]</p><p>Origin, ploidy and chromosome number from selected <i>Campanula</i> species.</p

SNPs showing significant associations with the three traits considered.

<p>SNPs showing significant associations with the three traits considered.</p

Phenotypic and Genotypic Analysis of Newly Obtained Interspecific Hybrids in the <i>Campanula</i> Genus

<div><p>Interspecific hybridisation creates new phenotypes within several ornamental plant species including the <i>Campanula</i> genus. We have employed phenotypic and genotypic methods to analyse and evaluate interspecific hybridisation among cultivars of four <i>Campanula</i> species, i.e. <i>C</i>. <i>cochleariifolia</i>, C. <i>isophylla</i>, <i>C</i>. <i>medium</i> and <i>C</i>. <i>formanekiana</i>. Hybrids were analysed using amplified fragment length polymorphism (AFLP), flow cytometry and biometrical measurements. Results of correlation matrices demonstrated heterogeneous phenotypes for the parental species, which confirmed our basic premise for new phenotypes of interspecific hybrids. AFLP assays confirmed the hybridity and identified self-pollinated plants. Limitation of flow cytometry analysis detection was observed while detecting the hybridity status of two closely related parents, e.g. <i>C</i>. <i>cochleariiafolia</i> × <i>C</i>. <i>isophylla</i>. Phenotypic characteristics such as shoot habitus and flower colour were strongly influenced by one of the parental species in most crosses. Rooting analysis revealed that inferior rooting quality occurred more often in interspecific hybrids than in the parental species. Only interspecific hybrid lines of <i>C</i>. <i>formanekiana</i> ‘White’ × <i>C</i>. <i>medium</i> ‘Pink’ showed a high rooting level. Phenotype analyses demonstrated a separation from the interspecific hybrid lines of <i>C</i>. <i>formanekiana</i> ‘White’ × <i>C</i>. <i>medium</i> ‘Pink’ to the other clustered hybrids of <i>C</i>. <i>formanekiana</i> and <i>C</i>. <i>medium</i>. In our study we demonstrated that the use of correlation matrices is a suitable tool for identifying suitable cross material. This study presents a comprehensive overview for analysing newly obtained interspecific hybrids. The chosen methods can be used as guidance for analyses for further interspecific hybrids in <i>Campanula</i>, as well as in other ornamental species.</p></div