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Publication The development of phenotypic protocols and adjustment of experimental designs in Pelargonium zonale breeding(2018) Molenaar, Heike; Piepho, Hans-PeterOrnamental plant variety improvement is limited by current phenotyping approaches and the lack of use of experimental designs. Robust phenotypic data obtained from experiments laid out to best control local variation by blocking allow adequate statistical analysis and are crucial for any breeding purpose, including MAS. Often experiments consist of multiple phases like in P. zonale breeding, where in the first phase stock plants are cultivated to obtain the stem cutting count and in the second phase the stem cuttings are further assess for root formation. The first analyses of rooting experiments raised questions regarding options for improving the two-phase experimental layout, for example whether there is a disadvantage to using exactly the same design in both phases. The other question was, whether a design can be optimized across both phases, such that the MVD can be decreased. Instead of generating a separate layout for each phase. Moreover, optimal selection methods that maximize selection gain in P. zonale breeding based on available data collected from unreplicated trials and containing pedigree information were sought. This thesis was conducted to evaluate the benefits of using two-phase experimental designs and corresponding analysis in P. zonale for production related traits, for which it was necessary to establish phenotyping protocols. To optimize the rooting experiments with their two-phase nature, alternative approaches were explored involving two-phase design generation either in phase wise order or across phases. Furthermore, selection methods considering pedigreeinformation (family-index selection) or not (individual selection), were evaluated to enhance selection efficiency in P. zonale breeding. The benefits of using experimental designs in P. zonale breeding was shown by the simulated response to selection. Alternative designs were evaluated by the MVD obtained by the intrablock analysis and the joint inter-block-intra-block analysis. The efficiency of individual and family-index selection was evaluated in terms of heritability obtained from linear mixed models implementing the selection methods. Simulated response to selection varied greatly, depending on the genotypic variances of the breeding population and traits. However, by using efficient designs allowing adequate analysis, a varietal improvement of over 20% of stock plant reduction is possible for stem cutting count, root formation, branch count and flower count. The smallest MVD for alternative designs was most frequently obtained for designs generated across phases rather than for each phase separately, in particular when both phases of the design were separated with a single pseudolevel. Family-index selection was superior to individual selection in P. zonale indicating that the pedigree-based BLUP procedure can further enhance selection efficiency in productionrelated traits in P. zonale. The quantification of genotypic variation by phenotypic protocols and the optimized two-phase designs for estimating genotypic values were necessary and successful steps in laying the foundation for effective MAS. Phenotypic protocols effectively characterized the genetic material on an observational unit level, while the two-phase experimental designs enabled effective characterization on a genotype level by adjusting entry means using linear mixed models. The resulting adjusted entry means are the basis for future genotype phenotype association for MAS.