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The Ecosystem Services (ES) concept highlights the varied contributions the environment provides to humans and there are a wide range of methods/tools available to assess ES. However, in real-world decision contexts a single tool is rarely sufficient and methods must be combined to meet practitioner needs. Here, results from the OpenNESS project are presented to illustrate the methods selected to meet the needs of 24 real-world case studies and better understand why and how methods are combined to meet practical needs. Results showed that within the cases methods were combined to: i) address a range of ES; ii) assess both supply and demand of ES; iii) assess a range of value types; iv) reach different stakeholder groups v) cover weaknesses in other methods used and vi) to meet specific decision context needs. Methods were linked in a variety of ways: i) as input-output chains of methods; ii) through learning; iii) through method development and iv) through comparison/triangulation of results. The paper synthesises these case study-based experiences to provide insight to others working in practical contexts as to where, and in what contexts, different methods can be combined and how this can add value to case study analyses.

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Production and use of doubled haploids (DH) is becoming an essential part of maize breeding programs worldwide as DH lines offer several advantages in line development and evaluation. One of the critical steps in maize DH line production is doubling the chromosomes of in vivo-derived haploids so that naturally sterile haploids become reproductively fertile diploids (DH) to produce seed. This step of artificially doubling the chromosomes is labor-intensive and costly; hence, optimizing protocols to improve the doubling success is critical for achieving efficiencies in the DH production pipelines. Immersion of 3–4-day old germinating haploid seedlings in colchicine solution is commonly used for chromosome doubling in large-scale maize DH line production. This manuscript presents a new method of colchicine application to haploid seedlings that showed superior doubling rates compared to other methods like standard seedling immersion, seed immersion, root immersion, and direct application of colchicine solution to the seedlings at V2 stage in the greenhouse trays. The new method involves immersing the crown region of the haploid seedlings along with all the seedling roots at V2 stage in the colchicine solution. Further experiments to optimize this method indicated that increasing colchicine concentration had a very positive effect on overall success rate in chromosomal doubling, while not drastically affecting survival rate. The optimized method showed on average 5.6 times higher overall success rate (OSR) compared to the standard haploid seedling immersion method which was the second-best method in our experiments. This improved method of colchicine application saves resources by reducing the number of haploids to be generated and handled in a maize DH production pipeline.