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We present the first national DNA barcode resource that covers the native flowering plants and conifers for the nation of Wales (1143 species). Using the plant DNA barcode markers rbcL and matK, we have assembled 97.7% coverage for rbcL, 90.2% for matK, and a dual-locus barcode for 89.7% of the native Welsh flora. We have sampled multiple individuals for each species, resulting in 3304 rbcL and 2419 matK sequences. The majority of our samples (85%) are from DNA extracted from herbarium specimens. Recoverability of DNA barcodes is lower using herbarium specimens, compared to freshly collected material, mostly due to lower amplification success, but this is balanced by the increased efficiency of sampling species that have already been collected, identified, and verified by taxonomic experts. The effectiveness of the DNA barcodes for identification (level of discrimination) is assessed using four approaches: the presence of a barcode gap (using pairwise and multiple alignments), formation of monophyletic groups using Neighbour-Joining trees, and sequence similarity in BLASTn searches. These approaches yield similar results, providing relative discrimination levels of 69.4 to 74.9% of all species and 98.6 to 99.8% of genera using both markers. Species discrimination can be further improved using spatially explicit sampling. Mean species discrimination using barcode gap analysis (with a multiple alignment) is 81.6% within 10×10 km squares and 93.3% for 2×2 km squares. Our database of DNA barcodes for Welsh native flowering plants and conifers represents the most complete coverage of any national flora, and offers a valuable platform for a wide range of applications that require accurate species identification.

This thesis reports the development of several new and/or improved pipelines for investigating nucleotide alignments as part of DNA barcoding and DNA metabarcoding. In addition, the existing barcode regions were investigated to discover if possible mini-barcodes could be used in order to improve identification in a wide range of plant species. The pipelines developed in the project have also been designed to help the general barcode community in a wide range of areas from forensic analysis, food identification, monitoring illicit trade in plants and animals and numerous metabarcoding projects. From the development of the improved alignment pipeline, users will be able to align the rbcL and matK barcode regions with confidence, with only a minimal amount of manual editing required for matK. The results achieved for the rbcL were roughly the same for the standalone alignment tools tested, and for the matK gene the results were better than the standalone alignment tools but not as good as the manually edited version of the matK alignment. From the development of the new metabarcoding analysis pipeline, the results showed that by implementing the pipeline the number of reads decreased but the sequences being lost are low quality, by removing these reads the average quality of the reads left increases. The result is significant as the user would like to keep as many reads as possible with also achieving the highest quality reads possible at the same time. From investigating the barcode regions to find mini-barcodes, it was possible to find regions that could be used as mini-barcode region for both the rbcL and the matK barcode region. To extract these regions primer combinations were designed for the rbcL and for APG-III classification groups for the matK region. One of the rbcL primer combinations achieved 86 % and several of the matK primer combinations achieved above 60%. This result is significant as these primers could now be used along with the metabarcoding approach to extract matK sequences.

Despite being well-positioned to promote healthy lifestyles in young children, early years practitioners often face barriers to supporting child weight management. This mixed-methods study aimed to assess the preliminary effectiveness and acceptability of an e-learning toolkit (HealthyWEY) designed to upskill and support multi-agency professionals to promote healthy weight in early childhood. A total of 54 health visitors/community nursery nurses, 38 children’s centre staff and 17 other health professionals engaged with the HealthyWEY e-learning, which drew on self-determination theory and consisted of nine modules that were completed over 7–10 weeks. Non-parametric statistical analysis using Wilcoxon’s signed-rank tests were used to explore participants’ practice-based knowledge, psychological needs satisfaction and motivations for prioritising pre-school child weight from pre- to post-intervention. Focus groups (n = 11) were conducted with a sub-sample of multi-agency professionals (n = 39) to explore the process of implementation across sites, while interviews were also conducted with two parents/carers who took part in consultations with HealthyWEY-trained practitioners. After completing the HealthyWEY e-learning, participants perceived fewer barriers to pre-school child weight management (median change = −0.7; p < 0.001), greater autonomy (median change = 0.7, p < 0.001), competence (median change = 0.8, p < 0.001) and relatedness (median change = 0.4, p < 0.001) and a higher autonomous motivation towards promoting healthy weight (median change = 0.3, p < 0.001). E-learning was perceived to be highly relevant to participants’ roles and congruent with local child weight strategies. Challenges to implementation included time constraints and disruptions related to the COVID-19 pandemic. Recommendations for a better user experience were offered by enhancing the toolkit’s design and interactivity. Engagement with the HealthyWEY e-learning led to promising changes in perceived barriers and motivational variables. The toolkit was perceived to be acceptable amongst multi-agency workforces, albeit challenging to prioritise within time-pressured health and early years settings.