Development and validation of high-throughput and low-cost STARP assays for genes underpinning economically important traits in wheat.

KEY MESSAGE: We developed and validated 56 gene-specific semi-thermal asymmetric reverse PCR (STARP) markers for 46 genes of important wheat quality, biotic and abiotic stress resistance, grain yield, and adaptation-related traits for marker-assisted selection in wheat breeding. Development of high-throughput, low-cost, gene-specific molecular markers is important for marker-assisted selection in wheat breeding. In this study, we developed 56 gene-specific semi-thermal asymmetric reverse PCR (STARP) markers for wheat quality, tolerance to biotic and abiotic stresses, grain yield, and adaptation-related traits. The STARP assays were validated by (1) comparison of the assays with corresponding diagnostic STS/CAPS markers on 40 diverse wheat cultivars and (2) characterization of allelic effects based on the phenotypic and genotypic data of three segregating populations and 305 diverse wheat accessions from China and 13 other countries. The STARP assays showed the advantages of high-throughput, accuracy, flexibility, simple assay design, low operational costs, and platform compatibility. The state-of-the-art assays of this study provide a robust and reliable molecular marker toolkit for wheat breeding programs.

Development and validation of KASP assays for genes underpinning key economic traits in bread wheat.

KEY MESSAGE: We developed and validated a robust marker toolkit for high-throughput and cost-effective screening of a large number of functional genes in wheat. Functional markers (FMs) are the most valuable markers for crop breeding programs, and high-throughput genotyping for FMs could provide an excellent opportunity to effectively practice marker-assisted selection while breeding cultivars. Here we developed and validated kompetitive allele-specific PCR (KASP) assays for genes that underpin economically important traits in bread wheat including adaptability, grain yield, quality, and biotic and abiotic stress resistances. In total, 70 KASP assays either developed in this study or obtained from public databases were validated for reliability in application. The validation of KASP assays were conducted by (a) comparing the assays with available gel-based PCR markers on 23 diverse wheat accessions, (b) validation of the derived allelic information using phenotypes of a panel comprised of 300 diverse cultivars from China and 13 other countries, and (c) additional testing, where possible, of the assays in four segregating populations. All KASP assays being reported were significantly associated with the relevant phenotypes in the cultivars panel and bi-parental populations, thus revealing potential application in wheat breeding programs. The results revealed 45 times superiority of the KASP assays in speed than gel-based PCR markers. KASP has recently emerged as single-plex high-throughput genotyping technology; this is the first report on high-throughput screening of a large number of functional genes in a major crop. Such assays could greatly accelerate the characterization of crossing parents and advanced lines for marker-assisted selection and can complement the inflexible, high-density SNP arrays. Our results offer a robust and reliable molecular marker toolkit that can contribute towards maximizing genetic gains in wheat breeding programs.

A pilot study to determine whether using a lightweight, wearable micro-camera improves dietary assessment accuracy and offers information on macronutrients and eating rate.

A major limitation in nutritional science is the lack of understanding of the nutritional intake of free-living people. There is an inverse relationship between accuracy of reporting of energy intake by all current nutritional methodologies and body weight. In this pilot study we aim to explore whether using a novel lightweight, wearable micro-camera improves the accuracy of dietary intake assessment. Doubly labelled water (DLW) was used to estimate energy expenditure and intake over a 14-d period, over which time participants (n 6) completed a food diary and wore a micro-camera on 2 of the days. Comparisons were made between the estimated energy intake from the reported food diary alone and together with the images from the micro-camera recordings. There was an average daily deficit of 3912 kJ using food diaries to estimate energy intake compared with estimated energy expenditure from DLW (P=0·0118), representing an under-reporting rate of 34 %. Analysis of food diaries alone showed a significant deficit in estimated daily energy intake compared with estimated intake from food diary analysis with images from the micro-camera recordings (405 kJ). Use of the micro-camera images in conjunction with food diaries improves the accuracy of dietary assessment and provides valuable information on macronutrient intake and eating rate. There is a need to develop this recording technique to remove user and assessor bias.

Hierarchical HMM based learning of navigation primitives for cooperative robotic endovascular catheterization.

Despite increased use of remote-controlled steerable catheter navigation systems for endovascular intervention, most current designs are based on master configurations which tend to alter natural operator tool interactions. This introduces problems to both ergonomics and shared human-robot control. This paper proposes a novel cooperative robotic catheterization system based on learning-from-demonstration. By encoding the higher-level structure of a catheterization task as a sequence of primitive motions, we demonstrate how to achieve prospective learning for complex tasks whilst incorporating subject-specific variations. A hierarchical Hidden Markov Model is used to model each movement primitive as well as their sequential relationship. This model is applied to generation of motion sequences, recognition of operator input, and prediction of future movements for the robot. The framework is validated by comparing catheter tip motions against the manual approach, showing significant improvements in the quality of catheterization. The results motivate the design of collaborative robotic systems that are intuitive to use, while reducing the cognitive workload of the operator.