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Introduction: Growing grass-legume mixtures for forage production improves both yield productivity and nutritional quality, while also benefiting the environment by promoting species biodiversity and enhancing soil fertility (through nitrogen fixation). Consequently, assessing legume proportions in grass-legume mixed swards is essential for breeding and cultivation. This study introduces an approach for automated classification and mapping of species in mixed grass-clover swards using object-based image analysis (OBIA). Methods: The OBIA procedure was established for both RGB and ten band multispectral (MS) images capturedby an unmanned aerial vehicle (UAV). The workflow integrated structural (canopy heights) and spectral variables (bands, vegetation indices) along with a machine learning algorithm (Random Forest) to perform image segmentation and classification. Spatial k-fold cross-validation was employed to assess accuracy. Results and discussion: Results demonstrated good performance, achieving an overall accuracy of approximately 70%, for both RGB and MS-based imagery, with grass and clover classes yielding similar F1 scores, exceeding 0.7 values. The effectiveness of the OBIA procedure and classification was examined by analyzing correlations between predicted clover fractions and dry matter yield (DMY) proportions. This quantification revealed a positive and strong relationship, with R2 values exceeding 0.8 for RGB and MS-based classification outcomes. This indicates the potential of estimating (relative) clover coverage, which could assist breeders but also farmers in a precision agriculture context. [ABSTRACT FROM AUTHOR]

Breeding and selection of nursery plants require evaluation of a wide variety of traits. Characteristics that are visually scored in the field include aesthetic beauty as well as tolerance to (a)biotic stresses, among others. This work proposes methodologies based on vegetation indices and canopy height data derived from visual imagery captured using an RGB (red, green, blue) camera embedded in a drone to rank and select genotypes. This approach relies on quantitative evaluation standards that exclude breeder bias and speed up data collection. A proof of concept for nursery plants was developed in two woody ornamentals: sweet box (Sarcococca Lindl.) and garden rose (Rosa L.). This work aimed to compare methodologies and to propose how drones (unmanned aerial vehicles, UAV) and high throughput field phenotyping (HTFP) can be used in a commercial plant selection program. Data was gathered in 2019 in three well-established breeding trials, two for sweet box and one for rose. Characteristics discussed include plant architecture traits such as plant height and shape in the sweet box and rose as well as floribundity, continuous flowering and disease resistance in roses. Correlations were calculated between on-ground measurements and UAV-derived methods, obtaining significant results. The advantages and disadvantages of the methodology and the approach for each of the traits are discussed. [ABSTRACT FROM AUTHOR]

To induce new variation within the Escallonia genus, chromosome doubling was performed in E. rubra, E. rosea, and E. illinita, three important species within this genus of mainly evergreen woody ornamental species. Obtained tetraploids and diploid controls were analyzed for rooting capacity, leaf and flower characteristics, and plant architecture using image analysis and cold tolerance. In the present study, a breeders' collection of 23 accessions was characterized cytogenetically and described morphologically. All analyzed species and cultivars were diploid (2n = 2x = 24), with exception of E. pendula, a tetraploid. Today, breeding in Escallonia is limited to lucky finds in seedling populations and few efforts in interspecific hybridization. Three selected Escallonia species underwent an in vitro chromosome doubling with both oryzalin and trifluralin applied as either a continuous or shock treatment. The treatments successfully induced polyploids in all three species. Image analysis revealed that tetraploid E. rosea had decreased shoot length (from 3.8 to 1.3 cm), higher circularity and more dense growth habit compared to diploids. No significant changes in cold tolerance were seen. Tetraploid E. illinita did not differ in shoot length, but an increased outgrowth of axillary buds on the main axis led to denser plants. For tetraploid E. rubra, an increase in plant height (from 4.9 to 5.5 cm) was observed together with a large decrease in circularity and density due to a more polar outgrowth of branches on the main axis. E. rubra tetraploids bore larger flowers than diploids and had an increased cold tolerance (from -7.7 to -11.8°C). Leaf width and area of tetraploids increased for both E. illinita and E. rubra, while a decrease was seen in E. rosea genotypes. For all three species, the rooting capacity of the tetraploids did not differ from the diploids. We conclude that the effect of polyploidization on Escallonia was highly variable and species dependent. [ABSTRACT FROM AUTHOR]

Background: Genetic studies and breeding of agricultural crops frequently involve phenotypic characterization of large collections of genotypes grown in field conditions. These evaluations are typically based on visual observations and manual (destructive) measurements. Robust image capture and analysis procedures that allow phenotyping large collections of genotypes in time series during developmental phases represent a clear advantage as they allow non-destructive monitoring of plant growth and performance. A L. perenne germplasm panel including wild accessions, breeding material and commercial varieties has been used to develop a low-cost, high-throughput phenotyping tool for determining plant growth based on images of individual plants during two consecutive growing seasons. Further we have determined the correlation between image analysis-based estimates of the plant's base area and the capacity to regrow after cutting, with manual counts of tiller number and measurements of leaf growth 2 weeks after cutting, respectively. When working with field-grown plants, image acquisition and image segmentation are particularly challenging as outdoor light conditions vary throughout the day and the season, and variable soil colours hamper the delineation of the object of interest in the image. Therefore we have used several segmentation methods including colour-, texture- and edge-based approaches, and factors derived after a fast Fourier transformation. The performance of the procedure developed has been analysed in terms of effectiveness across different environmental conditions and time points in the season. Results: The procedure developed was able to analyse correctly 77.2 % of the 24,048 top view images processed. High correlations were found between plant's base area (image analysis-based) and tiller number (manual measurement) and between regrowth after cutting (image analysis-based) and leaf growth 2 weeks after cutting (manual measurement), with r values up to 0.792 and 0.824, respectively. Nevertheless, these relations depend on the origin of the plant material (forage breeding lines, current forage varieties, current turf varieties, and wild accessions) and the period in the season. Conclusions: The image-derived parameters presented here deliver reliable, objective data, complementary to the breeders' scores, and are useful for genetic studies. Furthermore, large variation was shown among genotypes for the parameters investigated. [ABSTRACT FROM AUTHOR]

In azalea breeding, flower colour is the most essential selection criterion, but leaf morphology also influences attractiveness of the plant. Despite extensive study of the inheritance of flower colour, no explanation has yet been found for the pink phenotype. We have used image analysis to quantify flower colour and leaf morphology. Flower colour was quantified in a whole population. Pink flower colour has been confirmed to be the result of a gene-dosage effect; two major QTLs were found for flower colour as well as some minor QTLs that seem to be related to pink coloration. Leaf morphology (colour and shape) was scored in four unrelated populations by means of image analysis. The image analysis generated continuous, highly informative data for QTL mapping. Both classical parameters and symmetrical elliptic Fourier descriptors successfully described leaf morphology. Image analysis resulted in large data sets that had to be combined in principal components. Only a limited number of QTLs were found for leaf colour, but we could discern some major QTLs for leaf shape and size that were consistent over the different mapping populations. These QTLs are the most interesting candidates for future analysis of the selected traits. The multi-population approach certainly proved to be valuable for QTL mapping of complex traits. For leaf morphology, however, more research is needed to identify the most valuable QTLs for future use in MAS. [ABSTRACT FROM AUTHOR]