Key messageExploitation of data from a ryegrass breeding program has enabled rapid development and implementation of genomic selection for sward-based biomass yield with a twofold-to-threefold increase in genetic gain.AbstractGenomic selection, which uses genome-wide sequence polymorphism data and quantitative genetics techniques to predict plant performance, has large potential for the improvement in pasture plants. Major factors influencing the accuracy of genomic selection include the size of reference populations, trait heritability values and the genetic diversity of breeding populations. Global diversity of the important forage species perennial ryegrass is high and so would require a large reference population in order to achieve moderate accuracies of genomic selection. However, diversity of germplasm within a breeding program is likely to be lower. In addition, de novo construction and characterisation of reference populations are a logistically complex process. Consequently, historical phenotypic records for seasonal biomass yield and heading date over a 18-year period within a commercial perennial ryegrass breeding program have been accessed, and target populations have been characterised with a high-density transcriptome-based genotyping-by-sequencing assay. Ability to predict observed phenotypic performance in each successive year was assessed by using all synthetic populations from previous years as a reference population. Moderate and high accuracies were achieved for the two traits, respectively, consistent with broad-sense heritability values. The present study represents the first demonstration and validation of genomic selection for seasonal biomass yield within a diverse commercial breeding program across multiple years. These results, supported by previous simulation studies, demonstrate the ability to predict sward-based phenotypic performance early in the process of individual plant selection, so shortening the breeding cycle, increasing the rate of genetic gain and allowing rapid adoption in ryegrass improvement programs.Electronic supplementary materialThe online version of this article (10.1007/s00122-018-3121-7) contains supplementary material, which is available to authorized users.

Background In-field measurement of yield and growth rate in pasture species is imprecise and costly, limiting scientific and commercial application. Our study proposed a LiDAR-based mobile platform for non-invasive vegetative biomass and growth rate estimation in perennial ryegrass ( Lolium perenne L.). This included design and build of the platform, development of an algorithm for volumetric estimation, and field validation of the system. The LiDAR-based volumetric estimates were compared against fresh weight and dry weight data across different ages of plants, seasons, stages of regrowth, sites, and row configurations. Results The project had three phases, the last one comprising four experiments. Phase 1: a LiDAR-based, field-ready prototype mobile platform for perennial ryegrassrecognition in single row plots was developed. Phase 2: real-time volumetric data capture, modelling and analysis software were developed and integrated and the resultant algorithm was validated in the field. Phase 3. LiDAR Volume data were collected via the LiDAR platform and field-validated in four experiments. Expt.1: single-row plots of cultivars and experimental diploid breeding populations were scanned in the southern hemisphere spring for biomass estimation. Significant ( P < 0.001) correlations were observed between LiDAR Volume and both fresh and dry weight data from 360 individual plots (R 2 = 0.89 and 0.86 respectively). Expt 2: recurrent scanning of single row plots over long time intervals of a few weeks was conducted, and growth was estimated over an 83 day period. Expt 3: recurrent scanning of single-row plots over nine short time intervals of 2 to 5 days was conducted, and growth rate was observed over a 26 day period. Expt 4: recurrent scanning of paired-row plots over an annual cycle of repeated growth and defoliation was conducted, showing an overall mean correlation of LiDAR Volume and fresh weight of R 2 = 0.79 for 1008 observations made across seven different harvests between March and December 2018. Conclusions Here we report development and validation of LiDAR-based volumetric estimation as an efficient and effective tool for measuring fresh weight, dry weight and growth rate in single and paired-row plots of perennial ryegrass for the first time, with a consistently high level of accuracy. This development offers precise, non-destructive and cost-effective estimation of these economic traits in the field for ryegrass and potentially other pasture grasses in the future, based on the platform and algorithm developed for ryegrass. Electronic supplementary material The online version of this article (10.1186/s13007-019-0456-2) contains supplementary material, which is available to authorized users.

In perennial ryegrass (Lolium perenne L), annual and seasonal dry matter yield (DMY) and nutritive quality of herbage are high-priority traits targeted for improvement through selective breeding. Genomic prediction (GP) has proven to be a valuable tool for improving complex traits and may be further enhanced through the use of multi-trait (MT) prediction models. In this study, we evaluated the relative performance of MT prediction models to improve predictive ability for DMY and key nutritive quality traits, using two different training populations (TP1, n = 463 and TP2, n = 517) phenotyped at multiple locations. MT models outperformed single-trait (ST) models by 24% to 59% for DMY and 67% to 105% for nutritive quality traits, such as low, high, and total WSC, when a correlated secondary trait was included in both the training and test set (MT-CV2) or in the test set alone (MT-CV3) (trait-assisted genomic selection). However, when a secondary trait was included in training set and not the test set (MT-CV1), the predictive ability was not statistically significant (p > 0.05) compared to the ST model. We evaluated the impact of training set size when using a MT-CV2 model. Using a highly correlated trait (r g = 0.88) as the secondary trait in the MT-CV2 model, there was no loss in predictive ability for DMY even when the training set was reduced to 50% of its original size. In contrast, using a weakly correlated secondary trait (r g = 0.56) in the MT-CV2 model, predictive ability began to decline when the training set size was reduced by only 11% from its original size. Using a ST model, genomic predictive ability in a population unrelated to the training set was poor (r p = −0.06). However, when using an MT-CV2 model, the predictive ability was positive and high (r p = 0.76) for the same population. Our results demonstrate the first assessment of MT models in forage species and illustrate the prospects of using MT genomic selection in forages, and other outcrossing plant species, to accelerate genetic gains for complex agronomical traits, such as DMY and nutritive quality characteristics.

A targeted amplicon-based genotyping-by-sequencing approach has permitted cost-effective and accurate discrimination between ryegrass species (perennial, Italian and inter-species hybrid), and identification of cultivars based on bulked samples. Perennial ryegrass and Italian ryegrass are the most important temperate forage species for global agriculture, and are represented in the commercial pasture seed market by numerous cultivars each composed of multiple highly heterozygous individuals. Previous studies have identified difficulties in the use of morphophysiological criteria to discriminate between these two closely related taxa. Recently, a highly multiplexed single nucleotide polymorphism (SNP)-based genotyping assay has been developed that permits accurate differentiation between both species and cultivars of ryegrasses at the genetic level. This assay has since been further developed into an amplicon-based genotyping-by-sequencing (GBS) approach implemented on a second-generation sequencing platform, allowing accelerated throughput and ca. sixfold reduction in cost. Using the GBS approach, 63 cultivars of perennial, Italian and interspecific hybrid ryegrasses, as well as intergeneric Festulolium hybrids, were genotyped. The genetic relationships between cultivars were interpreted in terms of known breeding histories and indistinct species boundaries within the Lolium genus, as well as suitability of current cultivar registration methodologies. An example of applicability to quality assurance and control (QA/QC) of seed purity is also described. Rapid, low-cost genotypic assays provide new opportunities for breeders to more fully explore genetic diversity within breeding programs, allowing the combination of novel unique genetic backgrounds. Such tools also offer the potential to more accurately define cultivar identities, allowing protection of varieties in the commercial market and supporting processes of cultivar accreditation and quality assurance.

Plant breeding has had, and continues to have, an important role in providing farmers with resilient pastures. Early breeding relied on improvement of ecotype populations and this was accelerated by crossing with selected introduced germplasm. The primary traits under selection have targeted speed of establishment, total and/or seasonal dry matter (DM) yield, nutritive value or feed quality, flowering time and reduced aftermath heading, disease resistance, persistence and seed yield. Continued improvement through plant breeding to meet environmental concerns and tolerances to both biotic and abiotic stresses will be achieved through ongoing plant introductions, exploiting heterosis, speed breeding, genomic selection, improvements in phenotyping, metabolomics, improved compatibility with beneficial microbes, and potentially the use of transgenic and gene editing technologies.

Key message Breeding target traits can be broadened to include nutritive value and plant breeder’s rights traits in perennial ryegrass by using in-field regression-based spectroscopy phenotyping and genomic selection. Abstract Perennial ryegrass breeding has focused on biomass yield, but expansion into a broader set of traits is needed to benefit livestock industries whilst also providing support for intellectual property protection of cultivars. Numerous breeding objectives can be targeted simultaneously with the development of sensor-based phenomics and genomic selection (GS). Of particular interest are nutritive value (NV), which has been difficult and expensive to measure using traditional phenotyping methods, resulting in limited genetic improvement to date, and traits required to obtain varietal protection, known as plant breeder’s rights (PBR) traits. In order to assess phenotyping requirements for NV improvement and potential for genetic improvement, in-field reflectance-based spectroscopy was assessed and GS evaluated in a single population for three key NV traits, captured across four timepoints. Using three prediction approaches, the possibility of targeting PBR traits using GS was evaluated for five traits recorded across three years of a breeding program. Prediction accuracy was generally low to moderate for NV traits and moderate to high for PBR traits, with heritability highly correlated with GS accuracy. NV did not show significant or consistent correlation between timepoints highlighting the need to incorporate seasonal NV into selection indexes and the value of being able to regularly monitor NV across seasons. This study has demonstrated the ability to implement GS for both NV and PBR traits in perennial ryegrass, facilitating the expansion of ryegrass breeding targets to agronomically relevant traits while ensuring necessary varietal protection is achieved.

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