Construction and genetic characterization of an interspecific raspberry hybrids panel aiming resistance to late leaf rust and adaptation to tropical regions.

Raspberries (Rubus spp) are temperate climate fruits with profitable high returns and have the potential for diversification of fruit growing in mid to low-latitude regions. However, there are still no cultivars adapted to climatic conditions and high pressure of diseases that occurs in tropical areas. In this context, our objective was to evaluate the genetic diversity from a 116 raspberry genotypes panel obtained from interspecific crosses in a testcross scheme with four cultivars already introduced in Brazil. The panel was genotyped via genotyping-by-sequencing. 28,373 and 27,281 SNPs were obtained, using the species R. occidentalis and R. idaeus genomes as references, respectively. A third marker dataset was constructed consisting of 41,292 non-coincident markers. Overall, there were no differences in the results when using the different marker sets for the subsequent analyses. The mean heterozygosity was 0.54. The average effective population size was 174, indicating great genetic variability. The other analyses revealed that the half-sibling families were structured in three groups. It is concluded that the studied panel has great potential for breeding and further genetic studies. Moreover, only one of the three marker matrices is sufficient for diversity studies.

On the usefulness of mock genomes to define heterotic pools, testers, and hybrid predictions in orphan crops.

The advances in genomics in recent years have increased the accuracy and efficiency of breeding programs for many crops. Nevertheless, the adoption of genomic enhancement for several other crops essential in developing countries is still limited, especially for those that do not have a reference genome. These crops are more often called orphans. This is the first report to show how the results provided by different platforms, including the use of a simulated genome, called the mock genome, can generate in population structure and genetic diversity studies, especially when the intention is to use this information to support the formation of heterotic groups, choice of testers, and genomic prediction of single crosses. For that, we used a method to assemble a reference genome to perform the single-nucleotide polymorphism (SNP) calling without needing an external genome. Thus, we compared the analysis results using the mock genome with the standard approaches (array and genotyping-by-sequencing (GBS)). The results showed that the GBS-Mock presented similar results to the standard methods of genetic diversity studies, division of heterotic groups, the definition of testers, and genomic prediction. These results showed that a mock genome constructed from the population's intrinsic polymorphisms to perform the SNP calling is an effective alternative for conducting genomic studies of this nature in orphan crops, especially those that do not have a reference genome.

Optimizing self-pollinated crop breeding employing genomic selection: From schemes to updating training sets.

Long-term breeding schemes using genomic selection (GS) can boost the response to selection per year. Although several studies have shown that GS delivers a higher response to selection, only a few analyze which stage GS produces better results and how to update the training population to maintain prediction accuracy. We used stochastic simulation to compare five GS breeding schemes in a self-pollinated long-term breeding program. Also, we evaluated four strategies, using distinct methods and sizes, to update the training set. Finally, regarding breeding schemes, we proposed a new approach using GS to select the best individuals in each F2 progeny, based on genomic estimated breeding values and genetic divergence, to cross them and generate a new recombination event. Our results showed that the best scenario was using GS in F2, followed by the phenotypic selection of new parents in F4. For TS updating, adding new data every cycle (over 768) to update the TS maintains the prediction accuracy at satisfactory levels for more breeding cycles. However, only the last three generations can be kept in the TS, optimizing the genetic relationship between TS and the targeted population and reducing the computing demand and risks. Hence, we believe that our results may help breeders optimize GS in their programs and improve genetic gain in long-term schemes.

Genotyping marker density and prediction models effects in long-term breeding schemes of cross-pollinated crops.

KEY MESSAGE: In genomic recurrent selection, the more markers, the better because they buffer the linkage disequilibrium losses caused by recombination over cycles, and consequently, provide higher responses to selection. Reductions of genotyping marker density have been extensively evaluated as potential strategies to reduce the genotyping costs of genomic selection (GS). Low-density marker panels are appealing in GS because they entail lower multicollinearity and computing time and allow more individuals to be genotyped for the same cost. However, statistical models used in GS are usually evaluated with empirical data, using "static" training sets and populations. This may be adequate for making predictions during a breeding program's initial cycles but not for the long-term. Moreover, studies that focus on long selective breeding cycles generally do not consider GS models with the effect of dominance, which is particularly important for breeding outcomes in cross-pollinated crops. Hence, dominance effects are important and unexplored in GS for long-term programs involving allogamous species. To address it, we employed two approaches: analysis of empirical maize datasets and simulations of long-term breeding applying phenotypic and genomic recurrent selection (intrapopulation and reciprocal schemes). In both schemes, we simulated twenty breeding cycles and assessed the effect of marker density reduction on the population mean, the best crosses, additive variance, selective accuracy, and response to selection with models [additive, additive-dominant, general (GCA), and this plus specific combining ability (GCA + SCA)]. Our results indicate that marker reduction based on linkage disequilibrium levels provides useful predictions only within a cycle, as accuracy significantly decreases over cycles. In the long-term, without training set updating, high-marker density provides the best responses to selection. The model to be used depends on the breeding scheme: additive for intrapopulation and additive-dominant or GCA + SCA for reciprocal.

Automated Machine Learning: A Case Study of Genomic "Image-Based" Prediction in Maize Hybrids.

Machine learning methods such as multilayer perceptrons (MLP) and Convolutional Neural Networks (CNN) have emerged as promising methods for genomic prediction (GP). In this context, we assess the performance of MLP and CNN on regression and classification tasks in a case study with maize hybrids. The genomic information was provided to the MLP as a relationship matrix and to the CNN as "genomic images." In the regression task, the machine learning models were compared along with GBLUP. Under the classification task, MLP and CNN were compared. In this case, the traits (plant height and grain yield) were discretized in such a way to create balanced (moderate selection intensity) and unbalanced (extreme selection intensity) datasets for further evaluations. An automatic hyperparameter search for MLP and CNN was performed, and the best models were reported. For both task types, several metrics were calculated under a validation scheme to assess the effect of the prediction method and other variables. Overall, MLP and CNN presented competitive results to GBLUP. Also, we bring new insights on automated machine learning for genomic prediction and its implications to plant breeding.

Population-tailored mock genome enables genomic studies in species without a reference genome.

Based on molecular markers, genomic prediction enables us to speed up breeding schemes and increase the response to selection. There are several high-throughput genotyping platforms able to deliver thousands of molecular markers for genomic study purposes. However, even though its widely applied in plant breeding, species without a reference genome cannot fully benefit from genomic tools and modern breeding schemes. We used a method to assemble a population-tailored mock genome to call single-nucleotide polymorphism (SNP) markers without an available reference genome, and for the first time, we compared the results with standard genotyping platforms (array and genotyping-by-sequencing (GBS) using a reference genome) for performance in genomic prediction models. Our results indicate that using a population-tailored mock genome to call SNP delivers reliable estimates for the genomic relationship between genotypes. Furthermore, genomic prediction estimates were comparable to standard approaches, especially when considering only additive effects. However, mock genomes were slightly worse than arrays at predicting traits influenced by dominance effects, but still performed as well as standard GBS methods that use a reference genome. Nevertheless, the array-based SNP markers methods achieved the best predictive ability and reliability to estimate variance components. Overall, the mock genomes can be a worthy alternative for genomic selection studies, especially for those species where the reference genome is not available.

Optimizing Genomic-Enabled Prediction in Small-Scale Maize Hybrid Breeding Programs: A Roadmap Review.

The usefulness of genomic prediction (GP) for many animal and plant breeding programs has been highlighted for many studies in the last 20 years. In maize breeding programs, mostly dedicated to delivering more highly adapted and productive hybrids, this approach has been proved successful for both large- and small-scale breeding programs worldwide. Here, we present some of the strategies developed to improve the accuracy of GP in tropical maize, focusing on its use under low budget and small-scale conditions achieved for most of the hybrid breeding programs in developing countries. We highlight the most important outcomes obtained by the University of São Paulo (USP, Brazil) and how they can improve the accuracy of prediction in tropical maize hybrids. Our roadmap starts with the efforts for germplasm characterization, moving on to the practices for mating design, and the selection of the genotypes that are used to compose the training population in field phenotyping trials. Factors including population structure and the importance of non-additive effects (dominance and epistasis) controlling the desired trait are also outlined. Finally, we explain how the source of the molecular markers, environmental, and the modeling of genotype-environment interaction can affect the accuracy of GP. Results of 7 years of research in a public maize hybrid breeding program under tropical conditions are discussed, and with the great advances that have been made, we find that what is yet to come is exciting. The use of open-source software for the quality control of molecular markers, implementing GP, and envirotyping pipelines may reduce costs in an efficient computational manner. We conclude that exploring new models/tools using high-throughput phenotyping data along with large-scale envirotyping may bring more resolution and realism when predicting genotype performances. Despite the initial costs, mostly for genotyping, the GP platforms in combination with these other data sources can be a cost-effective approach for predicting the performance of maize hybrids for a large set of growing conditions.

On the genetic architecture in a public tropical maize panel of the symbiosis between corn and plant growth-promoting bacteria aiming to improve plant resilience.

Exploring the symbiosis between plants and plant growth-promoting bacteria (PGPB) is a new challenge for sustainable agriculture. Even though many works have reported the beneficial effects of PGPB in increasing plant resilience for several stresses, its potential is not yet widely explored. One of the many reasons is the differential symbiosis performance depending on the host genotype. This opens doors to plant breeding programs to explore the genetic variability and develop new cultivars with higher responses to PGPB interaction and, therefore, have higher resilience to stress. Hence, we aimed to study the genetic architecture of the symbiosis between PGPB and tropical maize germplasm, using a public association panel and its impact on plant resilience. Our findings reveal that the synthetic PGPB population can modulate and impact root architecture traits and improve resilience to nitrogen stress, and 37 regions were significant for controlling the symbiosis between PGPB and tropical maize. In addition, we found two overlapping SNPs in the GWAS analysis indicating strong candidates for further investigations. Furthermore, genomic prediction analysis with genomic relationship matrix computed using only significant SNPs obtained from GWAS analysis substantially increased the predictive ability for several traits endorsing the importance of these genomic regions for the response of PGPB. Finally, the public tropical panel reveals a significant genetic variability to the symbiosis with the PGPB and can be a source of alleles to improve plant resilience. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-021-01257-6.

A novel way to validate UAS-based high-throughput phenotyping protocols using in silico experiments for plant breeding purposes.

KEY MESSAGE: It is possible to make inferences regarding the feasibility and applicability of plant high-throughput phenotyping via computer simulations. Protocol validation has been a key challenge to the establishment of high-throughput phenotyping (HTP) in breeding programs. We add to this matter by proposing an innovative way for designing and validating aerial imagery-based HTP approaches with in silico 3D experiments for plant breeding purposes. The algorithm is constructed following a pipeline composed of the simulation of phenotypic values, three-dimensional modeling of trials, and image rendering. Our tool is exemplified by testing a set of experimental setups that are of interest in the context of maize breeding using a comprehensive case study. We report on how the choice of (percentile of) points in dense clouds, the experimental repeatability (heritability), the treatment variance (genetic variability), and the flight altitude affect the accuracy of high-throughput plant height estimation based on conventional structure-from-motion (SfM) and multi-view stereo (MVS) pipelines. The evaluation of both the algorithm and the case study was driven by comparisons of the computer-simulated (ground truth) and the HTP-estimated values using correlations, regressions, and similarity indices. Our results showed that the 3D experiments can be adequately reconstructed, enabling inference-making. Moreover, it suggests that treatment variance, repeatability, and the choice of the percentile of points are highly influential over the accuracy of HTP. Conversely, flight altitude influenced the quality of reconstruction but not the accuracy of plant height estimation. Therefore, we believe that our tool can be of high value, enabling the promotion of new insights and further understanding of the events underlying the practice of high-throughput phenotyping.