Harnessing landrace diversity empowers wheat breeding.

Harnessing genetic diversity in major staple crops through the development of new breeding capabilities is essential to ensure food security1. Here we examined the genetic and phenotypic diversity of the A. E. Watkins landrace collection2 of bread wheat (Triticum aestivum), a major global cereal, by whole-genome re-sequencing of 827 Watkins landraces and 208 modern cultivars and in-depth field evaluation spanning a decade. We found that modern cultivars are derived from two of the seven ancestral groups of wheat and maintain very long-range haplotype integrity. The remaining five groups represent untapped genetic sources, providing access to landrace-specific alleles and haplotypes for breeding. Linkage disequilibrium-based haplotypes and association genetics analyses link Watkins genomes to the thousands of identified high-resolution quantitative trait loci and significant marker-trait associations. Using these structured germplasm, genotyping and informatics resources, we revealed many Watkins-unique beneficial haplotypes that can confer superior traits in modern wheat. Furthermore, we assessed the phenotypic effects of 44,338 Watkins-unique haplotypes, introgressed from 143 prioritized quantitative trait loci in the context of modern cultivars, bridging the gap between landrace diversity and current breeding. This study establishes a framework for systematically utilizing genetic diversity in crop improvement to achieve sustainable food security.

Genetic variation and heritability of grain protein deviation in European wheat genotypes.

There is a well-established negative relationship between the yield and the concentration of protein in the mature wheat grain. However, some wheat genotypes consistently deviate from this relationship, a phenomenon known as Grain Protein Deviation (GPD). Positive GPD is therefore of considerable interest in relation to reducing the requirement for nitrogen fertilization for producing wheat for breadmaking. We have carried out two sets of field experiments on multiple sites in South East England. The first set comprised 11 field trials of 6 cultivars grown over three years (2008-2011) and the second comprised 9 field trials of 40 genotypes grown over two years (2015-2017) and 5 field trials of 30 genotypes grown in a single year (2017-2018). All trials comprised three replicate randomized plots of each genotype and nutrient regime. These studies showed strong genetic variation in GPD, which also differed in stability between genotypes, with cultivars bred in the UK generally having higher GPD and higher stability than those bred in other European countries. The heritability of GPD was estimated as 0.44, based on data from the field trials of 30 and 40 genotypes. The largest component contributing to the genetic variance was genotype (0.30), with a smaller contribution of the interaction between genotype and year/site (0.11) and a small (but statistically significant) contribution of nitrogen level. These studies suggest that selection for GPD is a viable target for breeders.

Field phenotyping for African crops: overview and perspectives.

Improvements in crop productivity are required to meet the dietary demands of the rapidly-increasing African population. The development of key staple crop cultivars that are high-yielding and resilient to biotic and abiotic stresses is essential. To contribute to this objective, high-throughput plant phenotyping approaches are important enablers for the African plant science community to measure complex quantitative phenotypes and to establish the genetic basis of agriculturally relevant traits. These advances will facilitate the screening of germplasm for optimum performance and adaptation to low-input agriculture and resource-constrained environments. Increasing the capacity to investigate plant function and structure through non-invasive technologies is an effective strategy to aid plant breeding and additionally may contribute to precision agriculture. However, despite the significant global advances in basic knowledge and sensor technology for plant phenotyping, Africa still lags behind in the development and implementation of these systems due to several practical, financial, geographical and political barriers. Currently, field phenotyping is mostly carried out by manual methods that are prone to error, costly, labor-intensive and may come with adverse economic implications. Therefore, improvements in advanced field phenotyping capabilities and appropriate implementation are key factors for success in modern breeding and agricultural monitoring. In this review, we provide an overview of the current state of field phenotyping and the challenges limiting its implementation in some African countries. We suggest that the lack of appropriate field phenotyping infrastructures is impeding the development of improved crop cultivars and will have a detrimental impact on the agricultural sector and on food security. We highlight the prospects for integrating emerging and advanced low-cost phenotyping technologies into breeding protocols and characterizing crop responses to environmental challenges in field experimentation. Finally, we explore strategies for overcoming the barriers and maximizing the full potential of emerging field phenotyping technologies in African agriculture. This review paper will open new windows and provide new perspectives for breeders and the entire plant science community in Africa.

Machine Learning Methods for Automatic Segmentation of Images of Field- and Glasshouse-Based Plants for High-Throughput Phenotyping.

Image segmentation is a fundamental but critical step for achieving automated high- throughput phenotyping. While conventional segmentation methods perform well in homogenous environments, the performance decreases when used in more complex environments. This study aimed to develop a fast and robust neural-network-based segmentation tool to phenotype plants in both field and glasshouse environments in a high-throughput manner. Digital images of cowpea (from glasshouse) and wheat (from field) with different nutrient supplies across their full growth cycle were acquired. Image patches from 20 randomly selected images from the acquired dataset were transformed from their original RGB format to multiple color spaces. The pixels in the patches were annotated as foreground and background with a pixel having a feature vector of 24 color properties. A feature selection technique was applied to choose the sensitive features, which were used to train a multilayer perceptron network (MLP) and two other traditional machine learning models: support vector machines (SVMs) and random forest (RF). The performance of these models, together with two standard color-index segmentation techniques (excess green (ExG) and excess green-red (ExGR)), was compared. The proposed method outperformed the other methods in producing quality segmented images with over 98%-pixel classification accuracy. Regression models developed from the different segmentation methods to predict Soil Plant Analysis Development (SPAD) values of cowpea and wheat showed that images from the proposed MLP method produced models with high predictive power and accuracy comparably. This method will be an essential tool for the development of a data analysis pipeline for high-throughput plant phenotyping. The proposed technique is capable of learning from different environmental conditions, with a high level of robustness.

Modeling the spatial-spectral characteristics of plants for nutrient status identification using hyperspectral data and deep learning methods.

Sustainable fertilizer management in precision agriculture is essential for both economic and environmental reasons. To effectively manage fertilizer input, various methods are employed to monitor and track plant nutrient status. One such method is hyperspectral imaging, which has been on the rise in recent times. It is a remote sensing tool used to monitor plant physiological changes in response to environmental conditions and nutrient availability. However, conventional hyperspectral processing mainly focuses on either the spectral or spatial information of plants. This study aims to develop a hybrid convolution neural network (CNN) capable of simultaneously extracting spatial and spectral information from quinoa and cowpea plants to identify their nutrient status at different growth stages. To achieve this, a nutrient experiment with four treatments (high and low levels of nitrogen and phosphorus) was conducted in a glasshouse. A hybrid CNN model comprising a 3D CNN (extracts joint spectral-spatial information) and a 2D CNN (for abstract spatial information extraction) was proposed. Three pre-processing techniques, including second-order derivative, standard normal variate, and linear discriminant analysis, were applied to selected regions of interest within the plant spectral hypercube. Together with the raw data, these datasets were used as inputs to train the proposed model. This was done to assess the impact of different pre-processing techniques on hyperspectral-based nutrient phenotyping. The performance of the proposed model was compared with a 3D CNN, a 2D CNN, and a Hybrid Spectral Network (HybridSN) model. Effective wavebands were selected from the best-performing dataset using a greedy stepwise-based correlation feature selection (CFS) technique. The selected wavebands were then used to retrain the models to identify the nutrient status at five selected plant growth stages. From the results, the proposed hybrid model achieved a classification accuracy of over 94% on the test dataset, demonstrating its potential for identifying nitrogen and phosphorus status in cowpea and quinoa at different growth stages.

Genetic Diversity in Nitrogen Fertiliser Responses and N Gas Emission in Modern Wheat.

Crops assimilate nitrogen (N) as ammonium via the glutamine synthetase/glutamate synthase (GS/GOGAT) pathway which is of central importance for N uptake and potentially represents a bottle neck for N fertiliser-use efficiency. The aim of this study was to assess whether genetic diversity for N-assimilation capacity exists in wheat and could be exploited for breeding. Wheat plants rapidly, within 6 h, responded to N application with an increase in GS activity. This was not accompanied by an increase in GS gene transcript abundance and a comparison of GS1 and GS2 protein models revealed a high degree of sequence conservation. N responsiveness amongst ten wheat varieties was assessed by measuring GS enzyme activity, leaf tissue ammonium, and by a leaf-disc assay as a proxy for apoplastic ammonia. Based on these data, a high-GS group showing an overall positive response to N could be distinguished from an inefficient, low-GS group. Subsequent gas emission measurements confirmed plant ammonia emission in response to N application and also revealed emission of N2O when N was provided as nitrate, which is in agreement with our current understanding that N2O is a by-product of nitrate reduction. Taken together, the data suggest that there is scope for improving N assimilation capacity in wheat and that further investigations into the regulation and role of GS-GOGAT in NH3 emission is justified. Likewise, emission of the climate gas N2O needs to be reduced, and future research should focus on assessing the nitrate reductase pathway in wheat and explore fertiliser management options.

Determination of wheat spike and spikelet architecture and grain traits using X-ray Computed Tomography imaging.

BACKGROUND: Wheat spike architecture is a key determinant of multiple grain yield components and detailed examination of spike morphometric traits is beneficial to explain wheat grain yield and the effects of differing agronomy and genetics. However, quantification of spike morphometric traits has been very limited because it relies on time-consuming manual measurements. RESULTS: In this study, using X-ray Computed Tomography imaging, we proposed a method to efficiently detect the 3D architecture of wheat spikes and component spikelets by clustering grains based on their Euclidean distance and relative positions. Morphometric characteristics of wheat spikelets and grains, e.g., number, size and spatial distribution along the spike can be determined. Two commercial wheat cultivars, one old, Maris Widgeon, and one modern, Siskin, were studied as examples. The average grain volume of Maris Widgeon and Siskin did not differ, but Siskin had more grains per spike and therefore greater total grain volume per spike. The spike length and spikelet number were not statistically different between the two cultivars. However, Siskin had a higher spikelet density (number of spikelets per unit spike length), with more grains and greater grain volume per spikelet than Maris Widgeon. Spatial distribution analysis revealed the number of grains, the average grain volume and the total grain volume of individual spikelets varied along the spike. Siskin had more grains and greater grain volumes per spikelet from spikelet 6, but not spikelet 1-5, compared with Maris Widgeon. The distribution of average grain volume along the spike was similar for the two wheat cultivars. CONCLUSION: The proposed method can efficiently extract spike, spikelet and grain morphometric traits of different wheat cultivars, which can contribute to a more detailed understanding of the sink of wheat grain yield.

Impacts of G x E x M on Nitrogen Use Efficiency in Wheat and Future Prospects.

Globally it has been estimated that only one third of applied N is recovered in the harvested component of grain crops. This represents an incredible waste of resource and the overuse has detrimental environmental and economic consequences. There is substantial variation in nutrient use efficiency (NUE) from region to region, between crops and in different cropping systems. As a consequence, both local and crop specific solutions will be required for NUE improvement at local as well as at national and international levels. Strategies to improve NUE will involve improvements to germplasm and optimized agronomy adapted to climate and location. Essential to effective solutions will be an understanding of genetics (G), environment (E), and management (M) and their interactions (G x E x M). Implementing appropriate solutions will require agronomic management, attention to environmental factors and improved varieties, optimized for current and future climate scenarios. As NUE is a complex trait with many contributing processes, identifying the correct trait for improvement is not trivial. Key processes include nitrogen capture (uptake efficiency), utilization efficiency (closely related to yield), partitioning (harvest index: biochemical and organ-specific) and trade-offs between yield and quality aspects (grain nitrogen content), as well as interactions with capture and utilization of other nutrients. A long-term experiment, the Broadbalk experiment at Rothamsted, highlights many factors influencing yield and nitrogen utilization in wheat over the last 175 years, particularly management and yearly variation. A more recent series of trials conducted over the past 16 years has focused on separating the key physiological sub-traits of NUE, highlighting both genetic and seasonal variation. This perspective describes these two contrasting studies which indicate G x E x M interactions involved in nitrogen utilization and summarizes prospects for the future including the utilization of high throughput phenotyping technology.

Novel sources of variation in grain Zinc (Zn) concentration in bread wheat germplasm derived from Watkins landraces.

A diverse panel of 245 wheat genotypes, derived from crosses between landraces from the Watkins collection representing global diversity in the early 20th century and the modern wheat cultivar Paragon, was grown at two field sites in the UK in 2015-16 and the concentrations of zinc and iron determined in wholegrain using inductively coupled plasma-mass spectrometry (ICP-MS). Zinc concentrations in wholegrain varied from 24-49 mg kg-1 and were correlated with iron concentration (r = 0.64) and grain protein content (r = 0.14). However, the correlation with yield was low (r = -0.16) indicating little yield dilution. A sub-set of 24 wheat lines were selected from 245 wheat genotypes and characterised for Zn and Fe concentrations in wholegrain and white flour over two sites and years. White flours from 24 selected lines contained 8-15 mg kg-1 of zinc, which was positively correlated with the wholegrain Zn concentration (r = 0.79, averaged across sites and years). This demonstrates the potential to exploit the diversity in landraces to increase the concentration of Zn in wholegrain and flour of modern high yielding bread wheat cultivars.