Rapid and High Throughput Hydroponics Phenotyping Method for Evaluating Chickpea Resistance to Phytophthora Root Rot.

Phytophthora root rot (PRR) is a major constraint to chickpea production in Australia. Management options for controlling the disease are limited to crop rotation and avoiding high risk paddocks for planting. Current Australian cultivars have partial PRR resistance, and new sources of resistance are needed to breed cultivars with improved resistance. Field- and glasshouse-based PRR resistance phenotyping methods are labour intensive, time consuming, and provide seasonally variable results; hence, these methods limit breeding programs' abilities to screen large numbers of genotypes. In this study, we developed a new space saving (400 plants/m2), rapid (<12 days), and simplified hydroponics-based PRR phenotyping method, which eliminated seedling transplant requirements following germination and preparation of zoospore inoculum. The method also provided post-phenotyping propagation all the way through to seed production for selected high-resistance lines. A test of 11 diverse chickpea genotypes provided both qualitative (PRR symptoms) and quantitative (amount of pathogen DNA in roots) results demonstrating that the method successfully differentiated between genotypes with differing PRR resistance. Furthermore, PRR resistance hydroponic assessment results for 180 recombinant inbred lines (RILs) were correlated strongly with the field-based phenotyping, indicating the field phenotype relevance of this method. Finally, post-phenotyping high-resistance genotypes were selected. These were successfully transplanted and propagated all the way through to seed production; this demonstrated the utility of the rapid hydroponics method (RHM) for selection of individuals from segregating populations. The RHM will facilitate the rapid identification and propagation of new PRR resistance sources, especially in large breeding populations at early evaluation stages.

A Multi-Environment Trial Analysis of Frost Susceptibility in Wheat and Barley Under Australian Frost-Prone Field Conditions.

Low temperatures during the flowering period of cereals can lead to floret sterility, yield reduction, and economic losses in Australian crops. In order to breed for improved frost susceptibility, selection methods are urgently required to identify novel sources of frost tolerant germplasm. However, the presence of genotype by environment interactions (i.e. variety responses to a change in environment) is a major constraint to select the most appropriate varieties in any given target environment. An advanced method of analysis for multi-environment trials that includes factor analytic selection tools to summarize overall performance and stability to a specific trait across the environments could deliver useful information to guide growers and plant breeding programs in providing the most appropriate decision making-strategy. In this study, the updated selection tools approached in this multi-environment trials (MET) analysis have allowed variety comparisons with similar frost susceptibility but which have a different response to changes in the environment or vice versa. This MET analysis included a wide range of sowing dates grown at multiple locations from 2010 to 2019, respectively. These results, as far as we are aware, show for the first-time genotypic differences to frost damage through a MET analysis by phenotyping a vast number of accurate empirical measurements that reached in excess of 557,000 spikes. This has resulted in a substantial number of experimental units (10,317 and 5,563 in wheat and barley, respectively) across a wide range of sowing times grown at multiple locations from 2010 to 2019. Varieties with low frost overall performance (OP) and low frost stability (root mean square deviation -RMSD) were less frost susceptible, with performance more consistent across all environments, while varieties with low OP and high RMSD were adapted to specific environmental conditions.

Genomic selection in multi-environment plant breeding trials using a factor analytic linear mixed model.

Genomic selection (GS) is a statistical and breeding methodology designed to improve genetic gain. It has proven to be successful in animal breeding; however, key points of difference have not been fully considered in the transfer of GS from animal to plant breeding. In plant breeding, individuals (varieties) are typically evaluated across a number of locations in multiple years (environments) in formally designed comparative experiments, called multi-environment trials (METs). The design structure of individual trials can be complex and needs to be modelled appropriately. Another key feature of MET data sets is the presence of variety by environment interaction (VEI), that is the differential response of varieties to a change in environment. In this paper, a single-step factor analytic linear mixed model is developed for plant breeding MET data sets that incorporates molecular marker data, appropriately accommodates non-genetic sources of variation within trials and models VEI. A recently developed set of selection tools, which are natural derivatives of factor analytic models, are used to facilitate GS for a motivating data set from an Australian plant breeding company. The power and versatility of these tools is demonstrated for the variety by environment and marker by environment effects.

Increased accuracy of starch granule type quantification using mixture distributions.

BACKGROUND: The proportion of granule types in wheat starch is an important characteristic that can affect its functionality. It is widely accepted that granule types are either large, disc-shaped A-type granules or small, spherical B-type granules. Additionally, there are some reports of the tiny C-type granules. The differences between these granule types are due to its carbohydrate composition and crystallinity which is highly, but not perfectly, correlated with the granule size. A majority of the studies that have considered granule types analyse them based on a size threshold rather than chemical composition. This is understandable due to the expense of separating starch into different types. While the use of a size threshold to classify granule type is a low-cost measure, this results in misclassification. We present an alternative, statistical method to quantify the proportion of granule types by a fit of the mixture distribution, along with an R package, a web based app and a video tutorial for how to use the web app to enable its straightforward application. RESULTS: Our results show that the reliability of the genotypic effects increase approximately 60% using the proportions of the A-type and B-type granule estimated by the mixture distribution over the standard size-threshold measure. Although there was a marginal drop in reliability for C-type granules. The latter is likely due to the low observed genetic variance for C-type granules. CONCLUSIONS: The determination of the proportion of granule types from size-distribution is better achieved by using the mixing probabilities from the fit of the mixture distribution rather than using a size-threshold.

Multiple Factors Affect Socioeconomics and Wellbeing of Artisanal Sea Cucumber Fishers.

Small-scale fisheries are important to livelihoods and subsistence seafood consumption of millions of fishers. Sea cucumbers are fished worldwide for export to Asia, yet few studies have assessed factors affecting socioeconomics and wellbeing among fishers. We interviewed 476 men and women sea cucumber fishers at multiple villages within multiple locations in Fiji, Kiribati, Tonga and New Caledonia using structured questionnaires. Low rates of subsistence consumption confirmed a primary role of sea cucumbers in income security. Prices of sea cucumbers sold by fishers varied greatly among countries, depending on the species. Gender variation in landing prices could be due to women catching smaller sea cucumbers or because some traders take advantage of them. Dissatisfaction with fishery income was common (44% of fishers), especially for i-Kiribati fishers, male fishers, and fishers experiencing difficulty selling their catch, but was uncorrelated with sale prices. Income dissatisfaction worsened with age. The number of livelihood activities averaged 2.2-2.5 across countries, and varied significantly among locations. Sea cucumbers were often a primary source of income to fishers, especially in Tonga. Other common livelihood activities were fishing other marine resources, copra production in Kiribati, agriculture in Fiji, and salaried jobs in New Caledonia. Fishing other coastal and coral reef resources was the most common fall-back livelihood option if fishers were forced to exit the fishery. Our data highlight large disparities in subsistence consumption, gender-related price equity, and livelihood diversity among parallel artisanal fisheries. Improvement of supply chains in dispersed small-scale fisheries appears as a critical need for enhancing income and wellbeing of fishers. Strong evidence for co-dependence among small-scale fisheries, through fall-back livelihood preferences of fishers, suggests that resource managers must mitigate concomitant effects on other fisheries when considering fishery closures. That is likely to depend on livelihood diversification programs to take pressure off co-dependent fisheries.

Ppd-1 is a key regulator of inflorescence architecture and paired spikelet development in wheat.

The domestication of cereal crops such as wheat, maize, rice and barley has included the modification of inflorescence architecture to improve grain yield and ease harvesting(1). Yield increases have often been achieved through modifying the number and arrangement of spikelets, which are specialized reproductive branches that form part of the inflorescence. Multiple genes that control spikelet development have been identified in maize, rice and barley(2-5). However, little is known about the genetic underpinnings of this process in wheat. Here, we describe a modified spikelet arrangement in wheat, termed paired spikelets. Combining comprehensive QTL and mutant analyses, we show that Photoperiod-1 (Ppd-1), a pseudo-response regulator gene that controls photoperiod-dependent floral induction, has a major inhibitory effect on paired spikelet formation by regulating the expression of FLOWERING LOCUS T (FT)(6,7). These findings show that modulated expression of the two important flowering genes, Ppd-1 and FT, can be used to form a wheat inflorescence with a more elaborate arrangement and increased number of grain producing spikelets.

Factor analytic mixed models for the provision of grower information from national crop variety testing programs.

KEY MESSAGE: Factor analytic mixed models for national crop variety testing programs have the potential to improve industry productivity through appropriate modelling and reporting to growers of variety by environment interaction. Crop variety testing programs are conducted in many countries world-wide. Within each program, data are combined across locations and seasons, and analysed in order to provide information to assist growers in choosing the best varieties for their conditions. Despite major advances in the statistical analysis of multi-environment trial data, such methodology has not been adopted within national variety testing programs. The most commonly used approach involves a variance component model that includes variety and environment main effects, and variety by environment (V × E) interaction effects. The variety predictions obtained from such an analysis, and subsequently reported to growers, are typically on a long-term regional basis. In Australia, the variance component model has been found to be inadequate in terms of modelling V × E interaction, and the reporting of information at a regional level often masks important local V × E interaction. In contrast, the factor analytic mixed model approach that is widely used in Australian plant breeding programs, has regularly been found to provide a parsimonious and informative model for V × E effects, and accurate predictions. In this paper we develop an approach for the analysis of crop variety evaluation data that is based on a factor analytic mixed model. The information obtained from such an analysis may well be superior, but will only enhance industry productivity if mechanisms exist for successful technology transfer. With this in mind, we offer a suggested reporting format that is user-friendly and contains far greater local information for individual growers than is currently the case.

Effects of mesh size and escape gaps on discarding in an Australian giant mud crab (Scylla serrata) trap fishery.

In response to concerns over excessive discarding from Australian recreational round traps (with four funnel entrances) used to target giant mud crabs, Scylla serrata, an experiment was done to assess the independent and cumulative utility of paired, bottom-located horizontal escape gaps (46×120 mm) and increasing mesh size (from 51 to 101 mm). Compared to conventional traps comprising 51-mm mesh throughout, those with the same mesh size and escape gaps caught significantly fewer (by 95%) undersize (<85 mm carapace length--CL) crabs while maintaining legal catches. Traps made from 101-mm mesh (but with the same funnel entrances as conventional designs) and with and without escape gaps similarly retained fewer undersize crabs and also yellowfin bream Acanthopagrus australis (the key bycatch species) by up to 94%, but there were concomitant reductions in fishing power for legal sizes of S. serrata. Although there were no immediate mortalities among any discarded crabs, there was a greater bias towards wounding among post molts than late inter-molts and less damage to individuals in the 101-mm conventional than 51-mm conventional traps (without escape gaps). The results support retrospectively fitting escape gaps in conventional S. serrata traps as a means for reducing discarding, but additional work is required to determine appropriate mesh sizes/configurations that maximize species and size selectivity.

Factor analytic and reduced animal models for the investigation of additive genotype-by-environment interaction in outcrossing plant species with application to a Pinus radiata breeding programme.

KEY MESSAGE: Modelling additive genotype-by-environment interaction is best achieved with the use of factor analytic models. With numerous environments and for outcrossing plant species, computation is facilitated using reduced animal models. The development of efficient plant breeding strategies requires a knowledge of the magnitude and structure of genotype-by-environment interaction. This information can be obtained from appropriate linear mixed model analyses of phenotypic data from multi-environment trials. The use of factor analytic models for genotype-by-environment effects is known to provide a reliable, parsimonious and holistic approach for obtaining estimates of genetic correlations between all pairs of trials. When breeding for outcrossing species the focus is on estimating additive genetic correlations and effects which is achieved by including pedigree information in the analysis. The use of factor analytic models in this setting may be computationally prohibitive when the number of environments is moderate to large. In this paper, we present an approach that uses an approximate reduced animal model to overcome the computational issues associated with factor analytic models for additive genotype-by-environment effects. The approach is illustrated using a Pinus radiata breeding dataset involving 77 trials, located in environments across New Zealand and south eastern Australia, and with pedigree information on 315,581 trees. Using this approach we demonstrate the existence of substantial additive genotype-by-environment interaction for the trait of stem diameter measured at breast height. This finding has potentially significant implications for both breeding and deployment strategies. Although our approach has been developed for forest tree breeding programmes, it is directly applicable for other outcrossing plant species, including sugarcane, maize and numerous horticultural crops.

Multivariate whole genome average interval mapping: QTL analysis for multiple traits and/or environments.

A major aim in some plant-based studies is the determination of quantitative trait loci (QTL) for multiple traits or across multiple environments. Understanding these QTL by trait or QTL by environment interactions can be of great value to the plant breeder. A whole genome approach for the analysis of QTL is presented for such multivariate applications. The approach is an extension of whole genome average interval mapping in which all intervals on a linkage map are included in the analysis simultaneously. A random effects working model is proposed for the multivariate (trait or environment) QTL effects for each interval, with a variance-covariance matrix linking the variates in a particular interval. The significance of the variance-covariance matrix for the QTL effects is tested and if significant, an outlier detection technique is used to select a putative QTL. This QTL by variate interaction is transferred to the fixed effects. The process is repeated until the variance-covariance matrix for QTL random effects is not significant; at this point all putative QTL have been selected. Unlinked markers can also be included in the analysis. A simulation study was conducted to examine the performance of the approach and demonstrated the multivariate approach results in increased power for detecting QTL in comparison to univariate methods. The approach is illustrated for data arising from experiments involving two doubled haploid populations. The first involves analysis of two wheat traits, α-amylase activity and height, while the second is concerned with a multi-environment trial for extensibility of flour dough. The method provides an approach for multi-trait and multi-environment QTL analysis in the presence of non-genetic sources of variation.

Estimation in a multiplicative mixed model involving a genetic relationship matrix.

Genetic models partitioning additive and non-additive genetic effects for populations tested in replicated multi-environment trials (METs) in a plant breeding program have recently been presented in the literature. For these data, the variance model involves the direct product of a large numerator relationship matrix A, and a complex structure for the genotype by environment interaction effects, generally of a factor analytic (FA) form. With MET data, we expect a high correlation in genotype rankings between environments, leading to non-positive definite covariance matrices. Estimation methods for reduced rank models have been derived for the FA formulation with independent genotypes, and we employ these estimation methods for the more complex case involving the numerator relationship matrix. We examine the performance of differing genetic models for MET data with an embedded pedigree structure, and consider the magnitude of the non-additive variance. The capacity of existing software packages to fit these complex models is largely due to the use of the sparse matrix methodology and the average information algorithm. Here, we present an extension to the standard formulation necessary for estimation with a factor analytic structure across multiple environments.

The analysis of QTL by simultaneous use of the full linkage map.

An extension of interval mapping is presented that incorporates all intervals on the linkage map simultaneously. The approach uses a working model in which the sizes of putative QTL for all intervals across the genome are random effects. An outlier detection method is used to screen for possible QTL. Selected QTL are subsequently fitted as fixed effects. This screening and selection approach is repeated until the variance component for QTL sizes is not statistically significant. A comprehensive simulation study is conducted in which map uncertainty is included. The proposed method is shown to be superior to composite interval mapping in terms of power of detection of QTL. There is an increase in the rate of false positive QTL detected when using the new approach, but this rate decreases as the population size increases. The new approach is much simpler computationally. The analysis of flour milling yield in a doubled haploid population illustrates the improved power of detection of QTL using the approach, and also shows how vital it is to allow for sources of non-genetic variation in the analysis.

Joint modeling of additive and non-additive (genetic line) effects in multi-environment trials.

A statistical approach for the analysis of multi-environment trials (METs) is presented, in which selection of best performing lines, best parents, and best combination of parents can be determined. The genetic effect of a line is partitioned into additive, dominance and residual non-additive effects. The dominance effects are estimated through the incorporation of the dominance relationship matrix, which is presented under varying levels of inbreeding. A computationally efficient way of fitting dominance effects is presented which partitions dominance effects into between family dominance and within family dominance line effects. The overall approach is applicable to inbred lines, hybrid lines and other general population structures where pedigree information is available.

The analysis of longitudinal data using mixed model L-splines.

L-splines are a large family of smoothing splines defined in terms of a linear differential operator. This article develops L-splines within the context of linear mixed models and uses the resulting mixed model L-spline to analyze longitudinal data from a grassland experiment. In the spirit of time-series analysis, a periodic mixed model L-spline is developed, which partitions data into a smooth periodic component plus smooth long-term trend.