Advancements in high-throughput genotyping and sequencing technologies are enabling the development of a vast range of genomic tools and resources for a new revolution in plant breeding. Several genotyping-by-sequencing (GBS) methods including capture-based, genome complexity reduction and sequencing of cDNA (GBS-t) are available for application in trait dissection, association mapping, and genomic selection (GS) in crop plants. The aims of this study were to identify genomic regions conferring resistance to Ascochyta blight (AB) introgressed from the wild Cicer echinospernum into the domesticated C. arietinum, through a conventional recombinant inbred population genotyped using a variety of GBS methods. Evaluation of GBS methods revealed that capture-based approaches are robust and reproducible while GBS-t is rapid and flexible. A genetic linkage map consisting of 5886 polymorphic loci spanning 717.26 cM was generated. Using field phenotyping data from two years, a single genomic region on LG4 was identified with quantitative trait loci (QTL) mapping. Both GBS methods reported in this study are well suited for applications in genomics assisted plant breeding. Linked markers for AB resistance, identified in the current study, provide an important resource for the deployment into chickpea breeding programs for marker-assisted selection (MAS).

Chickpea production in Australia is constrained by both waterlogging and the root disease Phytophthora root rot (PRR). Soil saturation is an important pre-condition for significant disease development for many soil-borne Phytophthora spp. In wet years, water can pool in low lying areas within a field, resulting in waterlogging, which, in the presence of PRR, can result in a significant yield loss for Australian chickpea varieties. In these circumstances, the specific cause of death is often difficult to discern, as the damage is rapid and the spread of PRR can be explosive in nature. The present study describes the impact of soil waterlogging on oxygen availability and the ability of P. medicaginis to infect chickpea plants. Late waterlogging in combination with PRR reduced the total plant biomass by an average of 94%; however, waterlogging alone accounted for 88% of this loss across three reference genotypes. Additional experiments found that under hypoxic conditions associated with waterlogging, P. medicaganis did not proliferate as determined by zoospore counts and DNA detection using qPCR. Consequently, minimizing waterlogging damage through breeding and agronomic practices should be a key priority for integrated disease management, as waterlogging alone results in plant stunting, yield loss and a reduced resistance to PRR.

Phytophthora root rot (PRR) caused by the soilborne oomycete Phytophthora medicaginis is a significant constraint to chickpea (Cicer arietinum) production across the northern grains region of Australia. In flooded soil, which is conducive to PRR disease development, up to 70% yield loss can occur in the most resistant Australian cultivars. Incorporating waterlogging tolerance in soybean (Glycine max) has been shown to improve quantitative resistance to Phytophthora sojae. Root growth of three chickpea genotypes was assessed at the seedling stage under waterlogging, PRR, and the combination of these abiotic and biotic constraints. Levels of waterlogging tolerance in chickpea are inherently low, yet selected genotypes displayed variability in root traits linked to improved waterlogging tolerance. The PRR moderately susceptible chickpea cultivar Yorker and PRR very susceptible Rupali demonstrated an eightfold increase in early adventitious root growth from the epicotyl region under waterlogging stress, compared with the PRR resistant interspecific backcross genotype 04067-81-2-1-1 (C. echinospermum × C. arietinum*2). Selection for primary root depth, which was significantly greater in 04067-81-2-1-1 under waterlogging, appears to improve PRR resistance compared with root replacement traits. Soilborne Phytophthora spp. are reportedly attracted to branch sites and leached exudates. We propose that compromised root barriers at emergence sites of adventitious roots under waterlogging conditions hasten hyphal entry, potentially increasing susceptibility to PRR. Hence, screening for root depth and absence of adventitious root development under waterlogged conditions may offer a novel proxy phenotyping method for PRR resistance traits at early stages of chickpea breeding.

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.