The CIMMYT Australia ICARDA Germplasm Evaluation concept: a model for international cooperation and impact.

Bread wheat germplasm is accessed from the International Maize and Wheat Improvement Centre (CIMMYT) and the International Centre for Agricultural Research in the Dry Areas (ICARDA) by Australian wheat breeders and researchers through the CIMMYT Australia ICARDA Germplasm Evaluation (CAIGE) program. The CAIGE program coordinates the selection, importation, quarantine, dissemination, and evaluation of the imported bread wheat germplasm and the management of associated data and information. This paper describes the CAIGE model and assesses both the genetic and economic impacts of these materials on the Australian wheat industry after commercialisation of wheat breeding in the early 21st century and the establishment of CAIGE. The CAIGE concept was validated using data collected and analysed from multi-environment trials between 2017 and 2020. The impact of cultivars with and without CAIGE contribution to pedigree on yield was estimated using production-by-variety statistics. Net gain in yield, estimated as the yield difference between CAIGE and Non-CAIGE varieties, was multiplied by the percentage contribution to pedigree to estimate the additional yield. The CAIGE bread wheat program identified diverse, high-yielding, and disease-resistant germplasm and significantly improved the capture and dissemination of information. The benefit-cost ratio, calculated as the sum of benefits divided by investments, indicated that, for every dollar invested in CAIGE, a further $20 was generated in benefits. The internal rate of return was estimated at 163% and the modified rate at 18%. The benefits of these international materials to Australian wheat breeding remained significant.

Characterization of Leaf Rust Resistance in International Barley Germplasm Using Genome-Wide Association Studies.

A panel of 114 genetically diverse barley lines were assessed in the greenhouse and field for resistance to the pathogen Puccinia hordei, the causal agent of barley leaf rust. Multi-pathotype tests revealed that 16.6% of the lines carried the all-stage resistance (ASR) gene Rph3, followed by Rph2 (4.4%), Rph1 (1.7%), Rph12 (1.7%) or Rph19 (1.7%). Five lines (4.4%) were postulated to carry the gene combinations Rph2+9.am, Rph2+19 and Rph8+19. Three lines (2.6%) were postulated to carry Rph15 based on seedling rust tests and genotyping with a marker linked closely to this gene. Based on greenhouse seedling tests and adult-plant field tests, 84 genotypes (73.7%) were identified as carrying APR, and genotyping with molecular markers linked closely to three known APR genes (Rph20, Rph23 and Rph24) revealed that 48 of the 84 genotypes (57.1%) likely carry novel (uncharacterized) sources of APR. Seven lines were found to carry known APR gene combinations (Rph20+Rph23, Rph23+Rph24 and Rph20+Rph24), and these lines had higher levels of field resistance compared to those carrying each of these three APR genes singly. GWAS identified 12 putative QTLs; strongly associated markers located on chromosomes 1H, 2H, 3H, 5H and 7H. Of these, the QTL on chromosome 7H had the largest effect on resistance response to P. hordei. Overall, these studies detected several potentially novel genomic regions associated with resistance. The findings provide useful information for breeders to support the utilization of these sources of resistance to diversify resistance to leaf rust in barley and increase resistance durability.

Late-Maturity Alpha-Amylase in Wheat (Triticum aestivum) and Its Impact on Fresh White Sauce Qualities.

When wheat experiences a cold-temperature 'shock' during the late stage of grain filling, it triggers the abnormal synthesis of late-maturity α-amylase (LMA). This increases the enzyme content in affected grain, which can lead to a drastic reduction in falling number (FN). By commercial standards, a low FN is taken as an indication of inferior quality, deemed unsuitable for end-product usage. Hence, LMA-affected grains are either rejected or downgraded to feed grade at the grain receiving point. However, previous studies have found no substantial correlation between low FN-LMA and bread quality. The present study extends previous investigations to semi-solid food, evaluating the physical quality of fresh white sauce processed from LMA-affected flour. Results show that high-LMA flours had low FNs and exhibited poor pasting characteristics. However, gelation occurred in the presence of other components during fresh white sauce processing. This demonstrates that LMA-affected flours may have new applications in low-viscosity products.

Lessons from a GWAS study of a wheat pre-breeding program: pyramiding resistance alleles to Fusarium crown rot.

Much has been published on QTL detection for complex traits using bi-parental and multi-parental crosses (linkage analysis) or diversity panels (GWAS studies). While successful for detection, transferability of results to real applications has proven more difficult. Here, we combined a QTL detection approach using a pre-breeding populations which utilized intensive phenotypic selection for the target trait across multiple plant generations, combined with rapid generation turnover (i.e. "speed breeding") to allow cycling of multiple plant generations each year. The reasoning is that QTL mapping information would complement the selection process by identifying the genome regions under selection within the relevant germplasm. Questions to answer were the location of the genomic regions determining response to selection and the origin of the favourable alleles within the pedigree. We used data from a pre-breeding program that aimed at pyramiding different resistance sources to Fusarium crown rot into elite (but susceptible) wheat backgrounds. The population resulted from a complex backcrossing scheme involving multiple resistance donors and multiple elite backgrounds, akin to a MAGIC population (985 genotypes in total, with founders, and two major offspring layers within the pedigree). A significant increase in the resistance level was observed (i.e. a positive response to selection) after the selection process, and 17 regions significantly associated with that response were identified using a GWAS approach. Those regions included known QTL as well as potentially novel regions contributing resistance to Fusarium crown rot. In addition, we were able to trace back the sources of the favourable alleles for each QTL. We demonstrate that QTL detection using breeding populations under selection for the target trait can identify QTL controlling the target trait and that the frequency of the favourable alleles was increased as a response to selection, thereby validating the QTL detected. This is a valuable opportunistic approach that can provide QTL information that is more easily transferred to breeding applications.

Speed breeding is a powerful tool to accelerate crop research and breeding.

The growing human population and a changing environment have raised significant concern for global food security, with the current improvement rate of several important crops inadequate to meet future demand 1 . This slow improvement rate is attributed partly to the long generation times of crop plants. Here, we present a method called 'speed breeding', which greatly shortens generation time and accelerates breeding and research programmes. Speed breeding can be used to achieve up to 6 generations per year for spring wheat (Triticum aestivum), durum wheat (T. durum), barley (Hordeum vulgare), chickpea (Cicer arietinum) and pea (Pisum sativum), and 4 generations for canola (Brassica napus), instead of 2-3 under normal glasshouse conditions. We demonstrate that speed breeding in fully enclosed, controlled-environment growth chambers can accelerate plant development for research purposes, including phenotyping of adult plant traits, mutant studies and transformation. The use of supplemental lighting in a glasshouse environment allows rapid generation cycling through single seed descent (SSD) and potential for adaptation to larger-scale crop improvement programs. Cost saving through light-emitting diode (LED) supplemental lighting is also outlined. We envisage great potential for integrating speed breeding with other modern crop breeding technologies, including high-throughput genotyping, genome editing and genomic selection, accelerating the rate of crop improvement.

Application of Multiple Imputation for Missing Values in Three-Way Three-Mode Multi-Environment Trial Data.

It is a common occurrence in plant breeding programs to observe missing values in three-way three-mode multi-environment trial (MET) data. We proposed modifications of models for estimating missing observations for these data arrays, and developed a novel approach in terms of hierarchical clustering. Multiple imputation (MI) was used in four ways, multiple agglomerative hierarchical clustering, normal distribution model, normal regression model, and predictive mean match. The later three models used both Bayesian analysis and non-Bayesian analysis, while the first approach used a clustering procedure with randomly selected attributes and assigned real values from the nearest neighbour to the one with missing observations. Different proportions of data entries in six complete datasets were randomly selected to be missing and the MI methods were compared based on the efficiency and accuracy of estimating those values. The results indicated that the models using Bayesian analysis had slightly higher accuracy of estimation performance than those using non-Bayesian analysis but they were more time-consuming. However, the novel approach of multiple agglomerative hierarchical clustering demonstrated the overall best performances.

Simulated breeding with QU-GENE graphical user interface.

Comparing the efficiencies of breeding methods with field experiments is a costly, long-term process. QU-GENE is a highly flexible genetic and breeding simulation platform capable of simulating the performance of a range of different breeding strategies and for a continuum of genetic models ranging from simple to complex. In this chapter we describe some of the basic mechanics behind the QU-GENE user interface and give a simplified example of how it works.

Origin of leaf rust adult plant resistance gene Rph20 in barley.

Rph20 is the only reported, simply inherited gene conferring moderate to high levels of adult plant resistance (APR) to leaf rust (Puccinia hordei Otth) in barley (Hordeum vulgare L.). Key parental genotypes were examined to determine the origin of Rph20 in two-rowed barley. The Dutch cultivar 'Vada' (released in the 1950s) and parents, 'Hordeum laevigatum' and 'Gull' ('Gold'), along with the related cultivar 'Emir' (a derivative of 'Delta'), were assessed for APR to P. hordei in a disease screening nursery. The marker bPb-0837-PCR, co-located with Rph20 on the short arm of chromosome 5H (5HS), was used to screen genotypes for the resistance allele, Rph20.ai. Results from phenotypic assessment and DNA analysis confirmed that Rph20 originated from the landrace 'H. laevigatum' (i.e., Hordeum vulgare subsp. vulgare). Tracing back this gene through the pedigrees of two-rowed barley cultivars, indicated that Rph20 has contributed APR to P. hordei for more than 60 years. Although there have been no reports of an Rph20-virulent pathotype, the search for alternative sources of APR should continue to avoid widespread reliance upon a single resistance factor.

Occurrence of LINE, gypsy-like, and copia-like retrotransposons in the clonally propagated sweet potato (Ipomoea batatas L.).

Retrotransposons are a class of transposable elements that represent a major fraction of the repetitive DNA of most eukaryotes. Their abundance stems from their expansive replication strategies. We screened and isolated sequence fragments of long terminal repeat (LTR), gypsy-like reverse transcriptase (rt) and gypsy-like envelope (env) domains, and two partial sequences of non-LTR retrotransposons, long interspersed element (LINE), in the clonally propagated allohexaploid sweet potato (Ipomoea batatas (L.) Lam.) genome. Using dot-blot hybridization, these elements were found to be present in the ~1597 Mb haploid sweet potato genome with copy numbers ranging from ~50 to ~4100 as observed in the partial LTR (IbLtr-1) and LINE (IbLi-1) sequences, respectively. The continuous clonal propagation of sweet potato may have contributed to such a multitude of copies of some of these genomic elements. Interestingly, the isolated gypsy-like env and gypsy-like rt sequence fragments, IbGy-1 (~2100 copies) and IbGy-2 (~540 copies), respectively, were found to be homologous to the Bagy-2 cDNA sequences of barley (Hordeum vulgare L.). Although the isolated partial sequences were found to be homologous to other transcriptionally active elements, future studies are required to determine whether they represent elements that are transcriptionally active under normal and (or) stressful conditions.

Impact of depth of pedigree and inclusion of historical data on the estimation of additive variance and breeding values in a sugarcane breeding program.

Sugarcane breeders in Australia combine data across four selection programs to obtain estimates of breeding value for parents. When these data are combined with full pedigree information back to founding parents, computing limitations mean it is not possible to obtain information on all parents. Family data from one sugarcane selection program were analysed using two different genetic models to investigate how different depths of pedigree and amount of data affect the reliability of estimating breeding value of sugarcane parents. These were the parental and animal models. Additive variance components and breeding values estimated from different amounts of information were compared for both models. The accuracy of estimating additive variance components and breeding values improved as more pedigree information and historical data were included in analyses. However, adding years of data had a much larger effect on the estimation of variance components of the population, and breeding values of the parents. To accurately estimate breeding values of all sugarcane parents, a minimum of three generations of pedigree and 5 years of historical data were required, while more information (four generations of pedigree and 7 years of historical data) was required when identifying top parents to be selected for future cross pollination.