The development of commercial varieties that are resistant or tolerant to crown rot caused by Fusarium species is an important goal for cereal breeding programs internationally. Ideally, this research requires experimental sites that are initially free from Fusarium in order to establish treatment plots that compare growth in the presence and absence of these soil- and stubble-borne pathogens. Specifically, the assessment of tolerance requires control plots free of disease to determine the reduction in crop yield in plots where the disease is present. The ability of soil solarisation to reduce the background Fusarium pseudograminearum level occurring at experimental sites in comparison to current stubble management techniques was investigated across three field trials at Wellcamp in Queensland. Stubble from a susceptible durum (Triticum turgidum ssp. durum) cultivar inoculated with F. pseudograminearum was incorporated by cultivation into the trial sites to establish a significant background level of inoculum prior to the application of all subsequent treatment plots. In these trials, solarisation over a period of twelve weeks reduced the presence of F. pseudograminearum to low detection levels when compared to the traditional crown rot management techniques of cultivation or growth of the non-host cover crops mungbean (Vigna radiata) and soybean (Glycine max). No negative effects of solarisation were observed on a subsequent crop of bread wheat (Triticum aestivum), with significantly higher yields observed in the solarised treatments. Solarisation has the potential to deliver near zero level crown rot reference sites for experimental purposes in one short application between cropping seasons.

Hexaploid spring wheat ( Triticum aestivum ) may exhibit significant crown rot disease responses to infection by Fusarium pseudograminearum , with a range of susceptibility levels available in commercial cultivars. Dry conditions during grain-fill may lead to the expression of prematurely senescing culms, which typically fail to set grain. Assessment of hexaploid spring wheat plants exhibiting both non-senescent and prematurely senescent culms was performed using visual discolouration, F. pseudograminearum biomass and vascular colonisation in culm sections sampled at three different heights above the crown and at the peduncle. Samples from six commercial cultivars were collected at milk development in 2014 from Narrabri and Tamworth, New South Wales and Wellcamp, Queensland. Prematurely senescent culms exhibited greater visual discolouration, F. pseudograminearum biomass and vascular colonisation than non-senescent culms in each cultivar. A further comparison of the parameters across four time points from milk development, when senescent culms are first observed, to maturity was conducted for cultivar Sunlin. F. pseudograminearum biomass and vascular colonisation were significantly different between culm conditions prior to maturation, while visual discolouration was not significantly different between culm conditions at any single time point. Colonisation of xylem and phloem tissue was extensive in the basal portions of prematurely senescent culms (36 to 99%) prior to maturation, and suggests significant impacts on water and nutrient movement during crown rot disease. Despite a range of resistance ratings being assessed, cultivars were not reliably separated based upon responses to severe disease pressure, demonstrating the continuing need for improved crown rot resistance in commercial wheat.

Genetic variability information, which is a heritable difference among maize cultivars, is important to a long-term plant breeding program. In the last decade a number of researchers have studied the correlation between phenotypic and genetic variability in maize by using Deoxyribonucleic Acid (DNA) markers. Maize landraces as maize cultivars grown 'on farm' by farmers who reproduce their seed every year, and they can be distinguished by specific traits, so maize landraces are composed of widely variable populations, where the variation can be seen between and within populations. Five white maize landrace seed samples were obtained from five farmers in each of three villages, Nunmafo, Amol and Ajaobaki; five yellow landrace maize seed samples were obtained from five farmers in Nunmafo village only; while the five certified improved open pollinated varieties (Lamuru, Piet kuning, Bisma, Harapan, Kalingga) were obtained from the Indonesia Cereal Research Institute (ICRI) Maros. The banding patterns of the samples' fingerprint were evaluated in the scoring range of 50-700bp fragment weight using fragment size standard ladder (Figure 6.3). Image data was viewed on the computer screen and saved in the SAGA computer program. The gel images were scored manually in Excel 2010 program for presence (score 1) or absence (score 0) of bands, and the binary data used for further analysis. The results of this study for AFLP analysis of West Timor maize landrace populations from three villages and varieties have confirmed that there are extensivegenetic variations within West Timor yellow and white maize landrace populations and varieties. There were two clusters of genetic variation among all samples. The first cluster was a general West Timor maize landrace population, with more than 80% similarity among individuals, and the second cluster was amuch more diverse grouping with less than 80% similarity. It suggests that even though a high molecular diversity had been found among West Timor maize landrace populations most maize genotypes belong to a West Timor maize landrace cluster. In addition the analysis indicated not only significant difference between maize landraces and maize varieties but also maize landraces from the three villages were genetically significantly different from each other. The molecular marker characterization of West Timor maize landrace populations will be a key step for obtaining an improved understanding of the population so they can be managed carefully for the future. Moreover, due to the genetic diversity captured within local landraces, farmers have the opportunity to perform the phenotypic selections based on plant type and seed yield which are likely to give them significant genetic gain in yield in subsequent seasons. Plant breeders have the additional opportunity of applying molecular marker techniques to assist selection.

Net form of net blotch caused by Pyrenophora teres f. teres (Ptt) is a major foliar disease of barley (Hordeum vulgare) worldwide. Knowledge of the evolution of Ptt pathogen populations is important for development of durable host‐plant resistance. This study was conducted to investigate changes in genetic structure of a Ptt population within a barley field during three cropping years. The susceptible barley cultivar Henley was inoculated with Ptt isolate NB050. Leaf samples were collected during the years 2013–15 and 174 single spore Ptt isolates stored. Genotyping using Diversity Arrays Technology markers identified that 25% of isolates were clones of the inoculated isolate and 75% of isolates were multilocus genotypes (MLGs) differing from the original inoculated genotype. The novel genotypes probably originated from a combination of windborne spores from neighbouring fields, infected seed and sexual recombination in the field. The rapid change in the genotypic composition of the Ptt population in this study suggests adaptive potential of novel genotypes and demonstrates the need for barley breeders to use multiple sources of host‐plant resistance to safeguard against resistance being overcome.

Pyrenophora teres f. teres ( Ptt ) and P. teres f. maculata ( Ptm ) cause net and spot form of net blotch of barley ( Hordeum vulgare L. ), respectively. Both pathogens co-exist in barley fields and each can reproduce sexually, resulting in hybridisation and potential generation of novel virulences that could overcome barley host resistances. In this study, three field experiments were conducted during three successive years to investigate the occurrence of hybridisation. Susceptible barley was sown and inoculated with Ptt and Ptm . Form-specific PCR markers were used to analyse 822 conidia and 223 ascospores sampled from infected leaf tissue and 317 P. teres isolates collected across Australia during 1976–2015. None of the isolates were hybrids. Investigation of ascospores indicated that hybridisation had taken place within the forms, demonstrating preference for recombination within forms. Possible contributions of reproductive barriers have been appraised but further investigation is required to explore the rare hybridisation between the forms.

The 2G Triticum timopheevii introgression harbours genes for multiple disease resistance and quality traits in bread wheat. In order to transfer this segment from bread wheat into durum, the bread wheat line Sunguard, which carries this introgressed 2G segment was crossed with three tetraploid durum parents. A significant difference was observed in the segregation ratio of the 2G segment in the different crosses at the F2 generation with two of the three populations indicating segregation distortion against the hexaploid 2G segment. In these populations, the presence of the 2G segment was strongly correlated with the presence of D-genome chromosomes. These results were confirmed in the F4 generation of these populations. Six plants were identified in the F4 generation, which had retained the introgressed 2G segment in a homozygous condition and did not have a complete D-genome set. Two of these lines only had two non-homologous D-genome chromosomes in the F5 generation. Thus, the 2G segment and possibly other translocations can be transferred into durum wheat through hexaploid/tetraploid hybridisation.

Rising atmospheric carbon dioxide concentration ([CO2]) significantly influences plant growth, development, and biomass. Increased photosynthesis rate, together with lower stomatal conductance, has been identified as the key factors that stimulate plant growth at elevated [CO2] (e[CO2]). However, variations in photosynthesis and stomatal conductance alone cannot fully explain the dynamic changes in plant growth. Stimulation of photosynthesis at e[CO2] is always associated with post-photosynthetic secondary metabolic processes that include carbon and nitrogen metabolism, cell cycle functions, and hormonal regulation. Most studies have focused on photosynthesis and stomatal conductance in response to e[CO2], despite the emerging evidence of e[CO2]'s role in moderating secondary metabolism in plants. In this review, we briefly discuss the effects of e[CO2] on photosynthesis and stomatal conductance and then focus on the changes in other cellular mechanisms and growth processes at e[CO2] in relation to plant growth and development. Finally, knowledge gaps in understanding plant growth responses to e[CO2] have been identified with the aim of improving crop productivity under a CO2 rich atmosphere.

Both hexaploid bread wheat (AABBDD) (Triticum aestivum L.) and tetraploid durum wheat (AABB) (T. turgidum spp. durum) are highly significant global food crops. Crossing these two wheats with different ploidy levels results in pentaploid (AABBD) F1 lines. This study investigated the differences in the retention of D chromosomes between different hexaploid × tetraploid crosses in subsequent generations by using molecular and cytological techniques. Significant differences (P < 0.05) were observed in the retention of D chromosomes in the F2 generation depending on the parents of the original cross. One of the crosses, 2WE25 × 950329, retained at least one copy of each D chromosome in 48% of its F2 lines. For this cross, the retention or elimination of D chromosomes was determined through several subsequent self-fertilised generations. Cytological analysis indicated that D chromosomes were still being eliminated at the F5 generation, suggesting that in some hexaploid × tetraploid crosses, D chromosomes are unstable for many generations. This study provides information on the variation in D chromosome retention in different hexaploid × tetraploid wheat crosses and suggests efficient strategies for utilising D genome retention or elimination to improve bread and durum wheat, respectively.

Fusarium crown rot is a significant disease of durum wheat (Triticum turgidum L. var. durum), which exhibits high levels of disease susceptibility. The most extreme symptom of crown rot is a prematurely senescing culm that typically fails to set grain. Individual crown rot-affected durum wheat plants displaying both nonsenescent and prematurely senescent culms were harvested to compare visual discoloration, Fusarium pseudograminearum biomass, and vascular colonization in culm sections sampled at three different heights above the crown. Field samples of EGA Bellaroi were collected at Wellcamp, QLD, in 2011, 2012, 2013, and 2014, and of Hyperno at Narrabri, NSW, in 2014. Prematurely senescent culms exhibited greater visual discoloration, F. pseudograminearum biomass, and vascular colonization than nonsenescent culms in each year they were examined. The extent of these differences varied between environments and timing of collection in each year. Vascular colonization initially occurred in xylem vessels and spread into phloem tissues as disease severity increased. The increased presence of hyphae in vascular bundles of prematurely senescing culms provides strong evidence for the hypothesis that restriction of water and nutrient movement in a diseased culm is a key factor in crown rot severity.

Fusarium crown rot is a major disease of wheat and barley worldwide, with the most frequently isolated causal agents being Fusarium pseudograminearum and F. culmorum. This study has successfully designed a quantitative polymerase chain reaction assay that is specific for F. culmorum, which has been used in conjunction with a previously established F. pseudograminearum-specific assay to compare the location and extent of infection by each fungus across a range of potential hosts, including six winter and three summer cereal species. All common winter cereals, excluding oat, demonstrated a similar range of visual and fungal biomass results when inoculated with either F. pseudograminearum or F. culmorum. Oat exhibited the lowest visual disease ratings and fungal biomass values of the winter cereals, while the sorghum, maize, and rice cultivars returned the lowest values overall. The ranking of host species according to visual discoloration was strongly correlated for both pathogens. Visual reactions to F. pseudograminearum were greater than those caused by F. culmorum in all potential hosts trialed; however, fungal biomass results only indicated this trend for barley. These results demonstrate significant variation in the ability of these pathogens to colonize the range of cereal species examined and also suggest differences between the pathogens in their patterns of host colonization.

Pyrenophora teres f. teres and P. teres f. maculata cause net form and spot form, respectively, of net blotch on barley (Hordeum vulgare). The two forms reproduce sexually, producing hybrids with genetic and pathogenic variability. Phenotypic identification of hybrids is challenging because lesions induced by hybrids on host plants resemble lesions induced by either P. teres f. teres or P. teres f. maculata. In this study, 12 sequence-specific polymerase chain reaction markers were developed based on expressed regions spread across the genome. The primers were validated using 210 P. teres isolates, 2 putative field hybrids (WAC10721 and SNB172), 50 laboratory-produced hybrids, and 7 isolates collected from barley grass (H. leporinum). The sequence-specific markers confirmed isolate WAC10721 as a hybrid. Only four P. teres f. teres markers amplified on DNA of barley grass isolates. Amplified fragment length polymorphism markers suggested that P. teres barley grass isolates are genetically different from P. teres barley isolates and that the second putative hybrid (SNB172) is a barley grass isolate. We developed a suite of markers which clearly distinguish the two forms of P. teres and enable unambiguous identification of hybrids.

Interspecific hybridisation between hexaploid and tetraploid wheat species leads to the development of F1 pentaploid hybrids with unique chromosomal constitutions. Pentaploid hybrids derived from bread wheat (Triticum aestivum L.) and durum wheat (Triticum turgidum spp. durum Desf.) crosses can improve the genetic background of either parent by transferring traits of interest. The genetic variability derived from bread and durum wheat and transferred into pentaploid hybrids has the potential to improve disease resistance, abiotic tolerance, and grain quality, and to enhance agronomic characters. Nonetheless, pentaploid wheat hybrids have not been fully exploited in breeding programs aimed at improving crops. There are several potential barriers for efficient pentaploid wheat production, such as low pollen compatibility, poor seed set, failed seedling establishment, and frequent sterility in F1 hybrids. However, most of the barriers can be overcome by careful selection of the parental genotypes and by employing the higher ploidy level genotype as the maternal parent. In this review, we summarize the current research on pentaploid wheat hybrids and analyze the advantages and pitfalls of current methods used to assess pentaploid-derived lines. Furthermore, we discuss current and potential applications in commercial breeding programs and future directions for research into pentaploid wheat.

Net blotch, caused by Pyrenophora teres, is a major barley (Hordeum vulgare) leaf disease worldwide. P. teres occurs as two forms-P. teres f. teres, and P. teres f. maculata-inducing net and spot-like symptoms, respectively. An intact-seedling assay, where entire seedlings are inoculated by spraying with a conidial suspension, is frequently used for phenotyping net blotch. However, this presents a biosecurity risk in the glasshouse when nonlocal isolates are being screened. Alternatively, a detached-leaf assay (DLA-droplet method) can be used in which leaf segments laid out in a covered tray are inoculated with droplets of a conidial suspension, confining the inoculum. However, using this method, net and spot form symptoms cannot be distinguished from each other. We have developed an improved DLA (DLA-spray method) in which detached whole leaves are sprayed with the inoculum to produce distinct lesions. We compare the results for the three phenotyping methods above using four isolates from both net and spot forms of the disease to inoculate a standard set of eight barley genotypes. Results indicate that the DLA-spray method is a functional, informative and rapid test that readily differentiates the two forms of the pathogen in a biosecure environment.

Hexaploid/tetraploid and tetraploid/hexaploid wheat hybrids were established using the hexaploid (Triticum aestivum L.) bread wheat LRC2010-150 and the tetraploid durum wheat (T. turgidum spp. durum) WID802. Thirty F2 progeny from each cross were characterised using Diversity Arrays Technology (DArTseq™) markers to determine whether there are differences between the crosses in the proportion of A, B and D genomic material inherited from each parent. Inheritance of the A and B genome from the tetraploid durum parent varied from 32 to 63% among the 60 lines assessed, and results indicated significant differences between the two F2 populations in the mean overall proportion of chromosomes A and B inherited from each parent. Significant differences were also observed between the crosses in the proportion of chromosomal segments on 2B, 3A, 3B and 4A inherited from the tetraploid parent. The F2 populations also showed significant differences in the average retention of D chromosomes per line with the tetraploid/hexaploid cross retaining a mean of 2.83 chromosomes while the reciprocal cross retained a mean of 1.8 chromosomes per line. A strong negative correlation was observed in individual lines from both populations between the proportion of the A and B genome inherited from the tetraploid durum parent and the retention of the D genome. The implication of these results for the design of efficient crossing strategies between hexaploid and tetraploid wheats is discussed.

Disease severity of crown rot (Fusarium pseudograminearum) in winter cereal culms is typically assessed at harvest maturity by recording incidence of infection, whitehead production and discolouration of basal internodes. This study has investigated the relationship between the proportion of discolouration of individual internodes and the extent of host tissue colonisation by the pathogen, based on species-specific quantitative PCR (qPCR). Field-grown plots were used to compare visual ratings and F. pseudograminearum DNA content of eight cereal genotypes (six hard white spring wheats, one durum wheat and one barley) at 16 and 22 weeks after planting (WAP) in 2009 and 2013. At 16 WAP (post anthesis – early milk development) strong correlations were present between visual discolouration and fungal DNA content per unit of tissue weight for internodes one, two and three in 2009 and 2013. At 22 WAP (harvest maturity) these relationships were only significant for internode two in 2009 and internode three in the 2013 trial. Differences in both visual symptoms and fungal DNA content between the genotypes occurred at both sample times, but the most significant distinction between genotypes was at 16 WAP. We conclude that visible discolouration of cereal culms is a useful indicator of fungal biomass in culm tissue, as estimated by qPCR. Furthermore, assessment of diseased culms at 16 WAP provides a stronger correlation with fungal colonisation and discriminates more clearly between disease reactions in different host genotypes, in comparison to assessment at harvest maturity.

Radiant frost is a significant production constraint to wheat (Triticum aestivum) and barley (Hordeum vulgare), particularly in regions where spring-habit cereals are grown through winter, maturing in spring. However, damage to winter-habit cereals in reproductive stages is also reported. Crops are particularly susceptible to frost once awns or spikes emerge from the protection of the flag leaf sheath. Post-head-emergence frost (PHEF) is a problem distinct from other cold-mediated production constraints. To date, useful increased PHEF resistance in cereals has not been identified. Given the renewed interest in reproductive frost damage in cereals, it is timely to review the problem. Here we update the extent and impacts of PHEF and document current management options to combat this challenge. We clarify terminology useful for discussing PHEF in relation to chilling and other freezing stresses. We discuss problems characterizing radiant frost, the environmental conditions leading to PHEF damage, and the effects of frost at different growth stages. PHEF resistant cultivars would be highly desirable, to both reduce the incidence of direct frost damage and to allow the timing of crop maturity to be managed to maximize yield potential. A framework of potential adaptation mechanisms is outlined. Clarification of these critical issues will sharpen research focus, improving opportunities to identify genetic sources for improved PHEF resistance.

Histopathological assessment of the crown rot pathogen Fusarium pseudograminearum was performed using fluorescence microscopy of culm tissues of six cereal genotypes grown in inoculated field conditions. Tissue samples were collected at 10, 16, and 22 weeks after planting (WAP). Colonization of culm tissues was initiated through epidermal penetration, most distinctly through stomatal apertures, and progressed into the parenchymatous hypoderm, which exhibited the discoloration used as the basis for visual assessment of disease. Hyphae spread from the culm base vertically through the tissues, initially via the hypoderm and pith cavity. Colonization of sclerified cells occurred later in the disease process. Both xylem and phloem tissues became colonized by 16 WAP in all host genotypes, with colonization being less extensive in the more resistant genotypes. Culms displaying dead head symptoms revealed dense colonization in at least the first three internodes, with frequent xylem vessel and phloem cell occlusions. Paired living culms from the same plants exhibited less extensive colonization. These observations have revealed the ability of F. pseudograminearum to colonize all cell types of nodal and internodal sections, including vascular tissues, across all host genotypes. This study is the first detailed examination of the pattern of F. pseudograminearum colonization in adult hosts and indicates a potential vascular mechanism by which the effects of crown rot are produced.

QTL identified for seedling and adult plant crown rot resistance in four partially resistant hexaploid wheat sources. PCR-based markers identified for use in marker-assisted selection.

A recently developed spot form of blotch differential set of 16 barley lines was tested for reaction response to 60 Pyrenophora teres f. maculata isolates from geographically disperse barley crops of Australia. Twelve barley lines (Arimont, Barque, Chebec, CI5286, CI5791, CI9214, CII6150, Dairokkaku, Esperance Orge 289, Galleon, Keel, Skiff, Torrens and TR250) provided differential response between the isolates. The susceptible controls Gairdner and Kombar provided indication of isolate virulence or avirulence. Abundant pathogenic diversity was revealed with 33 designated pathotypes, some of which related to geographic region. AFLP analysis also revealed abundant diversity with each of the isolates representing a unique genotype and one isolate that contained both AFLP bands unique to P. teres f. maculata and P. teres f. teres, the cause of spot form and net form of net blotch respectively, suggesting that sexual recombination between the net form and spot form isolates may have occurred naturally in the field.

Inoculation of wheat seedlings by the crown rot pathogen Fusarium pseudograminearum is commonly used to screen for resistance based on leaf sheath discolouration. While leaf sheath discolouration is effective for describing disease reactions, the degree to which the pathogen colonises the full range of seedling host tissues is an important aspect of the disease process, which has not been systematically examined. After single point droplet inoculation of the coleoptile, quantitative PCR was used to determine the spread of F. pseudograminearum into different host tissues during crown rot pathogenesis at 14 and 28 days after inoculation. Quantitative PCR and visual assessment indicated that while most seedling tissues (leaf sheath, leaf blade, sub-crown internode, primary root and secondary root) were colonised, the basal portion of leaf sheath tissue supported the highest density of F. pseudograminearum . Other significant sites of colonisation were the upper leaf sheath and sub-crown internode. By comparison, the quantity of mycelium detected in the primary and secondary roots was significantly lower, while the infection of leaf blade tissues was severely restricted.