R. Loughman

Mapping and validation of the genes for resistance to Pyrenophora teres f. teres in barley (Hordeum vulgare L.)

Identification and deployment of disease resistance genes are key objectives of Australian barley breeding programs. Two doubled haploid (DH) populations derived from Tallon × Kaputar (TK) and VB9524 × ND11231 (VN) crosses were used to identify markers for net type net blotch (NTNB) (Pyrenophora teres f. teres). The maps included 263 and 250 markers for TK and VN populations, respectively. The TK population was screened with 5 pathotypes and the VN population with 1 pathotype of NTNB as seedlings in the glasshouse. In addition, the TK population was subjected to natural infection in the field at Hermitage Research Station, Qld. Analyses of the markers were performed using the software packages MapManager and Qgene. One region on chromosome 6H was strongly associated with resistance to NTNB in both populations (R2 = 83% for TK and 66% for VN). In the TK population, 2 more quantitative trait loci (QTLs) were identified on chromosomes 2H and 3H, with R2 values of 30% and 31%, respectively. These associations were consistent over all pathotypes studied during the seedling stage. The same QTL on chromosome 6H was also found to be highly significantly associated (R2 = 65%) with the adult plant (field) response in the TK population. There are several very closely linked markers showing strong associations in these regions. Association of the 4 markers on chromosome 6H QTL with resistance to the NTNB has been validated in 2 other DH populations derived from barley crosses Pompadour × Stirling and WPG8412 × Stirling. These markers present an opportunity for marker assisted selection of lines resistant to NTNB in barley breeding programs.

Current virulence of Pyrenophora teres on barley in Western Australia

Studies on variation, occurrence, and distribution of virulence in Pyrenophora teres are helpful to identify effective sources of resistance that can be used for barley breeding in Western Australia. Seventy-nine isolates of Pyrenophora teres were collected from different barley fields of Western Australia in 1995-96. Seventy-four induced net type symptoms (P. teres f. teres) and five induced spot type symptoms (P. teres f. maculata). Net type isolate responses on 47 barley lines were similar to the range of responses induced by nine historical isolates collected in the region between 1975 and 1985. These net type isolates were classified into two distinct groups based on virulence to the cultivar Beecher. Isolates were further classified into eight groups based on minor pathogenic variation among the population. The virulence phenotype present in an eastern Australian isolate was not observed in any isolates collected from Western Australia. An analysis of variance on a subset of 12 net type isolates indicated a significant line × isolate interaction (P < 0.001), with the interaction term variance component four times larger than the error variance. Based on these studies, the virulence among net type isolates has remained stable in Western Australia for the last 19 years. Spot type isolates were collected from a wider geographic area than previously reported and varied in virulence based on response to barley line Herta. Variation in spot-type isolates is reported for the first time from the region. The results from this study are being used in the development of resistant varieties.

Quantitative trait loci for seedling and adult plant resistance to Stagonospora nodorum in wheat

Stagonospora nodorum blotch (SNB) caused by Stagonospora nodorum is a severe disease of wheat (Triticum aestivum) in many areas of the world. S. nodorum affects both seedling and adult plants causing necrosis of leaf and glume tissue, inhibiting photosynthetic capabilities, and reducing grain yield. The aims of this study were to evaluate disease response of 280 doubled haploid (DH) individuals derived from a cross between resistant (6HRWSN125) and susceptible (WAWHT2074) genotypes, compare quantitative trait loci (QTL) for seedling and adult plant resistance in two consecutive years, and assess the contribution of QTL on grain weight. Flag leaves and glumes of individuals from the DH population were inoculated with mixed isolates of S. nodorum at similar maturity time to provide accurate disease evaluation independent of morphological traits and identify true resistance for QTL analysis. Fungicide protected and inoculated plots were used to measure relative grain weight (RGW) as a yield-related trait under pathogen infection. The lack of similar QTL and little or no correlation in disease scores indicate different genes control seedling and adult plant disease and independent genes control flag leaf and glume resistance. This study consistently identified a QTL on chromosome 2DL for flag leaf resistance (QSnl.daw-2D) and 4BL for glume resistance (QSng.daw-4B) from the resistant parent, 6HRWSN125, explaining 4 to 19% of the phenotypic variation at each locus. A total of 5 QTL for RGW were consistently detected, where two were in the same marker interval for QSnl.daw-2D and QSng.daw-4B indicating the contribution of these QTL to yield related traits. Therefore, RGW measurement in QTL analysis could be used as a reliable indicator of grain yield affected by S. nodorum infection.

Resistance to septoria nodorum blotch in the Aegilops tauschii accession RL5271 is controlled by a single gene

Septoria nodorum blotch is the most important leaf disease of wheat in Western Australia. A potentially useful source of resistance has been identified in an accession of Aegilops tauschii. To study the genetics of resistance of this source a cross was made between the resistant Ae. tauschii accession, RL5271, and a susceptible accession, CPI110889. The resistant parent took significantly longer to develop symptoms, developed significantly fewer lesions and expressed significantly lower levels of disease than the susceptible parent. The F1 mean response for disease severity indicated there was no complete dominance. The F3 families were classified using three approaches. In the first approach the individual F3 plant response was used to classify the F3 families. In the second approach the F3 family means and standard errors were used to classify the F3 families. In the final approach Best Linear Unbiased Predictors of disease score and standard error for each F3 family derived from a REML analysis were used to classify the F3 families. The genotypic ratios generated by each of the approaches suggested that resistance is controlled by a single gene. The effectiveness of the resistance and its simple genetic control in the Ae. tauschii, accession RL5271 may be a useful resistance source for use in a bread wheat breeding program.

Resistance in cultivated barleys to Pyrenophora teres f. teres and prospects of its utilisation in marker identification and breeding

Net type net blotch (NTNB) is a prevalent disease in Australia, causing significant losses in barley yield and quality. Its impact can be reduced with the identification and utilisation of effective sources of resistance. Sixty-nine cultivated barley lines were screened as seedlings against 9 isolates of Pyrenophora teres f. teres from Australia, and in the field in Western Australia. Resistance expressed in seedlings was frequently expressed in adult plants in the field, indicating that these sources are potentially useful for resistance breeding. Of these lines, 24 with the best overall resistance were identified, which could be used against virulence diversity present in P. teres f. teres in Australia. As a prelude to the evaluation of established mapping populations in the Australian Barley Molecular Marker Program, 42 parental lines were screened against a range of Australian isolates of P. teres f. teres. Variation in net blotch responses was observed among parents of the mapping populations. Ten principal mapping populations appear to provide opportunities to map resistances and identify molecular markers linked to NTNB resistance genes effective against Australian pathotypes.

Septoria nodorum blotch resistance in Aegilops tauschii and its expression in synthetic amphiploids

A collection of 433 Aegilops tauschii was screened for response to infection with Stagonospora nodorum. Resistance similar or marginally superior to the range observed in spring wheat genotypes was readily identified. Three lines, RL5271, Aus18911, and Aus21712, were resistant to a range of pathogen isolates and were similar in resistance to a highly resistant French line, No.33. Accessions of Ae. tauschii assessed as resistant or susceptible as seedlings had corresponding reactions when tested as adult plants, with resistance being commonly expressed as restricted lesion development. Infection frequency differed between some Ae. tauschii lines. Seedling resistance in synthetic bread wheats was expressed partially or not at all depending on both the tetraploid and the tetraploid/Ae. tauschii combination. Assessment of adult responses among a range of synthetics showed occasional expression of moderate resistance around the level observed among reference bread wheat cultivars of similar maturity. Disease escape associated with late maturity was common. One synthetic wheat, #231, derived from a resistant Ae. tauschii, exhibited low adult disease expression associated with late maturity. This line was resistant in seedling tests and seedling resistance was dominant in F1 hybrids to bread wheat.

Characterisation and diversity of Pyrenophora teres f. maculata isolates in Western Australia

Spot type net blotch caused by Pyrenophora teres f. maculata (Ptm) has become a prominent disease in Western Australia, as has also occurred elsewhere. The disease has a negative impact on both grain yield and quality resulting from reduced grain size. Lack of resistance and stubble retention are the likely factors in the increased severity of the disease in barley growing areas of Western Australia. Because of the increasing importance of spot type net blotch and the need to improve barley resistance, understanding pathogen virulence is a high priority as this has direct impact on the identification and utilization of resistance genes in breeding programs. Ninety nine isolates of Ptm were collected from geographically dispersed barley fields of Western Australia during 2001 and 2002. Forty nine sporulating isolates of Ptm were classified into seven isolate groups (IGs) on the basis of their infection responses on 26 differential barley lines. The 26 lines were likewise classified into four line groups (LGs) based on their distinguishing response to the spot type net blotch isolates. The varied infection responses among the differential barley lines demonstrated a wide geographic dispersal of IGs as well as previously undetected virulence in Ptm in Western Australia. The commercially grown barley cultivars Baudin and Gairdner are regarded as susceptible to spot type net blotch, but showed a range of reactions to the various Ptm isolates as seedling plants. The variability in the pathogen and the resistance identified in some genotypes used in this study are being investigated further to develop superior, adapted germplasm for use in barley breeding programs in Australia.

A single gene controls resistance to septoria nodorum blotch in the Aegilops tauschii accession AUS21712

A potential source of resistance to septoria nodorum blotch had been identified in an accession of the wild wheat, Aegilops tauschii. A cross was made between the resistant Ae. tauschii accession, AUS21712, and a susceptible accession, CPI110889, to study the genetics of resistance. The parental accessions and the F1, F3, and F4 progeny were screened in the glasshouse as seedlings. The resistant parent took significantly longer to develop symptoms, developed significantly fewer lesions, and expressed significantly lower levels of disease than the susceptible parent. The F1 mean response for disease severity indicated resistance was dominant. The genotypic ratios generated from the screening of the F3 and F4 generations were not significantly different from the genotypic ratio expected for a single gene. The efficacy of the resistance and its simple genetic control in the Ae. tauschii accession AUS21712 means that the potential exists to use this Ae. tauschii resistance gene in a bread wheat breeding program.

Loci on chromosomes 1A and 2A affect resistance to tan (yellow) spot in wheat populations not segregating for tsn1

Key messageQTL for tan spot resistance were mapped on wheat chromosomes 1A and 2A. Lines were developed with resistance alleles at these loci and at the tsn1 locus on chromosome 5B. These lines expressed significantly higher resistance than the parent withtsn1only.AbstractTan spot (syn. yellow spot and yellow leaf spot) caused by Pyrenophora tritici-repentis is an important foliar disease of wheat in Australia. Few resistance genes have been mapped in Australian germplasm and only one, known as tsn1 located on chromosome 5B, is known in Australian breeding programs. This gene confers insensitivity to the fungal effector ToxA. The main aim of this study was to map novel resistance loci in two populations: Calingiri/Wyalkatchem, which is fixed for the ToxA-insensitivity allele tsn1, and IGW2574/Annuello, which is fixed for the ToxA-sensitivity allele Tsn1. A second aim was to combine new loci with tsn1 to develop lines with improved resistance. Tan spot severity was evaluated at various growth stages and in multiple environments. Symptom severity traits exhibited quantitative variation. The most significant quantitative trait loci (QTL) were detected on chromosomes 2A and 1A. The QTL on 2A explained up to 29.2% of the genotypic variation in the Calingiri/Wyalkatchem population with the resistance allele contributed by Wyalkatchem. The QTL on 1A explained up to 28.1% of the genotypic variation in the IGW2574/Annuello population with the resistance allele contributed by Annuello. The resistance alleles at both QTL were successfully combined with tsn1 to develop lines that express significantly better resistance at both seedling and adult plant stages than Calingiri which has tsn1 only.

A major QTL controlling adult plant resistance for barley leaf rust

Race-specific resistance genes (Rph) for leaf rust (Puccinia hordei) are often overcome by new pathotypes with matching virulence. Adult plant resistance (APR) is considered potentially more durable for controlling barley leaf rust. Previous studies established that the cultivar Pompadour carried APR to leaf rust. A doubled haploid population (DH) of 200 lines developed from a cross Pompadour/Stirling, and the parents were phenotyped for leaf rust resistance at five field experimental sites in three agricultural zones in Australia. Using a linkage map of SSR and DArT molecular markers, a major QTL associated with the leaf rust resistance was identified on the short arm of chromosome 5H. This QTL explained between 31% and 86% of the phenotypic variation for the APR at different sites. A PCR-based molecular marker was developed and mapped at 1.6 cM to the APR gene. The present study provides new genetic material and a molecular tool for breeding new varieties with adult plant leaf rust resistance using marker-assisted selection.