Lesley A. Boyd

QTL identification and microphenotype characterisation of the developmentally regulated yellow rust resistance in the UK wheat cultivar Guardian

Yellow rust (causal agent: Puccinia striiformis f.sp. tritici) resistance in the UK wheat cultivar Guardian is developmentally regulated, resistance increasing as the plant matures. Yellow rust resistance was assessed under field conditions on plants after ear emergence to ensure maximum expression of resistance. Three quantitative trait loci (QTL) for yellow rust resistance were identified, being located on chromosomes 1B (QPst.jic-1B), 2D (QPst.jic-2D) and 4B (QPst.jic-4B). The largest resistance effect, QPst.jic-1B located to the same position on the long arm of chromosome 1B as the known durable source of yellow rust resistance, Yr29. Microscopic studies were carried out to determine what effect the resistance in Guardian had on the development of P. striiformis f.sp. tritici. While the adult plant resistance in Guardian did not prevent germinated urediniospores from establishing an effective infection site, the growth of hyphae within flag leaf tissue was significantly inhibited, slowing the development of microcolonies. 3,3-diaminabenzadine (DAB) and trypan blue staining indicated that this inhibition of hyphal growth was not associated with hydrogen peroxide accumulation or extensive plant cell death.

Identification of adult plant resistance to stripe rust in the wheat cultivar Cappelle-Desprez

Following the appearance of stripe rust in South Africa in 1996, efforts have been made to identify new sources of durable resistance. The French cultivar Cappelle-Desprez has long been considered a source of durable, adult plant resistance (APR) to stripe rust. As Cappelle-Desprez contains the seedling resistance genes Yr3a and Yr4a, wheat lines were developed from which Yr3a and Yr4a had been removed, while selecting for Cappelle-Desprez derived APR effective against South African pathotypes of the stripe rust fungus, Puccinia striiformis f. sp. tritici. Line Yr16DH70, adapted to South African wheat growing conditions, was selected and crossed to the stripe rust susceptible cultivar Palmiet to develop a segregating recombinant inbred line mapping population. A major effect QTL, QYr.ufs-2A was identified on the short arm of chromosome 2A derived from Cappelle-Desprez, along with three QTL of smaller effect, QYr.ufs-2D, QYr.ufs-5B and QYr.ufs-6D. QYr.ufs-2D was located within a region on the short arm of chromosome 2D believed to be the location of the stripe rust resistance gene Yr16. An additional minor effect QTL, QYr.ufs-4B, was identified in the cv. Palmiet. An examination of individual RILs carrying single or combinations of each QTL indicated significant resistance effects when QYr.ufs-2A was combined with the three minor QTL from Cappelle-Desprez, and between QYr.ufs-2D and QYr.ufs-5B.

Cellular and transcriptional responses of wheat during compatible and incompatible race‐specific interactions with Puccinia striiformis f. sp. tritici

The initial stages of Puccinia striiformis f. sp. tritici (the causal agent of yellow rust in wheat) infection triggered a hypersensitive cell death (HCD) response in both compatible and Yr1-mediated incompatible interactions, although the response was earlier and more extensive in the incompatible interaction. Later stages of fungal development were only associated with an HCD response in the incompatible interaction, the HCD response being effectively suppressed in the compatible interaction. Cell autofluorescence was seen in mesophyll cells in direct contact with fungal infection hyphae (primary HCD) and in adjacent mesophyll cells (secondary HCD), indicating the activation of cell-to-cell signalling. Microarray analysis identified a number of defence-related transcripts implicated in Yr1-mediated resistance, including classical pathogenesis-related (PR) transcripts and genes involved in plant cell defence responses, such as the oxidative burst and cell wall fortification. A quantitative reverse transcriptase-polymerase chain reaction time course analysis identified a number of defence-related genes, including PR2, PR4, PR9, PR10 and WIR1 transcripts, associated with the latter stages of Yr1-mediated resistance. A meta-analysis comparison of the Yr1-regulated transcriptome with the resistance transcriptomes of the race-specific resistance gene Yr5 and the race-nonspecific adult plant resistance gene Yr39 indicated limited transcript commonality. Common transcripts were largely confined to classic PR and defence-related genes.

The development of a STS marker linked to a yellow rust resistance derived from the wheat cultivar Moro

Abstract.A sequence-tagged-site (STS) marker has been developed for a gene conferring yellow rust resistance originating from the wheat cultivar Moro. The single, dominant, seedling yellow rust resistance gene, designated YrMoro, was mapped to the group 1 chromosomes. The STS marker was developed from an AFLP band which cosegregated with the YrMoro gene. Sequence-specific primers were made which incorporated the selective bases of the AFLP primers, plus 16 and 17 additional bases extending into the AFLP band. This simple, PCR-based marker will allow wheat breeders to pyramid this resistance gene, along with other resistance genes, into a single wheat genotype.

The genetic characterisation of stripe rust resistance in the German wheat cultivar Alcedo

Stripe rust resistance in the German winter wheat cv. Alcedo has been described as durable, the resistance having remained effective when grown extensively in Germany and Eastern Europe between 1975 and 1989. Genetic characterisation of field resistance in a cross between Alcedo and the stripe rust susceptible UK winter wheat cv. Brigadier identified two major QTL in Alcedo located on the long arms of chromosomes 2D (QPst.jic-2D) and 4B (QPst.jic-4B). Stripe rust resistance was evaluated by measuring the extent of fungal growth, percentage infection (Pi) and the necrotic/chlorotic response of the plant to infection, infection type (IT). Both QPst.jic-2D and QPst.jic-4B contributed significantly to the reduction in stripe rust infection (Pi), with QPst.jic-2D explaining up to 36.20% and QPst.jic-4B 28.90% of the phenotypic variation measured for Pi. Both QTL were identified by the IT phenotypic scores, with QPst.jic-2D in particular being associated with a strong necrotic phenotype (low IT), QPst.jic-2D explaining up to 53.10% of IT phenotypic variation and QPst.jic-4B 22.30%. In addition, two small effect QTL for field stripe rust resistance were identified in Brigadier, QPst.jic-1B on the long arm of chromosome 1B and QPst.jic-5A on the short arm of chromosome 5A. The influence of QPst.jic-1B was primarily seen with the Pi phenotype, contributing up to 13.10% of the explained phenotypic variation. QPst.jic-5A was only detected using an approximate multiple-QTL model and selecting markers linked to the major effect QTL, QPst.jic-2D and QPst.jic-4B as co-factors. Seedling stripe rust resistance was also mapped in the cross, which confirmed the location of Yr17 from Brigadier to the short arm of chromosome 2A. A seedling expressed QTL was also located in Alcedo that mapped to the same location as the field stripe rust resistance QPst.jic-2D.

Wheat mutants showing altered adult plant disease resistance

Wheat mutants, selected on an altered resistance phenotype to Puccinia striiformis f.sp. tritici, the causal agent of yellow rust, were assessed in greenhouse tests to determine the growth stage at which the mutant phenotype was expressed and which components of yellow rust resistance were altered. Four of the mutants showed reduced levels of yellow rust infection, while three exhibited increased levels of infection. The infection characters altered were those seen after the establishment of hyphal growth, i.e. days after inoculation to sporulation, the percentage of inoculated leaf tissue producing sporulating colonies and the number of spores produced per cm2 of inoculated leaf tissue. The altered resistance phenotype was developmentally regulated in all of the mutants. The changes seen in the infection characters differed for each mutant, suggesting that different genes may have been altered. An altered resistance phenotype to brown rust and/or powdery mildew was also seen for some of the yellow rust mutants.

Molecular and cellular expression of quantitative resistance in barley to powdery mildew

The induction of five defence-related genes was examined in two barley cvs, P-03 and S-03, in direct relation to the growth and development of two near-isogenic Erysiphe graminis f.sp. hordei isolates, CC142 ( Aa6 ) and CC143 ( Va6 ). The barley cvs contained the mildew resistance gene Mla6 , while the E. graminis f.sp. hordei isolates differed only in virulence to Mla6 . Differences in the development of the two isolates on the two cvs were reflected in the patterns of defence gene induction. The defence genes examined included a chitinase, a peroxidase, a phenylalanine ammonia lyase, a leaf-specific thionin and a gene showing homology to the pathogenesis-related R protein from tobacco. The two isolates showed similar patterns of development up to 16 h after inoculation. At this time, a retardation in the development of the avirulent isolate, CC142, became apparent on S-03. However, inhibition of CC142 on P-03 was not seen until 48 h after inoculation. This difference in CC142 development was paralleled by an earlier, specific defence gene response to the avirulent isolate in S-03. By 16 h after inoculation higher transcript levels of the chitinase and the peroxidase genes were seen in S-03, while in P-03, a specific defence gene response to CC142 was only seen after 24 h. The earlier defence gene response, seen in S-03 to the avirulent isolate, may contribute to the earlier inhibition of CC142 development seen on this cultivar.

Development of a doubled haploid mapping population and linkage map for the bread wheat cross Kariega × Avocet S

A doubled haploid wheat population of 150 lines was constructed from the F1 of a cross between cultivars Kariega and Avocet S using the wheat-maize technique. Segregation and linkage analysis of 203 DNA markers, two storage protein markers and the stem rust resistance gene Sr26 yielded 31 linkage groups of which 28 were chromosome-anchored. These linkage groups covered the entire genome of 21 chromosomes. SSR markers were advantageous for chromosome anchoring and AFLP markers for consolidation of the various linkage groups. A significant feature of the linkage map is a relatively low level of polymorphism for markers on the D genome, viz, 18.5% of all markers mapped. The Kariega × Avocet S doubled haploid population and linkage maps have emerged as valuable resources for further genetic studies of qualitative and quantitative traits of economic importance in bread wheat.

An LRR/Malectin Receptor-Like Kinase Mediates Resistance to Non-adapted and Adapted Powdery Mildew Fungi in Barley and Wheat

Pattern recognition receptors (PRRs) belonging to the multigene family of receptor-like kinases (RLKs) are the sensing devices of plants for microbe- or pathogen-associated molecular patterns released from microbial organisms. Here we describe Rnr8 (for Required for non-host resistance 8) encoding HvLEMK1, a LRR-malectin domain-containing transmembrane RLK that mediates non-host resistance of barley to the non-adapted wheat powdery mildew fungus Blumeria graminis f.sp. tritici. Transgenic barley lines with silenced HvLEMK1 allow entry and colony growth of the non-adapted pathogen, although sporulation was reduced and final colony size did not reach that of the adapted barley powdery mildew fungus B. graminis f.sp. hordei. Transient expression of the barley or wheat LEMK1 genes enhanced resistance in wheat to the adapted wheat powdery mildew fungus while expression of the same genes did not protect barley from attack by the barley powdery mildew fungus. The results suggest that HvLEMK1 is a factor mediating non-host resistance in barley and quantitative host resistance in wheat to the wheat powdery mildew fungus.

Stem Rust Resistance in a Geographically Diverse Collection of Spring Wheat Lines Collected from Across Africa

Following the emergence of the Ug99 lineage of Puccinia graminis f. sp. tritici (Pgt) a collective international effort has been undertaken to identify new sources of wheat stem rust resistance effective against these races. Analyses were undertaken in a collection of wheat genotypes gathered from across Africa to identify stem rust resistance effective against the Pgt races found in Eastern and Southern Africa. The African wheat collection consisted of historic genotypes collected in Kenya, South Africa, Ethiopia, Sudan, Zambia, Morocco, and Tunisia, and current South African breeding lines. Both Bayesian cluster and principal coordinate analyses placed the wheat lines from Sudan in a distinct group, but indicated a degree of genetic relatedness among the other wheat lines despite originating from countries across Africa. Seedling screens with Pgt race PTKST, pedigree information and marker haplotype analysis confirmed the presence of Sr2, Sr36, Sr24, Sr31, and Lr34/Yr18/Sr57 in a number of the lines. A genome-wide association study (GWAS) undertaken with Diversiry Arrays Technology (DArT) and stem rust (Sr) gene associated markers and Stem Area Infected (SAI) and Reaction Type (RT) field phenotypes, collected from trials carried out across two seasons in Kenya in 2009 and in South Africa in 2011, identified 29 marker-trait associations (MTA). Three MTA were in common between SAI and RT, with the biggest effect MTA being found on chromosome 6AS. Two wheat lines, W1406 and W6979 that exhibited high levels of adult plant stem rust resistance were selected to generate bi-parental mapping populations. Only the MTA on chromosomes 6AS and 3BS, and the locus Lr34/Yr18/Sr57 were confirmed following QTL mapping. Additional stem rust resistance QTL, not detected by the GWAS, were found on chromosomes 2BS, 2DL, 3DL, and 4D.