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Barley, host to the wheat and rye attacking forms of the stem rust pathogen, Puccinia graminis (Pg), is considered inherently more resistant to Pg than wheat. To investigate whether this enhanced basal defence is associated with early infection or colonisation processes, adult plants of selected barley and wheat entries were inoculated with Pg f. sp. tritici (Pgt) races BNGSC and PTKST, and Pg f. sp. secalis (Pgs) isolate UVPgs1. Flag leaf sheaths on the last stem internode were sampled for analysis. Using scanning electron microscopy and epidermal stripping, early Pg infection structure development was compared between barley and wheat. Sub-stomatal vesicle appearance and production of haustorial mother cells were similar at 24 and 48 h post inoculation (hpi), respectively. Significant variation occurred among host entries (P ≤ 0.01) for colony size and fungal biomass at 120 hpi. Differences between the Pgt races and the Pgs isolate as well as the host entry-Pg isolate interaction proved insignificant (P ≥ 0.05). The period between 120 and 240 hpi may hold some explanation for the perceived lower stem rust receptivity in barley, since a steep increase in accumulated fungal biomass observed in the susceptible wheat Line 37 − 07 was not seen in the susceptible barley entries at the same time interval.

Race 2SA55 of the wheat stem rust pathogen, Puccinia graminis f. sp. tritici, was described for the first time during 2000 in South Africa. This race is of particular interest as it was the first local report of increased virulence towards barley cultivars by wheat stem rust. Using three original accessions of 2SA55 from the rust collection of the University of the Free State, nine single pustule isolates were established. Phenotyping of isolates to characterise 2SA55 according to the North American nomenclature system revealed variation in virulence for Sr9g on the tester lines Kubanka and Acme, which was not detected in the initial description of 2SA55. Seven isolates coded as BNGSC, which except for virulence on Sr9b resembled the initial avirulence/virulence description for 2SA55. The remaining two isolates with Sr9g virulence coded as BPGSC. The seedling infection types for the two 2SA55 derived races BNGSC and BPGSC on the Stakman differential set as well as on an extended set of Sr lines and 144 South African wheat cultivars and advanced breeding lines revealed no further distinction in reaction between them. Microsatellite analysis indicated that while race BN/PGSC shares phenotypic similarity with several of the South African non-Ug99 races, it was genetically distinct.

Optimized inoculation procedures are an important consideration in achieving repeatable plant infection when working with biotrophic rust fungi. Several plant pathology laboratories specializing in rust research employ a system where the collection and application of fungal spores are accomplished using an exchangeable gelatin capsule. Urediniospores are collected from erumpent pustules on plant surfaces into a capsule fitted to a cyclone collector controlled by a vacuum pump. By adding light mineral oil to the same capsule, the spore suspension is then sprayed onto plants by means of a dedicated atomizer (inoculator) connected to an air pressure source. Although devices are not commercially available, modern day technologies provide an opportunity to efficiently design and manufacture collectors and inoculators. Using a process called Additive Manufacturing (AM), also known as “3D printing,” the bodies of a collector and inoculator were digitally designed and then laser-sintered in nylon. Depending on availability, copper or aluminum tubes were fitted to the bodies of both devices afterward to either facilitate directed collection of spores from rust pustules on plant surfaces or act as a siphon tube to deliver the spore suspension contained in the capsule. No statistical differences were found between AM and metal inoculators for spray delivery time or spore deposition per unit area. In replicated collection and inoculation tests of wheat seedlings with urediniospore bulks or single pustule collections of Puccinia triticina and P. graminis f. sp. tritici, the causal organisms of leaf rust and stem rust, consistent and satisfactory infection levels were achieved. Immersing used devices in acetone for 60 s followed by a 2 h heat treatment at 75°C produced no contaminant infection in follow-up tests.

The release of commercial wheat cultivars resistant to stripe rust, caused by Puccinia striiformis f. sp. tritici (Pst), remains one of the primary objectives in many breeding programs. Previous studies of adult plant resistance derived from the winter wheat cultivar Cappelle-Desprez identified the quantitative trait loci (QTL) QYr.ufs-2A, QYr.ufs-2D, QYr.ufs-5B and QYr.ufs-6D to affect stripe rust under South African conditions. Phenotypic field assessment, fluorescence microscopy and molecular analysis were used to characterise recombinant inbred lines differing in number and combinations of these QTL. Besides the confirmation of enhanced resistance through co-occurring resistance loci, varying levels of defence, conditioned by different QTL combinations were observed. Carriers of QYr.ufs-2A or QYr.ufs-2D, accompanied by at least one other QTL, exhibited higher resistance levels than lines with a single QTL. The knowledge gained in this study will help wheat breeders to develop cultivars with more diverse combinations and potentially more durable sources of stripe rust resistance.