Additive Manufacturing of Devices Used for Collection and Application of Cereal Rust Urediniospores

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

Additive Manufacturing of Devices Used for Collection and Application of Cereal Rust Urediniospores

Published ArticleOptimized 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

Fig rust caused by Phakopsora nishidana in South Africa

Fig rust, caused by Cerotelium fici, was first recorded in South Africa in 1927. Recent observations have revealed high incidence of rust and untimely defoliation of fig trees (Ficus carica) in residential gardens and commercial orchards. Using phylogenetic analysis, the causal organism of a fig rust isolate (PREM63073) collected in 2020 was confirmed as Phakopsora nishidana. Inoculation and microscope studies showed that mulberry plants were immune to P. nishidana isolate PREM63073. Infection of fig leaves occurred through stomata on the abaxial leaf surfaces. Very long germ tubes were observed for P. nishidana, often with no clear contact with the leaf surfaces and an apparent lack of directional growth towards stomata. Inoculated plants from 15 fig cultivars varied in their severity of leaf infection, whereas fruit of the cultivar Kadota developed reddish-brown blemishes without sporulation. Currently, C. fici and P. nishidana are recognised as occurring on F. carica in South Africa. This suggests a need to resolve the worldwide distribution and identity of the rust species involved

SMSS J130522.47-293113.0: A high-latitude stellar X-ray source with pc-scale outflow relics?

We report the discovery of an unusual stellar system SMSS J130522.47-293113.0. The optical spectrum is dominated by a blue continuum together with emission lines of hydrogen, neutral, and ionized helium, and the NIII, CIII blend at ~4640-4650 Å. The emission-line profiles vary in strength and position on time-scales as short as 1 d, while optical photometry reveals fluctuations of as much as ~0.2mag in g on time-scales as short as 10-15 min. The system is a weak X-ray source (f 0.3-10 = (1.2 ± 0.1) × 10 -13 ergs cm 2 s -1 in the 0.3-10 keV band) but is not detected at radio wavelengths (3s upper limit of 50 µJy at 5.5 GHz). The most intriguing property of the system, however, is the existence of two 'blobs', a few arcsec in size, that are symmetrically located 3'. 8 (2.2 pc for our preferred system distance of ~2 kpc) each side of the central object. The blobs are detected in optical and near-IR broad-band images but do not show any excess emission in Ha images. We discuss the interpretation of the system, suggesting that the central object is most likely a nova-like CV, and that the blobs are relics of a pc-scale accretion-powered collimated outflow

Potential for re-emergence of wheat stem rust in the United Kingdom

Wheat stem rust, a devastating disease of wheat and barley caused by the fungal pathogen Puccinia graminis f. sp. tritici, was largely eradicated in Western Europe during the mid-to-late twentieth century. However, isolated outbreaks have occurred in recent years. Here we investigate whether a lack of resistance in modern European varieties, increased presence of its alternate host barberry and changes in climatic conditions could be facilitating its resurgence. We report the first wheat stem rust occurrence in the United Kingdom in nearly 60 years, with only 20% of UK wheat varieties resistant to this strain. Climate changes over the past 25 years also suggest increasingly conducive conditions for infection. Furthermore, we document the first occurrence in decades of P. graminis on barberry in the UK. Our data illustrate that wheat stem rust does occur in the UK and, when climatic conditions are conducive, could severely harm wheat and barley production.</p

Accomplishments in wheat rust research in South Africa

Rust diseases, although seasonal, have been severe constraints in wheat production in South Africa for almost 300 years. Rust research gained momentum with the institution of annual surveys in the 1980s, followed by race identification, an understanding of rust epidemiology, and eventually a focused collaboration amongst pathologists, breeders and geneticists. Diversity in South African populations of Puccinia triticina, P. graminis f. sp. tritici and P. striiformis f. sp. tritici has been described and isolates are available to accurately phenotype wheat germplasm and study pathogen populations at national, regional and global levels. Sources of resistance have been, and still are, methodically analysed and molecular marker systems were developed to incorporate, stack and verify complex resistance gene combinations in breeding lines and cultivars. Vigilance, capacity, new technologies, collaboration and sustained funding are critical for maintaining and improving the current research impetus for future management of these important diseases. Significance: Rust diseases threaten wheat crops worldwide, including in South Africa. Management of rusts includes regular surveillance, pathogen diversity studies, rigorous screening of wheat germplasm, and efficient breeding and selection for resistance. Collaboration among plant pathologists, geneticists and breeders has provided momentum in rust research and control in South Africa in recent years

Accomplishments in wheat rust research in South Africa

CITATION: Pretorius, Z. A. et al. 2020. Accomplishments in wheat rust research in South Africa. South African Journal of Science, 116(11/12),:7688, doi:10.17159/sajs.2020/7688.The original publication is available at https://sajs.co.zaRust diseases, although seasonal, have been severe constraints in wheat production in South Africa for almost 300 years. Rust research gained momentum with the institution of annual surveys in the 1980s, followed by race identification, an understanding of rust epidemiology, and eventually a focused collaboration amongst pathologists, breeders and geneticists. Diversity in South African populations of Puccinia triticina, P. graminis f. sp. tritici and P. striiformis f. sp. tritici has been described and isolates are available to accurately phenotype wheat germplasm and study pathogen populations at national, regional and global levels. Sources of resistance have been, and still are, methodically analysed and molecular marker systems were developed to incorporate, stack and verify complex resistance gene combinations in breeding lines and cultivars. Vigilance, capacity, new technologies, collaboration and sustained funding are critical for maintaining and improving the current research impetus for future management of these important diseases.https://sajs.co.za/article/view/7688Publisher's versio

Genome Targeted Introgression of Resistance to African Stem Rust from Aegilops sharonensis into Bread Wheat

Many accessions of the wheat wild relative Sharon goatgrass ( Eig., ) are resistant to African races of the stem rust pathogen (i.e., Ug99 group races), which currently threaten wheat production worldwide. A procedure was designed to introgress the respective resistances to specific bread wheat genomes by producing plants homozygous for the A and B genomes and hemizygous for the D and S genomes or homozygous for the A and D genomes and hemizygous for the B and S genomes. In these genotypes, which lack the allele, homeologous pairing was expected mainly between chromosomes of the D and S genomes or B and S genomes, respectively. An antigametocidal (AG) wheat mutant () was used to overcome gametocidal effects. Wheat lines initially found resistant at the seedling stage were also highly resistant at the adult plant stage in rust nurseries established in the field. DNA of 41 selected homozygous resistant lines, analyzed by the Axiom wheat 820K SNP array, showed alien chromatin mainly in wheat chromosomes 1B, 1D, and 5B. This work suggests that, in most cases, it is possible to target introgressions into the homeologous chromosome of a selected genome of bread wheat

Assessing the Individual and Combined Effects of QTL for Adult Plant Stripe Rust Resistance Derived from Cappelle-Desprez

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

Dissection of the multigenic wheat stem rust resistance present in the Montenegrin spring wheat accession PI 362698

Abstract Background Research to identify and characterize stem rust resistance genes in common wheat, Triticum aestivum, has been stimulated by the emergence of Ug99-lineage races of the wheat stem rust pathogen, Puccinia graminis f. sp. tritici (Pgt), in Eastern Africa. The Montenegrin spring wheat landrace PI 362698 was identified as a source of Pgt resistance. This accession exhibits resistance to multiple Ug99-lineage and North American Pgt races at seedling and adult-plant stages. A recombinant inbred population was developed by crossing the susceptible line LMPG-6 with a single plant selection of PI 362698. A genetic map was constructed using the Illumina iSelect 90 K wheat assay and the markers csLv34, NB-LRR3, and wMAS000003 and quantitative trait locus (QTL) analysis was performed. Results QTL analysis identified five significant QTLs (α = 0.05) on chromosomes 2B, 3B, 6A, 6D, and 7A associated with wheat stem rust resistance. The QTL on chromosome 3B was identified using both field data from Kenya (Pgt Ug99-lineage races) and seedling data from Pgt race MCCF. This QTL potentially corresponds to Sr12 or a new allele of Sr12. The multi-pathogen resistance gene Sr57 located on chromosome 7D is present in PI 362698 according to the diagnostic markers csLv34 and wMAS000003, however a significant QTL was not detected at this locus. The QTLs on chromosomes 2B, 6A, and 6D were identified during seedling trials and are thought to correspond to Sr16, Sr8a, and Sr5, respectively. The QTL identified on chromosome 7A was detected using MCCF seedling data and may be Sr15 or a potentially novel allele of recently detected Ug99 resistance QTLs. Conclusions The combination of resistance QTLs found in PI 362698 is like the resistance gene combination present in the broadly resistant cultivar Thatcher. As such, PI 362698 may not be a landrace as previously thought. PI 362698 has been crossed with North Dakota wheat germplasm for future breeding efforts. Additional work is needed to fully understand why the combination of genes present in PI 362698 and ‘Thatcher’ provide such durable resistance