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Spongospora subterranea f. sp. subterranea is an important pathogen of potato responsible for major losses in most potato growing regions of the world. Infection is initiated by bi-flagellated motile zoospores released from long-lived resting spores. Zoospore chemotaxis to the host plant root is widely believed to be stimulated by host root exudate compounds, although direct evidence is lacking. This study refined the traditional chemotaxis capillary assay. With this we provided the first empirical evidence of S. subterranea zoospore chemotaxis. Individual potato root exudate metabolites were either taxis neutral, inhibitory, or attractant to the zoospores. L-glutamine was the strongest chemoattractant, while spermine was the most inhibitory. Zoospore motility and chemotaxis were constrained by strongly acidic or alkaline solutions of pH < 5.3 and > 8.5, respectively. Beyond pH, ionic constituents of the test solution affected zoospore motility as Sorensen’s phosphate buffer stalled zoospore motility, but HEPES buffer at the same concentration and pH had little or no negative motility effect. Zoospore motility, as characterized by several parameters, influenced chemotaxis. Among the parameters measured, total distance travelled was the best predictor of zoospore chemotaxis. The characterization of environmental and ecological effects on zoospore motility and chemotaxis highlights useful targets for S. subterranea disease control through manipulation of zoospore taxis or selection of host resistance traits.

This study examined the natural and experimental host range and aphid and graft transmission of the tentative polerovirus phasey bean mild yellows virus (PBMYV). Eleven complete coding sequences from PBMYV isolates were determined from a range of hosts and locations. We found two genetically distinct variants of PBMYV. PBMYV-1 was the originally described variant, and PBMYV-2 had a large putative recombination in open reading frame 5 such that PBMYV-1 and PBMYV-2 shared only 65-66% amino acid sequence identity in the P5 protein. The virus was transmitted by a clonal colony of cowpea aphids (Aphis craccivora) and by grafting with infected scions but was not transmitted by a clonal colony of green peach aphids (Myzus persicae). PBMYV was found in natural infections in 11 host species with a range of symptoms and severity, including seven important grain legume crops from across a wide geographic area in Australia. PBMYV was common and widespread in the tropical weed phasey bean (Macroptilium lathyroides), but it is likely that there are other major alternative hosts for the virus in temperate regions of Australia. The experimental host range of PBMYV included the Fabaceae hosts chickpea (Cicer arietinum), faba bean (Vicia faba), pea (Pisum sativum), and phasey bean, but transmissions failed to infect several other members of the families Asteraceae, Cucurbitaceae, Fabaceae and Solanaceae. PBMYV was commonly found in grain legume crops in eastern and western Australia, sometimes at greater than 90% incidence. This new knowledge about PBMYV warrants further assessments of its economic impact on important grain legume crops.

Tuber greening is one of the major causes of quality loss in the potato industry, however, the impact of nitrogen in-field fertilisation on this visual defect remains unknown. Two field experiments, one in Tasmania with final N treatment rates of 40, 100 and 190 kg N ha-1 for Nicola and 35, 95 and 185 Kg N ha-1 for Maranca, and another in South Australia with N rates of 100, 200 and 300 kg ha-1 for Maranca, showed a positive linear relationship (R2 = 0.99, 0.86 and 0.90, respectively) between tuber N content and tuber greening, as determined by chlorophyll content. However, increased N fertilisation rates did not directly translate to increased tuber N content, with responses appearing to be variety specific. We conclude that tuber N content is a risk factor associated with tuber greening, but the manipulation of tuber N content through N fertilisation rate is not straightforward and may depend on variety and location.

Light-induced tuber greening is one of the most important quality defects of potato. Although varietal and maturity factors are known to affect greening resistance, physiological mechanisms of resistance are poorly understood. We proposed that physiological and biochemical factors within the tuber periderm provide resistance and hypothesised that resistance is primarily related to suberin content. We investigated differences in the tuber periderm between genotypes and tuber maturities that varied in greening propensity. We examined suberin and light-induced pigment accumulation, and phellem cell development and studied greening propensity in mutant and chemically treated tubers with enhanced suberisation. Resistance to greening was strongly linked to increased suberin in the periderm, which varied with variety and tuber maturity. Furthermore, greening was reduced in mutant and chemically treated tubers with enhanced suberisation. Increases in phellem cell layers and light-induced carotenoids and anthocyanins were identified as secondary resistance factors. Our work represents the first physiological mechanism of varietal and tuber maturity resistance to greening, expanding the known functionality of suberin and providing for the first time a biomarker that will aid producers and breeders in selection and improvement of potato varieties for greening resistance.

The pathogen Spongospora subterranea infects potato roots and developing tubers resulting in tuber yield and quality losses. Currently, there are no fully effective treatments for disease control. Host resistance is an important tool in disease management and understanding the molecular mechanisms of defence responses in roots of potato plants is required for the breeding of novel resistant cultivars. Here, we integrated transcriptomic and proteomic datasets to uncover these mechanisms underlying S. subterranea resistance in potato roots. This multi-omics approach identified upregulation of glutathione metabolism at the levels of RNA and protein in the resistant cultivar but not in the susceptible cultivar. Upregulation of the lignin metabolic process, which is an important component of plant defence, was also specific to the resistant cultivar at the transcriptome level. In addition, the inositol phosphate pathway was upregulated in the susceptible cultivar but downregulated in the resistant cultivar in response to S. subterranea infection. We provide large-scale multi-omics data of Spongospora-potato interaction and suggest an important role of glutathione metabolism in disease resistance.

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Simple Summary Infections of potato tubers and roots by Spongospora subterranea result in powdery scab and root diseases. Losses due to infections with S. subterranea are substantial in most potato-growing regions of the world with no fully effective treatments available. Understanding the gene regulation of pathogens in their host is dependent on multidimensional datasets. In this study, we profiled the transcriptome and proteome of S. subterranea within the susceptible and resistant host. Enzyme activity and nucleic acid metabolism appear to be important to the virulence of S. subterranea. Our results provide a good resource for future functional studies of powdery scab and might be useful in S. subterranea inoculum management. Abstract Spongospora subterranea is an obligate biotrophic pathogen, causing substantial economic loss to potato industries globally. Currently, there are no fully effective management strategies for the control of potato diseases caused by S. subterranea. To further our understanding of S. subterranea biology during infection, we characterized the transcriptome and proteome of the pathogen during the invasion of roots of a susceptible and a resistant potato cultivar. A total of 7650 transcripts from S. subterranea were identified in the transcriptome analysis in which 1377 transcripts were differentially expressed between two cultivars. In proteome analysis, we identified 117 proteins with 42 proteins significantly changed in comparisons between resistant and susceptible cultivars. The functional annotation of transcriptome data indicated that the gene ontology terms related to the transportation and actin processes were induced in the resistant cultivar. The downregulation of enzyme activity and nucleic acid metabolism in the resistant cultivar suggests a probable influence of these processes in the virulence of S. subterranea. The protein analysis results indicated that the majority of differentially expressed proteins were related to the metabolic processes and transporter activity. The present study provides a comprehensive molecular insight into the multiple layers of gene regulation that contribute to S. subterranea infection and development in planta and illuminates the role of host immunity in affecting pathogen responses.

For soilborne pathogens, germination of the resting or dormant propagule that enables persistence within the soil environment is a key point in pathogenesis. Spongospora subterranea is an obligate soilborne protozoan that infects the roots and tubers of potato causing root and powdery scab disease for which there are currently no effective controls. A better understanding of the molecular basis of resting spore germination of S. subterranea could be important for development of novel disease interventions. However, as an obligate biotroph and soil dwelling organism, the application of new omics techniques for the study of the pre-infection process in S. subterranea has been problematic. Here, RNA sequencing was used to analyse the reprogramming of S. subterranea resting spores during the transition to zoospores in an in-vitro model. More than 63 million mean high-quality reads per sample were generated from the resting and germinating spores. By using a combination of reference-based and de novo transcriptome assembly, 6,664 unigenes were identified. The identified unigenes were subsequently annotated based on known proteins using BLAST search. Of 5,448 annotated genes, 570 genes were identified to be differentially expressed during the germination of S. subterranea resting spores, with most of the significant genes belonging to transcription and translation, amino acids biosynthesis, transport, energy metabolic processes, fatty acid metabolism, stress response and DNA repair. The datasets generated in this study provide a basic knowledge of the physiological processes associated with spore germination and will facilitate functional predictions of novel genes in S. subterranea and other plasmodiophorids. We introduce several candidate genes related to the germination of an obligate biotrophic soilborne pathogen which could be applied to the development of antimicrobial agents for soil inoculum management.

The soil-borne and obligate plant-associated nature of S. subterranea has hindered a detailed study of this pathogen and in particular, the regulatory pathways driving the germination of S. subterranea remain unknown. To better understand the mechanisms that control the transition from dormancy to germination, protein profiles between dormant and germination stimulant-treated resting spores were compared using label-free quantitative proteomics. Among the ~680 proteins identified 20 proteins were found to be differentially expressed during the germination of S. subterranea resting spores. Elongation factor Tu, histones (H2A and H15), proteasome and DJ-1_PfpI, involved in transcription and translation, were upregulated during the germination of resting spores. Downregulation of both actin and beta-tubulin proteins occurred in the germinating spores, indicating that the changes in the cell wall cytoskeleton may be necessary for the morphological changes during the germination of the resting spore in S. subterranea. Our findings provide new approaches for the study of these and similar recalcitrant micro-organisms provide the first insights into the basic protein components of S. subterranea spores. A better understanding of S. subterranea biology may lead to the development of novel approaches for the management of persistent soil inoculum.

Attempts at management of diseases caused by protozoan plant parasitic Phytomyxea have often been ineffective. The dormant life stage is characterised by long-lived highly robust resting spores that are largely impervious to chemical treatment and environmental stress. This review explores some life stage weaknesses and highlights possible control measures associated with resting spore germination and zoospore taxis. With phytomyxid pathogens of agricultural importance, zoospore release from resting spores is stimulated by plant root exudates. On germination, the zoospores are attracted to host roots by chemoattractant components of root exudates. Both the relatively metabolically inactive resting spore and motile zoospore need to sense the chemical environment to determine the suitability of these germination stimulants or attractants respectively, before they can initiate an appropriate response. Blocking such sensing could inhibit resting spore germination or zoospore taxis. Conversely, the short life span and the vulnerability of zoospores to the environment require them to infect their host within a few hours after release. Identifying a mechanism or conditions that could synchronise resting spore germination in the absence of host plants could lead to diminished pathogen populations in the field.

Spongospora subterranea, which causes powdery scab of potato, infects a diverse range of plant species. Crop rotation as a powdery scab management tool will be compromised if pathogen hosts exist between potato crops. Opium poppy (Papaver somniferum) and pyrethrum (Tanacetum cinerariifolium) are important crops within intensive vegetable production rotations in NW Tasmania. Measurements of S. subterranea soil inoculum within a commercial field showed pathogen amounts were substantially elevated following an opium poppy crop, which suggested host status. In glasshouse testing, opium poppy and pyrethrum were confirmed as hosts of S. subterranea, with opium poppy the more susceptible of the two. Both species were less susceptible than tomato, a known host. Observations of early growth suggested inoculation impacts on all three plant species, although at 16 (tomato and opium poppy) or 26 (pyrethrum) weeks postinoculation, only tomato had significantly reduced shoot and root development. The role of rotation crops in inoculum persistence and the possible role of S. subterranea as a minor pathogen of nonpotato crops are discussed.

Tomato plants are highly susceptible to root infection by Spongospora subterranea and are commonly used as bioassay hosts. The impacts of root infection with S. subterranea on plant productivity and yield have been debated. Recent experiments with potato, the major economic host of S. subterranea, have indicated significantly reduced plant growth and potato yield following heavy infection. However, there have been very few similar studies that have examined the possible impacts of S. subterranea infection on tomato plant growth. Three tomato cultivars, "Grape," "Roma" and "Truss," were challenged with S. subterranea inoculum in hydroponic culture. Moderate to severe zoosporangial infections were observed with minor but statistically significant differences in susceptibility among the three tomato cultivars. Zoosporangial root infection in the absence of root gall formation resulted in significantly diminished shoot lengths and plant fresh weights in pathogen challenge tests conducted both in hydroponic culture and glasshouse-grown plants in potting mix. Root lengths were reduced, but the differences were statistically significant in a single trial only. The findings from this study demonstrate that, as with potato, root infection by S. subterranea can result in reduced tomato plant growth and that root gall production associated with root infection was not necessary for this retardation of growth response. This further suggests that possible yield impacts in other crop species that are hosts for S. subterranea root infection are worthy of examination.

Greening is a major cause of quality loss in potato tubers. As underground stems, potato tubers are non-photosynthetic plant organs that lack photosynthetic machinery. However, after light exposure, amyloplasts convert to chloroplasts in tuber peripheral cell layers, which cause the accumulation of the green photosynthetic pigment, chlorophyll. Tuber greening can be impacted by genetic, cultural, physiological and environmental factors including planting depth, tuber physiological age, temperature, atmospheric oxygen levels, and lighting conditions. Numerous studies have been devoted to understand and control this costly defect for the potato industry. This review brings together the available knowledge on light-induced greening, from causes to solutions and suggestions on further research with a focus on identifying the underlying mechanisms of tuber greening.

Hoagland’s solution (HS), a defined nutrient supplement for plants, has been previously reported to stimulate zoospore release from resting spores of the potato pathogen Spongospora subterranea f. sp. subterranea. This study obtained direct empirical evidence for an increase in zoospore release with HS treatment, and identified Fe-EDTA as the stimulant component of HS. Stimulation of resting spores by HS and Fe-EDTA resulted in greater and earlier zoospore release compared to a distilled water control, and in the presence of a susceptible tomato host plant resulted in enhanced root infection. Given the labile nature of S. subterranea zoospores, it was postulated that stimulation of premature release of zoospores from the dormant resting spores in absence of susceptible hosts could reduce soil inoculum levels. In two glasshouse trials in the absence of host plants, both Fe-EDTA and HS soil treatments reduced S. subterranea soil inoculum levels, providing proof of concept for the ‘germinate to exterminate’ approach to inoculum management.

A series of experiments was conducted under glasshouse and hydroponic conditions to determine the specific window period for common scab disease infection in potato tubers. The study was performed in a glasshouse system where separate tubers from the root zone were inoculated at different intervals during plant growth along with a novel hydroponic system to inoculate individual tubers at specific times of development growth allowing non-destructive observations of common scab symptoms developing. The window of tuber susceptibility to common scab disease infection was shown to vary with the season or conditions under which the plants were grown. Different internodes on tubers were found susceptible to infection at different times during tuber development. Basal internodes, which are the first sections of the tuber to expand, were susceptible to infection in the beginning of tuber development, whereas apical internodes only became susceptible later in tuber growth when the basal internodes were no longer susceptible.Journal of Nepal Agricultural Research Council Vol.3 2017: 19-26

The soil-borne potato pathogen Spongospora subterranea persists in soil as sporosori, which are aggregates of resting spores. Resting spores may germinate in the presence of plant or environmental stimuli, but direct evidence for resting spore dormancy is limited. A soilless tomato bait plant bioassay and microscopic examination were used to examine features of S. subterranea resting spore dormancy and infectivity. Dried sporosori inocula prepared from tuber lesions and root galls were infective after both short- and long-term storage (1 week to 5 years for tuber lesions and 1 week to 1 year for root galls) with both young and mature root galls inocula showing infectivity. This demonstrated that a proportion of all S. subterranea resting spores regardless of maturity exhibit characteristics of stimuli-responsive dormancy, germinating under the stimulatory conditions of the bait host plant bioassay. However, evidence for constitutive dormancy within the resting spore population was also provided as incubation of sporosorus inoculum in a germination-stimulating environment did not fully exhaust germination potential even after 2.4 years. We conclude that S. subterranea sporosori contain both exogenous (stimuli-responsive) and constitutively dormant resting spores, which enables successful host infection by germination in response to plant stimuli and long-term persistence in the soil.

The incidence of Groundnut rosette assistor virus (GRAV) in farmers’ fields and sequence diversity of groundnut rosette disease (GRD) agents were assessed in the three northern groundnut production regions of Ghana. GRAV incidence was high (69.5 to 75.0%) but not significantly different between the regions. Nucleotide sequencing of GRAV coat protein (CP) gene revealed 99–100% identity among the Ghanaian isolates and 97–100% similarity to GRAV sequences from Nigeria and Malawi for both nucleotide and predicted amino acids. Nucleotide sequence identities of partial ORF3 and 4 of Groundnut rosette virus (GRV) among the Ghanaian isolates were more variable (89–100%). Ghanaian GRV isolates were more closely related in nucleotide sequence identity to Nigerian isolates (95–98%) than Malawian isolates (88–90%). Similarly, nucleotide identity within Ghanaian GRV-sat RNA’s were close (94–100%), but distinct from Nigerian (82–87%) and Malawian (82–86%) GRV-sat RNAs. Ghanaian isolates of all three agents of GRD showed no obvious isolate diversity patterns based on the regions from where they were collected. We present the first report on the distribution and genetic diversity of GRD agents in Ghana.

Twelve cultivars of groundnut were screened in field trials for resistance to groundnut rosette disease (GRD), caused by coinfection with Groundnut rosette assistor virus (GRAV), Groundnut rosette virus (GRV), and its satellite RNA in the coastal savannah of Ghana. ‘Oboshie’ groundnut was rated as highly resistant; ‘Bremaowuo’, ‘Nkatefufuo’, and ‘Behenase’ as resistant; and ‘Nkosuor’, ‘Kumawu’, and ‘Otuhia’ as moderately resistant. GRAV infection rates of 11.8 to 61.8% (dry season) and 13.9 to 100% (wet season) were found, which included symptomless plants, suggesting that some lacked coinfection with GRV and its satellite. Chlorotic ringspot and line-pattern symptoms were observed, suggesting infection with Groundnut ringspot virus (GRSV). Virus identity was confirmed by enzyme-linked immunosorbent assay, reverse-transcription polymerase chain reaction, and amplicon sequencing. This is the first report of GRSV in Ghana. GRSV infection rates were 0.0 to 69.5% (dry season) and 26.1 to 69.5% (wet season). Mixed infections of GRAV and GRSV were common in all cultivars except Nkosuor and Bremaowuo in the dry season. Most cultivars graft inoculated with GRD showed significantly reduced height, leaf area, chlorophyll content, dry haulm weight, and seed yield compared with healthy plants. The sources of resistance to GRD and possibly GRAV and GRSV identified in this study could be exploited in groundnut breeding programs.

Common scab and powdery scab are major soilborne diseases of potato (Solanum tuberosum). The agricultural cost of both diseases is due to discounting or rejection of infected seed tubers or the formation of lesions on the surface of the tubers that make them unmarketable or, for powdery scab, yield loss. There are no widely accessible and efficacious disease management options for either common scab or powdery scab. In prior work, low-dose foliar treatment with the auxin analog herbicide 2,4-D has been shown to reduce common scab severity of select S. tuberosum cultivars in field and greenhouse settings. In the present study, we address the broad applicability of 2,4-D in disease management by testing the efficacy of 2,4-D treatment across diverse potato cultivars and in suppressing multiple scab diseases of potato. Specifically, we tested whether low-dose 2,4-D treatment is broadly efficacious on multiple white potato cultivars used in Eastern United States potato production. Additionally, we sought to determine whether 2,4-D is efficacious for mitigating powdery scab for Russet Burbank potatoes in a commercial field setting. In two years of field trials, low-dose foliar 2,4-D treatment significantly reduced common scab of all tested potato cultivars with no impact on total tuber yield. In two years of field trials in Tasmania, Australia, low-dose foliar 2,4-D treatment also reduced powdery scab of Russet Burbank potato with no impact on total tuber yield while stimulating potato plant root growth. The ability of 2,4-D to suppress two major tuber diseases and stimulate potato root growth warrants further investigation and optimization for integration into commercial growing systems.

Common scab, a globally important potato disease, is caused by infection of tubers with pathogenic Streptomyces spp. Previously, disease-resistant potato somaclones were obtained through cell selections against the pathogen’s toxin, known to be essential for disease. Further testing revealed that these clones had broad-spectrum resistance to diverse tuber-invading pathogens, and that resistance was restricted to tuber tissues. The mechanism of enhanced disease resistance was not known. Tuber periderm tissues from disease-resistant clones and their susceptible parent were examined histologically following challenge with the pathogen and its purified toxin. Relative expression of genes associated with tuber suberin biosynthesis and innate defense pathways within these tissues were also examined. The disease-resistant somaclones reacted to both pathogen and toxin by producing more phellem cell layers in the tuber periderm, and accumulating greater suberin polyphenols in these tissues. Furthermore, they had greater expression of genes associated with suberin biosynthesis. In contrast, signaling genes associated with innate defense responses were not differentially expressed between resistant and susceptible clones. The resistance phenotype is due to induction of increased periderm cell layers and suberization of the tuber periderm preventing infection. The somaclones provide a valuable resource for further examination of suberization responses and its genetic control.

Utilising certified seed is an important principle in providing a potato crop with the greatest opportunity to grow to its full potential free of disease. Within Australia economically important tuber blemish diseases affecting potato tuber quality include Black scurf, Common scab, Powdery scab and Root knot nematode, caused by infections with the pathogens, Rhizoctonia solani , Streptomyces scabies, Spongospora subterranea f. sp. subterranea , and Meloidogyne spp. respectively. All of these pathogens may be carried on seed-tubers providing inoculum for disease in the subsequent crops, and are assessed in potato seed-tuber certification schemes world-wide. Typically seedlots are certified by visual subjective assessments of disease but recently developed quantitative polymerase chain reaction (qPCR) tools that detect and enumerate pathogen DNA provide an additional measure of seed tuber health. We compared both visual and qPCR assessments of potato seedlots passing through the Tasmanian potato certification scheme over a three year period. Further, selected tubers carrying tuber-borne inoculum loads of the three pathogens were grown in pathogen-free potting soil to establish whether the concentration of pathogen DNA from the tuber peel was associated with progeny crop disease. Outcomes suggest that visual assessment, in most circumstances provides an accurate measure of tuber seed health for certification; however qPCR was able to clearly differentiate difficult to distinguish diseases and identify levels of pathogen DNA present on symptomless tubers. Additionally, it was shown that planting tubers with high pathogen inoculum loads led to enhanced progeny disease compared to planting 'certified' tubers.

Proximate and elemental composition of four peanut genotypes infected with groundnut rosette disease (GRD) was examined. Moisture and ash content generally decreased while fat and energy content increased in seeds from diseased plants. Protein and carbohydrate varied between seeds of diseased and healthy plants of the different varieties with no consistent pattern. Instrumental neutron activation analysis of 10 elements within leaves, stems and seeds showed elevated levels of K, Al and Cl in leaves, stems and seeds in at least three of the four varieties infected with GRD while Na was decreased in stems but increased in seeds. While significant differences were found, Mg, Mn, Ca and Zn did not show any consistent change with respect to plant part or genotype, between diseased and healthy plants. V and Fe were found at low levels in leaves and stems and not detected in seeds. This represents the first report on the effect of GRD on the nutritive quality of peanuts.

Grading seed tubers is a common practice to remove visually diseased tubers and meet certification standards. This study sought to quantify Spongospora subterranea inocula, the causal agent of powdery scab in eight commercial seedlots prior to and following grading. Pathogen levels generally decreased although the extent of reduction was dependant on the mean disease level within each seedlot, prior to grading. Where mean seedlot disease incidence was high (20 to 40 %) grading was not effective, reducing pathogen levels by only 2-fold (P > 0.05) on the visually disease-free seed following grading which remained a high risk for disease in progeny plants. However, where mean seedlot disease incidence was low (5 to 7 %) grading substantially reduced S. subterranea inoculum (12–23-fold, P

Spongospora subterranea (Wallr.) Lagerh. is the causative agent of powdery scab and root diseases of potato. Diseases induced by S. subterranea causes substantial economic losses to the global potato industry. The process of disease development in the S. subterranea-potato pathosystem have long been studied, but critical events prior to infection and disease development remain poorly understood because the subject have received relatively little attention. Nonetheless, new knowledge of host-pathogen-environment interactions have been gained in recent years. This paper provides the current knowledge of the key events which leads to the development of potato diseases caused by S. subterranea and highlights future research to address identified knowledge gaps. This will further our understanding of the interactions between S. subterranea and its potato host and contribute to improved disease control measures.

Australian hazelnuts are a new and emerging industry with the potential annual value in Tasmania alone estimated to be in the order of $ 10-15 million. Currently tree propagation is a slow and inefficient process with suckers collected from mature trees and grown on in nurseries. This parent material may harbour diseases and pests and surviving progeny may have inconsistent growth rates, undesirable suckering and non-uniform growth habits, making orchard establishment challenging. The development of tissue culture technologies for hazels has been pioneered in Oregon, USA, resulting in new cultivars and improved planting stock underpinning successful industry establishment in the USA. There has been little success with tissue culture production of local cultivars in Australia. This project examined strategies to optimise the transfer of glasshouse and field grown cuttings (explants) into axenic tissue culture, by testing different disinfestation treatments with sodium hypochlorite (NaOCl). Further, once in tissue culture, we tested the impact of varying agar content in media on key growth parameters. Outcomes indicated fungal and bacterial contamination from field-sourced explants was greater than those from glasshousesourced and that a greater NaOCl concentration (2.1% compared to 1.05%) was required to obtain axenic tissue cultures. Also, low agar content (0.2-0.4%) in tissue culture media produced improved hazel explant growth with greater shoot fresh weights and node numbers than with high agar content. These findings may help underpin the development of micro-propagation as a useful source of quality planting material for the hazelnut industry in Australia.

The Arabidopsis mutant ( ucu2 - 2/gi - 2 ) is thaxtomin A, isoxaben and NPA-sensitive indicated by root growth and ion flux responses providing new insights into these compounds mode of action and interactions. Thaxtomin A (TA) is a cellulose biosynthetic inhibitor (CBI) that promotes plant cell hypertrophy and cell death. Electrophysiological analysis of steady-state K+ and Ca2+ fluxes in Arabidopsis thaliana roots pretreated with TA for 24 h indicated a disturbance in the regulation of ion movement across the plant cell membrane. The observed inability to control solute movement, recorded in rapidly growing meristematic and elongation root zones, may partly explain typical root toxicity responses to TA treatment. Of note, the TA-sensitive mutant (ucu2-2/gi-2) was more susceptible with K+ and Ca2+ fluxes altered between 1.3 and eightfold compared to the wild-type control where fluxes altered between 1.2 and threefold. Root growth inhibition assays showed that the ucu2-2/gi-2 mutant had an increased sensitivity to the auxin 2,4-D, but not IAA or NAA; it also had increased sensitivity to the auxin efflux transport inhibitor, 1-naphthylphthalamic acid (NPA), but not 2,3,5- Triiodobenzoic acid (TIBA), when compared to the WT. The NPA sensitivity data were supported by electrophysiological analysis of H+ fluxes in the mature (but not elongation) root zone. Increased sensitivity to the CBI, isoxaben (IXB), but not dichlobenil was recorded. Increased sensitivity to both TA and IXB corresponded with higher levels of accumulation of these toxins in the root tissue, compared to the WT. Further root growth inhibition assays showed no altered sensitivity of ucu2-2/gi-2 to two other plant pathogen toxins, alternariol and fusaric acid. Identification of a TA-sensitive Arabidopsis mutant provides further insight into how this CBI toxin interacts with plant cells.

Light conditions in retail stores may contribute to potato greening. In this study, we aimed to develop a potato tuber greening risk rating model for retail stores based on light quality and intensity parameters. This was achieved by firstly exposing three potato varieties (Nicola, Maranca and Kennebec) to seven specific light wavelengths (370, 420, 450, 530, 630, 660 and 735 nm) to determine the tuber greening propensity. Detailed light quality and intensity measurements from 25 retail stores were then combined with the greening propensity data to develop a tuber greening risk rating model. Our study showed that maximum greening occurred under blue light (450 nm), while 53%, 65% and 75% less occurred under green (530 nm), red (660 nm) and orange (630 nm) light, respectively. Greening risk, which varied between stores, was found to be related to light intensity level, and partially explained potato stock loss in stores. Our results from this study suggested that other in-store management practices, including lighting duration, average potato turnover, and light protection during non-retail periods, likely influence tuber greening risk.

Spongospora subterranea is a soil-borne plant pathogen responsible for the economically significant root and powdery scab diseases of potato. However, the obligate biotrophic nature of S. subterranea has made the detailed study of the pathogen problematic. Here, we first compared the benefits of sporosori partial purification utilizing Ludox&reg, gradient centrifugation. We then undertook optimization efforts for protein isolation comparing the use of a urea buffer followed by single-pot solid-phase-enhanced sample preparation (SP3) and a sodium dodecyl sulphate (SDS) buffer followed by suspension-trapping (S-Trap). Label-free, quantitative proteomics was then used to evaluate the efficiency of the sporosori purification and the protein preparation methods. The purification protocol produced a highly purified suspension of S. subterranea sporosori without affecting the viability of the spores. The results indicated that the use of a combination of SDS and S-Trap for sample clean-up and digestion obtained a significantly higher number of identified proteins compared to using urea and SP3, with 218 and 652 proteins identified using the SP3 and S-Trap methods, respectively. The analysis of proteins by mass spectrometry showed that the number of identified proteins increased by approximately 40% after the purification of spores by Ludox&reg, These results suggested a potential use of the described spore purification and protein preparation methods for the proteomics study of obligate biotrophic pathogens such as S. subterranea.

Maintaining the health, and therefore productivity, of agricultural soils is vital for continued sustainable agricultural production to support the world’s growing population. Potatoes are grown in a variety of agro-ecological systems and are one of the most important food crops worldwide. Potato crops are demanding on the soil with significant heavy machinery traffic, intensive tillage operations and high inputs of fertiliser, pesticides and water. Maintaining or improving soil health can therefore be challenging for growers. This review considers the different aspects of soil health in a potato production context, how to measure them and how they can be influenced by management practices. Soil health is a complex concept encompassing the physical, chemical and biological properties of the soil and their role in ecosystem services and the growth of plants. Although our understanding of soil health and its impact on crop productivity has improved in the last 30 years, many knowledge gaps remain.

Measurement of pathogens on seed tubers is essential for informing likelihood of subsequent potato disease. Here we utilized quantitative PCR assessment of pathogen DNA and visual assessment of disease to measure seed tuber inoculum and used this to model development of disease in potato grown in pathogen-free soil. Analysis by recursive partitioning and modeling using receiver operating curves indicated both abundance of Rhizoctonia solani AG3 and Streptomyces scabies DNA, and disease symptoms associated with these pathogens on seed tubers could predict subsequent disease in progeny tubers and for R. solani, stolons. In contrast, abundance of Spongospora subterranea DNA and disease symptoms on seed tubers were not consistently associated with powdery scab in progeny tubers. The relationship between S. subterranea DNA and seed tuber symptoms on root galling was stronger. Symptomless seed tubers that carried high levels of S. subterranea DNA were also associated with greater root galling than those with low pathogen DNA levels. There was a modest association between root galling and powdery scab in progeny tubers. These results highlight the importance of using certified seed tubers, and demonstrate a statistical tool for measuring the impact of seed tuber-borne inoculum.

The tomato potato psyllid, Bactericera cockerelli (Sulc, 1909), is a major biosecurity threat to Australian solanaceous crop industries. Native to North and Central America, B. cockerelli was accidentally introduced into New Zealand in the mid-2000s. The psyllid is a vector of a phloem-limited, alpha-proteobacterium, ‘Candidatus Liberibacter solanacearum’ (= ‘Ca. L. psyllaurous’), which is associated with psyllid yellows disease in tomatoes, potatoes, capsicums, eggplants and tamarillos, and zebra chip disease in potatoes. Both the vector and pathogen have had a devastating impact on the solanaceous crop industries they affect, resulting in millions of dollars of losses annually through increased pest control and surveillance, reduced yields and disruption to commodity export markets. In April 2014, B. cockerelli and ‘Ca. L. solanacearum’ were reported on Norfolk Island, and it is feared that they could enter other Australian States or Territories either through the accidental importation of infested plant material or by the dispersal of psyllids on easterly airflows from New Zealand. In this paper, we give an overview of the threat of the vector and pathogen to Australian commodities. We report on an ongoing surveillance program, initiated in February 2011, designed to detect incursions of B. cockerelli in eastern Australian potato fields using yellow sticky traps. During the last 3 years, over 2300 traps were placed in the major potato growing regions of Tasmania, Victoria, South Australia and Queensland. No B. cockerelli were detected but over 9600 other Psylloidea were trapped, mainly Ctenarytaina spp. and Acizzia spp. (Psyllidae). Only 0.3% of all Psylloidea caught belonged to the Triozidae allowing the rapid differentiation of the majority of trapped psyllids from B. cockerelli based on wing venation. Recommendations are made for further research and to extend the surveillance program to include other Australian solanaceous crop industries.

Applications of the synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D) to the foliage of potato plants can reduce common scab, a tuber disease. However, in prior research effective applications at 200 mg L−1 2,4-D resulted in phytotoxic side effects with reduced tuber yield and quality. This study showed that minimal significant threshold rates from 8.3 to 23.6 mg L−1 2,4-D reduced disease incidence in pot trials, and from 10.8 to 41.0 mg L−1 minimised disease severity in both pot and field trials. In only one pot trial, significant phytotoxicity was found with rates of 100 mg L−1 or greater, reducing mean total tuber mass per plot and 38 mg L−1 or greater, reducing mean mass per tuber. Notably, within the field trial, a more reliable plant growth system for estimation of yield, no significant impacts were observed. Disease control was associated with decreased sensitivity of tubers to thaxtomin A, the phytotoxin produced by the common scab pathogen essential for disease induction. The amount of residual 2,4-D in tubers at harvest varied with cultivar, Russet Burbank accumulating more 2,4-D than Desiree. Application rates less than 100 mg L−1 resulted in levels of 2,4-D below the Australian standard maximum residue limit. These data suggest that applications of 2,4-D at low rates could provide a commercially suitable control strategy for common scab.

Potato seed certification is a disease management tool that minimises the risk of spreading seed tuber-borne inoculum of infectious diseases. Traditionally, certification sampling strategies have relied upon visual assessment of a seedlot from samples taken at one or two points within the load of seed tubers. However methodologies in selection of tuber samples have not been critically assessed for their precision in estimating disease load. This study presents an analysis of 37 potato seedlots over a 3 year period. Analysis of sample data using receiver operating curves (ROCs) indicates that point sampling taking two samples of 100 tubers at the beginning and end of a seedlot gives equivalent disease estimation as a continuous sampling strategy taking ten samples of 20 tubers randomly throughout the seedlot, although at lower statistical precision. This was confirmed both by visual assessment of tuber-borne disease and by analysis of pathogen DNA content from tuber peel. Across the 3 years of study, powdery scab and black scurf were the major seed tuber-borne diseases recognised and this corresponded with high levels of pathogen DNA from peel analysis for both Spongospora subterranea and Rhizoctonia solani AG3 respectively.

Variant somaclones of potato cultivar Russet Burbank, selected for resistance to common scab using in vitro cell selection techniques, were tested for resistance to powdery scab, another important disease of potato caused by Spongospora subterranea. This pathogen also invades roots, producing root galls. Most variants consistently showed increased resistance to powdery scab, both in field and glasshouse challenge, when compared to the parental cultivar, several significantly so. On average, the best variant reduced powdery scab incidence by 51% and severity (tuber surface coverage) by 64%. In contrast, no improvement in the extent of root infection and root galling was seen. These results suggest host interactions during root and tuber infection are distinct. Correlation analyses of disease indices amongst the selected variants showed no association between Sp. subterranea root infection and gall scores, nor between root infection and tuber disease severity. However, a weak positive association was found between root gall score and tuber disease, and a strong correlation between tuber disease incidence and severity scores. Interestingly, positive correlations were also found between the extent of powdery and common scab resistance expressed and both incidence and severity of these diseases within the variants, suggesting a common defence mechanism. The role of thaxtomin A in selecting for concurrent resistance to both diseases is discussed.

Root exudation has importance in soil chemical ecology influencing rhizosphere microbiota. Prior studies reported root exudates from host and nonhost plants stimulated resting spore germination of Spongospora subterranea, the powdery scab pathogen of potato, but the identities of stimulatory compounds were unknown. This study showed that potato root exudates stimulated S. subterranea resting spore germination, releasing more zoospores at an earlier time than the control. We detected 24 low molecular weight organic compounds within potato root exudates and identified specific amino acids, sugars, organic acids, and other compounds that were stimulatory to S. subterranea resting spore germination. Given that several stimulatory compounds are commonly found in exudates of diverse plant species, we support observations of nonhost-specific stimulation. We provide knowledge of S. subterranea resting spore biology and chemical ecology that may be useful in formulating new disease management strategies.

Production of the phytotoxin thaxtomin A by pathogenic Streptomyces spp. is essential for induction of common scab disease in potato. The disease can be significantly reduced by a range of chemicals applied as foliar sprays before tuber initiation. We tested a range of chemicals that had previously demonstrated varying capacities to reduce common scab for both disease suppression and their ability to inhibit thaxtomin A toxicity in both ‘Desiree’ and ‘Russet Burbank’ potato. Our results for disease suppression generally supported previous studies. Our tuber slice assays with thaxtomin A showed a strong correlation between the ability of the chemical to suppress common scab symptom development and the ability of the chemical to inhibit thaxtomin A toxicity. A Bayesian measurement error linear regression model was derived for each cultivar and trial and demonstrated a clear positive relationship between disease and thaxtomin-A-induced necrosis. The relationships obtained were much stronger than would have been obtained without adjustment for measurement error. This demonstrates that disease mitigation using chemical foliar sprays is strongly correlated with the ability of the chemical to inhibit thaxtomin A toxicity, suggesting this mechanism as a key mode of action for understanding this novel disease control strategy.