Molecular Characterization of Defense of Brassica napus (Oilseed Rape) to Rhizoctonia solani AG2-1 Confirmed by Functional Analysis in Arabidopsis thaliana.

Rhizoctonia solani is a necrotrophic, soilborne fungal pathogen associated with significant establishment losses in Brassica napus (oilseed rape; OSR). The anastomosis group (AG) 2-1 of R. solani is the most virulent to OSR, causing damping-off, root and hypocotyl rot, and seedling death. Resistance to R. solani AG2-1 in OSR has not been identified, and the regulation of OSR defense to its adapted pathogen, AG2-1, has not been investigated. In this work, we used confocal microscopy to visualize the progress of infection by sclerotia of AG2-1 on B. napus varieties with contrasting disease phenotypes. We defined their defense response using gene expression studies and functional analysis with Arabidopsis thaliana mutants. Our results showed existing variation in susceptibility to AG2-1 and plant growth between OSR varieties, and differential expression of genes of hormonal and defense pathways related to auxin, ethylene, jasmonic acid, abscisic acid, salicylic acid, and reactive oxygen species regulation. Auxin, abscisic acid signaling, and the MYC2 branch of jasmonate signaling contributed to the susceptibility to AG2-1, while induced systemic resistance was enhanced by NAPDH RBOHD, ethylene signaling, and the ERF/PDF branch of jasmonate signaling. These results pave the way for future research, which will lead to the development of Brassica crops that are more resistant to AG2-1 of R. solani and reduce dependence on chemical control options.

Yield Loss and Integrated Disease Control of Rhizoctonia solani AG2-1 Using Seed Treatment and Sowing Rate of Oilseed Rape.

Rhizoctonia solani anastomosis group (AG) 2-1 is an ubiquitous soilborne pathogen causing severe damping-off of oilseed rape (OSR). In the absence of varietal resistance to AG2-1, there are limited methods for integrated disease management. The objectives of these field studies were to quantify yield losses due to AG2-1 and to determine the effectiveness of integrated control using sedaxane, fludioxonil, and metalaxyl-M applied as seed treatment on two OSR genotypes at a sowing rate of 40 (low) or 80 (high) seeds m-2. Crop assessments of green area index (GAI), vigor, and cabbage stem flea beetle (CSFB) Psylliodes chrysocephala damage were carried out at GS16, while pathogen DNA in soil was quantified using real-time PCR at GS32. Yield and seed weight losses of 41 and 18%, respectively, were associated with reduced establishment, GAI, vigor, and delayed development and flowering of OSR. Seed treatment reduced AG2-1 DNA in soil by 80%, resulting in a 94, 16, and 64% increase of establishment, thousand seed weight (TSW), and yield, respectively. Seed treatment also mitigated the effects of AG2-1 on delaying plant development, resulting in increased uniformity of crop flowering. OSR plants infected with AG2-1 suffered 27% more damage by the CSFB, indicating positive pathogen-pest interaction at the expense of the OSR host. Optimum control of AG2-1 infection was achieved by integrating low sowing rate and seed treatment. However, under dual pest and pathogen attack, high sowing rates should be combined with the use of seed treatment to mitigate seedling death and delayed development caused by AG2-1 and CSFB damage.

Yield Losses and Control by Sedaxane and Fludioxonil of Soilborne Rhizoctonia, Microdochium, and Fusarium Species in Winter Wheat.

Soilborne Rhizoctonia, Microdochium, and Fusarium species are major causal agents of seedling and stem-base diseases of wheat. Currently, seed treatments are considered the most effective solution for their control. Rhizoctonia solani anastomosis groups (AGs) 2-1 and 5, R. cerealis, Microdochium, and Fusarium spp., were used in series of field experiments to determine their capability to cause soilborne and stem-base disease and to quantify their comparative losses in the establishment and yield of wheat. The effectiveness and response to seed treatment formulated with 10 g sedaxane and 5 g fludioxonil 100 kg-1 against these soilborne pathogens were also determined. Our results showed that damping-off caused by soilborne R. cerealis was associated with significant reductions in the emergence and establishment, resulting in stunted growth and low plant numbers. The pathogen also caused sharp eyespot associated with reductions in the ear partitioning index. R. solani AG 2-1 and AG 5 were weakly pathogenic and failed to cause significant damping-off, root rot, and stem-base disease in wheat. Fusarium graminearum and F. culmorum applied as soilborne inoculum failed to cause severe disease. Microdochium spp. caused brown foot rot disease and soilborne M. nivale reduced wheat emergence. Applications of sedaxane and fludioxonil increased plant emergence and reduced damping-off, early stem-base disease, and brown foot rot, thus providing protection against multiple soilborne pathogens. R. cerealis reduced the thousand grain weight by 3.6%, whereas seed treatment including fludioxonil and sedaxane against soilborne R. cerealis or M. nivale resulted in a 4% yield increase.

Distinct branches of the N-end rule pathway modulate the plant immune response.

The N-end rule pathway is a highly conserved constituent of the ubiquitin proteasome system, yet little is known about its biological roles. Here we explored the role of the N-end rule pathway in the plant immune response. We investigated the genetic influences of components of the pathway and known protein substrates on physiological, biochemical and metabolic responses to pathogen infection. We show that the glutamine (Gln) deamidation and cysteine (Cys) oxidation branches are both components of the plant immune system, through the E3 ligase PROTEOLYSIS (PRT)6. In Arabidopsis thaliana Gln-specific amino-terminal (Nt)-amidase (NTAQ1) controls the expression of specific defence-response genes, activates the synthesis pathway for the phytoalexin camalexin and influences basal resistance to the hemibiotroph pathogen Pseudomonas syringae pv tomato (Pst). The Nt-Cys ETHYLENE RESPONSE FACTOR VII transcription factor substrates enhance pathogen-induced stomatal closure. Transgenic barley with reduced HvPRT6 expression showed enhanced resistance to Ps. japonica and Blumeria graminis f. sp. hordei, indicating a conserved role of the pathway. We propose that that separate branches of the N-end rule pathway act as distinct components of the plant immune response in flowering plants.

Altered gene expression by sedaxane increases PSII efficiency, photosynthesis and growth and improves tolerance to drought in wheat seedlings.

Succinate dehydrogenase inhibitor (SDHI) fungicides have been shown to increase PSII efficiency and photosynthesis under drought stress in the absence of disease to enhance the biomass and yield of winter wheat. However, the molecular mechanism of improved photosynthetic efficiency observed in SDHI-treated wheat has not been previously elucidated. Here we used a combination of chlorophyll fluorescence, gas exchange and gene expression analysis, to aid our understanding of the basis of the physiological responses of wheat seedlings under drought conditions to sedaxane, a novel SDHI seed treatment. We show that sedaxane increased the efficiency of PSII photochemistry, reduced non-photochemical quenching and improved the photosynthesis and biomass in wheat correlating with systemic changes in the expression of genes involved in defense, chlorophyll synthesis and cell wall modification. We applied a coexpression network-based approach using differentially expressed genes of leaves, roots and pregerminated seeds from our wheat array datasets to identify the most important hub genes, with top ranked correlation (higher gene association value and z-score) involved in cell wall expansion and strengthening, wax and pigment biosynthesis and defense. The results indicate that sedaxane confers tolerant responses of wheat plants grown under drought conditions by redirecting metabolites from defense/stress responses towards growth and adaptive development.

Aphid Infestation Increases Fusarium langsethiae and T-2 and HT-2 Mycotoxins in Wheat.

Fusarium langsethiae is a fungal pathogen of cereal crops that is an increasing problem in northern Europe, but much of its epidemiology is poorly understood. The species produces the mycotoxins T-2 and HT-2, which are highly toxic. It was hypothesized that grain aphids, Sitobion avenae, may transmit F. langsethiae inoculum between wheat plants, and a series of transmission experiments and volatile chemical analyses was performed to test this. Manual translocation of aphids from inoculated to uninfected hosts resulted in pathogen DNA accumulation in hosts. However, the free movement of wingless aphids from infected to healthy plants did not. The addition of winged aphids reared on F. langsethiae-inoculated wheat seedlings to wheat plants also did not achieve successful pathogen transfer. While our data suggested that aphid transmission of the pathogen was not very efficient, we observed an increase in disease when aphids were present. After seedling inoculation, an increase in pathogen DNA accumulation in seedling leaves was observed upon treatment with aphids. Furthermore, the presence of aphids on wheat plants with F. langsethiae-inoculated ears not only led to a rise in the amount of F. langsethiae DNA in infected grain but also to an increase in the concentrations of T-2 and HT-2 toxins, with more than 3-fold higher toxin levels than diseased plants without aphids. This work highlights that aphids increase the susceptibility of wheat host plants to F. langsethiae and that aphid infestation is a risk factor for accumulating increased levels of T-2 and HT-2 in wheat products. IMPORTANCE: Fusarium langsethiae is shown here to cause increased contamination levels of grain with toxins produced by fungus when aphids share the host plant. This effect has also recently been demonstrated with Fusarium graminearum, yet the two fungal species show stark differences in their effect on aphid populations. In both cases, aphids improve the ability of the pathogens to cause and initiate Fusarium head blight (FHB) disease in wheat, but F. langsethiae may be able to act as a dispersal agent. F. langsethiae contributes harmful toxins to wheat grain that need to be controlled, but as yet, its epidemiology is unresolved. This work reveals insights into the role aphids play in promoting the successful colonization of this species in wheat and the benefit of controlling aphid populations on crops that are at high risk of FHB.

Sharing a Host Plant (Wheat [Triticum aestivum]) Increases the Fitness of Fusarium graminearum and the Severity of Fusarium Head Blight but Reduces the Fitness of Grain Aphids (Sitobion avenae).

We hypothesized that interactions between fusarium head blight-causing pathogens and herbivores are likely to occur because they share wheat as a host plant. Our aim was to investigate the interactions between the grain aphid, Sitobion avenae, and Fusarium graminearum on wheat ears and the role that host volatile chemicals play in mediating interactions. Wheat ears were treated with aphids and F. graminearum inoculum, together or separately, and disease progress was monitored by visual assessment and by quantification of pathogen DNA and mycotoxins. Plants exposed to both aphids and F. graminearum inoculum showed accelerated disease progression, with a 2-fold increase in disease severity and 5-fold increase in mycotoxin accumulation over those of plants treated only with F. graminearum. Furthermore, the longer the period of aphid colonization of the host prior to inoculation with F. graminearum, the greater the amount of pathogen DNA that accumulated. Headspace samples of plant volatiles were collected for use in aphid olfactometer assays and were analyzed by gas chromatography-mass spectrometry (GC-MS) and GC-coupled electroantennography. Disease-induced plant volatiles were repellent to aphids, and 2-pentadecanone was the key semiochemical underpinning the repellent effect. We measured aphid survival and fecundity on infected wheat ears and found that both were markedly reduced on infected ears. Thus, interactions between F. graminearum and grain aphids on wheat ears benefit the pathogen at the expense of the pest. Our findings have important consequences for disease epidemiology, because we show increased spread and development of host disease, together with greater disease severity and greater accumulation of pathogen DNA and mycotoxin, when aphids are present.

Foliar application of isopyrazam and epoxiconazole improves photosystem II efficiency, biomass and yield in winter wheat.

A range of fungicides including epoxiconazole, azoxystrobin and isopyrazam, were applied to winter wheat at GS 31/32 to determine their effect on photosystem II (PSII) efficiency, biomass and yield. Frequent, repeated measurements of chlorophyll fluorescence were carried on plants grown under different water regimes in controlled environment and in the field to establish the transiency of fluorescence changes in relation to fungicide application. Application of the succinate dehydrogenase inhibitor isopyrazam in a mixture with the triazole epoxiconazole increased PSII efficiency associated with a 28% increase in biomass in the controlled environment and 4% increase in grain yield in the field in the absence of disease pressure. Application of isopyrazam and epoxiconazole increased efficiency of PSII photochemistry (Fv'/Fm') as early as 4h following application associated with improved photosynthetic gas exchange and increased rates of electron transport. We reveal a strong, positive relationship between Fv'/Fm' and CO2 assimilation rate, stomatal conductance and transpiration rate in controlled environment and Fv'/Fm' detected just after anthesis on the flag leaf at GS 73 and grain yield in field. We conclude that application of a specific combination of fungicides with positive effects of plant physiology in the absence of disease pressure results in enhanced biomass and yield in winter wheat. Additionally, an accurate and frequent assessment of photosynthetic efficiency of winter wheat plants can be used to predict yield and biomass in the field.

Chlorophyll Fluorescence on the Fast Timescale.

Chlorophyll fluorescence is a rapid and noninvasive tool used for probing the activity of photosynthesis that can be used in vivo and in the field. It is highly relevant to the demands of high-throughput crop phenotyping and can be automated or manually applied. In this chapter, we describe protocols and advice for making fast timescale fluorescence measurements using handheld equipment in the laboratory or in the field in the context of phenotyping. While interpretation of some measured parameters requires caution for the purpose of identifying underlying mechanisms, we demonstrate this technique is appropriate for some applications where convenience, rapidity, and sensitivity are required.

Rapid screening for resistance to Sitobion avenae (F.) and Rhopalosiphum padi (L.) in winter wheat seedlings and selection of efficient assessment methods.

BACKGROUND: Sitobion avenae (F.) and Rhopalosiphum padi (L.) are harmful pests of wheat [Triticum aestivum (L.)]. No genetic resistance against the aphids has been identified in commercial wheat varieties and resistance phenotyping can be time-consuming and laborious. Here, we tested a high-throughput phenotyping method to screen 29 commercial winter wheat varieties for alate antixenosis and antibiosis. We validated this method using comprehensive behavioural analyses, including alate attraction to volatile organic compounds (VOCs) and a feeding bioassay using an electrical penetration graph (EPG), subsequently highlighting possible sources of resistance. RESULTS: We observed differences in alate behaviour upon assessing alate settlement on wheat seedlings and attraction towards VOCs, revealing the importance of visual and early post-alighting cues for alate host selection. Aphid settlement was four times higher on the most preferred variety than on the least preferred variety. Using an EPG bioassay, we identified phloem feeding and stylet derailment parameters linked to resistance. We found antibiosis assessment on detached leaves to be an inadequate screen because it produced results inconsistent with intact leaves assessment. Alate and nymph mortality were identified as key traits signifying antibiosis, showing significant positive relationships with alate reproduction and nymph mean relative growth rate. CONCLUSIONS: Overall, antixenosis and antibiosis varietal responses were consistent for both aphid species. Alate settlement on wheat seedlings was a more efficient antixenosis screen than an olfactometer assay using VOCs. In addition to assessing alate and nymph survival for antibiosis, this allows for more rapid phenotyping of large numbers of genotypes to identify novel aphid resistance genes for varietal improvement. © 2024 The Author(s). Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Aggressiveness and mycotoxin profile of Fusarium avenaceum isolates causing Fusarium seedling blight and Fusarium head blight in UK malting barley.

Introduction: Fusarium avenaceum causing Fusarium seedling blight (FSB) and Fusarium head blight (FHB) on barley is associated with economic losses of crop yield and quality, and the accumulation of mycotoxins including the enniatins (ENNs) A, A1, B and B1. Although F. avenaceum is the main producer of ENNs, studies on the ability of isolates to cause severe Fusarium diseases or produce mycotoxins in barley are limited. Methods: In this work, we investigated the aggressiveness of nine isolates of F. avenaceum to two cultivars of malting barley, Moonshine and Quench, and defined their ENN mycotoxin profiles in in vitro and in planta experiments. We assessed and compared the severity of FSB and FHB caused by these isolates to disease severity by F. graminearum, F. tricinctum and F. poae. Quantitative real-time polymerase chain reaction and Liquid Chromatography Tandem Mass Spectrometry assays were used to quantify pathogen DNA and mycotoxin accumulation, respectively, in barley heads. Results: Isolates of F. avenaceum were equally aggressive to barley stems and heads and caused the most severe FSB symptoms resulting in up to 55% reductions of stem and root length. Fusarium graminearum caused the most severe FHB disease, followed by the isolates of F. avenaceum with the most aggressive F. avenaceum isolates capable of causing similar bleaching of barley heads as F. avenaceum. Fusarium avenaceum isolates produced ENN B as the predominant mycotoxin, followed by ENN B1 and A1 in vitro. However, only the most aggressive isolates produced ENN A1 in planta and none produced ENN A or beauvericin (BEA) either in planta or in vitro. Discussion: The capacity of F. avenaceum isolates to produce ENNs was related to the accumulation of pathogen DNA in barley heads, whilst FHB severity was related to the synthesis and accumulation of ENN A1 in planta. Cv. Moonshine was significantly more resistant than Quench to FSB or FHB, caused by any Fusarium isolate, and to the accumulation of pathogen DNA, ENNs or BEA. In conclusion, aggressive F. avenaceum isolates are potent ENN producers causing severe FSB and FHB with ENN A1 requiring further investigation as potential virulence factor for F. avenaceum in cereals.

Fungal plant pathogen "mutagenomics" reveals tagged and untagged mutations in Zymoseptoria tritici and identifies SSK2 as key morphogenesis and stress-responsive virulence factor.

"Mutagenomics" is the combination of random mutagenesis, phenotypic screening, and whole-genome re-sequencing to uncover all tagged and untagged mutations linked with phenotypic changes in an organism. In this study, we performed a mutagenomics screen on the wheat pathogenic fungus Zymoseptoria tritici for altered morphogenetic switching and stress sensitivity phenotypes using Agrobacterium-mediated "random" T-DNA mutagenesis (ATMT). Biological screening identified four mutants which were strongly reduced in virulence on wheat. Whole genome re-sequencing defined the positions of the T-DNA insertion events and revealed several unlinked mutations potentially affecting gene functions. Remarkably, two independent reduced virulence mutant strains, with similarly altered stress sensitivities and aberrant hyphal growth phenotypes, were found to have a distinct loss of function mutations in the ZtSSK2 MAPKKK gene. One mutant strain had a direct T-DNA insertion affecting the predicted protein's N-terminus, while the other possessed an unlinked frameshift mutation towards the C-terminus. We used genetic complementation to restore both strains' wild-type (WT) function (virulence, morphogenesis, and stress response). We demonstrated that ZtSSK2 has a non-redundant function with ZtSTE11 in virulence through the biochemical activation of the stress-activated HOG1 MAPK pathway. Moreover, we present data suggesting that SSK2 has a unique role in activating this pathway in response to specific stresses. Finally, dual RNAseq-based transcriptome profiling of WT and SSK2 mutant strains revealed many HOG1-dependent transcriptional changes in the fungus during early infection and suggested that the host response does not discriminate between WT and mutant strains during this early phase. Together these data define new genes implicated in the virulence of the pathogen and emphasise the importance of a whole genome sequencing step in mutagenomic discovery pipelines.

A large bioassay identifies Stb resistance genes that provide broad resistance against Septoria tritici blotch disease in the UK.

Introduction: Septoria tritici blotch (STB) is one of the most damaging fungal diseases of wheat in Europe, largely due to the paucity of effective resistance genes against it in breeding materials. Currently dominant protection methods against this disease, e.g. fungicides and the disease resistance genes already deployed, are losing their effectiveness. Therefore, it is vital that other available disease resistance sources are identified, understood and deployed in a manner that maximises their effectiveness and durability. Methods: In this study, we assessed wheat genotypes containing nineteen known major STB resistance genes (Stb1 through to Stb19) or combinations thereof against a broad panel of 93 UK Zymoseptoria tritici isolates. Seedlings were inoculated using a cotton swab and monitored for four weeks. Four infection-related phenotypic traits were visually assessed. These were the days post infection to the development of first symptoms and pycnidia, percentage coverage of the infected leaf area with chlorosis/necrosis and percentage coverage of the infected leaf area with pycnidia. Results: The different Stb genes were found to vary greatly in the levels of protection they provided, with pycnidia coverage at four weeks differing significantly from susceptible controls for every tested genotype. Stb10, Stb11, Stb12, Stb16q, Stb17, and Stb19 were identified as contributing broad spectrum disease resistance, and synthetic hexaploid wheat lines were identified as particularly promising sources of broadly effective STB resistances. Discussion: No single Z. tritici isolate was found to be virulent against all tested resistance genes. Wheat genotypes carrying multiple Stb genes were found to provide higher levels of resistance than expected given their historical levels of use. Furthermore, it was noted that disease resistance controlled by different Stb genes was associated with different levels of chlorosis, with high levels of early chlorosis in some genotypes correlated with high resistance to fungal pycnidia development, potentially suggesting the presence of multiple resistance mechanisms.The knowledge obtained here will aid UK breeders in prioritising Stb genes for future breeding programmes, in which optimal combinations of resistance genes could be pyramided. In addition, this study identified the most interesting Stb genes for cloning and detailed functional analysis.

Impact of Initial Population Density of the Dubas Bug, Ommatissus lybicus (Hemiptera: Tropiduchidae), on Oviposition Behaviour, Chlorophyll, Biomass and Nutritional Response of Date Palm (Phoenix dactylifera).

The Dubas bug (Ommatissus lybicus) is an economically significant pest of date palms. In this study, the effect of the population density of O. lybicus on chlorophyll, measured by the soil plant analysis development (SPAD) chlorophyll meter, palm biomass, and the nutritional composition of date palms, were investigated. A further objective was to determine significant relationships between the population density of O. lybicus, the number of honeydew droplets, and oviposited eggs. Reductions of up to 8-11% and 29-34% in chlorophyll content and plant biomass, respectively, were caused by infestations exceeding 300 nymphs per palm seedling. Increasing the population density of O. lybicus to 600 insects per palm decreased oviposition by females, suggesting intraspecific competition for resources. There was a significant relationship between honeydew droplets produced by the pest population and chlorophyll content in the rachis, suggesting that treatment can be triggered at 3-6 nymphs/leaflet. Egg oviposition was preferentially on the rachis. Ca, Mg, K, and P were the main nutrients affected by the activity of the pest. Mg content was associated with reduced chlorophyll content under increasing pest density, suggesting that supplemental nutrition can be potentially utilized to sustain chlorophyll and increase palm tolerance to pest infestation.

The Arabidopsis thaliana N-recognin E3 ligase PROTEOLYSIS1 influences the immune response.

N-degron pathways of ubiquitin-mediated proteolysis (formerly known as the N-end rule pathway) control the stability of substrate proteins dependent on the amino-terminal (Nt) residue. Unlike yeast or mammalian N-recognin E3 ligases, which each recognize several different classes of Nt residues, in Arabidopsis thaliana, N-recognin functions of different N-degron pathways are carried out independently by PROTEOLYSIS (PRT)1, PRT6, and other unknown proteins. PRT1 recognizes type 2 aromatic Nt-destabilizing residues and PRT6 recognizes type 1 basic residues. These two N-recognin functions diverged as separate proteins early in the evolution of plants, before the conquest of the land. We demonstrate that loss of PRT1 function promotes the plant immune system, as mutant prt1-1 plants showed greater apoplastic resistance than WT to infection by the bacterial hemi-biotroph Pseudomonas syringae pv tomato (Pst) DC3000. Quantitative proteomics revealed increased accumulation of proteins associated with specific components of plant defense in the prt1-1 mutant, concomitant with increased accumulation of salicylic acid. The effects of the prt1 mutation were additional to known effects of prt6 in influencing the immune system, in particular, an observed over-accumulation of pipecolic acid (Pip) in the double-mutant prt1-1 prt6-1. These results demonstrate a potential role for PRT1 in controlling aspects of the plant immune system and suggest that PRT1 limits the onset of the defense response via degradation of substrates with type 2 Nt-destabilizing residues.

Chlorophyll Fluorescence on the Fast Timescale.

Chlorophyll fluorescence is a rapid and non-invasive tool used for probing the activity of photosynthesis that can be used in vivo and in the field. It is highly relevant to the demands of high-throughput crop phenotyping and can be automated or manually applied. Here we describe protocols and advice for making fast timescale fluorescence measurements using handheld equipment in the laboratory or in the field. While interpretation of some measured parameters requires caution, we demonstrate that this technique is appropriate for some applications where convenience, rapidity, and sensitivity are required.

Infestation by Myzus persicae Increases Susceptibility of Brassica napus cv. "Canard" to Rhizoctonia solani AG 2-1.

Activation of plant defense pathways can be influenced by the presence of different species of attacking organisms. Understanding the complicated interactions triggering plant defense mechanisms is of great interest as it may allow the development of more effective and sustainable disease control methods. Myzus persicae and Rhizoctonia solani anastomosis group (AG) 2-1 are two important organisms attacking oilseed rape (OSR), causing disease and reduced yields. At present, is unclear how these two interact with each other and with OSR defenses and therefore the aim of the present study was to gain a better insight into the indirect interaction between aphids and pathogen. In separate experiments, we assessed the effect of AG 2-1 infection on aphid performance, measured as growth rate and population increase and then the effect of aphid infestation on AG 2-1 by quantifying disease and the amount of fungal DNA in plant stems and compost for two OSR varieties, "Canard" and "Temple." Additionally, we examined the expression of genes related to jasmonic acid (JA) and salicylic acid (SA) defense pathways. There was no significant effect of AG 2-1 infection on M. persicae performance. However, aphid infestation in one of the varieties, "Canard," resulted in significantly increased disease symptoms caused by AG 2-1, although, the amount of fungal DNA was not significantly different between treatments. This meant that "Canard" plants had become more susceptible to the disease. Expression of LOX3 and MYC2 was elevated under AG 2-1 treatment but downregulated in plants with both aphids and pathogen. Therefore it seems plausible that alterations in the JA signaling due to aphid infestation resulted in the increased susceptibility to AG 2-1.

Canopy and Ear Traits Associated With Avoidance of Fusarium Head Blight in Wheat.

Doubled haploid and elite wheat genotypes were ground inoculated in three field experiments and head spray inoculated in two glasshouse experiments, using mixed Fusarium and Microdochium species, to identify crop canopy and ear traits associated with Fusarium head blight (FHB) disease. In all experiments, flag leaf length and tiller number were consistently identified as the most significant canopy traits contributing to progression of FHB caused by Fusarium graminearum, F. culmorum, and F. avenaceum. The influence of ear traits was greater for F. poae that may possess more diverse routes for transmission and spread. Consistently, spikelet density was associated with increased disease severity in the field. F. graminearum, F. culmorum, and F. langsethiae were the main mycotoxin producers and their respective toxins were significantly related to fungal biomass and number of spikelets per ear. Genotypes with lower tiller numbers, shorter flag leaves and less dense ears may be able to avoid FHB disease caused by F. graminearum, F. culmorum, F. avenaceum, or Microdochium species however selection for these canopy and ear architectural traits to enable disease avoidance in wheat is likely to result in a potential trade-off with grain yield and therefore only moderately advantageous in susceptible genotypes.

Development of high-throughput methods to screen disease caused by Rhizoctonia solani AG 2-1 in oilseed rape.

BACKGROUND: Rhizoctonia solani (Kühn) is a soil-borne, necrotrophic fungus causing damping off, root rot and stem canker in many cultivated plants worldwide. Oilseed rape (OSR, Brassica napus) is the primary host for anastomosis group (AG) 2-1 of R. solani causing pre- and post-emergence damping-off resulting in death of seedlings and impaired crop establishment. Presently, there are no known resistant OSR genotypes and the main methods for disease control are fungicide seed treatments and cultural practices. The identification of sources of resistance for crop breeding is essential for sustainable management of the disease. However, a high-throughput, reliable screening method for resistance traits is required. The aim of this work was to develop a low cost, rapid screening method for disease phenotyping and identification of resistance traits. RESULTS: Four growth systems were developed and tested: (1) nutrient media plates, (2) compost trays, (3) light expanded clay aggregate (LECA) trays, and (4) a hydroponic pouch and wick system. Seedlings were inoculated with virulent AG 2-1 to cause damping-off disease and grown for a period of 4-10 days. Visual disease assessments were carried out or disease was estimated through image analysis using ImageJ. CONCLUSION: Inoculation of LECA was the most suitable method for phenotyping disease caused by R. solani AG 2-1 as it enabled the detection of differences in disease severity among OSR genotypes within a short time period whilst allowing measurements to be conducted on whole plants. This system is expected to facilitate identification of resistant germplasm.

Chlorophyll fluorescence parameters allow the rapid detection and differentiation of plant responses in three different wheat pathosystems.

The present study was undertaken to identify chlorophyll fluorescence (CF) parameters that can quantify changes in PSII associated with plant responses in three different wheat pathosystems of foliar, stem-base and ear diseases. The pathosystems included powdery mildew caused by Blumeria graminis, eyespot caused by Oculimacula yallundae or Oculimacula acuformis and Fusarium head blight (FHB) caused by Fusarium culmorum, F. avenaceum or F. langsethiae. Fast CF transients (OJIP) were analysed with the JIP-test to determine changes in PSII photochemistry. Measurements on asymptomatic leaves showed that electron transport related parameters (ETo/RC, ψo and ϕEo) were important to identify varietal differences in resistance to powdery mildew during early stages of infection. The same parameters also allowed differentiation between F. langsethiae and other Fusarium spp. Where infections were caused by the necrotrophic pathogens, Oculimacula spp., F. culmorum or F. avenaceum, changes related to maximum efficiency of PSII photochemistry (Fv'/Fm') as well as flux of dissipated (DIo/RC), trapped (TRo/RC), or absorbed (ABS/RC) energy per active reaction centers were significant in detecting biotic stress and the effectiveness of fungicide treatment for disease control. Our results demonstrated that Fv'/Fm' correlated significantly with visual disease and pathogen DNA of different wheat pathosystems. OJIP was shown as a sensitive technique that can be explored as diagnostic tool in future crop disease management and varietal breeding programs.