Evaluating the importance of the tan spot ToxA–Tsn1 interaction in Australian wheat varieties

The necrotrophic fungal pathogen Pyrenophora tritici‐repentis (Ptr) causes the major wheat disease tan spot, and produces multiple necrotrophic effectors that contribute to virulence. The proteinaceous effector ToxA induces necrosis in wheat genotypes possessing the Tsn1 gene, although the importance of the ToxA–Tsn1 interaction itself in varietal disease development has not been well studied. Here, 40 Australian spring wheat varieties were assessed for ToxA sensitivity and disease response to a race 1 wildtype Ptr isolate and ToxA‐deleted strain at both seedling and tillering growth stages. ToxA sensitivity was generally associated with disease susceptibility, but did not always predict spreading necrotic symptoms. Whilst the majority of Tsn1 varieties exhibited lower disease scores following toxa mutant infection, several exhibited no distinct differences between wildtype and toxa symptoms. This implies that ToxA is not the major determinant in tan spot disease development in some host backgrounds and indicates the presence of additional effectors. Unexpectedly, several tsn1 varieties exhibited a reduction in disease severity following toxa mutant inoculation, which may suggest an indirect role for ToxA in pathogen fitness. Additionally, increased chlorosis was observed following toxa mutant infection in three varieties, and further work is required to determine whether this is likely to be due to ToxA epistasis of ToxC symptoms. Taken together, these observations demonstrate that Ptr interacts with the host in a complex and intricate manner, leading to a variety of disease reactions that are dependent or independent of the ToxA–Tsn1 interaction.     

Protection against bacterial soft rot by olive extracts is related to general defence induction in potato tubers

Controlling bacterial plant diseases remains a challenge, as direct chemical control is usually not possible. Obtaining new biocontrol methods, in particular efficient biomolecules able to boost defence reactions and limit infection or symptom development, is therefore of major importance. This study screened extracts from leaves of two Mediterranean plants rich in bioactive phenolic compounds, olive and carob, for their ability to reduce soft rot severity and to trigger phenylalanine ammonia‐lyase (PAL) activity in potato tuber slices. Extracts from olive leaves significantly reduced disease severity caused by Pectobacterium atrosepticum or Pectobacterium carotovorum, whereas carob leaf extracts significantly increased it. Olive extracts and its main phenolic components, oleuropein and hydroxytyrosol, also significantly increased PAL activity 7.5 h after application. None of the extracts or purified molecules reduced bacterial growth in vitro. Furthermore, the effect of these extracts varied according to potato cultivars. These data therefore open new ways for the biological control of soft rot bacteria, but stress the importance of understanding the causes of response difference in different potato varieties in order to obtain optimal efficacy.     

The effect of silicon on antioxidant metabolism of wheat leaves infected by Pyricularia oryzae

This study investigated whether the increase in wheat resistance to blast, caused by Pyricularia oryzae, potentiated by silicon (Si) is linked to changes in the activity of antioxidative enzymes. Wheat plants (cv. BR 18) were grown in hydroponic culture with either 0 (–Si) or 2 mm (+Si) Si and half of the plants in each group were inoculated with P. oryzae. Blast severity in the +Si plants was 70% lower compared to the ?Si plants at 96 h after inoculation (hai). Superoxide dismutase (SOD), catalase (CAT), peroxidase (POX), ascorbate peroxidase (APX) and glutathione‐S‐transferase (GST) activities were higher in the leaves of the ?Si plants compared with the +Si plants at 96 hai. This indicates that other mechanisms may have limited P. oryzae infection in the +Si plants restricting the generation of reactive oxygen species, obviating the need for increased antioxidative enzyme activity. In contrast, glutathione reductase (GR) activity at 96 hai was higher in the +Si plants than in the ?Si plants. Although the inoculated plants showed significantly higher concentration of malondialdehyde (MDA) than the non‐inoculated plants, lower MDA concentrations were observed in the +Si plants compared with the ?Si plants. The lower MDA concentration associated with decreased activities of SOD, CAT, POX, APX and GST, suggest that the amount of reactive oxygen species was lower in the +Si plants. However, GR appears to play a pivotal role in limiting oxidative stress caused by P. oryzae infection in +Si plants.     

Photosynthesis and sugar concentration are impaired by the defective active silicon uptake in rice plants infected with Bipolaris oryzae

The effect of soluble silicon (Si) on photosynthetic parameters and soluble sugar concentrations was determined in leaves of rice cv. Oochikara and mutant plants of Oochikara defective in active Si uptake [low silicon 1 (lsi1)]. Plants were grown in hydroponic culture amended with 0 (?Si) or 2 mm Si (+Si), under either low or high photon flux density (PFD) and with or without inoculation with Bipolaris oryzae, the causal agent of brown spot of rice. Leaf Si concentration increased by 141 and 435% in +Si cv. Oochikara and by 119 and 251% in +Si lsi1 mutant plants under high and low PFD, respectively, compared with ?Si plants. Plant biomass accumulation was improved by Si regardless of PFD, especially plants for cv. Oochikara. Brown spot severity was highest in ?Si plants for cv. Oochikara and lsi1 mutant plants under low PFD. In the presence of Si, disease severity in plants grown under both low and high PFD was reduced, except for lsi1 mutant plants under high PFD. Plant inoculation reduced the photosynthetic parameters measured regardless of plant material or Si supply. A decrease of net carbon assimilation rate (A) of inoculated plants under low PFD compared with non‐inoculated plants was associated with damage in the photosynthetic apparatus, except for +Si cv. Oochikara in which stomatal restriction [low water vapour conductance (g_s)] contributed to A reduction. Under high PFD, damage to the photosynthetic apparatus of inoculated plants was the main reason for the reduction in A for +Si and ?Si lsi1 mutant plants. In addition, for ?Si cv. Oochikara, a reduction in g_s contributed to reduced A. However, for +Si cv. Oochikara, g_s was the limiting factor for A. Inoculated plants of +Si cv. Oochikara had higher A values than +Si lsi1 mutant plants, regardless of environmental conditions. Soluble sugars were not detected in leaf tissues of plants under low PFD. For high PFD, Si improved the hexose concentration in non‐inoculated plants at 144 h after inoculation (hai) for lsi1 mutant plants and from 96 hai onwards for cv. Oochikara compared with ?Si plants. However, plant inoculation reduced hexose concentration compared with non‐inoculated plants, mainly in +Si plants, regardless of plant material. Sucrose concentration increased in leaves of cv. Oochikara in the presence of Si whether inoculated or not. For +Si lsi1 mutant plants, sucrose concentration increased only at 48 hai compared with ?Si plants, whether inoculated or not. The results of this study show that a minimum Si concentration is needed in leaf tissues of rice plants to avoid the negative impact of B. oryzae infection on photosynthesis and sugar concentration. High leaf Si concentration resulted in an increased soluble sugar concentration and together, but in independent ways, soluble sugar and Si reduced brown spot severity of rice.     

Exploring de novo specificity: the Pyrenophora tritici‐repentis–barley interaction

Pyrenophora tritici‐repentis (Ptr) is a destructive fungal pathogen of wheat worldwide. In addition to wheat, Ptr has been isolated from various other hosts in the family Poaceae, yet the nature of its interaction with those hosts is unknown. The Ptr–barley relationship was explored and the existence of a specific interaction between Ptr and barley is described for the first time; symptom development on several barley genotypes was evaluated in bioassays and by toxin infiltration into barley leaves. Ptr ToxB‐producing isolates of the fungus were able to cause significant damage when inoculated onto certain barley genotypes, and Ptr ToxB was able to induce chlorosis in a highly selective manner when infiltrated into those same genotypes. Ptr–barley specificity is subtle and can break with slight changes in temperature after infection. To understand the infection process in barley, a cytological analysis and in planta fungal biomass estimation using quantitative PCR were performed. The fungus penetrates through the host epidermal cells and advances to colonize the mesophyll layer intercellularly, with the infection process on barley closely resembling that on wheat. Here, evidence is provided for a specific interaction between barley and Ptr, expanding understanding of Ptr host specificity and breaking the assumption that the highest level of specificity seen with Ptr is restricted to particular genotypes of the wheat host.     

Absence of detectable yield penalty associated with insensitivity to Pleosporales necrotrophic effectors in wheat grown in the West Australian wheat belt

Genetic disease resistance is widely assumed, and occasionally proven, to cause host yield or fitness penalties due to inappropriate activation of defence response mechanisms or diversion of resources to surplus preformed defences. The study of resistance gene trade‐offs has so far been restricted to biotrophic pathogens. In some Pleosporales necrotrophic interactions, quantitative resistance is positively associated with insensitivity to effectors. Host lines that differ in sensitivity can easily be identified amongst current cultivars and advanced breeding lines. Large wheat cultivar trials were used to test whether lines sensitive or insensitive to three necrotrophic effectors from Pyrenophora tritici‐repentis and Parastagonospora nodorum differed in yield when subjected to natural disease and stress pressures in the West Australian wheat belt. There was no significant yield penalty associated with insensitivity to the fungal effectors ToxA, SnTox1 and SnTox3. Some yield gains were associated with insensitivity and some of these gains could be attributed to increased disease resistance. It is concluded that insensitivity to these effectors does not render such plants more vulnerable to any relevant biotic or abiotic stress present in these trials. These results suggest that the elimination of sensitivity alleles for necrotrophic effectors is a safe and facile strategy for improving disease resistance whilst maintaining or improving other desirable traits.     

Molecular and biochemical mechanisms of defence induced in pea by Rhizobium leguminosarum against Orobanche crenata

Orobanche species (broomrapes) are parasitic weeds which dramatically decrease the yields of many economically important dicotyledonous crops, including pea (Pisum sativum), in Mediterranean areas. Previously, we identified some Rhizobium leguminosarum strains, including P.SOM, which could both promote pea development and significantly reduce infection by Orobanche crenata, notably through induction of necrosis of attached parasites. In the present study, induced resistance against broomrape in the nodulated pea was shown to be associated with significant changes in rates of oxidative lipoxygenase (Lox) and phenylpropanoid/isoflavonoid pathways and in accumulation of derived toxins, including phenolics and pisatin (pea phytoalexin). Changes were followed for 5 weeks after inoculation and attack by Orobanche. In contrast to non‐inoculated plants or Orobanche only infected plants, polyphenoloxidase (PPO) activity and hydrogen peroxide content increased in response to bacteria inoculation indicating the involvement of oxidative processes. In parallel, the nodulated roots displayed high Lox activity related to the overexpression of the lox1 gene. Similarly, the expression of phenylalanine ammonia lyase (PAL) and 6a‐hydroxymaackiain 3‐O‐methyltransferase (Hmm6a) genes were induced early during nodule development, suggesting the central role of the phenylpropanoid/isoflavonoid pathways in the elicited defence. As a consequence, the derived products, phenolics and pisatin, accumulated in response to rhizobacteria and conferred mechanical and chemical barriers to the invading parasite. These results highlight the likely role of signalling pathways in induced resistance and suggest these mechanisms should be enhanced through integrated Orobanche management practices.     

Revue bibliographique: la maladie de la tache auréolée du blé

Tan spot caused by Pyrenophora tritici‐repentis is a wheat disease found worldwide which can cause significant losses. This disease is characterized by typical symptoms: a necrotic spot surrounded by chlorosis halo. On the basis of its ability to produce chlorosis and/or necrosis symptoms on a differential host set. Eight races of this pathogen are currently recognized. These symptoms are the result of a specific interaction between the host and at least three host specific toxins Ptr ToxA, Ptr ToxB and Ptr ToxC. This interaction seems to be a mirror image of the classical gene‐for‐gene described by Flore. This paper presents a first literature review in the French language, identifying the major aspects of this disease, its epidemiology and diversity of its causal agent.     

Comparison of root and foliar applications of potassium silicate in potentiating post‐infection defences of melon against powdery mildew

The application of silicon to the roots or leaves reduces the severity of powdery mildew (Podosphaera xanthii) in melon but the latter treatment is less effective. This study compared key biochemical defence responses of melon triggered by P. xanthii after root or foliar treatment with potassium silicate (PS). Treatments consisted of pathogen‐inoculated or mock‐inoculated plants supplied with PS via roots or foliarly, as well as a non‐treated control. The activity of defence enzymes and the concentration of phenolic compounds, lignin and malondialdehyde were determined from leaf samples at different time points after inoculation. Pathogen‐inoculated plants irrigated with PS showed both an accumulation of silicon and primed defence responses in leaves that were not observed in pathogen‐inoculated plants either sprayed with PS or not treated. These responses included the anticipated activity of peroxidase and accumulation of soluble phenols, the activation of chitinase and repression of catalase, and the stronger activation of superoxide dismutase, peroxidase and β‐1,3‐glucanase. Moreover, the lignin concentration increased in response to inoculation, whereas the malondialdehyde concentration decreased. For the foliar treatment, however, only an increase in lignin deposition was observed compared with the control plants. The results show that silicon strongly plays an active role in modulating the defence responses of melon against P. xanthii when supplied to the roots as opposed to the foliage.     

Spatial connectedness of plant species: potential links for apparent competition via plant diseases

This study evaluated the reactions of seven common C4 grasses of the tallgrass prairie of the USA Great Plains to the economically important wheat pathogens Pyrenophora tritici‐repentis and Gaeumannomyces graminis var. tritici (Ggt) isolated from wheat. The P. tritici‐repentis isolates (race 1) were pathogenic on all grasses tested, but symptom severity was markedly low. Three of the grass species inoculated with Ggt were highly susceptible, while four species exhibited no symptoms. Because measures of connectedness can provide a proxy for population processes, connectedness was evaluated within and among the seven grass species in representative tallgrass prairie environments for all potential pathogen‐sharing patterns. Andropogon gerardii was ubiquitous, so all plant species were well connected to it. Andropogon scoparius (= Schizachyrium scoparium), Sorghastrum nutans and Panicum virgatum were fairly common but specialized to particular environments. Bouteloua curtipendula was uncommon but occurred in all environments, while Buchloë dactyloides and Bouteloua gracilis were uncommon and only occurred in upland sites. Co‐occurrence of plant species was generally not reciprocal in that, for many species pairs, species A rarely occurred without potential exposure to inoculum from species B, while species B commonly occurred without species A. The three grass species susceptible to Ggt may act as sources of inoculum for each other within tallgrass prairie, with the potential to influence fitness, and tallgrass prairie and commercial wheat ecosystems in the Great Plains also have the potential to share both pathogens.     

Significant in vitro antagonism of the laurel wilt pathogen by endophytic fungi from the xylem of avocado does not predict their ability to control the disease

The ascomycete Raffaelea lauricola causes laurel wilt, a lethal vascular disease of avocado, Persea americana, and other members of the Lauraceae plant family. Few effective control measures for laurel wilt exist and new measures are needed. In this study, biological control of the disease with endophytic fungi from avocado was examined. Thirty‐two endophytes (24 operational taxonomic units or OTUs) isolated from the xylem of healthy trees (the infection court of R. lauricola) were evaluated against R. lauricola with in vitro dual‐culture assays. Nine OTUs that showed strong in vitro antagonism of the pathogen were tested in planta against laurel wilt. In three greenhouse experiments, grafted avocado plants of Simmonds or Russell cultivars, which are both susceptible to laurel wilt, were inoculated with endophytes and, after 10–16 days, inoculated with the same isolate of R. lauricola that was used in the in vitro assays. Within 14 days of inoculation with R. lauricola, laurel wilt developed in plants that were not treated with endophytes (positive controls) but also developed in endophyte‐treated plants to the extent observed in the positive controls (P = 0.05). The pathogen colonized plants rapidly and systemically, but endophytes generally did not colonize xylem more than 2 cm above the point at which plants were inoculated. Although the tested endophytes strongly antagonized the pathogen in vitro, this did not translate to an ability to reduce development of laurel wilt. The management of laurel wilt and other plant diseases with fungal endophytes is discussed.     

Early molecular signatures of responses of wheat to Zymoseptoria tritici in compatible and incompatible interactions

Zymoseptoria tritici, the causal agent of septoria tritici blotch, a serious foliar disease of wheat, is a necrotrophic pathogen that undergoes a long latent period. Emergence of insensitivity to fungicides, and pesticide reduction policies, mean there is a pressing need to understand septoria and control it through greater varietal resistance. Stb6 and Stb15, the most common qualitative resistance genes in modern wheat cultivars, determine specific resistance to avirulent fungal genotypes following a gene‐for‐gene relationship. This study investigated compatible and incompatible interactions of wheat with Z. tritici using eight combinations of cultivars and isolates, with the aim of identifying molecular responses that could be used as markers for disease resistance during the early, symptomless phase of colonization. The accumulation of TaMPK3 was estimated using western blotting, and the expression of genes implicated in gene‐for‐gene interactions of plants with a wide range of other pathogens was measured by qRT‐PCR during the presymptomatic stages of infection. Production of TaMPK3 and expression of most of the genes responded to inoculation with Z. tritici but varied considerably between experimental replicates. However, there was no significant difference between compatible and incompatible interactions in any of the responses tested. These results demonstrate that the molecular biology of the gene‐for‐gene interaction between wheat and Zymoseptoria is unlike that in many other plant diseases, indicate that environmental conditions may strongly influence early responses of wheat to infection by Z. tritici, and emphasize the importance of including both compatible and incompatible interactions when investigating the biology of this complex pathosystem.     

The Fusarium graminearum Xyr1 transcription factor regulates xylanase expression but is not essential for fungal virulence

The fungal pathogen Fusarium graminearum is the causal agent of fusarium head blight in wheat and other small grain cereals. This fungus is known to produce high amounts of cell wall‐degrading enzymes during infection of wheat spikes. In addition, wheat tissue is particularly rich in xylan, which can be hydrolysed by fungal xylanases. In order to establish the role of F. graminearum xylanase activity in pathogenicity, targeted gene disruption of the F. graminearum xyr1 gene, encoding the major regulator of xylanase gene expression, was performed. When grown on xylan as carbon source, the xylanase activity of the Δxyr1 mutant was dramatically reduced and fungal growth was significantly reduced compared to the wildtype fungus. When grown on carboxymethylcellulose, the cellulolytic activity of the mutant was also reduced and the mutant did not grow on wheat cell walls. The disruption of the xyr1 gene greatly reduced the expression of xylanase‐encoding genes both in vitro and during wheat spike infection, thus confirming the involvement of F. graminearum Xyr1 in the regulation of genes controlling xylan degradation. However, despite the deep impact caused by xyr1 gene disruption on the expression of xylanase genes and on total xylanase activity, the virulence of the Δxyr1 mutant appeared unaffected on Triticum aestivum and T. durum spikes and on soybean seedlings. In conclusion, although a possible role for residual xylanase activity in the virulence of F. graminearum cannot be conclusively excluded, the results question the importance of xylanase activity during the infection process.     

Nickel potentiates soybean resistance against infection by Phakopsora pachyrhizi

Asian soybean rust (ASR), caused by the fungus Phakopsora pachyrhizi, causes significant yield losses worldwide. Nickel (Ni) plays a key role in the metabolism of some profitable crops, such as soybeans, because it is a constituent of several biomolecules and is required for the catalytic process of several enzymes. This study investigated the effect of foliar Ni treatment on the potentiation of soybean cultivar TMG 135 resistance to P. pachyrhizi infection at the microscopic, biochemical, and molecular levels. The severity of ASR decreased by 35% in plants treated with Ni. The malondialdehyde concentration, an indicator of cellular oxidative damage, was high in the leaves of plants that were not treated with Ni and was linked to ASR severity and the extensive colonization of the palisade and spongy parenchyma cells by fungal hyphae. The lignin concentration, β-1,3-glucanase activity, and expression of the URE gene and the defence-related genes PAL1.1, PAL2.1, CHI1B1, and PR-1A were up-regulated in Ni-treated plants infected with P. pachyrhizi. The information provided by this study shows the great potential of Ni to increase the basal level of soybean resistance to ASR and to complement other control methods within the context of sustainable agriculture.     

Vavilov wheat accessions provide useful sources of resistance to tan spot (syn. yellow spot) of wheat

Host genetic resistance is the most effective and sustainable means of managing tan spot or yellow spot of wheat. The disease is becoming increasingly problematic due to the adoption of minimum tillage practices, evolution of effector‐mediated pathogenicity, and widespread cultivation of susceptible cultivars from a narrow genetic base. This highlights the importance of broadening the diversity of resistance factors in modern breeding germplasm. This study explored 300 genetically diverse wheat accessions, originally sourced from the N. I. Vavilov Institute of Plant Genetic Resources (VIR), St Petersburg, Russia. The collection was screened for resistance to tan spot at seedling and adult stage under controlled conditions, and in the field across 2 years. The phenotypic datasets, coupled with ToxA bioassay screening, identified a number of accessions with useful sources of resistance. Seedling disease response corresponded well with ToxA sensitivity (r = 0.49, P < 0.000), but not adult responses (r = ?0.02 to ?0.19, P < 0.002), and overall reactions to ToxA appeared to show poor correspondence with disease response at the adult stage. ToxA‐insensitive accessions were generally found resistant across different growth stages (all‐stage resistance, ASR) in all experiments (seedling and adult stage under controlled conditions and field). ToxA‐sensitive accessions that were susceptible at seedling stage, but resistant at both adult‐plant stages, were deemed to carry adult‐plant resistance (APR). This study provides detailed information on the degree of tan spot resistance in the Vavilov wheat collection and discusses strategies to harness these sources to boost the diversity of resistance factors in modern wheat breeding germplasm.     

Virulence assessment of Australian Pyrenophora tritici-repentis isolates

The virulence of 57 Australian isolates of Pyrenophora tritici-repentis (Ptr), a necrotrophic fungal pathogen responsible for the major wheat disease tan spot, was assessed through plant infection assays. Isolates collected from the northern, southern, and western wheat-cropping regions of Australia were evaluated against 16 Australian bread wheat cultivars under controlled growth conditions. Following infection, the wheat panel displayed varying disease symptoms ranging from tiny necrotic specks to spreading chlorotic and necrotic lesions. Analysis of variance indicated that the wheat cultivar exhibited a greater effect on the disease response, explaining 62.7% of the variation, in comparison to the isolate (10.4%). The interaction between the cultivar and the isolate was statistically significant and was attributed to 9.8% of the total variation. All Ptr isolates examined were able to cause disease, but did not display a clear distinction in virulence on the wheat panel investigated, instead showing subtle differences in aggressiveness. Based on the disease responses, there was no obvious pattern between isolate aggressiveness and cropping region. Some cultivars, such as Hydra, exhibited an effective level of resistance in relation to the panel of isolates tested. All 57 Ptr isolates were found to possess the ToxA effector gene and lack the ToxB effector gene. The gene expression level of ToxA was up-regulated at 3 days postinfection in both ToxA-sensitive and -insensitive cultivars, independent of ToxA–Tsn1 recognition.     

Potentiation of soybean resistance against Phakopsora pachyrhizi infection using phosphite combined with free amino acids

Considering the importance of Asian soybean rust (ASR), caused by Phakopsora pachyrhizi, in the decrease in soybean yield, this study investigated the potential of using phosphite combined with l -α-free amino acids (referred to as induced resistance [IR] stimulus hereafter) to boost defence responses of soybean plants against P. pachyrhizi infection. Plants were sprayed with water (control), acibenzolar-S-methyl (ASM) or IR stimulus and noninoculated or inoculated with P. pachyrhizi. Urediniospore germination was not affected by the IR stimulus in vitro. Reduced ASR severity, lower malondialdehyde concentration and less colonization of leaf tissues by P. pachyrhizi (lower TEF-1α expression from 1 to 15 days after inoculation [dai]) occurred for IR stimulus-sprayed plants. The pattern of gene expression for IR stimulus-sprayed and infected plants was strikingly similar but sometimes more remarkable than that in ASM-sprayed and infected plants. Higher production of phenolics and lignin along with stronger up-regulation of PAL1.3 (5 and 10 dai), PAL2.2 (3 dai), PAL3.1 (1, 3 and 5 dai), ICS1 (5 dai), CHIA1 (1, 5 and 10 dai), CHI1B1 (5 dai), PR-1A (5 and 10 dai), NR1-2 (5 and 10 dai) and INR-2 (5 and 10 dai) for IR stimulus-sprayed plants increased their resistance against ASR. In addition, IR stimulus-sprayed and infected plants showed less impairment of the photosynthetic apparatus and maintained high concentrations of chlorophyll a + b and carotenoids. These findings highlight the potential of using this IR stimulus for developing a well-tuned and effective defensive strategy in soybean plants against P. pachyrhizi infection.     

Histopathology of Sclerotinia sclerotiorum infection and oxalic acid function in susceptible and resistant soybean

The success of the necrotrophic fungus Sclerotinia sclerotiorum is largely dependent on its major virulence factor, oxalic acid (OA). Virulence is lost in transgenic plants that express OA degrading enzymes, e.g. oxalate oxidase (OxO). The histopathology of S. sclerotiorum infection and OA accumulation was examined in a transgenic soybean line over‐expressing OxO (OxO‐OE) and its isogenic parent (WT). In situ flower inoculation showed that the OxO‐OE plants were highly resistant to the pathogen while the WT parents were susceptible. This difference in resistance was not apparent in the floral tissues, as aggressive hyphal activity was similar on both hosts, showing that high OxO activity and low OA accumulation in OxO‐OE was not a deterrent. However, the process of fungal infection on excised leaf tissue differed on the two hosts. Primary lesions developed and showed similar severe ultrastructural damage on both hosts but rapid lesion expansion (colonization) proceeded only on the WT, concomitant with OA accumulation. Oxalic acid rose in OxO‐OE 1 day post‐inoculation and did not change over the following 3 days, showing that colonization can be blocked by maintaining low levels of OA. However, OxO degradation of OA did not deter initial host penetration and primary lesion formation. This shows that OA, the major virulence factor of S. sclerotiorum, is critical for host colonization but may not be required during primary lesion formation, suggesting that other factors are contributing to the establishment of the primary lesion.     

Fungal endophyte protects Atractylodes lancea from root rot caused by Fusarium oxysporum

Endophytic fungi, which stimulate a variety of defence reactions in host plants without causing visible disease symptoms, have been isolated from almost every plant. However, beneficial interactions between fungal endophytes and pathogens from the same habitat remain largely unknown. An inoculation of Atractylodes lancea plantlets with Gilmaniella sp. AL12 (AL12) prior to infection with Fusarium oxysporum prevented the necrotization of root tissues and plant growth retardation commonly associated with fusarium root rot. Quantification of F. oxysporum infections using real‐time PCR revealed a correlation between root rot symptoms and the relative amount of fungal DNA. Pretreatment with AL12 reduced the accumulation of reactive oxygen species stimulated by F. oxysporum. An in vitro analysis of their interactions under axenic culture conditions showed AL12 could inhibit F. oxysporum growth. Additionally, F. oxysporum infections were shown to decrease salicylic acid (SA) production compared with control plantlets. SA biosynthesis inhibitors, 2‐aminoindan‐2‐phosphonic acid and paclobutrazol, abolished the inhibition of F. oxysporum growth in A. lancea even after inoculation with AL12. The results indicated that the fungal endophyte protected A. lancea not only by direct antibiosis, but also by reversing the F. oxysporum‐mediated suppression of SA production.