Infection Cushions Research Articles

Transcription Factor SsSte12 Was Involved in Mycelium Growth and Development in Sclerotinia sclerotiorum.

Sclerotinia sclerotiorum is a challenging agricultural pathogen for management, causing large global economic losses annually. The sclerotia and infection cushions are critical for its long-term survival and successful penetration on a wide spectrum of hosts. The mitogen-activated protein kinase (MAPK) cascades serve as central signaling complexes that are involved in various aspects of sclerotia development and infection. In this study, the putative downstream transcription factor of MAPK pathway, SsSte12, was analyzed in S. sclerotiorum. Silencing SsSte12 in S. sclerotiorum resulted in phenotypes of delayed vegetative growth, reduced size of sclerotia, and fewer appressoria formation. Consequently, the SsSte12 RNAi mutants showed attenuated pathogenicity on the host plants due to the defect compound appressorium. Yeast two-hybrid (Y2H) and bimolecular fluorescence complementation assays demonstrated that the SsSte12 interacts with SsMcm1. However, the SsMcm1 expression is independent of the regulation of SsSte12 as revealed by qRT-PCR analysis in SsSte12 RNAi mutants. Together with high accumulation of SsSte12 transcripts in the early development of S. sclerotiorum, our results demonstrated that SsSte12 function was essential in the vegetative mycelial growth, sclerotia development, appressoria formation and penetration-dependent pathogenicity. Moreover, the SsSte12-SsMcm1 interaction might play a critical role in the regulation of the genes encoding these traits in S. sclerotiorum.

Histopathological Studies of Rhizoctonia solani Infection Process in Different Cultivars of Mungbean [Vigna radiata (L.) Wilczek

Aerial/web blight of mungbean is caused by Rhizoctonia solani Kuhn. The infection process of R. solani was studied on the leaves of susceptible (Pusa Vishal) and resistant (Hum-1) cultivars of mungbean. Leaves and stem of 21 days old cultivars of mungbean were inoculated with the mycelium of freshly growing culture (2 mm) of R. solani isolate (RDLM1) and incubated at 28 °C for different time intervals (3, 6, 18 and 24 h). On the susceptible genotype Pusa Vishal, infection cushions and necrotic lesions were developed after 18 h of inoculation. However, on the resistant genotype Hum-1 no lesions were visible till 24 h of inoculation. The hyphae of R. solani were observed penetrating through two ways (1) direct penetration-by swollen hyphae, lobate appressorium and infection cushion, (2) indirect-stomatal penetration. Differences were also observed in number of infection cushion formed in susceptible (45.37) and resistant (30.87) genotypes after 18 h of inoculation. Typical lesion formation after 18 h of inoculation on susceptible genotype was also validated in KPM-32, KPM-33, KPM-36, KPM-37, KPM-38, KPM-39, KPM-42, KPM-43, IPM-09-3 and IPM-02-3 genotypes of mungbean. Clear cut differences were also observed in transverse section of stem after 18 h of inoculation, where maximum colonization of stem was observed on cortex pith region.

Two Novel Hypovirulence-Associated Mycoviruses in the Phytopathogenic Fungus Botrytis cinerea: Molecular Characterization and Suppression of Infection Cushion Formation.

Botrytis cinerea is a necrotrophic fungus causing disease on many important agricultural crops. Two novel mycoviruses, namely Botrytis cinerea hypovirus 1 (BcHV1) and Botrytis cinerea fusarivirus 1 (BcFV1), were fully sequenced. The genome of BcHV1 is 10,214 nt long excluding a poly-A tail and possesses one large open reading frame (ORF) encoding a polyprotein possessing several conserved domains including RNA-dependent RNA polymerase (RdRp), showing homology to hypovirus-encoded polyproteins. Phylogenetic analysis indicated that BcHV1 may belong to the proposed genus Betahypovirus in the viral family Hypoviridae. The genome of BcFV1 is 8411 nt in length excluding the poly A tail and theoretically processes two major ORFs, namely ORF1 and ORF2. The larger ORF1 encoded polypeptide contains protein domains of an RdRp and a viral helicase, whereas the function of smaller ORF2 remains unknown. The BcFV1 was phylogenetically clustered with other fusariviruses forming an independent branch, indicating BcFV1 was a member in Fusariviridae. Both BcHV1 and BcFV1 were capable of being transmitted horizontally through hyphal anastomosis. Infection by BcHV1 alone caused attenuated virulence without affecting mycelial growth, significantly inhibited infection cushion (IC) formation, and altered expression of several IC-formation-associated genes. However, wound inoculation could fully rescue the virulence phenotype of the BcHV1 infected isolate. These results indicate the BcHV1-associated hypovirulence is caused by the viral influence on IC-formation-associated pathways.

Influence of Different Nitrogen Sources on Growth and Pathogenic Capability of Rhizoctonia solani Causing Root Rot of Faba Bean

The effect of different nitrogen sources (glucosamine sulfate, ammonium sulfate, aspartic acid, phenylalanine and peptone) in comparison to sodium nitrate, the major nitrogen compound in basal agar Czapek’s synthetic medium growth were studied on the linear growth of Rhizoctonia solani and its pathogenicity on faba bean germinated seeds. Ammonium sulfate exhibited faster liner growth and showed the same effect as the basal medium with sodium nitrate while glucosamine sulfate showed less growth rate compared with sodium nitrate. Glucosamine sulfate and ammonium sulfate showed a significant reduction in number of infection cushions which led to significant decrease of disease index in vitro. Under greenhouse conditions, glucosamine sulfate or peptone as a sole nitrogen sources in food requirements of Rhizoctonia solani inoculum depressed the virulence of the fungus. The effect of different amounts of glucosamine sulfate was determined on fungal growth rate, infection cushions, disease index in vitro and polyphenol oxidase activity. Increasing amount of glucosamine sulfate showed significant reduction of growth rate in comparison to the basal medium with sodium nitrate. All seeds subjected to R. solani grown on different amount of glucosamine sulfate showed the lower number of infection cushions, disease index and polyphenol oxidase activity compared with sodium nitrate. Under greenhouse conditions, disease index showed a significantly decreased effect when glucosamine sulfate used as soil applications and showed better effect on shoot weight and root weight compared with control plants treated with sodium nitrate. Our study proposes that glucosamine sulfate may act as controlling factor of pathogenicity genes of R. solani

Prediction of Cacao (Theobroma cacao) Resistance to Moniliophthora spp. Diseases via Genome-Wide Association Analysis and Genomic Selection.

Cacao (Theobroma cacao) is a globally important crop, and its yield is severely restricted by disease. Two of the most damaging diseases, witches’ broom disease (WBD) and frosty pod rot disease (FPRD), are caused by a pair of related fungi: Moniliophthora perniciosa and Moniliophthora roreri, respectively. Resistant cultivars are the most effective long-term strategy to address Moniliophthora diseases, but efficiently generating resistant and productive new cultivars will require robust methods for screening germplasm before field testing. Marker-assisted selection (MAS) and genomic selection (GS) provide two potential avenues for predicting the performance of new genotypes, potentially increasing the selection gain per unit time. To test the effectiveness of these two approaches, we performed a genome-wide association study (GWAS) and GS on three related populations of cacao in Ecuador genotyped with a 15K single nucleotide polymorphism (SNP) microarray for three measures of WBD infection (vegetative broom, cushion broom, and chirimoya pod), one of FPRD (monilia pod) and two productivity traits (total fresh weight of pods and % healthy pods produced). GWAS yielded several SNPs associated with disease resistance in each population, but none were significantly correlated with the same trait in other populations. Genomic selection, using one population as a training set to estimate the phenotypes of the remaining two (composed of different families), varied among traits, from a mean prediction accuracy of 0.46 (vegetative broom) to 0.15 (monilia pod), and varied between training populations. Simulations demonstrated that selecting seedlings using GWAS markers alone generates no improvement over selecting at random, but that GS improves the selection process significantly. Our results suggest that the GWAS markers discovered here are not sufficiently predictive across diverse germplasm to be useful for MAS, but that using all markers in a GS framework holds substantial promise in accelerating disease-resistance in cacao.

SsSm1, a Cerato-platanin family protein, is involved in the hyphal development and pathogenic process of Sclerotinia sclerotiorum

The filamentous fungus Sclerotinia sclerotiorum is an important plant pathogen with a worldwide distribution. It can infect a wide variety of plants, causing serious disease in many types of crops, such as rapeseed, sunflower and soybean. Sclerotinia stem rot caused by this fungus affects main crops and has led to great economic loss. Elicitors are a group of compounds that inspire the host plant to produce an immune response against invading pathogens. This study describes a protein that has high homology with the Trichoderma elicitor Sm1 and was found in the genome of S. sclerotiorum. We named this protein SsSm1. To determine whether this protein has an elicitor function like its homology protein, we constructed a heterologous expression vector for SsSm1 and expressed it in Escherichia coli. The protein of heterologous expression led to the formation of lesions in tobacco that closely resemble hypersensitive response lesions. Transient expression of the encoding gene of SsSm1 in tobacco leaves also caused hypersensitive response. Then, RNA silencing was used to identify the function of SsSm1. The hyphal growth and pathogenicity of silenced transformants were shown to be obviously lagging and branched abnormally. Transformants produced less infection cushions and deformed sclerotiorum. In addition, SsSm1 silencing caused weak tolerance to NaCl, sorbitol and SDS, and the sensitivity of mutants to carbendazim was also significantly decreased. Based on the above results, we speculate that this protein may be related to the development of hyphae, infection cushions and sclerotiorum, but the specific molecular mechanism needs to be studied further.

Infection cycle ofAlternaria brassicicolaonBrassica oleracealeaves under growth room conditions

Development of the necrotrophic fungusAlternaria brassicicolawas evaluated during infection of three cabbage varieties:Brassica oleraceavar. capitataf.alba‘Stone Head’ (white cabbage),B. oleraceavar. capitataf. rubra‘Langedijker Dauer’ (red cabbage) andB. oleraceavar. capitataf. sabauda‘Langedijker Dauerwirsing’ (Savoy cabbage). Following inoculation of cabbage leaves, conidial germination, germ tube growth, and appressorium formation were analysed during the first 24 h of infection. Differences in the dynamics of fungal development on leaves were observed, e.g. approximately 40% of conidia germinated on Savoy cabbage leaves at 4 h post‐inoculation (hpi) while only 20% germinated on red and white cabbage leaves. Leaf penetration on the three cabbage varieties mainly occurred through appressoria, rarely through stomata. Formation of infection cushions was found exclusively on red cabbage. Appressoria were first observed on red cabbage leaves at 6 hpi, and on white and Savoy cabbage leaves at 8 hpi. Conidiogenesis occurred directly from mature conidia at an early stage of fungal development (10 hpi), but later (48 hpi) it occurred through conidiophores. Disease progress and changes in the morphology of leaf surfaces were also observed. At the final 120 hpi measurement point, necroses on all investigated varieties were approximately the same size. Based on detailed light and electron microscopic analyses, an outline of theA. brassicicolainfection cycle onB. oleracealeaves under growth room conditions has been proposed. According to the authors’ knowledge, this report represents the most detailed up‐to‐date description of infection ofB. oleraceavarieties byA. brassicicola.

Responses of rice to Rhizoctonia solani and its toxic metabolite in relation to expression of Osmyb4 transcription factor

The reaction of IR 50, TRY 3, and IR 36 cultivars of rice to R. solani challenge, the causal agent of sheath blight, and its toxic metabolite was studied. Differential response of these cultivars to the pathogen and/or toxin inoculation was observed in detached leaf sheaths and greenhouse-grown plants. The observations were based on disease scoring, electrolyte leakage, and also microscopic views of infection cushions. The Osmyb4 gene expression was studied in the tissues from all these experiments and a correlation between the level of expression and disease response of the varieties was found at least in some experiments. The mechanisms regulated by Osmyb4 might have a lower but significant contribution to the tolerance of rice cultivars to sheath blight.

The Protein Disulfide Isomerase of Botrytis cinerea: An ER Protein Involved in Protein Folding and Redox Homeostasis Influences NADPH Oxidase Signaling Processes.

Botrytis cinerea is a filamentous plant pathogen, which infects hundreds of plant species; within its lifestyle, the production of reactive oxygen species (ROS) and a balanced redox homeostasis are essential parameters. The pathogen is capable of coping with the plant’s oxidative burst and even produces its own ROS to enhance the plant’s oxidative burst. Highly conserved NADPH oxidase (Nox) complexes produce the reactive molecules. The membrane-associated complexes regulate a large variety of vegetative and pathogenic processes. Besides their commonly accepted function at the plasma membrane, recent studies reveal that Nox complexes are also active at the membrane of the endoplasmic reticulum. In this study, we identified the essential ER protein BcPdi1 as new interaction partner of the NoxA complex in B. cinerea. Mutants that lack this ER chaperone display overlapping phenotypes to mutants of the NoxA signaling pathway. The protein appears to be involved in all major developmental processes, such as the formation of sclerotia, conidial anastomosis tubes and infection cushions (IC’s) and is needed for full virulence. Moreover, expression analyses and reporter gene studies indicate that BcPdi1 affects the redox homeostasis and unfolded protein response (UPR)-related genes. Besides the close association between BcPdi1 and BcNoxA, interaction studies provide evidence that the ER protein might likewise be involved in Ca2+ regulated processes. Finally, we were able to show that the potential key functions of the protein BcPdi1 might be affected by its phosphorylation state.

The septin protein Sep4 facilitates host infection by plant fungal pathogens via mediating initiation of infection structure formation.

Many phytopathogenic fungi use infection structures (IFSs, i.e., appressoria and infection cushions) to penetrate host cuticles. However, the conserved mechanisms that mediate initiation of IFS formation in divergent pathogens upon sensing the presence of host plants remain obscure. Here, we demonstrate that a conserved septin gene SEP4 plays crucial roles in this process. Disruption of SEP4 in the plant grey mould fungus Botrytis cinerea completely blocked IFS formation and abolished the virulence of ΔBcsep4 mutants on unwounded hosts. During IFS formation, mutants lacking SEP4 could produce reactive oxygen species (ROS) normally. Inhibition of ROS production in strains harbouring the SEP4 gene resulted in disordered assembly of Sep4 and the subsequent failure to form infection cushions, suggesting that proper Sep4 assembly regulated by ROS is required for initiation of IFS formation and infection. Moreover, loss of SEP4 severely impaired mutant conidiation, melanin and chitin accumulation in hyphal tips and lesion expansion on wounded hosts, but significantly promoted germ tube elongation and sclerotium production. SEP4-mediated fungal pathogenic development, including IFS formation, was validated in the hemibiotroph Magnaporthe oryzae. Our findings indicate that Sep4 plays pleiotropic roles in B. cinerea development and specifically facilities host infection by mediating initiation of IFS formation in divergent plant fungal pathogens in response to ROS signaling.

Screening and identification of new sources of resistance to sheath blight in wild rice accessions

A total of 218 wild rice accessions including O. rufipogon and O. nivara were evaluated under artificial inoculation for resistance to sheath blight disease for two years under field conditions. The germplasm identified as resistant to sheath blight were further evaluated through artificial inoculation for an additional year under glass house conditions. Based on three years of testing, two Oryza rufipogon accessions, namely, IC336719 and IC336721 were identified as resistant to sheath blight disease. In vitro characterization of resistant genotypes revealed that the size of sheath blight lesion formation was small along with comparatively lesser number of infection cushions and penetration pegs as compared to the susceptible checks. The O. rufipogon accessions identified in the present study are very valuable genetic resource, which can be utilized in the development of introgression lines and mapping QTL(s) governing resistance to sheath blight of rice.

Botrytis pyriformis sp. nov., a novel and likely saprophytic species of Botrytis

A novel species of Botrytis from Sedum sarmentosum was described based on morphology and analyses of DNA sequences of nuc rDNA ITS regions and three nuclear genes (G3PDH, HSP60, RPB2). Meanwhile pathogenicity in 32 plant species, response to temperature for growth and conidial germination for the species were determined. The Botrytis species was named Botrytis pyriformis sp. nov. It was characterized by formation of grayish mycelia, brownish conidia and melanized sclerotia on PDA. The conidia are pear-shaped, melanized and covered with abundant villiform appendages on the conidial surface. Comparison of the ITS sequences confirmed its placement in the genus Botrytis. Phylogenetic analysis based on DNA sequences of G3PDH, HSP60 and RPB2 genes indicated that B. pyriformis and other 30 Botrytis species form a monophyletic clade, which was further divided into three subclades. Subclade I comprised B. pyriformis alone, whereas subclades II and III comprised six and 24 Botrytis species, respectively. Botrytis pyriformis could not infect 32 plant species including S. sarmentosum, possibly due to deficiency in formation of infection cushions. This study presents a formal description and illustrations for B. pyriformis and provides experimental evidence, indicating that B. pyriformis might be a saprophytic species.

Histopathological assessment of the infection of maize leaves by Fusarium graminearum, F. proliferatum, and F. verticillioides

Young maize plants were inoculated on unfolded mature leaves and on folded immature leaves with Fusarium graminearum, Fusarium proliferatum, and Fusarium verticillioides suspensions. Infection and symptom development of disease on these asymptomatic mature leaves and immature leaves were then documented. Subcuticular infection was found by the three Fusarium species on both symptomatic and symptomless leaves. The three Fusarium species penetrated the stomata of immature leaves by the formation of appressoria-like structures, infection cushions or by direct penetration. Infection by the three species of Fusarium via stomata is reported here for the first time. The superficial hyphae and re-emerging hyphae of the three species produced conidia. The macroconidia of F. graminearum produced secondary macroconidia and F. proliferatum formed microconidia inside the leaf tissues that sporulated through stomata and trichomes. The infection of maize leaves by the three species of Fusarium and their sporulation may contribute inoculum to cob and kernel infection.

The Sclerotinia sclerotiorum Slt2 mitogen-activated protein kinase ortholog, SMK3, is required for infection initiation but not lesion expansion

Mitogen-activated protein kinases (MAPKs) play a central role in transferring signals and regulating gene expression in response to extracellular stimuli. An ortholog of the Saccharomyces cerevisiae cell wall integrity MAPK was identified in the phytopathogenic fungus Sclerotinia sclerotiorum. Disruption of the S. sclerotiorum Smk3 gene severely reduced virulence on intact host plant leaves but not on leaves stripped of cuticle wax. This was attributed to alterations in hyphal apical dominance leading to the inability to aggregate and form infection cushions. The mutation also caused loss of the ability to produce sclerotia, increased aerial hyphae formation, and altered hyphal hydrophobicity and cell wall integrity. Mutants had slower radial expansion rates on solid media but more tolerance to elevated temperatures. Loss of the SMK3 cell wall integrity MAPK appears to have impaired the ability of S. sclerotiorum to sense its surrounding environment, leading to misregulation of a variety of functions. Many of the phenotypes were similar to those observed in S. sclerotiorum adenylate cyclase and SMK1 MAPK mutants, suggesting that these signaling pathways co-regulate aspects of fungal growth, physiology, and pathogenicity.

BcIqg1, a fungal IQGAP homolog, interacts with NADPH oxidase, MAP kinase and calcium signaling proteins and regulates virulence and development in Botrytis cinerea.

NADPH oxidases (Nox) produce reactive oxygen species (ROS) in multicellular eukaryotic organisms. They trigger defense reactions ('oxidative burst') - in phagocytes and plant cells -, and are involved in a broad range of differentiation processes. Fungal Nox-complexes play a central role in vegetative, sexual and pathogenic processes. In contrast to mammalian systems, knowledge is limited about composition, localisation and connection to major signaling cascades in fungi. Here, we characterize a fungal homolog of the RasGAP scaffold protein IQGAP, which links several major signaling processes, including Nox in mammalian cell lines. We show that BcIqg1 interacts directly with a cytosolic, regulatory component (BcRac) and a membrane-associated subunit (BcNoxD) of a Nox-complex in the pathogen Botrytis cinerea. Thus, this protein may be a scaffold that mediates interaction of the catalytic subunits with the regulator BcNoxR. The protein interacts with modules of the MAP kinase- and calcium-dependent signaling pathways. Functional analysis of BcIqg1 substantiated its involvement in different signaling pathways. It mediates the Ca(2+) -triggered nuclear translocation of - BcCRZ1 and the MAP kinase BcBmp1. BcIqg1 is involved in resistance against oxidative and membrane stress and is required for several developmental processes including formation of sclerotia, conidial anastomosis tubes and infection cushions as well as for virulence.

Differential behaviour of sheath blight pathogen Rhizoctonia solani in tolerant and susceptible rice varieties before and during infection

This study characterized the early infection and establishment of the sheath blight pathogen Rhizoctonia solani on a tolerant rice variety, Swarnadhaan (IET 5656), and a susceptible variety, Swarna (MTU 7029). Assays using whole plants showed that disease severity was higher in Swarna than Swarnadhaan. In a detached leaf assay, Swarnadhaan showed a disease index that was 50% less than that with Swarna. Rhizoctonia solani exhibited different growth behaviour in the tolerant and susceptible varieties. The pathogen showed more hyphal growth in the susceptible host than in the tolerant variety. It also showed profuse branching, making intimate contact with the host surface to form more inter‐ and intracellular structures, and greater sclerotial development in the susceptible host compared to the tolerant one. Using light and scanning electron microscopy, it was observed for the first time that the pathogen could intercept host surface structures and use these for anchorage or penetration. Transformed R. solani, expressing green fluorescent protein, was observed using confocal laser scanning microscopy to investigate pathogen behaviour, including the formation of infection cushions and subsequent colonization of the host tissues. This is the first ultrastructural report to characterize the differential behaviour of the sheath blight pathogen in the vicinity and within tolerant and susceptible rice plants.

Histological observation of potato in response to Rhizoctonia solani infection

Rhizoctonia solani is the causal agent of stem canker and black scurf in potato, resulting in significant yield and economic losses. To understand the infection process of R. solani and potato responses to the pathogen, histological observation of inoculated potato was performed under light and electron microscopes, under laboratory conditions. R. solani was observed at the invasion site, for the initial infection time, and for the presence of infection structures (appressoria, infection cushions). These attributes were compared when R. solani affected different parts of the host: aboveground stems, underground stems, tubers; as well as in different potato cultivars: ‘Desiree’ (resistant to R. solani) and ‘Atlantic’ (susceptible). In aboveground stems of potato, the invasion of R. solani was mainly limited to intercellular spaces. In underground stems and tubers, the primary invasion sites were epidermal cracks and lenticels. Initial infection time was 8 to 12 h post inoculation (HPI) in aboveground stems, 8 HPI in underground stems, and 4 HPI in tubers. The hyphae of R. solani produced various infection structures, with a few infection cushions, and a large number of appressoria with different morphologies. In aboveground stems, more infection structures were found than in underground stems and tubers. ‘Desiree’ had fewer numbers and smaller sizes of lenticels, thicker cuticle and periderm, and fewer epidermal cracks compared to ‘Atlantic’. Fewer numbers of infection structure of R. solani were observed in ‘Desiree’ than in ‘Atlantic’. It was suggested that the epidermal and perithelial structures of potato were related to disease resistance, such as number and size of lenticels, thickness of cuticle and periderm, and epidermal cracks.

Attack modes and defence reactions in pathosystems involving Sclerotinia sclerotiorum, Brassica carinata, B. juncea and B. napus.

Sclerotinia stem rot (SSR, Sclerotinia sclerotiorum) is a damaging disease of oilseed brassicas world-wide. Host resistance is urgently needed to achieve control, yet the factors that contribute to stem resistance are not well understood. This study investigated the mechanisms of resistance to SSR. Stems of 5-week-old Brassica carinata, B. juncea and B. napus of known resistance were infected via filter paper discs impregnated with S. sclerotiorum mycelium under controlled conditions. Transverse sections of the stem and portions of the stem surface were examined using optical and scanning electron microscopy. The association of anatomical features with the severity of disease (measured by mean lesion length) was determined. Several distinct resistance mechanisms were recorded for the first time in these Brassica-pathogen interactions, including hypersensitive reactions and lignification within the stem cortex, endodermis and in tissues surrounding the lesions. Genotypes showing a strong lignification response 72 h post-infection (hpi) tended to have smaller lesions. Extensive vascular invasion by S. sclerotiorum was observed only in susceptible genotypes, especially in the vascular fibres and xylem. Mean lesion length was negatively correlated with the number of cell layers in the cortex, suggesting progress of S. sclerotiorum is impeded by more cell layers. Hyphae in the centre of lesions became highly vacuolate 72 hpi, reflecting an ageing process in S. sclerotiorum hyphal networks that was independent of host resistance. The infection process of S. sclerotiorum was analogous in B. carinata and B. napus. Infection cushions of the highly virulent isolate of S. sclerotiorum MBRS-1 were grouped together in dense parallel bundles, while hyphae in the infection cushions of a less aggressive isolate WW-3 were more diffuse, and this was unaffected by host genotype. A variety of mechanisms contribute to host resistance against S. sclerotiorum across the three Brassica species. These complex interactions between pathogen and host help to explain variable expressions of resistance often observed in the field.

SsPemG1 encodes an elicitor-homologous protein and regulates pathogenicity in Sclerotinia sclerotiorum

Elicitors are molecules that stimulate plant defense responses and are valuable in disease control strategies. Here, we report a gene named SsPemG1 encoding an elicitor-homologous protein. RNA silencing was used to identify the function of SsPemG1. SsPemG1 silencing induced strong virulence in both detached leaves and intact plants. A higher growth rate, reduced sodium chloride (NaCl) and sodium dodecyl sulfate (SDS) susceptibility, more infection cushions and greater cell-wall-degrading enzymes (CWDEs) activities all contributed to this virulence. These data will help us better understand the function of SsPemG1 in Sclerotinia sclerotiorum, provide a new method to study elicitors and a theoretical foundation for disease control of this fungus.

The signalling mucin Msb2 regulates surface sensing and host penetration via BMP1 MAP kinase signalling in Botrytis cinerea.

Botrytis cinerea is a necrotrophic fungus that infects a wide range of fruit, vegetable and flower crops. Penetration of the host cuticle occurs via infection structures that are formed in response to appropriate plant surface signals. The differentiation of these structures requires a highly conserved mitogen-activated protein (MAP) kinase cascade including the MAP kinase BMP1. In yeast and several plant-pathogenic fungi, the signalling mucin Msb2 has been shown to be involved in surface recognition and MAP kinase activation. In this study, a B. cinerea msb2 mutant was generated and characterized. The mutant showed normal growth, sporulation, sclerotia formation and stress resistance. In the absence of nutrients, abnormal germination with multiple germ tubes was observed. In the presence of sugars, normal germination occurred, but msb2 germlings were almost unable to form appressoria or infection cushions on hard surfaces. Nevertheless, the msb2 mutant showed only a moderate delay in lesion formation on different host plants, and formed expanding lesions similar to the wild-type. Although the wild-type showed increasing BMP1 phosphorylation during the first hours of germination on hard surfaces, the phosphorylation levels in the msb2 mutant were strongly reduced. Several genes encoding secreted proteins were found to be co-regulated by BMP1 and Msb2 during germination. Taken together, B. cinerea Msb2 is likely to represent a hard surface sensor of germlings and hyphae that triggers infection structure formation via the activation of the BMP1 MAP kinase pathway.