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Cultivation of apple trees in the highlands of Ethiopia began in 1955. In 2014, blistering of the bark due to cankers on the main stems mostly below the grafting points, followed by dieback and eventually death of apple trees, was observed in apple orchards in the Hadiya Zone in Ethiopia. This study aimed to identify the causal agent of canker and dieback symptoms on the apple trees. Symptomatic trunks from 20 trees (ten per cultivar) were sampled. Isolations were performed from ten trunks (five per cultivar). Fungus colonies with similar cultural features were obtained from all the samples, and the morphology of a representative isolate was characterized. Phylogenetic analyses of the concatenated internal transcribed spacers 1 and 2 and 5.8S rRNA gene, large subunit and actin gene regions confirmed the identity of two isolates as Didymosphaeria rubi-ulmifolii. Pathogenicity was confirmed for one isolate by inoculations of healthy branches of ‘Anna’ and ‘Dorsett Golden’ apple trees resulting in lesion formation, and subsequent re-isolation of the inoculated fungus. This study is the first report of D. rubi-ulmifolii associated with dieback of apple trees. This pathogen caused death of more than 26% of apple trees in one commercial orchard, and could cause severe losses for smallholder apple growers in Ethiopia. Future studies are required to assess the magnitude, distribution and management options of this economically important canker disease in Ethiopia.

Analysis of Causative Factors for Canker Disease Epidemic on Rubber Plants in South Sumatra. Lasiodiplodia theobromae is the cause of stem canker in the rubber plant (Hevea brasiliensis), this is a new disease and has never been reported in Indonesia. Currently, data on environmental factors and cultivation techniques related to the development of stem canker are not available so research needs to be done on this subject. The objective of the study was to analyze environmental factors and cultivation techniques related to stem canker on rubber trees in southern Sumatra. The study was conducted by collecting data on environmental factors and cultivation techniques and then analyzed the association with stem canker. Field observation was carried out at 21 locations in PT. Perkebunan Nusantara VII Business Unit Padang Plawi Bengkulu, Tulung Buyut Lampung, Musilandas Palembang and Experimental Garden of Sembawa Research Center, Indonesian Rubber Research Institute at Palembang to measure the severity and incidence of this disease. Besides that, the data of cultivation technique and the condition of the garden were taken from the officer at those locations. Soil sample from each location was taken in a composite way, then its physical and chemical components were analyzed. The analysis was conducted to find out the correlation between cultivation and disease severity using chi-square test. The relationship between chemical and physical factors of the soil with the disease severity was analyzed using a multivariate test of principal component analysis/PCA. Regression analysis was conducted to show the possible relation to the incidence and severity of this disease. The location, clones, weed control and the number of plants per hectare showed a correlation with the disease severity. There was a positive correlation between incidence and severity of the disease. The highest disease severity occurred at the Sembawa location on clone BPM 24, with the number of plant population per ha was ≥ 555 trees and weed control using herbicide. Content of soil nitrogen, dust, clay and water as well as cation exchange capacity (CEC)were suggested to significantly contribute to the severity of stem canker.

Almond (Prunus dulcis) is an important crop for Greece grown on 15.130 ha in 2019. In September 2019, a severe stem canker disease was observed in 6-year-old trees of cv Marta grafted on the rootstock 'F675C14', in a new almond grove of cvs Marta, Soleta, Antonela, Belona and Laurete, in Vlachiana, Heraklion, Crete, Greece. Only cv Marta trees were affected. Diseased trees exhibited cankers on trunks and branches with pale yellow to red-colored gum excreting from cankers, yellowing, leaf fall, twig and branch dieback, bark and wood tissue discoloration. Severely affected trees were killed. A Fusarium-like fungus was consistently isolated from symptomatic wood tissue previously surface-disinfested with 95% ethanol, on acidified potato dextrose agar (APDA). Emerging colonies were transferred to new PDA and the growth rate of the fungus was 7.86 mm/day at 24 °C in the dark. The abundant aerial mycelium was initially white, turning into pale orange in the centre after 7 days of growth on PDA. Microscopic observations revealed hyaline conidiophores measuring 26.74 ± 20.44 μm in length, developing microconidia 5.00 to 9.50 × 2.50 to 4.75 μm (average 6.64 × 3.50 μm) and macroconidia 10.00 to 23.25 × 3.75 to 5.50 μm (average 16.42 × 4.50 μm) in size. DNA from one representative single-spore isolate (code KOUB.AM.VR1) was extracted and the internal transcribed spacer region (ITS) of ribosomal DNA and translation elongation factor 1-alpha (EF 1-a) genes were amplified using the primer pairs ITS1/ITS4 (White et al. 1990) and EF1-F/EF2-R (O'Donnell et al. 1998), respectively. The PCR products were sequenced and deposited in GenBank (accession Nos. MW547397 and MW554492). Based on morphological characteristics (Leslie and Summerell 2006) and a BLAST search with 100.00% and 99.38% identity to published F. solani ITS and EF 1-a sequences in GenBank (KX034335.1, DQ247636.1) the fungus was identified as F. solani. Eight 3-year-old almond trees of cv. Marta were artificially inoculated in March 2020 by making a 6.0-mm-diameter hole into the trunk, inserting a 6-mm-diameter mycelial disc taken from a 10-day-old PDA culture, sealing the hole with cellophane membrane and covering with adhesive paper tape. Another eight trees of the same cultivar were mock-inoculated with sterilized PDA discs and served as controls. Potted trees were kept under ambient conditions. One month post inoculation, yellow gum was evident excreting around the inoculation point in F. solani-treated trees but not in the controls. Seven months post inoculation, longitudinal and transverse sections of inoculated trunks revealed internal and external symptoms similar to those observed under natural infection conditions and F. solani was steadily re-isolated from symptomatic wood tissue and identified by colony morphology. Neither symptoms nor positive isolations were observed in control trunks. Pathogenicity tests were repeated twice. Fusarium solani has been reported as the causal agent of stem canker or wood decay diseases in several woody hosts including bitternut hickory, black walnut, mulberry and pistachio trees (Crespo et al. 2019; Markakis et al. 2017; Park and Juzwik 2012; Tisserat 1987). To the best of our knowledge, this is the first worldwide report of stem canker caused by F. solani on almond tree. This disease could potentially be an increasing problem in almond growing areas and result in severe crop losses. Hence, effective management practices should be investigated and applied.

Phytophthora palmivora (Butler) is an hemibiotrophic oomycete capable of infecting over 200 plant species including one of the most economically important crops, Theobroma cacao L. commonly known as cocoa. It infects many parts of the cocoa plant including the pods, causing black pod rot disease. This review will focus on P. palmivora’s ability to infect a plant host to cause disease. We highlight some current findings in other Phytophthora sp. plant model systems demonstrating how the germ tube, the appressorium and the haustorium enable the plant pathogen to penetrate a plant cell and how they contribute to the disease development in planta. This review explores the molecular exchange between the oomycete and the plant host, and the role of plant immunity during the development of such structures, to understand the infection of cocoa pods by P. palmivora isolates from Papua New Guinea.

In the United Arab Emirates (UAE), royal poinciana (Delonix regia) trees suffer from stem canker disease. Symptoms of stem canker can be characterized by branch and leaf dryness, bark lesions, discoloration of xylem tissues, longitudinal wood necrosis and extensive gumming. General dieback signs were also observed leading to complete defoliation of leaves and ultimately death of trees in advanced stages. The fungus, Neoscytalidium dimidiatum DSM 109897, was consistently recovered from diseased royal poinciana tissues, this was confirmed by the molecular, structural and morphological studies. Phylogenetic analyses of the translation elongation factor 1-a (TEF1-&alpha, ) of N. dimidiatum from the UAE with reference specimens of Botryosphaeriaceae family validated the identity of the pathogen. To manage the disease, the chemical fungicides, Protifert&reg, Cidely&reg, Top and Amistrar&reg, Top, significantly inhibited mycelial growth and reduced conidial numbers of N. dimidiatum in laboratory and greenhouse experiments. The described &ldquo, apple bioassay&rdquo, is an innovative approach that can be useful when performing fungicide treatment studies. Under field conditions, Cidely&reg, Top proved to be the most effective fungicide against N. dimidiatum among all tested treatments. Our data suggest that the causal agent of stem canker disease on royal poinciana in the UAE is N. dimidiatum.

Fungi in the Cryphonectriaceae are important canker pathogens of plants in the Melastomataceae and Myrtaceae (Myrtales). These fungi are known to undergo host jumps or shifts. In this study, fruiting structures resembling those of Cryphonectriaceae were collected and isolated from dying branches of Syzygium cordatum and root collars of Heteropyxis natalensis in South Africa, and from cankers on the bark of Tibouchina grandifolia in La Réunion. A phylogenetic species concept was used to identify the fungi using partial sequences of the large subunit and internal transcribed spacer regions of the nuclear ribosomal DNA, and two regions of the β-tubulin gene. The results revealed a new genus and species in the Cryphonectriaceae from South Africa that is provided with the name Myrtonectria myrtacearum gen. et sp. nov. Two new species of Celoporthe (Cel.) were recognised from La Réunion and these are described as Cel. borbonica sp. nov. and Cel. tibouchinae sp. nov. The new taxa were mildly pathogenic in pathogenicity tests on a clone of Eucalyptus grandis. Similar to other related taxa in the Cryphonectriaceae, they appear to be endophytes and latent pathogens that could threaten Eucalyptus forestry in the future.

To manage stem canker disease on royal poinciana, actinobacterial isolates were used as biological control agents (BCAs) based on their strong in vitro inhibitory effects against Neoscytalidium dimidiatum. Streptomyces griseorubens UAE2 and Streptomyces&nbsp, wuyuanensis UAE1 had the ability to produce antifungal compounds and cell-wall-degrading enzymes (CWDEs). Only S. griseorubens, however, restored the activity of 1-aminocyclopropane-1-carboxylate (ACC) deaminase (ACCD). In vivo apple fruit bioassay showed that lesion development was successfully constrained by either isolates on fruits inoculated with N. dimidiatum. In our greenhouse and container nursery experiments, S. griseorubens showed almost complete suppression of disease symptoms. This was evident when the preventive treatment of S. griseorubens significantly (p &lt, 0.05) reduced the numbers of conidia of N. dimidiatum and defoliated leaves of royal poinciana seedlings to lesser levels than when S. wuyuanensis was applied, but comparable to control treatments (no pathogen). The disease management of stem canker was also associated with significant (p &lt, 0.05) decreases in ACC levels in royal poinciana stems when S. griseorubens was applied compared to the non-ACCD-producing S. wuyuanensis. This study is the first to report the superiority of antagonistic actinobacteria to enhance their effectiveness as BCAs not only for producing antifungal metabolites and CWDEs but also for secreting ACCD.

Canker and needle blight of Araucaria heterophylla (Norfolk Island pine) trees were observed during the surveys conducted in Addis Ababa and Adama cities (Ethiopia) from November to December 2016. The main objective of this study was to investigate fungal pathogens that cause diseases on Araucaria heterophylla trees. Six localities with Araucaria heterophylla plantings were purposively surveyed for disease symptoms. Samples from symptomatic parts of trees were collected, surface sterilized, cultured on PDA and morphologically identified for genus and species. A total of 36 isolates of fungi were identified. Based on macro- and microscopic morphological features of the colonies, the fungal isolates were found to be the genus Diplodia and species Lasiodiplodia theobromae (Pat.) Griffon & Maubl (syn. Botryodiplodia theobromae), the anamorph of Botryosphaeria rhodina (Berk. & M.A. Curtis) Arx. The pathogenicity test showed that the isolates of Lasiodiplodia theobromae, caused stem canker and needle blight on Araucaria heterophylla. This finding is important in the study of management options for future prevention and control of diseases in the country.

Oilseed rape residues are a crucial determinant of stem canker epidemiology as they support the sexual reproduction of the fungal pathogen Leptosphaeria maculans. The aim of this study was to characterize the impact of a resistance gene against L. maculans infection on residue microbial communities and to identify microorganisms interacting with this pathogen during residue degradation. We used near‐isogenic lines to obtain healthy and infected host plants. The microbiome associated with the two types of plant residues was characterized by metabarcoding. A combination of linear discriminant analysis and ecological network analysis was used to compare the microbial communities and to identify microorganisms interacting with L. maculans. Fungal community structure differed between the two lines at harvest, but not subsequently, suggesting that the presence/absence of the resistance gene influences the microbiome at the base of the stem whilst the plant is alive, but that this does not necessarily lead to differential colonization of the residues by fungi. Direct interactions with other members of the community involved many fungal and bacterial amplicon sequence variants (ASVs). L. maculans appeared to play a minor role in networks, whereas one ASV affiliated to Plenodomus biglobosus (synonym Leptosphaeria biglobosa) from the Leptosphaeria species complex may be considered a keystone taxon in the networks at harvest. This approach could be used to identify and promote microorganisms with beneficial effects against residue‐borne pathogens and, more broadly, to decipher the complex interactions between multispecies pathosystems and other microbial components in crop residues., Characterizing the impact of a resistance gene against Leptosphaeria maculans on the microbiome of oilseed rape residues using near‐isogenic lines helps to identify microorganisms interacting with this pathogen.

Carrots with different Rhizoctonia-like symptoms were found in the main Swedish carrot production areas from 2001–2020. The most commonly observed symptoms were a greyish-white felt-like mycelium and black scurf, the latter often associated with Rhizoctonia solani anastomosis group (AG) 3-PT on potato. An overall increase in disease incidence in all studied fields over time was observed for both symptoms. The majority of Rhizoctonia isolates sampled from carrot in the period 2015–2020 were identified as AG 3 (45%) and AG 5 (24%), followed by AG 1-IB (13%), AG 11 (5%), AG-E (5%), AG BI (3%), AG-K (3%) and AG 4-HGII (2%). To our knowledge, this is the first report describing AG 5 in Sweden as well as AG 3, AG 11 and AG-E inducing Rhizoctonia-like symptoms on carrot. Secondly, we report for the first time that R. solani AG 3, and the less observed AGs: AG 1-IB and AG 5 can induce black scurf symptoms on the taproot of carrots. Due to a widely used carrot-potato crop rotation in Swedish areas, a possible cross-over from potato to carrot is suggested. This information is of high importance to reduce Rhizoctonia inoculum in soils, since avoiding carrot-potato crop rotations needs to be considered.

Studies were conducted to determine the role of 3-methylthioproprionic acid (MTPA) in the pathogenicity of potato stem canker, Rhizoctonia solani, and the concentrations required to inhibit growth of R. solani under laboratory and plant house-based conditions. The experiments were laid out in a completely randomized design with five treatments and five replications. The treatments were 0, 1, 2, 4, and 8 mM concentrations of MTPA. The purified toxin exhibited maximal activity at pH 2.5 and 30°C. MTPA at 1, 2, 4, and 8 mM levels reduced plant height, chlorophyll content, haulm fresh weight, number of stolons, canopy development, and tuber weight of potato plants, as compared to the control. MTPA significantly affected mycelial growth with 8 mM causing the highest infection. The potato seedlings treated with MTPA concentrations of 1.0-8.0 mM induced necrosis of up to 80% of root system area. Cankers were resulted from the injection of potato seedling stems with 8.0 mM MTPA. The results showed the disappearance of cell membrane, rough mitochondrial and cell walls, change of the shape of chloroplasts, and swollen endoplasmic reticulum. Seventy-six (76) hours after toxin treatment, cell contents were completely broken, cytoplasm dissolved, and more chromatin were seen in the nucleus. The results suggested that high levels of the toxin concentration caused cell membrane and cytoplasm fracture. The integrity of cellular structure was destroyed by the phytotoxin. The concentrations of the phytotoxin were significantly correlated with pathogenicity and caused damage to the cell membrane of potato stem base tissue.

Soil microorganisms and soil fauna may have a large impact on the tuber yield of potato crops. The interaction between root-lesion nematodes and the pathogenic fungus Rhizoctonia solani K&uuml, hn was studied on potato plants grown in pots under controlled conditions. In two similar experiments, different combinations of nematodes and fungal mycelium were added to the pots at three occasions, at planting, after 14 days, and after 28 days. The nematodes reduced root biomass and the combination of nematodes and R. solani resulted in reduced tuber yield in both experiments, but the interaction was not synergistic. In contrast, the number of stem canker lesions decreased in the presence of nematodes compared to treatments with R. solani only. The time of inoculation influenced the severity of both fungal and nematode damage. The nematode damage on tubers was less severe if the nematodes were added at 28 days, while the number of severe stem canker lesions increased if the fungus was added at 28 days. However, the time of nematode inoculation did not affect the incidence of fungal damage, hence the nematodes did not assist R. solani to infect the plant. Our results highlight the underestimated importance of root-lesion nematodes, not resulting in obvious above ground symptoms or misshaped tubers yet affecting the performance of other pathogens.

En el 2011 y 2012 se realizó una investigación de los agentes causales de las enfermedades asociadas al cultivo de Pitaya (Hylocereus costaricensis) en Costa Rica. Una de las más importantes durante el periodo de estudio, debido a su alta incidencia (100%), fue el cáncer en el tallo. Los síntomas observados fueron pequeñas manchas anaranjadas y rojizas irregulares y lesiones de coloración beige-grisáceas con el centro abultado. Para determinar el agente causal se recolectaron 85 muestras, cladodios completos con diferentes estados de desarrollo de la enfermedad, a partir de los cuales se realizaron 40 aislamientos. De estos creció un organismo en forma consistente, con el cual se llevó a cabo la identificación, caracterización morfológica y molecular así como la verificación de los postulados de Koch. De acuerdo con la forma de crecimiento, coloración del micelio y la morfología de los artroconidios y de las características moleculares (secuenciación de la región ITS), el patógeno asociado a los síntomas presentó un 99% de similitud con Neoscytalidium dimidiatum (Penz.) (no. de accesión KJ513460, del hongo aislado en Costa Rica). En las pruebas de los postulados de Koch se obtuvo un 92% de infección y se presentó una sintomatología similar a la observada en el campo, además, se recuperó el mismo organismo que fue inoculado. Este es el primer reporte de este hongo causando daños en el tallo de Hylocereus costaricensis en Costa Rica. In 2011 infected samples were examined to identify the causal agent of stem rot of Hylocereus costaricensis in Costa Rica. During the rainy season, symptoms associated with the disease included a small yellowish rot that typically started from the edge of the cladodium and extended inward towards the stem. Eventually the whole stem decayed. During the dry season, orange circular spots appeared on the stem, turning dark brown. For determine the causal agent, were collected 25 samples of each type of decay, complete cladodes with different stages of symptom development, from which 15 isolations of each disease were made. A bacterium was obtained consistently from each symptomatology, ith which the identification, characterization based on biochemical tests and molecular identification was carried out, as well as the verification of Koch’s postulates. Results indicated that the symptoms were caused by the same bacteria. Based on sequencing of the 16S region, the bacterium showed 99.2% similarity with Enterobacter hormaechei (KJ999997). According to Koch’s postulates, a similar symptomatology was obtained on plants from the field and on inoculated plants. This is the first report of this pathogen causing damage to the stem of Hylocereus costarricensis in American continent. Universidad de Costa Rica/[ED-313]/UCR/Costa Rica UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias Agroalimentarias::Centro de Investigación en Protección de Cultivos (CIPROC) UCR::Vicerrectoría de Docencia::Ciencias Agroalimentarias::Facultad de Ciencias Agroalimentarias

Summary Background Teratosphaeria gauchensis and T. zuluensis are closely related fungi that cause Teratosphaeria (previously Coniothyrium) stem canker disease on Eucalyptus species propagated in plantations for commercial purposes. This disease is present in many countries in which Eucalyptus trees are planted, and continues to spread with the international trade of infected plant germplasm. Taxonomy Fungi, Ascomycota, Pezizomycotina, Dothideomycetes, Dothideomycetidae, Capnodiales, Teratosphaeriaceae, Teratosphaeria. Identification The causal agents form dark masses of pycnidia that are visible on the surface of distinct stem cankers that typically form on young green stem tissues. Accurate diagnosis of the causal agents requires DNA sequence data. Host range Nine species of Eucalyptus are known to be affected. Of these, E. grandis and its hybrids, which include some of the most important planting stock globally, appear to be particularly vulnerable. Disease symptoms Small necrotic lesions develop on young green stem tissue. These lesions coalesce to form large cankers that exude gum. Epicormic shoots develop below the girdling canker and, in severe cases, trees die. Useful websites Mycobank, https://www.mycobank.org; Publications of the Forestry and Agricultural Biotechnology Institute (FABI), https://www.fabinet.up.ac.za/index.php/journals.

Stem canker (blackleg) is economically the most important disease of oilseed rape world­wide. This disease is caused by two species of pathogenic fungi of the genus Leptosphaeria: Leptosphaeria maculans (Desm.) ces. and de Not anamorf Phoma lingam (Tode. Fr.) Desmas, that causes stem canker and blackleg root and Leptospaeria biglobosa Shoem and Brun, that causes symptoms such as cancers of the upper part of the stem usually causing less damage, and can cause serious damage in countries with higher summer temperatures. Pathogenic fungi from one area to another are transmitted by ascospores and contaminated (infected) seed. In order to prove the modes of transmission of parasites, the test is done transmitting the parasite by seeds. Disinfected seeds of oilseed rape cultivars Quinta were submerged in the suspension of pycnospores. Submerged seeds were kept at 20 º C ± 1 º C and 12 h photoperiod for 48 h. After planting in plastic containers seeds were kept at 25 º C ± 1 º C and 12h photoperiod. In this experiment the following isolates were used: C-5, L-5, K-7, LJ-3, S-11, St-1, GS-3 and Lm as a reference isolate (L. maculans) fol­lowed by K-113, K-115 and Lb as a reference isolate (L. biglobosa). The symptoms were as­sessed after 7 and 14 days after emergence. Ratings were made with + which meant visible symptoms on cotyledons or hypocotyl and - what referred to healthy plants. After 7 days, the isolates (Lm, C-5, L-5, K-7, LJ-3, S-11, St-1, GS-3) were caused disease symptoms on plants of oilseed rape. This pathogenicity is reflected in the number of seedlings, leaf spots and stalk lodging cotyledons. At izolate Lb (reference isolate L. biglobosa) K-113 and K-115 after 7 and 14 days was not observed pathogenicity on rapeseed plants. The control variety seeds were soaked in distilled water. The trial was set in 6 reps, and pathogenicity of fungal isolates in relation to the control was tested using Dunette's test. After 14 days re-isolation of pathogen was done. Rak stabla (crna noga) je najznačajnije oboljenje uljane repice širom sveta. Bolest mogu prouzrokovati dve vrste patogenih gljiva iz roda Leptosphaeria: Leptosphaeria maculans (Desm.) ces. i de Not, anamorf Phoma lingam (Tode. Fr.) Desmas, koja prouzrokuje rak stabla i crnu nogu na korenu i Leptospaeria biglobosa Shoem and Brun, koja prouzrokuje simptome slične raku ali na gornjim delovima stabla obično prouzrokujući manje štete. Ozbiljnije štete mogu nastati u zemljama sa višim letnjim temperaturama. Patogene gljive se iz jedne oblasti u drugu prenose askosporama i zaraženim semenom. Da bi se utvrdili načini prenošenja parazita testirana je mogućnost prenošenja semenom. Dezinfikovano seme uljane repice, kultivara Quinta je potopljeno u suspenziju piknospora. Ovakvo seme je držano na 20ºC ± 1ºC i 12h fotoperiodu tokom 48 sati. Posle setve seme je preneto u plastične posude a temperatura čuvanja je bila 25ºC ± 1º C i fotoperiod od 12h. U ovom eksperimentu korišćeni su sledeći izolati: C-5, L-5, K-7, LJ-3, S-11, St-1, GS-3 i Lm kao referentni izolat (L. maculans) kao i K-113, K-115 i Lb kao referentni izolat (L. biglobosa). Ocena prisustva simptoma je rađena posle 7 i 14 dana od nicanja. Sa znakom + su se obeležavale biljke sa vidljivim simptomima na kotiledonima ili hipokotilu a znakom - biljke bez pojave simptoma. Posle 7 dana izolati Lm, C-5, L-5, K-7, LJ-3, S-11, St-1 i GS-3 su izazvali simptome bolesti na biljkama uljane repice. Patogenost se ogledala kroz broj oštećenih klijanaca, lisnih pega i povijenih kotiledona. Kod izolata Lb (referentni izolat L. biglobosa) K-113 i K-115 posle 7 i 14 dana nije primećena patogenost na biljkama uljane repice. Kontrolna varijanta semena je potapana u destilovanu vodu. Ogled je postavljen u 6 ponavljanja, i patogenost izolata gljive u odnosu na kontrolu je izražena Danetovim testom. Posle 14 dana izvedena je ponovna izolacija patogena.

Summary Leptosphaeria maculans is the fungus responsible for the stem canker disease of oilseed rape (Brassica napus). AvrLm3 and AvrLm4-7, two avirulence effector genes of L. maculans, are involved in an unusual relationship: AvrLm4-7 suppresses the Rlm3-mediated resistance. Here, we assessed AvrLm3 polymorphism in a collection of 235 L. maculans isolates. No field isolates exhibited deletion or inactivating mutations in AvrLm3, as observed for other L. maculans avirulence genes. Eleven isoforms of the AvrLm3 protein were found. In isolates virulent towards both Rlm3 and Rlm7 (a3a7), the loss of the Rlm3-mediated resistance response was due to two distinct mechanisms. First, when AvrLm4-7 was inactivated (deletion or inactivating mutations), amino acid substitutions in AvrLm3 generated virulent isoforms of the protein. Secondly, when only point mutations were observed in AvrLm4-7, a3a7 isolates still contained an avirulent allele of AvrLm3. Directed mutagenesis confirmed that some point mutations in AvrLm4-7 were sufficient for the fungus to escape Rlm7-mediated resistance while maintaining the suppression of the AvrLm3 phenotype. Signatures of positive selection were also identified in AvrLm3. The complex evolutionary mechanisms enabling L. maculans to escape Rlm3-mediated resistance while preserving AvrLm3 integrity, along with observed reduced aggressiveness of isolates silenced for AvrLm3, serves to emphasize the importance of this effector in pathogenicity towards B. napus. While the common response to resistance gene pressure is local selection of isolates depleted in the cognate avirulence gene, this example contributes to complexify the gene-for-gene concept of plant-pathogen evolution with a “camouflaged” model allowing retention of non-dispensable avirulence effectors. This article is protected by copyright. All rights reserved.

Stem canker of brassicas is a severe disease of oilseed rape in Australia, Canada and Europe, including Poland. The disease is caused by Leptosphaeria maculans and L. biglobosa - two pathogens belonging to the class Dothideomycetes. The species differ in pathogenicity, but they have identical shape and size of fruiting bodies and spores of the generative and vegetative stages. Both pathogens are often found together in infected tissues of oilseed rape plants. The main goal of the experiments was to measure the rate of pseudothecial maturation and to monitor ascospore concentration of L. maculans and L. biglobosa in air samples. The paper is the first investigation on the generative stage development of these two species in south-east Poland. The studies were done for three consecutive years (2005-2007), for six most important months in pathogen development and plant infection, including 3 months in the spring (March - May) and 3 months in the autumn (September - November). The stage of pseudothecial maturation was assessed visually, based on the development of asci and ascospores. Monitoring of spore concentration in the air was performed using a Hirst-type 7-day volumetric trap. It was proved that differences in pseudothecial maturation rate in south-east Poland, encompassing the climatic regions of the Carpathian Foothills and Cracow, do not exceed two weeks within one season. The first and the highest ascospore concentration dates depended on weather conditions in a particular season. The total number of spores during the studied seasons varied from 9 to 12 spores/m3, which was from 70 to 90 times lower than the average from five other monitoring sites around Poland. The short exposition to spore showers and very small concentrations of L. maculans and L. biglobosa ascospores in air samples were the most probable reasons for relatively small damage of oilseed rape crops by stem canker in the south-east part of Poland.

Stem canker of brassicas, also known as blackleg is the most damaging disease of many Brassicaceae. The disease is caused by Leptosphaeria maculans (Desm.) Ces et de Not. and L. biglobosa sp. nov., Shoemaker & Brun, which coexist in plants and resulting in disease symptoms and decreased yield, quantity and quality of cultivated vegetables and oilseed rape. The paper presents taxonomic relationships between these coexisting pathogen species, describes particular stages of their life cycles, summarizes the differences between the species, and reviews methods for their identification.

A complex of soybean diseases is caused by species from the genus Diaporthe /Phomopsis. Diaporthe phaseolorum (anamorph Phomopsis phaseoli) and Phomopsis longicolla (teleomorph unknown) are described as soybean pathogens. The first species includes three varieties: D. phaseolorum var. sojae, anamorph Phomopsis sojae, the causal agent of pod and stem blight, and D. phaseolorum var. caulivora and D. phaseolorum var. meridionalis, agents of northern and southern stem canker. The species P. longicolla is the most common and most damaging agent of soybean seed decay. The diseases caused by parasites from this genus were first observed and described on soybean in the USA. Presently they are widespread in most soybean-growing regions around the world. Soybean in Serbia is attacked by all pathogens mentioned above, except for D. phaseolorum var. meridionalis, the causal agent of the southern stem canker. D. phaseolorum var. caulivora (northern stem canker) has the greatest economic importance because it causes wilt and drying of plants during pod development and grain filling. Most intensive outbreaks of the disease occurred in the 1980s in southern and southeastern Banat, southern Backa and Srem. Prematurely wilted plants yielded 50% to 62% (depending on variety) less than healthy plants. Such heavy losses raised the question of the profitability of soybean growing. P. sojae and P. longicolla are less important in Serbia at the moment. Intensive infections of soybean seed break out occasionally. It was found that P. longicolla is the primary agent of seed decay and latent infections of seed in our country, although the other members of this genus may cause identical symptoms. Sexual cycle of development (teleomorph stage) of the fungi of the genus Diaporthe /Phomopsis form perithecia with asci and ascospores, while in the asexual cycle (anamorph stage) various types of conidia (α and β) are formed in pycnidia. The species P. longicolla has no teleomorph stage. Infected harvest residues and soybean seeds are main sources of the inoculum. D. phaseolorum var. caulivora forms perithecia with ascospores on overwintered harvest residues. During growing season, ascospores infect leaf laminas, petioles, or injured plant parts. P. sojae and P. longicolla form pycnidia (P. sojae sometimes forms perithecia), and they infect plants by means of conidia. Seed infection by conidia or ascospores unfolds via pods. Infected crop residues are the main source of the inoculum, while infected seeds help the parasites spread over large distances. Humidity and temperature (soil and air) are the mains factors that affect the dynamics of fruiting body formation, spore release, establishment of infection and the development of disease symptoms on soybean plants. Considerable attention is devoted to finding effective sources of resistance to the disease complex caused by species from the genus Diaporthe/Phomopsis. Resistance to the agents of seed decay was found in several introduced (PI) genotypes. This resistance is controlled by one or two pairs of dominant genes. Some cultivars are resistant to D. phaseolorum var. meridionalis (southern stem canker). Their resistance is controlled by two or four pairs of genes (Rdc). There is no physiological resistance to D. phaseolorum var. caulivora, the agent of northern stem canker, but significant differences in susceptibility level exist among soybean genotypes. Molecular techniques (RAPD, PCR-RFLP) showed that significant variability was present within the population of D. phaseolorum var. sojae. Other species of fungi were suspected to exist within that population, which could not be distinguished on the basis of their morphological characteristics. This was subsequently confirmed by the sequencing of different regions of ribosomal DNA.

International audience; Stem canker of oilseed rape (canola, Brassica napus) is associated with a species complex of two closely related fungal species, Leptosphaeria maculans and L. biglobosa. Of these, L. maculans is the most damaging and develops gene-for-gene relationships with the host. Here, a wide scale analysis of the L. maculans - L. biglobosa species complex was performed throughout the American continent (23 locations from Chile to Canada) plus several locations inWestern Australia for comparison purposes, based on a collection of 1132 isolates from infected tissues of a susceptible cultivar. Fungal species were discriminated on the basis of morphological, phytopathological and molecular criteria and showed that L. biglobosa was closely associated with L. maculans in most of the locations. Multiple gene phylogeny using sequences of ITS, actin and b-tubulin confirmed the prevalence of the L. biglobosa ‘canadensis’ sub-clade in Canada, whereas up to three different sub-clades of L. biglobosa were found in Georgia (USA). Race structure of L. maculans was investigated using a combination of pathogenicity tests and PCR amplification of avirulence alleles AvrLm1, AvrLm4 and AvrLm6. Three contrasting situations were observed: (i) race structure in Ontario, Chile and Georgia was related to that of European and Western Australian populations, with a low race diversity; (ii) only one race was found in Mexico, and not found outside of this country; (iii) a large diversity of races was observed in central Canada (Manitoba, Alberta and Saskatchewan) with very specific features including maintenance of avirulence alleles absent from Europe, absence of the AvrLm7 allele common in Europe (or eastern Canada) and wide location-to-location variability.

In March 2006, diseased soybean plants (Glycine max) were found in the municipalities of Passo Fundo and Coxilha, state of Rio Grande do Sul, Brazil, showing withered, brown leaves and light to reddish-brown discoloration and necrosis (canker) of the lower half of the stem. Cultural characteristics in potato dextrose agar showed identical patterns to those described for Diaporthe phaseolorum varcaulivora (Dpc). The rDNA-ITS sequence data from the Brazilian strain CH 40/06 (GenBank accession number EU622854) were compared to homologous sequences from the NCBI GenBank database, and a high similarity with Dpc strains was verified. A phylogenetic reconstruction was performed to compare EU622854 with other related sequences and grouped with Dpc. This group was highly supported by the bootstrap test (99%). Morphology of the Brazilian specimen was also similar to that described in the literature for Dpc, but the ascospores were slightly narrower than the American original material of Dpc. In field surveys performed in the 2006/07 crop season, Dpc was also found in five additional municipalities in RS. Em março de 2006, plantas de soja (Glycine max) doentes foram encontradas em Passo Fundo e em Coxilha, Rio Grande do Sul, Brasil. O quadro sintomatológico caracterizava-se por plantas com folhas secas e de coloração marrom; necrose (cancro) na metade inferior da haste, de coloração marrom-clara a marrom-avermelhada, envolvendo a mesma, seguindo-se murcha e morte da planta. Características culturais em batata dextrose ágar mostraram padrões idênticos àqueles publicados para Diaporthe phaseolorum varcaulivora (Dpc). Os dados de seqüenciamento de DNA do isolado brasileiro CH 40/06 (número de acesso GenBank EU622854) foram comparados às seqüências homólogas de NCBI GenBank, e alta similaridade foi verificada com outros isolados de Dpc. Árvore filogenética foi elaborada a fim de comparar EU622854 com outras seqüências de Dpc, cujo agrupamento foi altamente confirmado pelo teste bootstrap (99%). Características morfológicas foram similares às descritas para Dpc na literatura, embora os ascosporos do espécime brasileiro fossem mais estreitos que o original americanoNa safra 2006/07, Dpc foi também encontrado em outros cinco municípios do RS.

International audience; Following Agrobacterium tumefaciens-mediated mutagenesis in Leptosphaeria maculans, we identified the mutant 210, displaying total loss of pathogenicity towards its host plant (Brassica napus). Microscopic observations showed that m210 is unable to germinate on the host leaf surface and is thus blocked at the pre-penetration stage. The pathogenicity phenotype is linked with a single T-DNA insertion into the promoter region of a typical plasma membrane H(+)-ATPase-encoding gene, termed Lmpma1, thus leading to a twofold reduction in Lmpma1 expression. Since LmPMA1 is involved in intracellular pH homeostasis, we postulate that reduction in LmPMA1 activity disturbs the electrochemical transmembrane gradient in m210, thus leading to conidia defective in turgor pressure generation on leaf surface. Whole genome survey showed that L. maculans possesses a second plasma membrane H(+)-ATPase-encoding gene, termed Lmpma2. Silencing experiments, expression analyses and phylogenetic studies allowed us to highlight the essential role assumed by the Lmpma1 isoform in L.maculans pathogenicity.

The sphinganine-analog mycotoxins (SAMs) fumonisin B1 and AAL toxins are inhibitors of eukaryotic sphinganine N-acyltransferase in vitro. Treatment of eukaryotes with SAMs generally results in an accumulation of sphingoid base precursors and a depletion of complex sphingolipids. The asc,asc genotypes of tomato (Lycopersicon esculentum) and Nicotiana umbratica are sensitive to SAMs and host of the AAL toxin-producing fungus Alternaria alternata f. sp. lycopersici. Codominant insensitivity to SAMs in tomato is mediated by the Asc-1 gene, and sensitivity is associated with a frame-shift mutation present in asc-1. We investigated the function of Asc-1 in mediating insensitivity to SAMs and resistance to the fungus by overexpression of asc-1 and Asc-1. In this study, it is shown that overexpression of these genes did not lead to visual symptoms in tomato hairy roots and N. umbratica plants. Overexpression of asc-1 did not influence the (in)sensitivity to SAMs. Overexpression of Asc-1 in SAM-sensitive hairy roots and N. umbratica plants, however, mediated a high insensitivity to SAMs and resistance to plant infection by Alternaria alternata f. sp. lycopersici.

The necrotrophic fungus Alternaria alternata f.sp. lycopersici produces AAL toxins that cause necrosis in tomato tissues with high specificity. Resistance or susceptibility of tomato to the fungus and insensitivity or sensitivity to AAL toxins are determined by a single locus, Asc. In order to investigate further the specificity of the host-fungus interaction, 200 species of Solanaceae were tested for their sensitivity to AAL toxins T-A and T-B. Twenty-five species were found to be sensitive to AAL toxins at a concentration of (0.2 mum) used for distinguishing sensitive and insensitive tomato plants. Three species were as sensitive as the sensitive tomato line, indicating that AAL toxins effectively act on a broader range of plant species within the Solanaceae.

LAMP is an innovative, simple, rapid, specific and cost-effective nucleic acid amplification method. Due to the use of a special enzyme – GspSSD polymerase, the reaction takes a short time and can be performed at isothermal conditions. The sensitivity and specificity of LAMP technique is significantly higher, than standard PCR techniques, as two or three specific primer pairs are used. The technique is regarded as a useful tool for the detection and identification of plant pathogens. In this work, LAMP was used to study the composition of the population of fungi of the genus Leptosphaeria, causing a damaging disease of oilseed rape, called blackleg or stem canker. The detection concerned DNA present in fungal spores contained in air samples obtained using Hirst-type volumetric trap, in Pomerania (north Poland) in 2010. The results achieved using the LAMP technique were similar to these obtained with previously used, highly specific method of Real-time PCR. Conducting LAMP reaction was much easier and less time-and cost-consuming, due to a simplified method of DNA isolation of pathogens from plant tissues. Then, the LAMP technique was used to assess the composition of the population of Leptosphaeria spp. in plants of oil- seed rape collected from the field in the Opole region (south-we- stern part of Poland) in 2013. In contrast to studies conducted in 2002–2003, the analysis of leaf symptoms showed a higher pro- portion of L. maculans compared to L. biglobosa, what reflects changes in the composition of pathogen population of fungi causing blackleg on oilseed rape in this part of Poland.

The anamorphic genus Phoma was subdivided into nine sections based on morphological characters, and included teleomorphs in Didymella, Leptosphaeria, Pleospora and Mycosphaerella, suggesting the polyphyly of the genus. Recent molecular, phylogenetic studies led to the conclusion that Phoma should be restricted to Didymellaceae. The present study focuses on the taxonomy of excluded Phoma species, currently classified in Phoma sections Plenodomus, Heterospora and Pilosa. Species of Leptosphaeria and Phoma section Plenodomus are reclassified in Plenodomus, Subplenodomus gen. nov., Leptosphaeria and Paraleptosphaeria gen. nov., based on the phylogeny determined by analysis of sequence data of the large subunit 28S nrDNA (LSU) and Internal Transcribed Spacer regions 1 & 2 and 5.8S nrDNA (ITS). Phoma heteromorphospora, type species of Phoma section Heterospora, and its allied species Phoma dimorphospora, are transferred to the genus Heterospora stat. nov. The Phoma acuta complex (teleomorph Leptosphaeria doliolum), is revised based on a multilocus sequence analysis of the LSU, ITS, small subunit 18S nrDNA (SSU), β-tubulin (TUB), and chitin synthase 1 (CHS-1) regions. Species of Phoma section Pilosa and allied Ascochyta species were determined to belong to Pleosporaceae based on analysis of actin (ACT) sequence data. Anamorphs that are similar morphologically to Phoma and described in Ascochyta, Asteromella, Coniothyrium, Plectophomella, Pleurophoma and Pyrenochaeta are included in this study. Phoma-like species, which grouped outside the Pleosporineae based on a LSU sequence analysis, are transferred to the genera Aposphaeria, Paraconiothyrium and Westerdykella. The genera Medicopsis gen. nov. and Nigrograna gen. nov. are introduced to accommodate the medically important species formerly known as Pyrenochaeta romeroi and Pyrenochaeta mackinnonii, respectively. Taxonomic novelties: New genera: Medicopsis Gruyter, Verkley & Crous, Nigrograna Gruyter, Verkley & Crous, Paraleptosphaeria Gruyter, Verkley & Crous, Subplenodomus Gruyter, Verkley & Crous. New species: Aposphaeria corallinolutea Gruyter, Aveskamp & Verkley, Paraconiothyrium maculicutis Verkley & Gruyter. New combinations: Coniothyrium carteri (Gruyter & Boerema) Verkley & Gruyter, C. dolichi (Mohanty) Verkley & Gruyter, C. glycines (R.B. Stewart) Verkley & Gruyter, C. multiporum (V.H. Pawar, P.N. Mathur & Thirum.) Verkley & Gruyter, C. telephii (Allesch.) Verkley & Gruyter, Heterospora (Boerema, Gruyter & Noordel.) Gruyter, Verkley & Crous, H. chenopodii (Westend.) Gruyter, Aveskamp & Verkley, H. dimorphospora (Speg.) Gruyter, Aveskamp & Verkley, Leptosphaeria errabunda (Desm.) Gruyter, Aveskamp & Verkley, L. etheridgei (L.J. Hutchison & Y. Hirats.) Gruyter, Aveskamp & Verkley, L. macrocapsa (Trail) Gruyter, Aveskamp & Verkley, L. pedicularis (Fuckel) Gruyter, Aveskamp & Verkley, L. rubefaciens (Togliani) Gruyter, Aveskamp & Verkley, L. sclerotioides (Sacc.) Gruyter, Aveskamp & Verkley, L. sydowii (Boerema, Kesteren & Loer.) Gruyter, Aveskamp & Verkley, L. veronicae (Hollos) Gruyter, Aveskamp & Verkley, Medicopsis romeroi (Borelli) Gruyter, Verkley & Crous, Nigrograna mackinnonii (Borelli) Gruyter, Verkley & Crous, Paraconiothyrium flavescens (Gruyter, Noordel. & Boerema) Verkley & Gruyter, Paracon. fuckelii (Sacc.) Verkley & Gruyter, Paracon. fusco-maculans (Sacc.) Verkley & Gruyter, Paracon. lini (Pass.) Verkley & Gruyter, Paracon. tiliae (F. Rudolphi) Verkley & Gruyter, Paraleptosphaeria dryadis (Johanson) Gruyter, Aveskamp & Verkley, Paralept. macrospora (Thum.) Gruyter, Aveskamp & Verkley, Paralept. nitschkei (Rehm ex G. Winter) Gruyter, Aveskamp & Verkley, Paralept. orobanches (Schweinitz: Fr.) Gruyter, Aveskamp & Verkley, Paralept. praetermissa (P. Karst.) Gruyter, Aveskamp & Verkley, Plenodomus agnitus (Desm.) Gruyter, Aveskamp & Verkley, Plen. biglobosus (Shoemaker & H. Brun) Gruyter, Aveskamp & Verkley, Plen. chrysanthemi (Zachos, Constantinou & Panag.) Gruyter, Aveskamp & Verkley, Plen. collinsoniae (Dearn. & House) Gruyter, Aveskamp & Verkley, Plen. confertus (Niessl ex Sacc.) Gruyter, Aveskamp & Verkley, Plen. congestus (M.T. Lucas) Gruyter, Aveskamp & Verkley, Plen. enteroleucus (Sacc.) Gruyter, Aveskamp & Verkley, Plen. fallaciosus (Berl.) Gruyter, Aveskamp & Verkley, Plen. hendersoniae (Fuckel) Gruyter, Aveskamp & Verkley, Plen. influorescens (Boerema & Loer.) Gruyter, Aveskamp & Verkley, Plen. libanotidis (Fuckel) Gruyter, Aveskamp & Verkley, Plen. lindquistii (Frezzi) Gruyter, Aveskamp & Verkley, Plen. lupini (Ellis & Everh.) Gruyter, Aveskamp & Verkley, Plen. pimpinellae (Lowen & Sivan.) Gruyter, Aveskamp & Verkley, Plen. tracheiphilus (Petri) Gruyter, Aveskamp & Verkley, Plen. visci (Moesz) Gruyter, Aveskamp & Verkley, Pleospora fallens (Sacc.) Gruyter & Verkley, Pleo. flavigena (Constantinou & Aa) Gruyter & Verkley, Pleo. incompta (Sacc. & Martelli) Gruyter & Verkley, Pyrenochaetopsis pratorum (P.R. Johnst. & Boerema) Gruyter, Aveskamp & Verkley, Subplenodomus apiicola (Kleb.) Gruyter, Aveskamp & Verkley, Subplen. drobnjacensis (Bubak) Gruyter, Aveskamp & Verkley, Subplen. valerianae (Henn.) Gruyter, Aveskamp & Verkley, Subplen. violicola (P. Syd.) Gruyter, Aveskamp & Verkley, Westerdykella capitulum (V.H. Pawar, P.N. Mathur & Thirum.) de Gruyter, Aveskamp & Verkley, W. minutispora (P.N. Mathur ex Gruyter & Noordel.) Gruyter, Aveskamp & Verkley. New names: Pleospora angustis Gruyter & Verkley, Pleospora halimiones Gruyter & Verkley.

Stem canker in blueberries (Vaccinium corymbosum L.) is a disease which is widely distributed around the world and of growing importance in Chile, associated with Neofusicoccum species. The purpose of the present work was to study the effects of water activity (aw) and temperature on mycelial growth in vitro of N. arbuti, N. australe and N. parvum, obtained from blueberries with symptoms of stem canker. According to the results obtained, 25 oC was the optimum temperature for mycelial growth of N. arbuti, N. australe and N. parvum. These species presented minimal growth at 35 oC and none at 0 and 5 oC. The effect of aw on mycelial growth was significant (P≤0.001) and varied with the species of Neofusicoccum and the temperature. At 25 oC, the optimum temperature, the three species of Neofusicoccum presented minimum growth with aw of 0.900 and 0.920, while optimum mycelial growth was found with aw equal to 0.990. Independent of the Neofusicoccum species, the effect of temperature (T) and aw on mycelial growth was best explained by y = -721.62 + 0.24T + 788.92aw. (R2 adj = 0.77, P≤0.001), suggesting that T and aw jointly explained 77% of the total variation on mycelial growth of Neofusicoccum spp. The results obtained provide important information for a better understanding of the biology of these plant pathogenic fungi, and suggest that temperature and aw could be important parameters for understanding factors that affect the inoculum production, colonization and infection by Neofusicoccum spp. in blueberry plantations. La cancrosis de la madera en arandanos (Vaccinium corymbosum L.) es una enfermedad de amplia distribucion en el mundo y de creciente importancia en Chile, asociada a la accion de especies de Neofusicoccum. Este trabajo tuvo como objetivo estudiar el efecto de la actividad de agua (aw) y de la temperatura sobre el crecimiento micelial in vitro de N. arbuti, N. australe y N. parvum, obtenidos de arandanos con sintomas de cancrosis de la madera. Segun los resultados obtenidos, 25 oC fue la temperatura optima para el crecimiento micelial de N. arbuti, N. australe y N. parvum. Estas especies crecieron minimamente a 35 oC y no crecieron a 0 y 5 oC. La aw afecto significativamente (P≤0.001) el crecimiento micelial y su efecto vario en funcion de la especie de Neofusicoccum y de la temperatura. A 25 oC, temperatura optima, las tres especies de Neofusicoccum crecieron minimamente con aw de 0,900 y 0,920, siendo optimo el crecimiento micelial con aw igual a 0,990. Independientemente de Neofusicoccum spp., el crecimiento micelial (y) entre 20 y 30 oC se explico a traves de y = -721,62 + 0,24T + 788,92aw. (R2 = 0,77, P≤0,001). Los resultados obtenidos aporta informacion para la mejor compresion de la biologia de estos hongos fitopatogenos y sugieren que la aw es un importante factor que podria afectar la produccion de inoculo, colonizacion y la infeccion por Neofusicoccum spp., en arandanos.

271-274 Stem canker caused by Diaporthe phaseolorum var. caulivora is an important disease of soybean in Argentina. The objective of this study was to determine its survival ability in artificially infested straw under laboratory and field conditions. In laboratory, stem pieces of soybean, maize, sorghum, sunflower, potato and wheat were autoclaved, placed in petri dishes on Potato Dextrose Agar and Water Agar, and inoculated with a 7-day-old pathogen culture. All crop residues were colonized and produced perithecia. Debris artificially infested with D. phaseolorum var. caulivora were placed in plastic net bags and transferred to an un-cropped area in a field plot at the University of Buenos Aires. Straws were left on the ground from winter to spring season in both 2007 and 2008 years. After 6 months abundant perithecia were developed in all straws. However, a higher number of perithecia on soybean and sunflower compared to maize, sorghum and potato was determined. These findings suggest that other crops, besides soybean, could maintain alive the inoculum of Diaporthe phaseolorum var. caulivora from soybean for at least 6-7 months.

Fungi are of primary ecological, biotechnological and economic importance. Many fundamental biological processes that are shared by animals and fungi are studied in fungi due to their experimental tractability. Many fungi are pathogens or mutualists and are model systems to analyse effector genes and their mechanisms of diversification. In this study, we report the genome sequence of the phytopathogenic ascomycete Leptosphaeria maculans and characterize its repertoire of protein effectors. The L. maculans genome has an unusual bipartite structure with alternating distinct guanine and cytosine-equilibrated and adenine and thymine (AT)-rich blocks of homogenous nucleotide composition. The AT-rich blocks comprise one-third of the genome and contain effector genes and families of transposable elements, both of which are affected by repeat-induced point mutation, a fungal-specific genome defence mechanism. This genomic environment for effectors promotes rapid sequence diversification and underpins the evolutionary potential of the fungus to adapt rapidly to novel host-derived constraints., Leptosphaeria maculans is a plant pathogen that causes stem canker of oilseed rape. Rouxel et al. sequence and describe the key features of the L. maculans genome, including partitioning into AT-rich blocks that are enriched in effector genes and transposable elements affected by repeat-induced point mutation.

International audience; Three avirulence genes, AvrLm1, AvrLm6, and AvrLm4-7, were recently identified in Leptosphaeria maculans and found to be localized as solo genes within large noncoding, heterochromatin-like regions mainly composed of retrotransposons, truncated and degenerated by repeat-induced point mutation (RIP). The Rlm6 resistance gene has been overcome within 3 years in outdoor experiments in France and, here, we investigate the molecular basis of evolution toward virulence at the AvrLm6 locus. A region of 235 kb was sequenced in a virulent isolate and showed the deletion of AvrLm6 and three divergent mosaics of retrotransposons. AvrLm6 was found to be absent from 66% of 70 virulent isolates, with multiple events of deletion. The sequencing of virulent alleles in 24 isolates revealed a few cases of point mutations that had created stop codons in the sequence. The most frequent mutation events, however, were RIP, leading to the modification of 4 to 9% of the bases compared with the avirulent allele and generating 2 to 4 stop codons. Thus, RIP is described for the first time as an efficient mechanism leading to virulence and the multiple patterns of mutation observed suggest that multiple RIP events could occur independently in a single field population during 1 year.

International audience; Random insertional mutagenesis was used to investigate pathogenicity determinants in Leptosphaeria maculans. One tagged nonpathogenic mutant, termed m20, was analysed in detail here. The mutant phenotype was investigated by microscopic analyses of infected plant tissues and in vitro growth assays. Complementation and silencing experiments were used to identify the altered gene. Its function was determined by bioinformatics analyses, cell biology experiments and functional studies. The mutant was blocked at the invasive growth phase after an unaffected initial penetration stage, and displayed a reduced growth rate and an aberrant hyphal morphology in vitro. The T-DNA insertion occurred in the intergenic region between two head-to-tail genes, leading to a complex deregulation of their expression. The unique gene accounting for the mutant phenotype was suggested to be the orthologue of the poorly conserved Saccharomyces cerevisiae gpi15, which encodes for one component of the glycosylphosphatidylinositol (GPI) anchor biosynthesis pathway. Consistent with this predicted function, a functional translational fusion with the green fluorescent protein (GFP) was targeted to the endoplasmic reticulum. Moreover, the mutant exhibited an altered cell wall and addition of glucosamine relieved growth defects. It is concluded that the GPI anchor biosynthetic pathway is required for morphogenesis, cell wall integrity and pathogenicity in Leptosphaeria maculans.

Map-based cloning of avirulence genes of the AvrLm1-2-6 cluster was recently undertaken in Leptosphaeria maculans and led to the identification of AvrLm1. The ensuing chromosome walk toward AvrLm6 resulted in the delineation of a 562-kb bacterial artificial chromosome (BAC) clone contig in an avirulent isolate. Following sequencing of the contig and sequence comparison with a virulent isolate, four AvrLm6 candidate genes were identified. Complementation of the virulent isolate with the four candidates was performed and one gene was found to fully restore the avirulent phenotype on Rlm6 oilseed rape genotypes. AvrLm6 was found to be located in the same genome context as AvrLm1, because it is a solo gene surrounded by 85 and 48 kb of degenerated repeats on its 5′ and 3′ sides, respectively. AvrLm6 is an orphan gene encoding a small, potentially secreted, cysteine-rich protein. Comparison of AvrLm1 and AvrLm6 expressions by quantitative reverse-transcription polymerase chain reaction revealed that both genes are highly overexpressed during primary leaf infection. Using RNA interference, decreasing expression of AvrLm6 was shown to result in virulence toward Rlm6 genotypes whenever the expression was reduced by more than 60% compared with the wild-type isolate.

A two year field experiment was conducted twice to assess effects of chemical soil disinfection at planting and methods of harvesting potatoes on stem infection withRhizoctonia solani in the subsequent year. In the first year of the experiments seven methods. including one with soil disinfection at planting, were applied in August. In the following year,R. solani stem and stolon infection (disease severity) on potato plants were assessed in June. Soil treatment at planting with pencycuron resulted in lowest disease severity in the following year. Compared with chemical haulm killing and haulm pulling. immature-crop-harvesting also resulted in a lower disease severity, but only when black scurf was scarce on tubers at harvest in the preceding year.

The infection process ofRhizoctonia solani AG-3 was studied on potato sprouts, cv. Bintje, in growth chamber trials at 15 °C. Initially hyphae ofR. solani grew predominantly in the longitudinal direction of the sprouts (runner hyphae). They tended to follow the junctions between epidermis cells as was observed by SEM. The hyphae formed side-branches mainly half-way of the subterranean parts of the sprouts. They branched several times with short swollen cells to form infection cushions. Lesions developed only underneath the infection cushions and were first observed five days after inoculation. The necrotic area was proportional to the area covered with infection cushions on the sprouts. Depth of the lesions could extend up to the vascular bundle. Sprouts were colonized only in healthy tissue in the epidermal layer underneath the infection cushion and in necrotic tissue. A few days after appearance of the lesions,R. solani formed brown, uninfective mycelium on and in the circumference of these lesions.

The effects of soil-borneRhizoctonia solani on yield and quality of potato were studied by an extensive individual plant sampling procedure. From 1983 to 1986, stem canker and stolon pruning were examined in 10 768 plants growing on a sandy soil. Tuber yield and quality and haulm yield per plant were also recorded.

Diaporthe aspalathi (Syn. Diaporthe phaseolorum var. meridionalis) is the causal agent of the southern stem canker (SSC) disease in soybean. This disease can kill plants from the middle to the end of the growing season resulting in severe yield loss. The mechanisms of SSC disease development and pathogen invasion of soybean are not fully understood. The genome sequence of D. aspalathi has not been described. In this article, we report the successful assembly of the draft genome sequence of a D. aspalathi isolate, designated MS-SSC91, that was isolated from the stem of a field-grown soybean plant in Mississippi, USA in 2006. This study represents the first reported genome sequence of D. aspalathi in the Diaporthe-Phomopsis complex. The whole genome shotgun sequence of the MS-SSC91 isolate has been deposited at DDBJ/EMBL/GenBank under the accession LJJS00000000 and the sequences could be found at the site http://www.ncbi.nlm.nih.gov/assembly/GCA_001447215.1/. The MS-SSC91 genome sequences will provide information on the genetic basis of fungal infection of the soybean stem. It is valuable for studying soybean-fungal interactions and developing new control strategies for this pathogen. Keywords: Draft genome, Diaporthe aspalathi, Stem Canker, Sequences, Soybean pathogen