Niebla ceruchis from Laguna Figueroa: dimorphic spore morphology and secondary compounds localized in pycnidia and apothecia

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Enzien, M.; Margulis, L.

1988-01-01

During and after the floods of 1979-80 Niebla ceruchis growing epiphytically on Lycium brevipes was one of the dominant aspects of the vegetation in the coastal dunal complex bordering the microbial mats at Laguna Figueroa, Baja California Norte, Mexico. The lichen on denuded branches of Lycium was far more extensively distributed than Lycium lacking lichen. Unusual traits of this Niebla ceruchis strain, namely localization of lichen compounds in the mycobiont reproductive structures (pycnidia and apothecia) and simultaneous presence of bilocular and quadrilocular ascospores, are reported. The abundance of this coastal lichen cover at the microbial mat site has persisted through April 1988.

Influence of culture media and environmental factors on mycelial growth and pycnidial production of Sphaeropsis pyriputrescens.

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Kim, Y K; Xiao, C L; Rogers, J D

2005-01-01

Sphaeropsis pyriputrescens, the causal agent of Sphaeropsis rot of pears and apples, is a recently described species. In this study the effects of culture media, temperature, water potential, pH and light on mycelial growth and pycnidial production of S. pyriputrescens were evaluated. Apple juice agar and pear juice agar were most suitable for mycelial growth of all six isolates tested. Cornmeal agar was not suitable for either mycelial growth or pycnidial production. The fungus grew from -3 to 25 C, with optimum growth at 20 C and no growth at 30 C. The fungus grew at water potential as low as -5.6 MPa on potassium chloride-amended potato-dextrose agar (PDA). Hyphal extension was not observed at -7.3 MPa after 10 d incubation, but growth resumed when the inoculum plugs were placed on PDA. The fungus grew at pH 3.3-6.3 and optimum growth was at pH 3.3-4.2. No mycelial growth was observed at pH above 7.2 after 10 d incubation, but growth resumed when the inoculum plugs were transferred onto PDA. Regardless of medium tested, few pycnidia formed at 20 C in the dark. Pycnidial production was enhanced significantly by fluorescent light, but continuous light appeared to reduce pycnidial production, depending on the medium. Oatmeal agar (OMA) was most suitable for production of pycnidia and conidia. Pycnidia that formed on 3 wk old OMA cultures at 20 C under 12 h light/12 h dark produced abundant conidia, and the technique is recommended for inoculum production.

Protein Kinase A subunits of the ascomycete pathogen Mycosphaerella graminicola regulate asexual fructification, filamentation, melanization and osmosensing

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Mehrabi, R.; Kema, G.H.J.

2006-01-01

As in many fungi, asexual reproduction of Mycosphaerella graminicola in planta is a complex process that requires proper differentiation of the infectious hyphae in the substomatal cavities of foliar tissue before pycnidia with conidia can be formed. In this study, we have investigated the role of

Phacidiopycnis washingtonensis--a new species associated with pome fruits from Washington State.

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Xiao, C L; Rogers, J D; Kim, Y K; Liu, Q

2005-01-01

A new species of Phacidiopycnis associated with pome fruits is described. The fungus causes fruit rot on apples during storage and is associated with a twig dieback and canker disease of crabapple trees and dead twigs of pear trees. To characterize the biology of the fungus and compare it with Ph. piri, the type species of the genus, effects of nine media and light on mycelial growth and pycnidial production, mycelial growth in response to temperature and mode of conidial germination in response to nutrient were determined. Apple-juice agar, pear-juice agar, prune-juice agar, potato-dextrose agar (PDA) and malt-extract agar, Czapek-Dox agar and oatmeal agar (OMA) favored mycelial growth. Cornmeal agar (CMA) did not favor mycelial growth. Light effect on pycnidial formation was medium dependent. Abundant pycnidia with mature conidia formed in 14 d old PDA and OMA cultures at 20 C, regardless of light, whereas none or very few pycnidia formed on other media in the dark. Fluorescent light stimulated formation of pycnidia except on CMA. The fungus grew at -3-25 C, with optimum growth at 15-20 C. Conidia germinated either by forming germ tubes or less often by budding. Budding of conidia occurred in 1 and 10% pear-juice solutions but not in 100% pear-juice solution. Six isolates of Ph. washingtonensis from different species of pome fruits had identical ITS sequences. The sizes of the ITS region were the same for both Ph. washingtonensis and Ph. piri, and four polymorphic nucleotide sites were found in the ITS region between Ph. washingtonensis and Ph. piri. The similarity in ITS sequences between these two taxa is confirmatory evidence for the erection of the new species of Phacidiopycnis associated with pome fruits we describe here.

Presencia de Phomopsis viticola (Saccardo Saccardo, agente causal de excoriosis, en viñedos de Córdoba, Argentina Presence of Phomopsis viticola (Saccardo Saccardo, causal agent of excoriosis, in vineyards of Córdoba, Argentina

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C.I. Cragnolini

2009-12-01

Full Text Available Desde 2001 se observan en viñedos de Córdoba, Argentina, hojas de Vitis vinifera y V. labrusca con manchas cloróticas y centro necrótico. Las plantas presentan nervaduras, pecíolos, raquis y entrenudos con puntos necróticos que al confluir agrietan el tejido cortical. Las bayas pueden marchitarse y desprenderse. En invierno los sarmientos toman color blanquecino con manchas negras, y en ellos y en órganos secos aparecen picnidios con cirros blanco-crema con conidios alfa y beta. El objetivo fue confirmar la presencia de Phomopsis viticola (excoriosis y cuantificar su incidencia. Se realizaron aislamientos a partir de pecíolos, brotes, zarcillos, bayas y sarmientos que produjeron micelio claro con picnidios y conidios alfa (7,5m±1,5 x 2,8m±2 y beta (19m±1 x 1,2m±0,5 o sólo alfa, típicos de P. viticola. La identificación fue confirmada mediante secuenciación molecular. En brotes de las variedades Malbec y Pinot noir inoculados con 3 10(4 conidios/mL, las hojas mostraron lesiones y picnidios 8 días después, y las bayas cirros amarillos a los 15-18 días. P. viticola (Sacc. Sacc. fue reaislado de los cirros y las colonias produjeron picnidios y conidios característicos 10-20 días después. Se detectó excoriosis en 80% de los viñedos evaluados, con mayor incidencia en viñedos añosos de las variedades Pinot noir e Isabella.Leaves with chlorotic spots and necrotic centers have been observed in Vitis vinifera and V. labrusca in vineyards of Córdoba, Argentina, since 2001. Plants exhibit veins, petioles, rachises, and internodes with necrotic spots that coalesce to form cracks in the cortical tissue. Grape berries can shrivel and fall off. Winter canes became whitish with black spots; pycnidia appear in canes and in dry organs, exuding whitish-cream cirri with alpha and beta conidia. The aim of this work was to confirm the presence of Phomopsis viticola (excoriosis and quantify its incidence. Isolates obtained from petioles

First report of shoot blight and dieback caused by Diplodia pinea on Pinus pinaster and P. radiata trees in Tunisia

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B.T. Linaldeddu

2009-01-01

Full Text Available Damage caused by Diplodia pinea on Pinus pinaster and P. radiata forests are reported for the fi rst time in Tunisia. The affected plants show shoot blight, canker and branch dieback. On decaying and dead branches pycnidia of D. pinea are observed. The fungus was repeatedly isolated from the bark of symptomatic branches. The results of pathogenicity tests confirm the virulence of D. pinea and the susceptibility of both Pinus species to infection.

Neofusicoccum luteum associated with leaf necrosis and fruit rot of olives in New South Wales, Australia

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V. Sergeeva

2009-09-01

Full Text Available Neofusicoccum luteum is reported for the first time from olives (Olea europaea, causing fruit rot and leaf necrosis. Affected fruits initially became brown with pycnidia developing on the surface, later drying out and becoming mummified. The fungus was shown to be pathogenic on both fruits and leaves. The association of Botryosphaeriaceae with rotting olive fruits in Mediterranean regions and in New South Wales, Australia indicates that these fungi play a significant role in fruit rots of olives and deserve greater attention.

Laboratory invesitgations of variability of Ascochyta fabae Speg. isolates obtained from horse bean

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Antoni Józef Filipowicz

2014-08-01

Full Text Available Fifty five isolates of Ascochyta fabae Speg. were investigated. They were selected from 1650 isolates of this fungus obtained from horse bean seeds in 1974-1976. All the isolates grew and sporulated on Potato Dextrose Agar, Malt Agar and Horse Bean Agar. The rate of their growth amounted to 1-4 mm per 24 hours. The variability of isolates in size of pycnidia and conidia and number of sepia was noticed. A few spores with untypical shapes were observed as well.

Saprophytic Activity and Sporulation of Cryphonectria parasitica on Dead Chestnut Wood in Forests with Naturally Established Hypovirulence.

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Prospero, S; Conedera, M; Heiniger, U; Rigling, D

2006-12-01

ABSTRACT Sustainable biological control of the chestnut blight fungus Crypho-nectria parasitica with hypovirulence depends on the production and dissemination of hypovirus-infected propagules of the pathogen. We investigated the ability of C. parasitica to sporulate and produce hypo-virus-infected spores on recently dead chestnut wood in coppice stands in southern Switzerland where hypovirulence has been naturally established. The number and type (active, inactive, or none) of cankers was assessed on experimentally cut and stacked stems, firewood stacks, and natural dead wood. Hypovirus-free and hypovirus-infected strains readily survived for more than 1 year in the chestnut blight cankers of the stacked stems. Sporulation of C. parasitica was observed on the surface of preexisting inactive and active cankers, as well as on newly colonized bark areas and was significantly more abundant than on comparable cankers on living stems. On all types of dead wood, we observed more stromata with perithecia than with pycnidia; however, a large proportion of the stromata was not differentiated. All perithecia examined yielded only hypovirus-free ascospores. The incidence of pycnidia that produced hypovirus-infected conidia ranged from 5% on natural dead wood to 41% on the experimental stacks. The mean virus transmission rate into conidia was 69%. Our study demonstrates a considerable saprophytic activity of C. parasitica on recently dead chestnut wood and supports the hypothesis of a role of this saprophytic phase in the epidemiology of hypovirulence.

Complete life cycle of the lichen fungus Calopadia puiggarii (Pilocarpaceae, Ascomycetes) documented in situ: propagule dispersal, establishment of symbiosis, thallus development, and formation of sexual and asexual reproductive structures.

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Sanders, William B

2014-11-01

The life histories of lichen fungi are not well known and cannot be readily studied in laboratory culture. This work documents in situ the complete life cycle of the widespread crustose lichen Calopadia puiggarii, which reproduces sexually and asexually on the surfaces of leaves. Plastic cover slips held in a mesh frame were placed over leaves in the field and successively removed for microphotography of colonizing lichens. Macroconidia produced within campylidia encircled photobiont cells and codispersed with them, a feature not reported previously for C. puiggarii. Dispersed macroconidia readily germinated and lichenized the photobionts. Algal cells were often dislodged from the encircling macroconidia, providing a likely source for the free-living populations observed. Aposymbiotically dispersed ascospores germinated and lichenized nearby algal cells soon after dispersal. Thallus areolae merged readily in early development, although adjacent mature thalli were often separated by growth inhibition zones. Pycnidia are reported for the first time in Calopadia; their pyriform microconidia probably function as male gametes (spermatia). Pycnidia, apothecia, and campylidia began development similarly as darkly pigmented primordia on the fungal prothallus. Abundant dispersal of ascospores, conidia, and photobionts allows C. puiggarii to quickly colonize leaves with the dual advantages of sexual and asexual reproduction, and with the added convenience of having its algal partner on hand. Fusions and prothallic capture of additional algae provide many opportunities for multiple mycobiont and photobiont genotypes to be combined in a single thallus, but the outcomes of such events remain to be explored. © 2014 Botanical Society of America, Inc.

A comparative analysis of the heterotrimeric G-protein Gα, Gβ and Gγ subunits in the wheat pathogen Stagonospora nodorum

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Gummer Joel P A

2012-07-01

Full Text Available Abstract Background It has been well established that the Gα subunit of the heterotrimeric G-protein in the wheat pathogen Stagonospora nodorum is required for a variety of phenotypes including pathogenicity, melanisation and asexual differentiation. The roles though of the Gγ and Gβ subunits though were unclear. The objective of this study was to identify and understand the role of these subunits and assess their requirement for pathogenicity and development. Results G-protein Gγ and Gβ subunits, named Gga1 and Gba1 respectively, were identified in the Stagonospora nodorum genome by comparative analysis with known fungal orthologues. A reverse genetics technique was used to study the role of these and revealed that the mutant strains displayed altered in vitro growth including a differential response to a variety of exogenous carbon sources. Pathogenicity assays showed that Stagonospora nodorum strains lacking Gba1 were essentially non-pathogenic whilst Gga1-impaired strains displayed significantly slower growth in planta. Subsequent sporulation assays showed that like the previously described Gα subunit mutants, both Gba1 and Gga1 were required for asexual sporulation with neither mutant strain being able to differentiate either pycnidia nor pycnidiospores under normal growth conditions. Continued incubation at 4°C was found to complement the mutation in each of the G-protein subunits with nearly wild-type levels of pycnidia recovered. Conclusion This study provides further evidence on the significance of cAMP-dependent signal transduction for many aspects of fungal development and pathogenicity. The observation that cold temperatures can complement the G-protein sporulation defect now provides an ideal tool by which asexual differentiation can now be dissected.

MORPHOLOGICAL IDENTIFICATION OF DIAPORTHE/PHOMOPSIS SP. ISOLATED FROM XANTHIUM ITALICUM

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Draženka Jurković

2007-12-01

Full Text Available Diaporthe/Phomopsis sp. was isolated from Xanthium italicum (Italian cockleburr for the first time in Eastern Croatia during 2004 year. As Diaporthe/Phomopsis species are known as pathogens of soybean, sunflower and other arable crops, it is important to study alternative hosts as a possible source of inoculum from an epidemiological point of view. This paper describes symptoms of infection on X. italicum, pathogen morphological and cultural characters on potato-dextrose agar (PDA, biometrical values of reproductive structures (pycnidia, beta conidia, perithecia, asci and ascospores of naturally infected plants and of cultures grown on PDA. Results obtained from undertaken studies indicate that our Phomopsis sp. isolates from X. italicum belong to the group of P. helianthi.

Morphological features of Camarosporium pini – the fungus associated to health state degradation in Austrian and Ponderosa pine

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Ivanová Helena

2017-06-01

Full Text Available The subject of this study is escalated occurrence of the pathogenic fungus Camarosporium pini in the needle tissue of symptomatic trees P. nigra and P. ponderosa var. jeffreyi growing in urbanized settings and parks. C. pini induces severe infections and initiates a blight and premature loss of second-year foliage in pine trees. The fungus was identified microscopically and on base of morphological keys. The affected needles displayed a distinct bluish-grey necrotic band in the centre. On the surface of infected needles, there were formed pycnidia producing brown, oval conidia with three transversal and one or two vertical walls. Disease symptoms, some important characteristics in pure culture, and distinctive morphological features of C. pini associated to the health state degradation in Austrian and Ponderosa pine are described and compared. Cumulative effects of these stressful biotic and various abiotic factors may explain the current situation concerning the decline in the P. nigra and P. ponderosa var. jeffreyi in Slovakia.

Outbreaks of phaeohyphomycosis in the chinook salmon (Oncorhynchus tshawytscha) caused by Phoma herbarum.

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Faisal, Mohamed; Elsayed, Ehab; Fitzgerald, Scott D; Silva, Victor; Mendoza, Leonel

2007-01-01

Phoma herbarum has been associated with two outbreaks of systemic mycosis in hatchery-reared chinook salmon (Oncorhynchus tshawytscha) fingerlings. Affected fish exhibited abnormal swimming behavior, exophthalmia, multiple rounded areas of muscle softening, protruded hemorrhagic vents, and abdominal swelling. In all affected fish, swimbladders were filled with whitish creamy viscous fungal mass, surrounded by dark red areas in swimbladder walls, kidneys, and musculature. Clinical and histopathological examinations suggest that the infection may have started primarily in the swimbladder and then spread to the kidneys, gastrointestinal tract, and surrounding musculature. Consistent microscopical findings included broad septate branched fungal hyaline hyphae, 5-12 microm in diameter within the swimbladder, stomach, and often within and adjacent to blood vessels. Profuse growths of woolly brown fungal colonies were obtained from swimbladders and kidneys on Sabouraud medium. On corn meal agar the formation of pycnidia, characteristic of Phoma spp., was detected within 10 days of incubation. Morphological and molecular analyses identified this fungus as Phoma herbarum. This report underscores systemic fungal infections as a threat to raceway-raised salmon.

Morphological and ecological variation of Gremmeniella abietina var. abietina in Pinus sylvestris, Pinus contorta and Picea abies sapling stands in northern Finland and the Kola Peninsula

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Kaitera, Juha; Seitamaeki, Leena; Jalkanen, Risto [Finnish Forest Research Inst., Rovaniemi (Finland). Rovaniemi Research Station

2000-07-01

The morphological and ecological variation of two types of Gremmeniella abietina var. abietina causing scleroderris canker on conifers was investigated in Pinus spp. and Picea sp. sapling stands in northern Finland and the Kola Peninsula. Small-tree type (STT or B type) of G. abietina was detected alone in 13 Scots pine, three lodgepole pine and two Norway spruce sapling stands out of 26 stands investigated, both STT and large-tree type (LTT or A type) were observed in six Scots pine stands, and LTT was detected alone in two Scots pine stands. For the first time, G. abietina was found to injure Norway spruce saplings in a respective plantation in northern Fennoscandia. STT isolates produced statistically significantly more conidia in vitro than LTT isolates. Morphological variation in conidia septation revealed that STT produced conidia with more than five septa more frequently than did LTT. There was a greater range in variation in septation in STT than in LTT, with overlapping between the types. Isolates of both types were equally associated with cankers, coloured wood, pycnidia or apothecia in the infected saplings.

Morphology, Pathogenicity and Management of Coniella Fruit Rot (Coniella granati on Pomegranate

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Aysun Uysal

2018-04-01

Full Text Available One of the objectives of the study was to identify the fungus involved in fruit rot on pomegranates in the Mediterranean Region of Turkey. The fungus designated as Coniella granati (Saccardo Petrak and Sydow based on morphological characteristics. The fungus colonized the fruit after 5 to 8 days, followed by the appearance of fruit rot symptoms leading to the formation of abundant pycnidia covering the peel. Secondly, the efficacy of fungicides against C. granati was evaluated by mycelial growth and conidial germination assays. Tebuconazole, boscalid+pyraclostrobin and iprodione at 1.0, 25, and 50 μgml-1 concentrations, respectively, completely inhibited mycelial growth. In the azoxystrobin and dodine, relatively higher concentrations required to inhibit mycelial growth. Tebuconazole exhibited the greatest inhibition (82.2% of mycelium growth. The EC50 values in mycelial growth of C. granati ranged from 0.13 to 151.9. The highest EC50 values occurred for tebuconazole (0.13μgml-1. Tebuconazole, boscalid+pyraclostrobin and iprodione at 200, 10 and 5 μgml-1 concentrations, respectively, were the highly effective in inhibiting conidial germination. Azoxystrobin exhibited a low effect (61% on conidial germination. The EC50 values on conidial germination of C. granati ranged from 0.2 to 28.7. Tebuconazole had the lowest EC50 value, while boscalid+pyraclostrobin exhibited the highest EC50 value.

Banana peel culture as an indigenous medium for easy identification of late-sporulation human fungal pathogens.

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Kindo, A J; Tupaki-Sreepurna, A; Yuvaraj, M

2016-01-01

Fungi are increasing in incidence as human pathogens and newer and rarer species are continuously being encountered. Identifying these species from growth on regular culture media may be challenging due to the absence of typical features. An indigenous and cheap medium, similar to the natural substrate of these fungi, was standardised in our laboratory as an aid to species identification in a conventional laboratory setting. Ripe banana peel pieces, sterilised in an autoclave at 121°C temperature and 15 lbs pressure for 15 min promoted good growth of hyphae and pycnidia or acervuli in coelomycetes, flabelliform and medusoid fruiting bodies of basidiomycetes and fruit bodies such as cleistothecium in ascomycetes. The growth from the primary isolation medium was taken and inoculated onto the pieces of double-autoclaved ripe banana peel pieces in a sterile glass Petri dish with some moisture (sprinkles of sterile distilled water). A few sterile coverslips were placed randomly inside the Petri dish for the growing fungus to stick on to it. The plates were kept at room temperature and left undisturbed for 15-20 days. At a time, one coverslip was taken out and placed on a slide with lactophenol cotton blue and focused under the microscope to look for fruit bodies. Lasiodiplodia theobromae, Macrophomina phaseolina, Nigrospora sphaerica, Chaetomium murorum, Nattrassia mangiferae and Schizophyllum commune were identified by characteristic features from growth on banana peel culture. Banana peel culture is a cheap and effective medium resembling the natural substrate of fungi and is useful for promoting characteristic reproductive structures that aid identification.

Bioecology of the fungus Sphaeropsis sapinea dyko & Sutton - agents of pinus species decline

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Milijašević Tanja

2002-01-01

months. The first visible symptoms of infection of the young shoots are in the form of brown necroses, resin bleeding and several short pairs of needles at the place. S. sapinea can infect male and female flowers, current year and second-year cones. The pycnidia with mature conidia can form on the shoots, needles, first-year and second-year cones during the year of infection. The first mature fructifications are mostly formed in the bark of young shoots, in the zone of infection, and they were recorded in late June at the earliest. They were recorded on the cones at the beginning of the third decade of July. Pycnidia are formed in the bark, on the needles and on the second-year and older cones, on male and female flowers, as well as on the cones in the first year of development. The size of S. sapinea conidia on different Pinus species is very variable throughout the world and ranges widely from 16.8-55.0×7.0-21.0 m. Our study shows that the size of pycnospores in our country is also variable and it ranges 14.8-51.4×9.7-20.2 m The optimal temperature for spore germination and germ tube growth is 27°C, and the optimal temperature for mycelial growth on the media PDA and MEA is 28°C. The formation of fruiting bodies in the culture is affected by light and the presence of sterilised needles on the mycelium The correct and timely control measures can contribute to reduce the consequences of this serious disease, by the combined application of silvicultural and chemical measures. Chemical control is recommended in the nurseries, severely infected plantations and in urban environments. The fungicides which showed the greatest efficiency are the preparations based on copper and Benomil.

Morphological characterization of fungi associated with the ascochyta blight complex and pathogenic variability of Mycosphaerella pinodes on field pea crops in central Alberta

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Hafiz Ahmed

2015-02-01

Full Text Available Field pea crops in central Alberta were surveyed for ascochyta blight from 2011 to 2012 and fungal isolates were recovered from foliar lesions on selected plants. Cultural and microscopic characterization of the 275 isolates obtained revealed that 272 were of Mycosphaerella pinodes and three were of Phoma medicaginis var. pinodella. Ascochyta pisi or Phoma koolunga were not identified. Isolates of M. pinodes were divided into two groups, GI and GII, based on visual assessment of culture characteristics. GI isolates (light to dark, mostly gray colony color; pycnidial distribution radial and concentric; conidia 10.5–14.5 × 4.2–6.2 μm most with one septum, occasionally two, constricted at the septum; spore mass light buff to flesh color were predominant (83%, while GII isolates (dark to gray colony color; pycnidia abundant; conidia 8–16 × 3.5–6.2 μm most with 1 septum, constricted at the septum; spore mass light buff to flesh color were less common (17%. The cultures of GII isolates were similar to recent descriptions of A. pisi, but they differed in spore color. In a host differential study, 13 pathotypes of M. pinodes were identified from 110 single-spore isolates. Pathotype I was predominant (88 isolates and virulent on all nine differential genotypes. The other pathotypes (pathotypes II–XIII were rare (1–6 isolates of each. Comparison of the present results with earlier studies suggests that pathotype I has been prevalent for many years, and that its aggressiveness on the host differentials has increased over time. Emphasis should be placed on breeding for resistance to M. pinodes in field pea cultivars intended for deployment in central Alberta.

The first occurrence of stem canker on oilseed rape caused by Leptosphaeria biglobosa in Serbia

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Mitrović Petar

2016-01-01

Full Text Available In October 2010 the occurrence of the stem canker symptoms in rapeseed was observed at the locality Rimski Šančevi in Serbia. Several strains of fungi were isolated from the stem. Morphological characteristics of the isolates were studied on a PDA medium at 25±1oC: growth rate, colour, shape and appearance of colonies and the edge of the colony, the pigment, size, colour and shape of pycnidiospores and pycnidia in 10 isolates (isolated in Serbia K-111, IK-112, K-113, K-114, K-115, K-116, IK-117, K-118, K-119, and K-120. All tested strains had fast growth, the regular form of colonies and poor sporulation on nutrient medium. Pycnidiospores are unicellular, hyaline, and mostly straight, with or without a drop of oil. Molecular identification was performed by the application the PCR technique using primers PN3/PN10. In addition to these 10 isolates, two reference strains obtained from the Centre for Agricultural Studies, Rothamsted, UK, which are marked with L. m (Leptosphaena maculanS, L. b (Leptosphaeria biglobosa and 7 reference isolates originating from Serbia, which are marked K-7, St-16, GS-25, L-5, C-3, LJ-2, S-1 were used. On the basis of DNA amplification with primers PN3 and PN10 tape length was about 580 bp for isolates (L.b, IK-111, K-112, K-113, K-114, K-115, K-116, K-117, K-118, K-119, K-120 while that was 560 bp for other group of isolates (L.m, K-7, St-16, GS-25, L-5, C-3, LJ- 2, S-1.This study showed that 10 isolates (K-111, K-112, K-113, IK- 114, K-115, K-116, K-117, K-118, K-119, K-120 isolated from the stem of rapeseed belong to the species Leptosphaeria biglobosa.