Morfološka, molekularna i patogena karakterizacija vrsta Diaporthe/Phomopsis na soji u Srbiji

Species of the genus Diaporthe/Phomopsis are endophytes on a very wide range of hosts and can be present as pathogens or saprophytes. Species of this genus cause a complex of diseases on the stem and seeds of soybean (Glycine max (L.) Merr.) and therefore belong to a group of important parasites of this plant. Diaporthe phaseolorum and Phomopsis longicolla were described as soybean pathogens. The first species includes three varieties: D. phaseolorum var. sojae, D. phaseolorum var. caulivora and D. phaseolorum var. meridionalis. Although several studies have extensively described the morphological and culture characteristics of these pathogens, their taxonomy has not fully been defined. The greatest taxonomic confusion is related to D. phaseolorum var. sojae, because this variety expressed variability of morphological characteristics. Since morphological variability of D. phaseolorum var. sojae was also observed in isolates from Serbia, there were needs for detailed examinations in order to exactly determine which Diaporthe/Phomopsis species are present on soybean in Serbia. By comparison of morphological, molecular and pathogenic data seven Diaporthe/Phomopsis species were identified. Besides the known parasites, D. neotheicola, D. eres, D. viticola and unidentified group Phomopsis sp. 9 were also detected on soybean seeds. Identification and phylogenetic grouping was performed based on sequences of ITS region and parts of LSU and TEF1-α genes. In order to find faster ways of detection and identification, ITS sequences of the studied species were used for the virtual digestion with restriction enzymes AluI, RsaI, HhaI, MseI, and StyI. Since different molecular profiles were generated for each studied species, it was concluded that PCR-RFLP technique can potentially be a useful method for rapid and reliable identification of Diaporthe/Phomopsis species that occur on soybean. By studying morphological characteristics of the identified species, it was found that the dimensions of reproductive organs overlapping, but also that they vary between isolates within a species, as well as within the isolates. Based on this, it was concluded that dimensions of reproductive organs are not reliable parameters for identification of Diaporthe/Phomopsis species isolated from soybeans...Endofitne vrste roda Diaporthe/Phomopsis imaju izuzetno širok spektar domaćina i u biljkama mogu biti prisutne kao patogeni ili saprofiti. Na stablu i semenu soje (Glycine max (L.) Merr.), vrste ovog roda prouzrokuju kompleks oboljenja i spadaju u grupu značajnih parazita ove biljne vrste. Kao paraziti soje opisane su vrste Diaporthe phaseolorum i Phomopsis longicolla. U okviru prvo pomenute vrste razlikuju se tri varijeteta, i to: D. phaseolorum var. sojae, D. phaseolorum var. caulivora i D. phaseolorum var. meridionalis. Iako su u mnogobrojnim istraživanjima detaljno opisane morfološke i odgajivačke karakteristike ovih patogena, njihova taksonomija još uvek nije u potpunosti definisana. Najveća taksonomska nedoumica odnosi se na D. phaseolorum var. sojae, jer je u okviru ovog varijeteta uočena izražena varijabilnost morfoloških karakteristika. S obzirom da je morfološka varijabilnost D. phaseolorum var. sojae uočena i kod izolata poreklom iz Srbije, ukazala se potreba za detaljnim ispitivanjem, kako bi se tačno utvrdile prisutne Diaporthe/Phomopsis vrste na soji u Srbiji. Uporednim proučavanjem morfoloških, molekularnih i patogenih karakteristika, identifikovano je ukupno sedam vrsta roda Diaporthe/Phomopsis. Pored poznatih parazita, P. longicolla, D. phaseolorum var. sojae i D. phaseolorum var. caulivora, na semenu soje su detektovane i vrste D. neotheicola, D. eres, D. viticola i neidentifikovana grupa Phomopsis sp. 9. Identifikacija i filogenetsko grupisanje Diaporthe/Phomopsis vrsta sprovedeno je na osnovu sekvenci ITS regiona i dela LSU i TEF1-α gena. U cilju iznalaženja bržeg načina detekcije i identifikacije, ITS sekvence proučavanih vrsta poslužile su za virtuelnu digestiju restrikcionim enzimima AluI, RsaI, HhaI, MseI i StyI. S obzirom da su za svaku proučavanu vrstu dobijeni različiti molekularni profili, konstatovano je da PCRRFLP tehnika potencijalno može biti korisna metoda za brzu i pouzdanu identifikaciju Diaporthe/Phomopsis vrsta koje se javljaju na soji. Proučavanjem morfoloških karakteristika identifikovanih vrsta, utvrđeno je da se dimenzije reproduktivnih organa preklapaju, ali i da veoma variraju između izolata unutar jedne vrste, kao i unutar samog izolata. Na osnovu toga, izveden je zaključak da dimenzije reproduktivnih organa nisu pouzdani pokazatelji pri identifikaciji Diaporthe/Phomopsis vrsta izolovanih iz soje..

Morphological, molecular and pathogenic characterization of species Diaporthe/Phomosis on soybean in Serbia

Endofitne vrste roda Diaporthe/Phomopsis imaju izuzetno širok spektar domaćina i u biljkama mogu biti prisutne kao patogeni ili saprofiti. Na stablu i semenu soje (Glycine max (L.) Merr.), vrste ovog roda prouzrokuju kompleks oboljenja i spadaju u grupu značajnih parazita ove biljne vrste. Kao paraziti soje opisane su vrste Diaporthe phaseolorum i Phomopsis longicolla. U okviru prvo pomenute vrste razlikuju se tri varijeteta, i to: D. phaseolorum var. sojae, D. phaseolorum var. caulivora i D. phaseolorum var. meridionalis. Iako su u mnogobrojnim istraživanjima detaljno opisane morfološke i odgajivačke karakteristike ovih patogena, njihova taksonomija još uvek nije u potpunosti definisana. Najveća taksonomska nedoumica odnosi se na D. phaseolorum var. sojae, jer je u okviru ovog varijeteta uočena izražena varijabilnost morfoloških karakteristika. S obzirom da je morfološka varijabilnost D. phaseolorum var. sojae uočena i kod izolata poreklom iz Srbije, ukazala se potreba za detaljnim ispitivanjem, kako bi se tačno utvrdile prisutne Diaporthe/Phomopsis vrste na soji u Srbiji. Uporednim proučavanjem morfoloških, molekularnih i patogenih karakteristika, identifikovano je ukupno sedam vrsta roda Diaporthe/Phomopsis. Pored poznatih parazita, P. longicolla, D. phaseolorum var. sojae i D. phaseolorum var. caulivora, na semenu soje su detektovane i vrste D. neotheicola, D. eres, D. viticola i neidentifikovana grupa Phomopsis sp. 9. Identifikacija i filogenetsko grupisanje Diaporthe/Phomopsis vrsta sprovedeno je na osnovu sekvenci ITS regiona i dela LSU i TEF1-α gena. U cilju iznalaženja bržeg načina detekcije i identifikacije, ITS sekvence proučavanih vrsta poslužile su za virtuelnu digestiju restrikcionim enzimima AluI, RsaI, HhaI, MseI i StyI. S obzirom da su za svaku proučavanu vrstu dobijeni različiti molekularni profili, konstatovano je da PCRRFLP tehnika potencijalno može biti korisna metoda za brzu i pouzdanu identifikaciju Diaporthe/Phomopsis vrsta koje se javljaju na soji. Proučavanjem morfoloških karakteristika identifikovanih vrsta, utvrđeno je da se dimenzije reproduktivnih organa preklapaju, ali i da veoma variraju između izolata unutar jedne vrste, kao i unutar samog izolata. Na osnovu toga, izveden je zaključak da dimenzije reproduktivnih organa nisu pouzdani pokazatelji pri identifikaciji Diaporthe/Phomopsis vrsta izolovanih iz soje...Species of the genus Diaporthe/Phomopsis are endophytes on a very wide range of hosts and can be present as pathogens or saprophytes. Species of this genus cause a complex of diseases on the stem and seeds of soybean (Glycine max (L.) Merr.) and therefore belong to a group of important parasites of this plant. Diaporthe phaseolorum and Phomopsis longicolla were described as soybean pathogens. The first species includes three varieties: D. phaseolorum var. sojae, D. phaseolorum var. caulivora and D. phaseolorum var. meridionalis. Although several studies have extensively described the morphological and culture characteristics of these pathogens, their taxonomy has not fully been defined. The greatest taxonomic confusion is related to D. phaseolorum var. sojae, because this variety expressed variability of morphological characteristics. Since morphological variability of D. phaseolorum var. sojae was also observed in isolates from Serbia, there were needs for detailed examinations in order to exactly determine which Diaporthe/Phomopsis species are present on soybean in Serbia. By comparison of morphological, molecular and pathogenic data seven Diaporthe/Phomopsis species were identified. Besides the known parasites, D. neotheicola, D. eres, D. viticola and unidentified group Phomopsis sp. 9 were also detected on soybean seeds. Identification and phylogenetic grouping was performed based on sequences of ITS region and parts of LSU and TEF1-α genes. In order to find faster ways of detection and identification, ITS sequences of the studied species were used for the virtual digestion with restriction enzymes AluI, RsaI, HhaI, MseI, and StyI. Since different molecular profiles were generated for each studied species, it was concluded that PCR-RFLP technique can potentially be a useful method for rapid and reliable identification of Diaporthe/Phomopsis species that occur on soybean. By studying morphological characteristics of the identified species, it was found that the dimensions of reproductive organs overlapping, but also that they vary between isolates within a species, as well as within the isolates. Based on this, it was concluded that dimensions of reproductive organs are not reliable parameters for identification of Diaporthe/Phomopsis species isolated from soybeans..

Otpornost soje prema patogenima

This paper presents an overview of the research on soybean resistance to pathogens. The review included most harmful agents of soybean diseases in Serbia, as well as those that are potentially harmful. Development and cultivation of resistant cultivars is the most efficient, economical and environmentally acceptable control measure for plant disease. It points to the variability in pathogenicity (physiological races) of parasites, especially expressed in Phytophthora sojae, Peronospora manshurica and Pseudomonas syringae pv. glycinea, which requires continuous breeding for resistance. Resistant, partially resistant and moderately susceptible genotypes, which are used as donors of resistance genes to different pathogens, are listed in this paper. Also, avirulent genes in the parasite and resistance genes in soybean are indicated. Gene mapping significantly contributes to better understanding of the mode of inheritance and consequently, more efficient breeding for disease resistance. Significant improvement is expected by using molecular techniques, especially in dealing with Sclerotinia sclerotiorum, Pseudomonas syringae pv. glycinea and Phomopsis longicolla. For these parasites only partial resistance has been reported but not complete resistance.Na osnovu višegodišnjih istraživanja utvrđeno je da su najštetniji i najrasprostranjeniji paraziti soje u Srbiji Peronospora manshurica (prouzrokovač plamenjače), Pseudomonas syringae pv. glycinea (bakteriozna pegavost), Sclerotinia sclerotiorum (bela trulež), Diaporthe phaseolorum var. caulivora (rak stabla), Macrophomina phaseolina (ugljenasta trulež) i Phomopsis spp. (trulež semena). Postoji realna opasnost da se na soji u našoj zemlji pojave i dva veoma destruktivna parazita; Phytophthora sojae, prouzrokovač fitoftorozne truleži i Diaporthe phaseolorum var. meridionalis prouzrokovač 'južnog' raka stabla. Unošenje otpornosti prema najznačajnijim parazitima u nove sorte soje je jedan od prioritetnih zadataka svih oplemenjivačkih programa. Najpre je potrebno iznaći odgovarajući izvor otpornosti, zatim obaviti povratna ukrštanja i u kasnijim generacijama identifikovati otporne genotipove. Uspeh u radu na stvaranju otpornih sorti soje uslovljen je postojanjem dobrih izvora rezistentnosti, kao i poznavanjem prirode otpornosti i načina nasleđivanja. Rezistentnost prema P. manshurica evidentirana je u više genotipova soje, a najčešće je uslovljena jednim dominantnim genom (Rpm). Ova otpornost je potpuna, ali nije trajna, jer se postepeno gubi usled pojave novih fizioloških rasa. Rasa br. 4 P. syringae pv. glycinea je dominantna na soji u svetu i kod nas. Ne postoje potpuno otporni genotipovi prema ovoj rasi, ali su prisutne značajne razlike u stepenu osetljivosti. Parcijalna otpornost kontrolisana je od strane jednog ili nekoliko Rpg gena. Mehanizmi izbegavanja oboljenja i parcijalna fiziološka rezistentnost uslovljavljaju razlike u reakciji sorti soje prema S. sclerotiorum. Mapirana su dva lokusa koji kontrolišu mehanizme izbegavanja bolesti i jedan lokus, najverovatnije odgovoran za parcijalnu fiziološku rezistentnost. Parcijalna rezistentnost utvrđena je kod nekoliko introdukovanih (PI) genotipova soje. U našim agroekološkim uslovima ranostasni genotipovi izbegavaju napad ovog oboljenja. Nije evidentirana potpuna otpornost prema D. phaseolorum var. caulivora, prouzrokovaču 'severnog' tipa raka stabla. Prouzrokovač 'južnog' tipa raka stabla D. phaseolorum var. meridionalis nije prisutan na soji u Srbiji. Otpornost prema ovoj gljivi nađena je u nekoliko genotipova soje, a uslovljavaju je dva (delimična otpornost), odnosno, četiri (potpuna) major gena. Pripadnici roda Diaporthe/Phomopsis su najštetniji paraziti semena soje, a vrsta P. longicolla je najzastupljenija. Zadovoljavajući nivo rezistentnosti prema ovom parazitu semena poseduje nekoliko genotipova soje. Priroda otpornosti nije u potpunosti razjašnjena, a novija istraživanja ukazuju da se radi o kvantitativnom svojstvu. Brojna istraživanja posvećena su proučavanju Phytophthora sojae (fitoftorozna trulež), veoma štetnog parazita soje. Opisano je više od 70 fizioloških rasa, identifikovani su avirulentni geni (Avr) u pojedinim rasama i pronađeni su geni rezistentnosti (Rps) u sortama soje. Tako je omogućeno uspešno oplemenjivanje soje na otpornost prema ovom parazitu

Tolerantnost genotipova pšenice prema sulfonilurei u kulturi 'zigotnog' embriona

Tolerance of wheat genotypes to the sulfonylurea herbicide metsulfuron-methyl was studied using in vitro culture. Six randomly selected wheat genotypes were used for isolation. Mature embryos were inoculated onto a modified MS medium to which three concentrations of metsulfuron-methyl were added: I-30 g l-1, II-60 g l-1, and III-90 g l-1. The control group of embryos was cultivated on a herbicide-free medium. After one month of cultivation, callus fresh weight was measured. There were significant differences among the genotypes regarding their tolerance to metsulfuron-methyl. The Pobeda cultivar expressed the highest and the Lr-12 genotype had the lowest level of herbicide tolerance.Tolerantnost genotipova pšenice prema metsulfuron-metilu, herbicidu iz grupe sulfonilurea preparat a, ispitana je u in vitro kulturi zigotnog embriona. Za izolaciju korišćeno je šest genotipova pšenice. Embrioni su inokulisani na modifikovanu MS podlogu, kojoj je metsulfuron-metil dodat u tri koncentracije: I - 30 g l-1, II - 60 g l-1 i III - 90 g l-1. Kontrolna grupa embriona gajena je na podlozi bez herbicida. Posle mesec dana gajenja izmerena je sveža masa kalusa. Rezultat i su pokazali da je prisustvo metsulfuron-metila u podlozi izazvalo inhibiciju porasta kalusa kod svih genotipova. Međutim, genotipovi su se značajno razlikovali među sobom u pogledu reakcije na ovaj herbicid. Na najnižoj koncentraciji metsulfuron-metila (I - 30 g l-1), genotip Lr-12 imao je najvišu redukciju sveže mase kalusa, u odnosu na kontrolu (80.5 %), dok je najnižu redukciju imao genotip Vel (63.3 %). Više koncentracije herbicida imale su još jače inhibitorno dejstvo kod svih genotipova. Rezultat i su pokazali da je sorta Pobeda bila najtolerantnija, dok je genotip Lr-12 bio najosetljiviji na prisustvo metsulfuron-metila u podlozi

The effects of the perturbated ionization potential and the magnetic component on the relativistic transition rate

In this paper we theoretically analyze the influence of the perturbated ionization potential and the component of the magnetic field on the transition rate, in a relativistic, linearly polarized laser field. The obtained results indicate that both effect play important roles during the ionization process, influence the rate, and must be considered. It is shown that a laser field influences the ionization potential of an electron strongly and causes that the rate curve shows significantly different behavior compared to the case with considered non-perturbated ionization potential.Publishe

Vrste roda Diaporthe/Phomopsis na soji u Srbiji

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 Bačka 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.Kompleks oboljenja na soji izazivaju gljive iz roda Diaporthe/Phomopsis. Kao paraziti soje opisana su tri varijeteta vrste Diaporthe phaseolorum (anamorf Phomopsis phaseoli), i to: D. phaseolorum var. sojae (syn. P. sojae), prouzrokovač paleži mahuna i stabla (pod and stem blight), zatim D. phaseolorum var. caulivora i D. phaseolorum var. meridionalis prouzrokovači 'severnog' i 'južnog' tipa raka stabla (soybean stem canker). Phomopsis longicolla (teleomorf nepoznat) je najčešći prouzrokovač truleži semena (soybean seed decay), mada ovo oboljenje izazivaju i sva tri varijeteta gljive D. phaseolorum. Svi pomenuti pripadnici roda, izuzev D. phaseolorum var. meridionalis, prisutni su na soji u Srbiji, a najveći značaj ima D. phaseolorum var. caulivora, prouzrokovač raka stabla. Poslednjih nekoliko decenija sprovedena su brojna, sveobuhvatna proučavanja parazita roda Diaporthe/ Phomopsis na soji u našoj zemlji. Nedavno započeta istraživanja na molekularnom nivou ukazuju da su pored pomenutih na soji prisutne i druge vrste ovog roda. U ovom radu su analizirani rezultati višegodišnjih istraživanja u svetu i kod nas

Microstructure and thermal characterization of Mo-doped Li6.87Nb2.34Ti5.78O21 solid-solution ceramics

In this work, the microstructure and thermal properties of lithium-niobium-titanium-oxide (Li-Nb-Ti-O) solid-solution ceramics were investigated using the X-ray diffraction (XRD), Raman spectroscopy, scanning electron microscopy (SEM), and differential scanning calorimetry (DSC) techniques. XRD and SEM analysis confirmed that Li-Nb-Ti-O ceramic sample synthesized by solid-state method reaches the desired composition of so-called Mphase of the Li2O-Nb2O5-TiO2 ternary system (Li6.87Nb2.34Ti5.78O21) that should have excellent microwave dielectric properties. The M-phase was obtained with lower than 1000ºC sintering temperature by doping with MoO3 as flux material, which makes this kind of ceramic material suitable for the low temperature co-fired ceramic (LTCC) technology applications. The heat flow data obtained from DSC measurement were used to calculate the specific heat capacity (Cp) of synthesized Li6.87Nb2.34Ti5.78O21 solid-solution ceramics, which is the property of a material that tells about its stability and functionality

TYPES OF TREATMENTS USED BY PARENTS OF CHILDREN WITH AUTISM

There is a large number of treatment types offered to parents of children diagnosed with autism spectrum disorders (ASD), but they are particularly vulnerable to the promoters of pseudoscience, which can lead to the use of ineffective types of treatment. This study represents an attempt to document the types of treatments used by parents of children with ASD in the Republic of Serbia. Secondary aim was to determine if certain parent or child characteristics influenced the types of treatments used. Modified version of survey constructed and conducted by Green, et al. (2006) was used. Nearly all (n=85) of the 86 treatments listed in our survey were being currently used or used in the past by at least one parent and the most commonly used treatment is speech and language therapy. The treatments were grouped into ten categories and the most commonly used is other treatments (80.6%), followed by standard therapies (79.2%) and vitamin supplements (65.3%).  It is also of great importance to educate the parents of children with ASD about evidence-based treatments and on how to distinguish them from treatments that do not have empirica

Importance of microbiological fertilizer used in soybean production: Agronomical and biological aspects

Inoculation is the best way to ensure good nodulation with the proper strain of nitrogen-fixing bacteria for increased yield in an environmentally safe manner. This investigation was conducted on three-year experiment which was set up in four replications with three-crop rotation (maize-soybean-wheat) at experimental field of the Institute of Field and Vegetable Crops, Novi Sad, Serbia. Effects of application of microbiological fertilizer (with nitrogen-fixing bacteria Bradyrhizobium japonicum) on root mass, number and mass of nodes per plant, soil biogeneity (number of ammonifiers, Azotobacter, actinomycetes), as well as the effect on soybean yield were studied. Obtained results indicated positive impact of inoculation on nitrogen fixation parameters. The higher amounts of mineral nitrogen had negative impact on atmospheric nitrogen fixation, that is, by increasing preplanting mineral nitrogen amount in the soil by 1 kg, the amount of calculated nitrogen from the air was decreased by 1.72 kg. Increasing abundance of studied microorganisms in the rhizosphere, as well as soybean yield using microbiological fertilizer was stimulated by plowing under crop residues. The impact of meteorogical conditions on the effects of inoclutaion was very significant

Raznolikost fitopatogenih gljiva na semenu soje u Srbiji

Soybean is one of the most important industrial crops in the world. The rich nutritional content of the seed is the reason for the increased cultivation of this crop all over the world. However, a large number of phytopathogenic fungi that exist in soybean seeds can reduce the nutritional content, germination, and seed vigor. Soybean seed samples were collected from experimental fields in seven locations in Vojvodina Province, Serbia in the period 2016-2018. From each sample, 100 seeds were randomly selected. Seeds were surface-disinfested in 4% sodium hypochlorite for 5 min, washed two times in sterile water, and dried on a filter paper. All 100 seeds were placed in 20 Petri dishes with standard potato dextrose agar medium (PDA), five seeds in each, and incubated at 22 ± 2°C for seven days, under 12 h of alternating light/dark conditions. The first assessment was performed on the fourth day after isolation. The health status analysis of soybean seed showed that Peronospora manshurica, Macrophominaphaseolina, Botrytis cinerea, Cercospora kikuchii as well as the species from genera Alternaria, Diaporthe, and Fusarium had colonized soybean seed in Serbia over three years. Species from the genus Alternaria and Peronospora manshurica were dominant in all three examined years. Furthermore, it has been noticed that weather conditions, location, and cultivar significantly influenced the intensity of the infection. The results of this study have shown which pathogens pose a threat to successful soybean seed production and help in finding preventive measures to control these pathogens during vegetation.Soja je jedna on najznačajnih gajenih industrijskih biljnih vrsta na svetu. Bogat nutritivni sadržaj zrna zaslužan je za sve veću učestalost gajenja ove industrijske biljke širom sveta. Međutim, veliki broj fitopatogenih gljiva egzistira u semenu soje i nepovoljno utiče na hranljivi sadržaj zrna pa samim tim i na smanjenu klijavost i životnu sposobnost. Zdravstveni pregled semena soje u trogodišnjem periodu od 2016. do 2018. godine, pokazao je da fitopatogene vrste iz rodova Alternaria, Diaporthe i Fusarium, kao i vrste Peronospora manshurica, Macrophominaphaseolina, Botrytis cinerea, i Cercospora kikuchii egzistiraju u semenu soje prikupljenom iz sedam lokaliteta u Srbiji. Ipak, P. manshurica i vrste iz roda Alternaria pokazale su se kao najdominantnije vrste u sve tri godine. Takođe, ustanovljeno je da vremenski uslovi, lokalitet gajenja i odabir sorte značajno utiču na intenzitet zaraze. Rezultati ovog istraživanja pokazuju koji patogeni predstavljaju pretnju uspešnoj semenskoj proizvodnji soje i pomažu u pronalaženju preventivnih mera za suzbijanje ovih patogena u toku vegetacije