Karakterizacija vrsta iz Fusarium fujikuroi kompleksa patogena zrna strnih žita u Srbiji
Characterization of Fusarium fujikuroi species complex pathogens of small grain kernels in Serbia
Докторанд
Savić, IvaМентор
Duduk, NatašaЧланови комисије
Stanković, SlavicaVico, Ivana
Kandić, Vesna
Lučev, Milica
Метаподаци
Приказ свих података о дисертацијиСажетак
Iz zrna obične pšenice, durum pšenice, ječma i tritikalea sakupljenih iz skladišta
sa 55 lokaliteta u Srbiji tokom četiri godine (2014.-2017.) dobijeni su izolati vrsta
Fusarium fujikuroi kompleksa. Odabrani izolati su identifikovani i okarakterisani na
osnovu patogenih, molekularnih, morfoloških i toksikoloških karakteristika. Na strnim
žitima identifikovane su tri vrste F. fujikuroi kompleksa - Fusarium verticillioides,
Fusarium proliferatum i Fusarium subglutinans. Ova ispitivanja su potvrdila prisustvo
ove tri vrste na zrnu pšenice i ječma u Srbiji. F. verticillioides, F. proliferatum i F.
subglutinans su po prvi put opisane na zrnu durum pšenice u Srbiji, kao i na zrnu
tritikalea u svetu.
Molekularna identifikacija vrsta F. fujikuroi kompleksa urađena je na osnovu tri
genska lokusa (ITS, TEF 1-α i RPB2). Sekvenciranjem tri genska lokusa dobijene su
sekvence na osnovu kojih su rekonstruisana filogenetska stabla za svaki region posebno.
Na osnovu TEF 1-α regiona uspešno su razdvojen...e sve tri vrste F. fujikuroi kompleksa,
dok na osnovu druga dva regiona to nije bilo moguće. Identifikacija vrsta urađena je i
primenom specifičnih prajmera (VER1/VER2, PRO1/PRO2, SUB1/SUB2, FV-F2/FV-FR,
FS-F1/FS-R i FS-F2/FS-R).
Na osnovu makroskopskih karakteristika kao što su izgled i boja micelije,
pigmentacija podloge uočena je varijabilnost kod sve tri vrste F. fujikuroi kompleksa, ali
na osnovu ovih karakteristika nije bilo moguće precizno razlikovati ove tri vrste.
Makrokonidije i mikrokonidije obrazovale su sve tri vrste, dok je prisustvo mezokonidija
zabeleženo samo kod izolata vrste F. subglutinans. Vrste F. fujikuroi kompleksa
obrazovale su makrokonidije ravnih ivica sa zakrivljenom apikalnom ćelijom i bazalnom
ćelijom u obliku stopala. Najduže makrokonidije obrazovala je vrsta F. subglutinans.
Mikrokonidije su bile bez septi, ovalnog oblika. Najkraće mikrokonidije obrazovala je
vrsta F. proliferatum. Vrsta F. verticillioides obrazovala je mikrokonidije u dugim lancima
na monofijalidama, dok je vrsta F. proliferatum obrazovala kraće lance mikrokonidija na
monofijalidama i polifijalidama. Kod vrste F. subglutinans mikrokonidije su se formirale
u lažnim glavicama na monofijalidama i polifijalidama. Načini obrazovanja
mikrokonidija mogu poslužiti u preliminarnoj identifikaciji ove tri vrste.
Virulentnost vrsta ispitana je na listovima ječma, klasu durum pšenice i klasu
pšenice, ječma i tritikalea. Na listovima ječma vrste F. proliferatum i F. subglutinans bile
su jednako virulentne, dok je vrsta F. verticillioides imala najmanju prosečnu ocenu
virulentnosti. Ispitivanja na klasu durum pšenice sprovedena su tokom dve godine.
Rezultati su pokazali da je virulentnost bila veća u drugoj godini ispitivanja. Tokom obe
godine najvirulentnija vrsta je bila F. verticillioides dok su vrste F. proliferatum i F.
subglutinans bile su jednako virulentne. Tokom jedne proizvodne godine ispitana je
virulentnost ovih vrsta na klasu pšenice, ječma i tritikalea, nakon čega je zaključeno da
su sve tri vrste F. fujikuroi kompleksa bile jednako virulentne.
Vrste ovog kompleksa su značajne zbog svoje sposobnosti da sintetišu
mikotoksine od kojih su najznačajniji fumonizini. Koncentracije sintetisanih fumonizina
ispitane su ELISA metodom. Dobijeni rezultati ukazali su na visoke koncentracije
fumonizina kod izolata vrsta F. verticillioides i F. proliferatum, dok su izolati vrste F.
subglutinans sintetisali fumonizine u niskim koncentracijama ili ispod praga detekcije.
Fusarium fujikuroi species complex isolates were obtained from grains of
common wheat, durum wheat, barley and triticale collected from storages from 55
locations in Serbia during four years (2014-2017). Selected isolates were identified and
characterized based on pathogenic, molecular, morphological and toxicological
characteristics. Three species of F. fujikuroi complex were identified on small grains -
Fusarium verticillioides, Fusarium proliferatum and Fusarium subglutinans. Results of
these researches confirmed the presence of these three species on wheat and barley
grains in Serbia. This is the first report of F. verticillioides, F. proliferatum and F.
subglutinans on durum wheat grain in Serbia, as well as on triticale grain in the world.
Molecular identification of the F. fujikuroi species complex was done on the basis
of three gene loci (ITS, TEF 1-α and RPB2). By sequencing three gene loci, sequences
were obtained on the basis of which phylogenetic trees were reconstructed fo...r each
region. All three species of the F. fujikuroi species complex were successfully separated
based on the TEF 1-α region, while on the other two regions it was not possible. Species
identification was also done using specific primers (VER1/VER2, PRO1/PRO2,
SUB1/SUB2, FV-F2/FV-FR, FS-F1/FS-R and FS-F2/FS-R).
Based on macroscopic characteristics such as appearance and color of the
mycelium, pigmentation of the substrate, variability was observed in all three species of
F. fujikuroi species complex, but based on these characteristics, it was not possible to
accurately distinguish these three species. All three species produced macroconidia and
microconidia, while the presence of mesoconidia was observed only in F. subglutinans
isolates. Species of the F. fujikuroi complex produced straight-edged macroconidia with a
curved apical cell and a foot-shaped basal cell. The species longest macroconidia were
produced by F. subglutinans. Microconidia were without septa, oval in shape. The
shortest microconidia were formed by F. proliferatum isolates. The species F.
verticillioides formed microconidia in long chains on monophialids, while F. proliferatum
formed shorter chains of microconidia on monophialids and polyphialids. Species F.
subglutinans formed microconidia in false heads on monophialids and polyphialids. The
methods of formation of microconidia can be useful in the preliminary identification of
these three species.
The virulence of the species was tested on barley leaves and spikes on durum
wheat, wheat, barley and triticale. On barley leaves, F. proliferatum and F. subglutinans
were equally virulent, while F. verticillioides had the lowest average virulence rating.
Research on the durum wheat spikes was conducted in two year period. The results
showed that virulence was higher in the second year of the trial. During both years, the
most virulent species was F. verticillioides, while the species F. proliferatum and F.
subglutinans were equally virulent. During one production year the virulence of these
species was tested on wheat, barley and triticale spikes, after which it was concluded
that all three species from F. fujikuroi complex were equally virulent.
The species of this complex are significant for their ability to synthesize
mycotoxins, of which the most important are fumonisins. The concentrations of
synthesized fumonisins were tested using the ELISA method. The obtained results
indicated high concentrations of fumonisins in isolates of F. verticillioides and F.
proliferatum, while isolates of F. subglutinans synthesized fumonisins in low
concentrations or below the detection rate.
Факултет:
Универзитет у Београду, Пољопривредни факултетДатум одбране:
07-09-2023Кључне речи:
Fusarium fujikuroi, Fusarium verticillioides, Fusarium proliferatum, Fusarium subglutinans, pšenica, durum pšenica, ječam, tritikale, fumonizini / Fusarium fujikuroi, Fusarium verticillioides, Fusarium proliferatum, Fusarium subglutinans, wheat, durum wheat, barley, triticale, fumonisinsRelated items
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