Rasprostranjenost PPV-D i PPV-Rec sojeva virusa šarke šljive (Plum pox virus) u Srbiji i dinamika njihovog širenja u zasadu šljive
Distribution of PPV-D and PPV-Rec strains of Plum pox virus in Serbia and the dynamics of their spread in plum orchard
Докторанд
Jevremović, Darko R.Ментор
Krstić, BrankaЧланови комисије
Paunović, SvetlanaIvanović, Mirko
Bulajić, Aleksandra
Petrović-Obradović, Olivera
Метаподаци
Приказ свих података о дисертацијиСажетак
Virus šarke šljive (Plum pox virus-PPV), prouzrokovač bolesti šarke šljive, je
prisutan u Srbiji skoro 80 godina i smatra se ekonomski najznačajnijim virusom
koštičavih voćaka. Od 7 opisanih sojeva PPV, u Srbiji je potvrđeno prisustvo tri
soja: PPV-M, PPV-D i PPV-Rec.
U cilju utvrđivanja raširenosti sojeva šarke u zasadima šljive i kajsije u
Srbiji, u periodu 2008‒2010. godine, standardizovanom procedurom je sakupljeno
ukupno 283 uzoraka lišća šljive i kajsije iz preko 90 zasada. Tipiziranje
detektovanih izolata sprovedeno je IC-RT-PCR metodom kroz 4 odvojene PCR
reakcije sa PPV-M i PPV-D specifičnim prajmerima sa ciljnim sekvencama u dva
regiona genoma. Za filogenetsku analizu i utvrđivanje stope diverziteta odabran je
41 izolat za sekvencioniranje fragmenata iz dva regiona genoma (C-ter NIb‒N-ter
CP i C-ter P3‒6K1‒N-ter CI).
U cilju ispitivanja dinamike širenja PPV-D i PPV-Rec sojeva u direktnoj
kompeticiji, tokom 2008. godine podignut je eksperimentalni zasad sa 400 stabala
šljive s...orte Čačanska lepotica. Osam stabala, koja su locirana u sredini zasada, su
inokulisana sa potpuno okarakterisanim PPV-D i PPV-Rec izolatima (4 stabla po
izolatu-interni izvor infekcije). Sva stabla Prunus vrsta u neposrednoj okolini
zasada su markirana, uzorkovana i testirana na prisustvo PPV (eksterni izvor
infekcije). Pozitivni uzorci su sekvencionirani u C-ter NIb‒N-ter CP regionu
genoma za dalju uporednu analizu sa sekvencama inokulisanih i novoinficiranih
stabala u zasadu. Svake godine, od 2008‒2011., sva stabla u eksperimentalnom
zasadu su vizuelno pregledana i ispitivana ELISA testom na prisustvo PPV.
Pozitivni uzorci su tipizirani IC-RT-PCR metodom i sekvencionirani u C-ter NIb‒Nter
CP regionu genoma. Tokom poslednje tri godine istraživanja vršeno je praćenje
prisustva i dinamike brojnosti lisnih vaši u eksperimentalnom zasadu...
Plum pox virus (PPV), the agent of Sharka disease, is present in Serbia
almost 80 years and it is considered as economically the most important virus of
stone fruits. Three strains, out of 7 described, are present in Serbia: PPV-M, PPV-D
and PPV-Rec.
In order to determine the prevalence of strains in plum and apricot
orchards in Serbia during the three-year period (2008-2010), a total of 283 plum
and apricot leaf samples from over 90 orchards were collected by standardized
procedure. Strain-typing of isolates was done by IC-RT-PCR in 4 separate PCR
reactions with PPV-M and PPV-D specific primers with target sequences in two
different genomic regions. For phylogenetic analysis and determination of
diversity rate, 41 isolates were selected for sequencing fragments from two
genomic regions (C-ter NIb‒N-ter CP and C-ter P3‒6K1‒N-ter CI).
In order to study the dynamics of spread of PPV-D and PPV-Rec strains in
direct competition, an experimental plum orchard of 400 trees of Čačanska
lepotic...a variety was built in 2008. Eight trees, which are located in the middle of
the orchard, were inoculated with a fully characterized PPV-D and PPV-Rec
isolates (4 trees per isolate-internal source of infection). All Prunus trees in the
vicinity of the experimental orchard were marked, sampled and tested for the
presence of PPV (external source of infection). Positive samples were sequenced in
C-ter NIb‒N-ter CP genomic region for further analysis with sequences of
inoculated and newly infected trees in the orchard. Every year, from 2008‒2011,
all trees from the orchard were visually inspected and tested by ELISA for the
presence of PPV. ELISA positive samples were typed by IC-RT-PCR and sequenced
in the C-ter NIb‒N-ter CP genomic region. During the last three years of research,
the presence and the number of aphids in the experimental orchard were
monitored...
Факултет:
Универзитет у Београду, Пољопривредни факултетДатум одбране:
12-03-2013Пројекти:
- Sharka Containment (EU-204429)
- Стварање и очување генетичког потенцијала континенталних врста воћака (RS-31064)