Eksperimentalna evolucija virusa mozaika pepina (PepMV) in virusa Y krompirja (PVY) v mešani in samostojnih okužbah
RNA virusi se zaradi nenatančnosti pri pomnoževanju in neučinkovitih ali odsotnih popravljalnih mehanizmov zelo hitro spreminjajo. V naravi so mešane okužbe z dvema virusoma pogoste, interakcija med njima pa lahko zanju, v primerjavi s samostojno okužbo, predstavlja spremenjene selekcijske pritiske, kar lahko vpliva tudi na njuno evolucijo. Virus mozaika pepina (PepMV) in virus Y krompirja (PVY) sta znana rastlinska škodljivca, prvi povzroča izgube pridelka paradižnika, drugi pa krompirja. Zanimalo nas je ali mešane okužbe virusov PepMV in PVY vplivajo na njuno evolucijo oziroma ali se zaradi neposrednih ali posrednih interakcij med virusoma v mešani okužbi lahko spremeni dinamika evolucije virusnih populacij v rastlinah. Da bi to preverili, smo uporabili pristop eksperimentalne evolucije na modelni rastlini Nicotiana benthamiana. Vzpostavili smo več evolucijskih linij mešanih in samostojnih okužb virusov PepMV in PVY in opravili več zaporednih prenosov virusov na nove generacije rastlin. Pri tem smo s pomočjo kvantitativne verižne reakcije s polimerazo spremljali koncentracije virusov tekom eksperimentalne evolucije v linijah z mešanimi in samostojnimi okužbami in opazovali bolezenska znamenja na rastlinah v različnih pasažah. Uspešno smo vzpostavili samostojne in mešane okužbe s PepMV in PVY v več evolucijskih linijah. Bolezenska znamenja so bila v mešanih okužbah izrazitejša kot v samostojnih. Namnoževanje in širjenje virusov PVY in PepMV poteka v rastlini N. benthamiana različno hitro. Opazili smo majhne spremembe v koncentracijah virusov tekom eksperimentalne evolucije, za katere predpostavljamo, da bi lahko bile posledica delovanja genetskega drsa in/ali adaptacije virusov na mešano okužbo.
RNA viruses mutate rapidly due to their error-prone replication and are one of the fastest-evolving biological entities. Mixed infections are common in nature and interaction between viruses in mixed infection can represent a change in selective pressures, in comparison to a single infection, and might have an effect on the evolution of viruses present in mixed infection. Pepino mosaic virus (PepMV) and Potato virus Y (PVY) are well-known plant pathogens, causing loss of tomato and potato yields, respectively. We were interested whether mixed infections of PepMV and PVY affect their evolution or whether due to direct or indirect interactions between viruses in a mixed infection, the dynamics of the evolution of viral populations in plants can change. In order to test this, we used an experimental evolution approach in Nicotiana benthamiana plants. Several evolutionary lines of mixed and single infections of PepMV and PVY were established and serial passages of virus populations onto non-infected plants were performed. Using quantitative polymerase chain reaction we monitored viral titers in evolutionary lines of mixed and single infections during the experimental evolution and observed symptoms of inoculated plants in different passages. Mixed and single infections of PepMV and PVY was successfully established in Nicotiana benthamiana plants. Disease symptoms were more pronounced in mixed infections, when compared with single infections. Multiplication and systemic infection formation of PVY and PepMV in N. benthamiana occurred at different pace for each of the viruses. We observed slight changes in viral titers during the course of experimental evolution and we hypothesise those could be attributed to the effect of genetic drift and/or adaptation of viruses in mixed infection.
2018
2018-09-16 07:46:59
1060
Virus mozaika pepina, virus Y krompirja, mešana okužba, sinergizem, eksperimentalna evolucija, inokulacija
Pepino mosaic virus, Potato virus Y, mixed infection, synergism, experimental evolution, inoculation
mb22
Tjaša
Pršin
70
Maja
Ravnikar
991
Denis
Kutnjak
994
VisID
16
156403
COBISS_ID
3
4930639
4133.pdf
3768367
Predstavitvena datoteka
2018-09-16 07:47:02