doctoral thesis
Incidence of viroses in autochthonous grapevine cultivars in Herzegovina vineyards

Ana Karačić (2015)
Sveučilište u Zagrebu
Agronomski fakultet
Metadata
TitlePojavnost viroza u autohtonim kultivarima vinove loze u hercegovačkomu vinogorju
AuthorAna Karačić
Mentor(s)Edyta Đermić
Abstract
Vinova loza važna je kultura kojoj infektivne bolesti virusne prirode mogu drastično skratiti razdoblje eksploatacije. Najvažnija je strateška metoda u kontroli virusnih bolesti vinove loze preventiva i provodi se korištenjem certificirana sadnog materijala pri podizanju novih vinograda. Da bi se uspješno kontroliralo rasprostranjivanje viroza, neophodno je provođenje specifičnih i pouzdanih dijagnostičkih metoda: enzimski imunotest na čvrstoj fazi (ELISA) i reverzna transkripcija uz lančanu reakciju polimeraze (RT-PCR). Primjenom navedenih dijagnostičkih metoda na ukupno 1332 testiranih trsova dobiven je uvid u pojavnost devet virusa na šest ekonomski najvažnijih autohtonih kultivara u hercegovačkim vinogorjima. U šest autohtonih kultivara serološkim metodama detektirani su: Grapevine leafroll-associated virus-3 (GLRaV-3) u 884 trsova (66,3 %), Grapevine fanleaf virus (GFLV) u 500 trsova (37,5 %), Grapevine leafroll-associated virus-1 (GLRaV-1) u 293 trsova (22,0 %), Grapevine fleck virus (GFkV) u 197 trsova (14,7 %), Grapevine leafrollassociated virus-4-9 (GLRaV-4-9) u 90 trsova (6,7 %), Arabis mosaic virus (ArMV) u 17 trsova (1,2 %), Grapevine leafroll-associated virus-2 (GLRaV-2) u 3 trsa (0,2 %), dok virusi: Grapevine virus A (GVA) i Grapevine virus B (GVB) nisu detektirani. Od svih je testiranih Vinova loza važna je kultura kojoj infektivne bolesti virusne prirode mogu drastično skratiti razdoblje eksploatacije. Najvažnija je strateška metoda u kontroli virusnih bolesti vinove loze preventiva i provodi se korištenjem certificirana sadnog materijala pri podizanju novih vinograda. Da bi se uspješno kontroliralo rasprostranjivanje viroza, neophodno je provođenje specifičnih i pouzdanih dijagnostičkih metoda: enzimski imunotest na čvrstoj fazi (ELISA) i reverzna transkripcija uz lančanu reakciju polimeraze (RT-PCR). Primjenom navedenih dijagnostičkih metoda na ukupno 1332 testiranih trsova dobiven je uvid u pojavnost devet virusa na šest ekonomski najvažnijih autohtonih kultivara u hercegovačkim vinogorjima. U šest autohtonih kultivara serološkim metodama detektirani su: Grapevine leafroll-associated virus-3 (GLRaV-3) u 884 trsova (66,3 %), Grapevine fanleaf virus (GFLV) u 500 trsova (37,5 %), Grapevine leafroll-associated virus-1 (GLRaV-1) u 293 trsova (22,0 %), Grapevine fleck virus (GFkV) u 197 trsova (14,7 %), Grapevine leafrollassociated virus-4-9 (GLRaV-4-9) u 90 trsova (6,7 %), Arabis mosaic virus (ArMV) u 17 trsova (1,2 %), Grapevine leafroll-associated virus-2 (GLRaV-2) u 3 trsa (0,2 %), dok virusi: Grapevine virus A (GVA) i Grapevine virus B (GVB) nisu detektirani. Od svih je testiranih trsova, 1153 (86,5 %) uzoraka nosilo barem jedan virus, dok je 179 (13,4 %) uzorka bilo zdravo, bez prisustva testiranih virusa. Zdravi trsovi predloženi su kao potencijalne biljke kandidati za proizvodnju sadnoga materijala ‘testiranog na viruse’. Analizom rezultata virusnih infekcija utvrđeno je da je 546 (40,9 %) uzoraka zaraženo samo s jednim virusom, 423 (31,7 %) s dva, 155 (11,6 %) s tri, 28 (2,1 %) s četiri, 3 (0,2 %) s pet što je ujedno i najveći broj analizom utvrđenih kombinacija. Od 90 uzoraka koji su serološki bili pozitivni na grupu virusa GLRaV-4-9 (GLRaV-4, -5, -6, -7, -9), izabrano je njih 27 kod kojih je metodom RT-PCR i korištenjem para početnica utvrđena prisutnost GLRaV-5 kod šest uzoraka kao i GLRaV-9 kod sedam uzoraka. Ostali virusi iz grupe GLRaV-4-9 nisu registrirani. Prisutnost GLRaV-2 u uzorku, u kojem je metodom ELISA detektiran, uspješno je potvrđena primjenom metode RT-PCR uz korištenje GLR2CP1/GLR2CP2 para početnica. Tijekom dvije vegetacijske sezone (2011/12 i 2012/13) vršeno je vizualno praćenje analiziranih trsova. Dominantni simptomi na šest autohtonih kultivara bili su: zastoj u kretanju vegetacije, skraćeni internodiji, različiti poremećaji u rastu i razvoju, uvijenost lišća, nejednolično odrvenjavanje i dozrijevanje grozdova, rehuljavost. Prema raspoloživim saznanjima, provedeno istraživanje predstavlja prvi nalaz GLRaV-2, GLRaV-5 i GLRaV-9 u Bosni i Hercegovini potvrđen molekularnim metodama, dok GFkV, GLRaV-2 i grupa virusa GLRaV-4-9 predstavlja prvi nalaz u Bosni i Hercegovini potvrđen serološkim metodama.
Keywordsgrapevine autochthonous cultivars detection ELISA Enzyme Linked Immunosorbent Assays RT-PCR
Parallel title (English)Incidence of viroses in autochthonous grapevine cultivars in Herzegovina vineyards
Committee MembersEdi Maletić (committee chairperson)
Marko Ivanković (committee member)
Dijana Škorić (committee member)
Ivan Pejić (committee member)
Mirna Ćurković-Perica (committee member)
GranterSveučilište u Zagrebu
Agronomski fakultet
PlaceZagreb
StateCroatia
Scientific field, discipline, subdisciplineBIOTECHNICAL SCIENCES
Agronomy
UDK631/632
APPLIED SCIENCES. MEDICINE. TECHNOLOGY
Agriculture. Plant diseases. Plant protection
Study programme typeuniversity
Study levelpostgraduate
Study programmeAgricultural Sciences
Academic title abbreviationdr. sc.
Genredoctoral thesis
Language Croatian
Defense date2015-12-11
Parallel abstract (English)
Grapevine is an important species in which infectious diseases caused by viruses can drastically reduce the period of exploitation. The most important control strategy of grapevine viroses is prevention and consists of the use of certified planting materials in the establishment of new vineyards. In order to successfully control the virus spread, the implementation of specific and reliable diagnostic methods is essential: enzyme-linked immunosorbent assay (ELISA) and reverse transcription and polymerase chain reaction (RTPCR). Using the above mentioned diagnostic techniques from the total number 1332 grapevine, an insight into the incidence of nine viruses in the six most important autochthonous cultivars grown in Herzegovina was gained. In the six autochthonous cultivars, using serological methods, the following viruses were detected: Grapevine leafroll-associated virus-3 (GLRaV-3) - in 884 grapevines (66,3 %), Grapevine fanleaf virus (GFLV) - in 500 grapevines (37,5%), Grapevine leafroll-associated virus-1 (GLRaV-1) - in 293 grapevines (22,0%), Grapevine fleck virus (GFkV) - in 197 grapevines (14,7 %), Grapevine leafrollassociated virus-4-9 (GLRaV-4-9) - in 90 grapevines (6,7 %), Arabis mosaic virus (ArMV) - in 17 grapevines (1,2%), Grapevine leafroll-associated virus-2 (GLRaV-2) - in 3 grapevines (0,2 %), while viruses Grapevine virus A (GVA) and Grapevine virus B (GVB) were not detected. Of all tested grapevines, 86,5 % specimens (1153 grapevines) had at least one virus, while 13,4 % specimens (179 grapevines) was free of tested viruses. The virus-free grapevines are proposed as a potential candidate plants for production of virus-tested planting material. The analysis of results showed that 546 grapevines (40,9%) were infected by only one virus, 423 (31,7 %) by two, 155 (11,6 %) by three, 28 (2,1 %) by four, 3 (0,2 %) by five, which presented the highest number of viruses in mixed infections. From 90 specimens positive on GLRaV-4-9 (GLRaV -4, -5, -6, -7, -9) viruses, 27 are chosen, and, by using RTPCR method employing pair of primers, the presence of GLRaV-5 in 6 specimens and GLRaV-9 in 7 specimens is detected. The other viruses belonging to GLRaV-4-9 group were not detected. The presence of GLRaV-2 in ELISA positive specimen is successfully confirmed using RT-PCR method with the use of GLR2CP1/ GLR2CP2 pair of primers. During two vegetation seasons (2011/12, 2012/13) visual observation of analysed grapevines was carried out. The dominant symptoms on 6 autochthonous cultivars: delayed start vegetation., reduced internodes, different disturbances in growth and development, leafroll, irregular lignification and ripening of grapes and, poor fruit set. According to our knowledge, this is the first report of GLRaV-2, GLRaV-5 and GLRaV-9 in Bosnia and Herzegovina on the genome (nucleic acids) based methods, and GFkV, GLRaV-2 and group of GLRaV-4-9 associated viruses on the basis of serological methods.
Parallel keywords (Croatian)vinova loza autohtoni kultivari detekcija ELISA RT-PCR
Versionaccepted version
Resource typetext
Access conditionAccess restricted to students and staff of home institution
Terms of usehttp://rightsstatements.org/vocab/InC/1.0/
Noteaccepted version
URN:NBNhttps://urn.nsk.hr/urn:nbn:hr:204:792684
CommitterVanja Masnov