Identifikacija i karakterizacija fitopatogenih gljiva prouzrokovača bolesti stabla kupine u Srbiji

Abstract

Tokom ispitivanja sprovedenog u periodu od 2013. do 2016. godine, pregledom pet komercijalno značajnijih sorti kupine na 24 lokaliteta u 13 okruga u Srbiji, ustanovljena je široka rasprostranjenost bolesti stabla kupine (incidenca 5 - 80%, prosečno 37,7%), a nekroza i pucanje kore bili su najčešći simptomi. U pojedinim lokalitetima zabeleženo je sušenje 40% biljaka uz direktno smanjenje prinosa od oko 50%. Rasvetljavanje etiologije bolesti stabla kupine u Srbiji obavljeno je kroz morfološku i molekularnu identifikaciju i karakterizaciju 468 izolata dobijenih iz 427 uzoraka. Nakon ispunjenja Kohovih postulata, na osnovu makroskopskih i mikroskopskih osobina dobijeni izolati izdvojeni su u 10 morfoloških grupa što je potvrđeno sekvenciranjem ITS regiona rDNA. Identifikovano je 10 vrsta gljiva prouzrokovača bolesti stabla kupine u Srbiji: Paraconiothyrium fuckelii, Gnomoniopsis idaeicola, Septoria spp., Botryosphaeria dothidea, Diplodia seriata, Botrytis cinerea, Colletotrichum kahawae, Diaporthe eres, Diaporthe betulae i Neofusicoccum parvum. Dodatna karakterizacija na osnovu filogenetskih analiza urađena je za vrste G. idaeicola, B. dothidea, D. seriata, C. kahawae, D. eres, D. betulae i N. parvum čime je određena njihova srodnost i međuodnos sa ostalim vrstama rodova Gnomoniopsis, Botryosphaeria, Diplodia, Colletotrichum, Diaporthe i Neofusicoccum. Izolate Septoria spp. moguće je bilo identifikovani do nivoa roda na osnovu morfoloških, patogenih i molekularnih odlika i biće predmet daljih istraživanja uz sekvenciranje dodatnih molekularnih markera. Tokom ovih istraživanja vrsta G. idaeicola prvi put je opisana na kupini u Srbiji, a kako se pokazalo da je rasprostranjena u skoro polovini ispitivanih zasada, to predstavlja i prvu masovnu pojavu na kupini u svetu kao i prvi uvid u moguću epidemiologiju nekroze stabla i sušenja kupine izazvane sa G. idaeicola. Izolati G. idaeicola iz Srbije ispoljili su morfološku, molekularnu i biološku varijabilnost što je ustanovljeno po prvi put u svetu. Utvrđena je različita brzina porasta i izgled kolonija, različito vreme i način sporulacije, kao i različita virulentnost. Filogenetske analize kombinovanih sekvenci za ITS region rDNA, β-tubulin, tef-1α i FG1093 otkriva postojanje dve odvojene grupe u okviru klastera G. idaeicola...

A Survey was conducted from 2013 to 2016 which included a total of 24 orchards planted with five of the most commonly grown blackberry cultivars in 13 administrative districts in Serbia. A wide distribution of blackberry cane diseases (incidence of 5 - 80%, with an average of 37,7%) was observed with the most frequent symptoms of bark necrosis and canker. In some of the localities, the wilting of up to 40% of plants was recorded with yield reduction of about 50%. The etiology of blackberry cane diseases was investigated through morphological and molecular identification and characterization of 468 isolates obtained from 427 samples. After fulfilling the Koch’s postulates, the isolates were grouped into 10 morphological groups based on macroscopic and microscopic features which were confirmed by the sequencing of the ITS region rDNA. Ten species of fungi causing blackberry cane diseases were identified including Paraconiothyrium fuckelii, Gnomoniopsis idaeicola, Septoria spp., Botryosphaeria dothidea, Diplodia seriata, Botrytis cinerea, Colletotrichum kahawae, Diaporthe eres, Diaporthe betulae and Neofusicoccum parvum. Additional characterization based on phylogenetic analysis was performed for G. idaeicola, B. dothidea, D. seriata, C. kahawae, D. eres, D. betulae and N. parvum and revealed their relationship to the additional species within the genera of Gnomoniopsis, Botryosphaeria, Diplodia, Colletotrichum, Diaporthe and Neofusicoccum. Isolates of Septoria spp. could be identified to the genus level based on morphological, pathogenic and molecular features and will be subjected to further research through sequencing of additional molecular markers. During this research G. idaeicola was detected on blackberries in Serbia for the first time. As it was found that G. idaeicola was distributed in almost half of the orchards, this is the first record of a massive outbreak of G. idaeicola on blackberries in the world as well as the first insight into the possible epidemiology of cane necrosis and wilting of blackberries caused by this fungus. The isolates G. idaeicola from Serbia exhibited morphological, molecular and biological variability which was recorded for the first time in the world. More specifically, different growth patterns and rates, sporulation patterns and virulence were documented...