Karakterizacija vrsta roda Monilinia patogena koštičavih voćaka u Srbiji i osetljivost na fungicide

Abstract

Prikupljanje obolelih delova koštičavih voćaka sa simptomima sušenja cvetova, grana i grančica i mrke truleži plodova, koje prouzrokuju vrste roda Monilinia, obavljeno je tokom trogodišnjeg perioda od 2010. do 2012. godine. Pregledom 119 lokaliteta iz 15 okruga u Srbiji, prikupljen je 321 uzorak obolelih biljnih delova kako iz komercijalnih zasada, tako i iz okućnica i sa zelenih pijaca. Dobijeno je ukupno 246 monosporijalnih izolata sa morfološkim odlikama vrsta roda Monilinia. Sa ciljem utvrđivanja rasprostranjenosti vrsta roda Monilinia i rasvetljavanja etiologije oboljenja u našoj zemlji, izvršena je identifikacija dobijenih izolata do nivoa vrste na osnovu patogenih, morfoloških, odgajivačkih i ekoloških karakteristika, sa jedne strane, i molekularnih karakteristika, sa druge strane. Na osnovu svih proučenih karakteristika utvrđeno je prisustvo tri različite vrste: M. laxa, M. fructigena i M. fructicola. Dominantan prouzrokovač mrke truleži plodova i sušenja cvetova, grana i grančica koštičavih voćaka u Srbiji je M. laxa (prisutna u 96,34% uzoraka), dok je zastupljenost ostalih vrsta ovog roda znatno manja – M. fructigena (2,44% uzoraka) i M. fructicola (1,22% uzoraka). Po prvi put u Srbiji, detektovano je prisustvo M. fructicola, patogena sa IA dela I Liste karantinskih štetnih organizama, na koštičavim voćkama u našoj zemlji. Analiza SSR markera pokazala je da izolati M. fructicola iz Srbije pripadaju haplotipu iz Italije, što ukazuje na mogući put introdukcije ovog karantinskog patogena u Srbiju. Tri vrste roda Monilinia mogu se jasno razdvojiti na osnovu kompleksa morfoloških karaktera koji obuhvata: izgled, oblik i boju kolonije, prisustvo spora i koncentričnog prstena spora u kulturi, brzinu porasta micelije, kao i veličinu konidija, načina klijanja konidija i dužinu kličine cevčice pre prvog grananja. Međutim, za pouzdanu i brzu detekciju može se preporučiti Multiplex PCR metoda koja je prilagođena i uspešno primenjena za specifično dokazivanje prisustva Monilinia spp. na koštičavim voćkama u Srbiji...

The diseased plant parts of stone fruits expressing symptoms of blossom, branch and twig blight and fruit brown rot, caused by the species from the genus Monilinia, were collected during a three-year period (from 2010 to 2012). Screening of 119 localities within 15 districts in Serbia was completed; 321 samples of diseased plant parts from commercial orchards, as well as from house yards and green markets, were collected. In total, 246 monosporial isolates with morphological characteristics resembling to those of Monilinia spp. were obtained. These isolates were identified at the species level based on their pathogenic, morphological, cultural and ecological characteristics; however, identification based on their molecular characteristics was also performed. This two-level identification was conducted in order to determine the distribution of the Monilinia species in Serbia and to clarify etiology of the disease they cause. According to their characteristics, the presence of three different species was determined: M. laxa, M. fructigena and M. fructicola. The dominant causal agent of fruit brown rot and blossom, branch and twig blight of stone fruits in Serbia was M. laxa (it was present in 96.34% samples) while the presence of M. fructigena (2.44% samples) and M. fructicola (1.22% samples) was smaller. It was the first time in Serbia that M. fructicola, a pathogen from IA part of the List I of quarantine pest organisms on stone fruit species in our country, was registered. Moreover, the analysis of SSR markers showed that M. fructicola isolates originating from Serbia belonged to the same haplotype as the isolates originating from Italy, which indicated a possible path of introduction of this species to Serbia. Three species of Monilinia genus could be clearly distinguished on the basis of the complexity of morphological characters (colony appearance, shape and colour, presence of spores and concentric ring of spores in a culture, mycelial growth rate, conidia size, mode of conidia germination and length of germ tube). However, Multiplex PCR method adjasted and successfully applied for specific attestation of presence of Monilinia spp. on stone fruit species in Serbia, can be recommended for reliable and rapid detection...