During 2012, an official survey was conducted for Monilinia species present on peach and nectarine in Croatia. In total, 169 Monilinia spp. isolates were collected from 24 commercial orchards and identified according to morphology in culture and PCR. Eighty of the isolates were identified as Monilinia laxa, 70 as M. fructigena and 19 as M. fructicola. M. fructicola was found only at one location in the Mediterranean part of the country, and this is the first record of this quarantine fungus in Croatia. PCR diagnostic tests using M. fructicola‐specific primer pair MO368‐5/MO368‐10R repeatedly gave false negatives for some isolates. PCR tests using primer pair ITS1‐Mfc1/ITS4‐Mfc1 amplified M. fructicola‐specific product in all isolates and was therefore shown to be more suitable for diagnostic purposes.
Trulež plodova pitomog kestena uzrokovana parazitskim gljivama može predstavljati značajan problem u plantažnoj proizvodnji te kulture. Tijekom 2018. godine, jaka pojava truleži plodova kestena zabilježena je u mješovitoj plantaži pitomog kestena i oraha u okolici Našica. U nasadu je sakupljeno 78 plodova (sorte Bouche de Betizac, Marsol i Maraval), među kojima su laboratorijskim pregledom u njih 63 utvrđeni simptomi truleži jezgre. Iz svih simptomatičnih plodova izolirane su gljive. Kombiniranom identifikacijom na temelju morfologije i analize sekvenci ITS1-5.8S-ITS2 ribosomske DNA, gljive iz simptomatičnih plodova determinirane su kao Gnomoniopsis smithogilvyi, Neofusicoccum parvum, Botryosphaeria dothidea, Diaporthe eres i Fusarium proliferatum. Vrsta G. smithogilvyi utvrđena je u 47 od 63 ploda (75 %), nakon koje su po zastupljenosti slijedile B. dothidea (11/63 ploda, 17 %), N. parvum (10/63 ploda, 16 %), D. eres (3/63 ploda, 5 %) i Fusarium sp. (1/63 ploda, 1,6 %). U 45 plodova utvrđena je samo jedna vrsta gljive, po dvije vrste nađene su u 11 plodova, a po tri vrste gljive u sedam plodova. Sve gljive utvrđene su na plodovima pitomog kestena u Hrvatskoj po prvi puta. Među njima ističe se prvi nalaz G. smithogilvyi, patogena koji u proteklih nekoliko godina uzrokuje velike štete na plodovima kestena u Europi.
In May 2021, a tomato producer reported an occurrence of a disease unknown so far in a greenhouse near Split, Croatia. About 30% of plants (cultivar Signora) have been affected. Symptoms resembled tomato pith necrosis, bacterial disease caused by Pseudomonas corrugata, known to occur sporadically in tomato greenhouse production in Croatia. Leaves on plants developed interveinal chlorosis, followed by necrosis and leaf collapse. When main stems were longitudinally cut, brown, disintegrated and water-soaked partly hollow pith was evident. Severely affected plants wilted. With suspicion on presence of P. corrugata, bacteria were isolated from surface-sterilized pith tissue of two tomato plants by plating onto sucrose peptone agar (SPA) and King’s B medium (KB). Colonies recovered were cream-colored on SPA and non-florescent on KB. Two isolates, assigned as 1-KB and 3A, were first identified by amplification of internal transcribed spacer (ITS1) between16S rRNA and 23S rRNA using primers D21 and D22 (Manceau and Horvais 1997). The 550-bp PCR products obtained were purified and sequenced. Subsequent BLAST search showed the sequences to have 100% identity with the strain DSM 16733 isolated from tomato in Italy (Accession No. LT629790.1) and 99.77% identity with the strain SM664-12 isolated from tomato in USA (Acc. No. KC405207.1) of Pseudomonas mediterranea from NCBI. ITS sequence for one isolate 3A was deposited in GenBank under the Accession No. OP765279.1. Further identification was performed by using species-specific primers PC1/1-PC1/2 for P. mediterranea (Catara et al. 2000, 2002). Amplification of 600 bp DNA fragment confirmed the identity of isolates 1-KB and 3A as P. mediterranea. For this region sequence of isolate 3A was deposited in GenBank under the Acc. No. OP068273.1. Pathogenicity was assessed on tomato plants (cultivar Moneymaker) grown in pots in bio-chamber. Plants were grown at 25/20 °C 12h/12h dark/light regime until 8-leaves stage (BBCH 18). P. mediterranea isolate 3A was used for the inoculation. Inoculum was prepared from the isolate grown on KB medium for 48 h and suspended in sterile distilled water (concentration of 109 CFU mL-1) by dilution plate counts. Ten plants were inoculated with 10 µl of bacterial suspension injected into the stem with a syringe. Five control plants were inoculated with sterile distilled water. After 40 days of plant growth, symptoms were visible on all plants inoculated with P. mediterranea isolate 3A. Although no wilting was observed and all plants were alive, chlorosis was observed on upper leaves, chlorosis and necrosis on middle leaves, while basal leaves wilted. Longitudinal cross-sections of stems revealed brownish pith tissue with longitudinal watery pits spreading from inoculation points (Fig.S1). Symptoms were not observed on control plants. Bacterium was reisolated from three plants showing the most severe symptoms and proved to be identical to the original using species-specific primer pair PC1/1-PC1/2. To our knowledge, this is the first confirmation of P. mediterranea causing tomato pith necrosis in Croatia. Tomato pith necrosis caused by P. mediterranea may become significant bacterial disease of greenhouse tomato in Croatia.
Posljednjih nekoliko godina, virusne bolesti tikvica i tikvi u Hrvatskoj javljaju se u sve jačem intenzitetu i uzrokuju gubitke u proizvodnji. Cilj rada bio je utvrditi prisutnost virusa mozaika krastavca (CMV), virusa žutog mozaika tikvice (ZYMV) i virusa mozaika lubenice (WMV) na tim kulturama. Tijekom 2019. godine, na 18 lokacija (polja na otvorenom) u šest županija provedeni su vizualni pregledi te je ukupno sakupljeno 43 simptomatičnih uzorka tikvice i 39 uzoraka tikve u svrhu laboratorijske analize na prisutnost CMV, ZYMV i WMV. Analiza je provedena imunoenzimskim testom DAS-ELISA korištenjem komercijalnih seroloških kompleta. Od 82 uzorka, CMV je potvrđen u njih 55 (67%), ZYMV u 50 (61%), a WMV u 10 uzoraka (12%). Pojedinačna zaraza s jednim od tri istraživana virusa utvrđena je u ukupno 38 uzoraka (46%). Mješovite zaraze s dva virusa zabilježene su u 36 uzoraka (44%): 32 uzorka (39%) s kombinacijom CMV + ZYMV te četiri uzorka (5%) s kombinacijom CMV + WMV. Mješovita zaraza sa sva tri virusa (CMV + ZYMV + WMV) utvrđena je u dva uzorka. U šest uzoraka (7%) nije zabilježen niti jedan od tri istraživana virusa. U osam nasada utvrđena je prisutnost jednog virusa, u devet nasada prisutnost dva virusa, a u jednom od 18 pregledanih nasada utvrđena je prisutnost sva tri istraživana virusa.
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