Tihomir Miličević

Wild vegetable mixes sold in the markets of Dalmatia (southern Croatia)

BackgroundDalmatia is an interesting place to study the use of wild greens as it lies at the intersection of influence of Slavs, who do not usually use many species of wild greens, and Mediterranean culinary culture, where the use of multiple wild greens is common. The aim of the study was to document the mixtures of wild green vegetables which are sold in all the vegetable markets of Dalmatia.MethodsAll vendors (68) in all 11 major markets of the Dalmatian coast were interviewed. The piles of wild vegetables they sold were searched and herbarium specimens taken from them.ResultsThe mean number of species in the mix was 5.7. The most commonly sold wild plants are: Sonchus oleraceus L., Allium ampeloprasum L., Foeniculum vulgare Mill., Urospermum picroides F.W.Schmidt, Papaver rhoeas L., Daucus carota L., Taraxacum sp., Picris echioides L., Silene latifolia Poir. and Crepis spp. Also the cultivated beet (Beta vulgaris L.) and a few cultivated Brassicaceae varieties are frequent components. Wild vegetables from the mix are usually boiled for 20–30 minutes and dressed with olive oil and salt. Altogether at least 37 wild taxa and 13 cultivated taxa were recorded.Apart from the mixes, Asparagus acutifolius L. and Tamus communis L. shoots are sold in separate bunches (they are usually eaten with eggs), as well as some Asteraceae species, the latter are eaten raw or briefly boiled.ConclusionsThe rich tradition of eating many wild greens may result both from strong Venetian and Greek influences and the necessity of using all food resources available in the barren, infertile land in the past. Although the number of wild-collected green vegetables is impressive we hypothesize that it may have decreased over the years, and that further in-depth local ethnobotanical studies are needed in Dalmatia to record the disappearing knowledge of edible plants.

Fusarium spp. Contamination of Wheat, Maize, Soybean, and Pea Grain in Croatia

Fusarium spp. Contamination of Wheat, Maize, Soybean, and Pea Grain in Croatia From 2002 to 2008, 203 samples of wheat, maize, soybean, and pea were analysed for the presence of Fusarium species. Contamination with Fusarium spp., expressed as the percentage of seeds with Fusarium colonies, ranged from 5 % to 69 % for wheat, from 25 % to 100 % for maize, from 4 % to 17 % for soybean, and from 3 % to 17 % for pea. 187 isolates were collected and the following 19 species determined: F. graminearum, F. poae, F. avenaceum, F. verticillioides, F. sporotrichioides, F. heterosporum, F. crookwellense, F. tricinctum, F. semitectum, F. oxysporum, F. proliferatum, F. solani, F. equiseti, F. pseudograminearum, F. chlamydosporum, F. sambucinum, F. compactum, F. scirpi, and F. culmorum. Dominant species were F. graminearum on wheat (27 % of isolates), F. verticillioides on maize (83 % of isolates), F. sporotrichioides on soybean (34 % of isolates), and F. proliferatum on pea (29 % of isolates). Among species identified, F. heterosporum, F. crookwellense, F. pseudograminearum, F. sambucinum, and F. compactum have been reported for the first time in Croatia. Kontaminacija zrna pšenice, kukuruza, soje i graška vrstama fusariuma u Hrvatskoj U periodu od 2002. do 2008. g. analizirana je prisutnost vrsta Fusariuma na 208 uzoraka zrna pšenice, kukuruza, soje i graška. Kontaminacija vrstama Fusariuma, izražena kao postotak sjemenki s kolonijama Fusarium spp., kretala se od 5 % do 69 % na pšenici, od 25 % do 100 % na kukuruzu, od 4 % do 17 % na soji te od 3 % do 17 % na grašku. Prosječna kontaminacija vrstama Fusariuma u različitim godinama varirala je od 10 % do 46 % na pšenici, od 50 % do 91 % na kukuruzu, od 5 % do 9 % na soji te od 7 % do 10 % na grašku. Vrste Fusariuma koje se javljaju na zrnu izolirane su i determinirane s odabranih uzoraka pšenice, kukuruza, soje i graška. Skupljeno je 187 izolata, a utvrđeno je 19 vrsta: F. graminearum, F. poae, F. avenaceum, F. verticillioides, F. sporotrichioides, F. heterosporum, F. crookwellense, F. tricinctum, F. semitectum, F. oxysporum, F. proliferatum, F. solani, F. equiseti, F. pseudograminearum, F. chlamydosporum, F. sambucinum, F. compactum, F. scirpi i F. culmorum. Dominantne vrste bile su F. graminearum na pšenici (27 % izolata), F. verticillioides na kukuruzu (83 % izolata), F. sporotrichioides na soji (34 % izolata) te F. proliferatum na grašku (29 % izolata). U Hrvatskoj su prvi put utvrđene vrste F. heterosporum, F. crookwellense, F. pseudograminearum, F. sambucinum i F. compactum.

Identification of Fusarium Species Isolated from Stored Apple Fruit in Croatia

Abstract Several species of the genus Fusarium can cause apple fruit to rot while stored. Since Fusarium taxonomy is very complex and has constantly been revised and updated over the last years, the aim of this study was to identify Fusarium species from rotten apples, based on combined morphological characteristics and molecular data. We identified 32 Fusarium isolates from rotten apple fruit of cultivars Golden Delicious, Jonagold, Idared, and Pink Lady, stored in Ultra Low Oxygen (ULO) conditions. Fusarium rot was detected in 9.4 % to 33.2 % of naturally infected apples, depending on the cultivar. The symptoms were similar in all four cultivars: a soft circular brown necrosis of different extent, with or without visible sporulation. Fusarium species were identified by the morphology of cultures grown on potato-dextrose agar (PDA) and carnation leaf agar (CLA). Twenty one isolates were identified as Fusarium avenaceum and confirmed as such with polymerase chain reaction (PCR) using specific primer pair FA-ITSF and FA-ITSR. F. pseudograminearum,F. semitectum, F. crookwellense, and F. compactum were identified by morphological characteristics. F.avenaceum can produce several mycotoxins and its dominance in Fusarium rot points to the risk of mycotoxin contamination of apple fruit juices and other products for human consumption. Pathogenicity tests showed typical symptoms of Fusarium rot in most of the inoculated wounded apple fruits. In this respect Fusarium avenaceum, as the dominant cause of Fusarium rot in stored apple fruits is a typical wound parasite. Fuzarijsku trulež ploda jabuke može uzrokovati veći broj vrsta roda Fusarium. Budući da je taksonomija roda Fusarium vrlo kompleksna te je podložna neprestanim promjenama posljednjih godina, cilj ovog rada bio je identificirati vrste roda Fusarium, izolirane sa zaraženih plodova jabuke na temelju morfoloških karakteristika i molekularnih analiza. Skupljena su ukupno 32 izolata roda Fusarium sa zaraženih plodova jabuke kultivara Golden Delicious, Jonagold, Idared i Pink Lady, nakon skladištenja u ULO („Ultra Low Oxygen“) uvjetima. Fuzarijska je trulež zabilježena na 9,4 % do 33,2 % plodova zahvaćenih skladišnim bolestima. Simptomi fuzarijske truleži bili su slični kod sva četiri kultivara, a javljali su se u obliku smeđe, mekane nekroze koja se postepeno širi, uz pojavu ili bez pojave vidljive sporulacije na površini ploda. Vrste su identificirane na temelju morfoloških karakteristika na krumpirsko-dekstroznom agaru (KDA) i agaru s listićem karanfila (LKA). Ukupno je 21 izolat determiniran kao vrsta Fusarium avenaceum, što je potvrđeno i lančanom reakcijom polimerazom (PCR) uz uporabu specifičnog para početnica FA-ITSF i FA-ITSR. Vrste F.pseudograminearum, F. semitectum, F. crookwellense i F. compactum identificirane su na temelju morfologije. Istraživanjem je utvrđeno da je F. avenaceum, vrsta koja je potencijalni proizvođač nekoliko mikotoksina, dominantan uzročnik fuzarijske truleži ploda jabuke nakon skladištenja. Visok postotak zaraze fuzarijskom truleži može izazvati kontaminaciju sokova jabuke i ostalih proizvoda mikotoksinima, sekundarnim metabolitima vrsta Fusarium štetnih za ljudsko zdravlje. Testovi patogenosti pokazali su da većina izolata uzrokuje pojavu tipičnih simptoma fuzarijske truleži nakon inokulacije na oštećeni plod. Navedeno upućuje na činjenicu da je F. avenaceum, kao dominantan uzročnik fuzarijske truleži ploda jabuke tipičan parazit rana

Grapevine Trunk Diseases Associated with Fungi from the Diaporthaceae Family in Croatian Vineyards

Abstract Grapevine trunk diseases (GTD) have a variety of symptoms and causes. The latter include fungal species from the family Diaporthaceae. The aim of our study was to determine Diaporthaceae species present in the woody parts of grapevines sampled from 12 vine-growing coastal and continental areas of Croatia. The fungi were isolated from diseased wood, and cultures analysed for phenotype (morphology and pathogenicity) and DNA sequence (ITS1, 5.8S, ITS2). Most isolates were identified as Phomopsis viticola, followed by Diaporthe neotheicola and Diaporthe eres. This is the first report of Diaporthe eres as a pathogen on grapevine in the world, while for Diaporthe neotheicola this is the first report in Croatia. Pathogenicity trials confirmed Phomopsis viticola as a strong and Diaporthe neotheicola as a weak pathogen. Diaporthe eres turned out to be a moderate pathogen, which implies that the species could have a more important role in the aetiology of GTD. Bolesti vinove loze koje se danas u općeprihvaćenoj engleskoj fitopatološkoj terminologiji označavaju kao grapevine trunk disease (GTD) obuhvaćaju bolesti drva vinove loze s različitom simptomatologijom i etiologijom. Kao jedan od uzroka GTD-a navode se i fitopatogene gljive iz porodice Diaporthaceae. Iz ove porodice na vinovoj lozi do sada je u Hrvatskoj utvrđena samo vrsta Phomopsis viticola kao uzročnik bolesti crna pjegavost i Phomopsis cotoneastri. U svijetu se navodi još 15 vrsta koje spadaju u rod Diaporthe/Phomopsis s različitom patogenosti na vinovoj lozi pa se stoga neke vrste smatraju endofitima, a neke patogenima. Radi utvrđivanja etiologije bolesti drva vinove loze povezanih s gljivama iz porodice Diaporthaceae uzimani su uzorci bolesnog drva i rozgve vinove loze iz različitih vinogorja unutar svih 12 vinogradarskih podregija kontinentalne i primorske Hrvatske. Gljive su iz zaraženog drva izolirane u čistu kulturu na hranjivu podlogu. Taksonomski status izolata utvrđen je na temelju njihove fenotipske karakterizacije (karakteristike kolonija i spora) i analizom DNA-sekvencija molekularnog markera ITS (ITS1, 5.8S, ITS2). Najveći broj izoliranih gljiva identificiran je kao vrsta Phomopsis viticola, dok je manji dio izoliranih gljiva pripadao vrstama Diaporthe neotheicola i Diaporthe eres. Za vrstu Diaporthe eres ovo je prvi nalaz na vinovoj lozi u svijetu, a za vrstu Diaporthe neotheicola prvi nalaz u Hrvatskoj. Testovi patogenosti potvrdili su da je Phomopsis viticola izrazito patogena vrsta na vinovoj lozi, dok se vrsta Diaporthe neotheicola pokazala slabim patogenom, a vrsta Diaporthe eres utvrđena je kao srednje jak patogen pa bi mogla imati važniju ulogu u etiologiji GTD-a.

FIRST REPORT OF FIG MOSAIC VIRUS AND FIG BADNAVIRUS 1 ON COMMON FIG TREES IN CROATIA

Common fig (Ficus carica L.) is one of the most common fruit trees in the Croatian coastal region. Fig mosaic symp- toms, first reported more than a half century ago (Perisic , 1952) are found on most cultivars. In 2014, leaf samples were taken from 30 autochthonous fig trees from two locations of the Istrian peninsula for laboratory analyses. Twenty six trees exhibited typical symptoms of foliar mosaic, one tree exhibited also fasciations, while the remaining four trees were asymptomatic. Extraction of total RNA was done us- ing an RNeasy plant mini kit (Qiagen, Germany) and ex- traction of DNA using a GenElute Plant Genomic DNA Miniprep Kit (Sigma-Aldrich, USA). Plants were tested by PCR for the presence of Fig mosaic virus (FMV) and Fig badnavirus 1 (FBV-1), applying previously described condi- tions and diagnostic primers (Elbeaino et al., 2009; Laney et al., 2012). A symptomless fig tree was used as negative control. The infection with FMV was confirmed in 26 symp- tomatic out of 30 sampled trees (87%), while all 30 plants were positive for FBV-1. One amplicon each of FMV and FBV-1 (isolates FMV-Cro-19S and FBV-1-Cro-17S, respec- tively), were sequenced in both directions (GenBank acces- sion Nos KT312843 and KT312844). A BLASTN program search showed 97% identity at the nucleotide level of the FMV-Cro-19S RdRp gene with the comparable sequence of FMV SB2-2 isolate from Serbia (AB697838), while the poly- protein gene from FBV-1-Cro-17S had a sequence identical to that of several FBV-1 isolates from the USA (JN050864, JN050875, JN112365). To the best of our knowledge, this is the first report of FMV and FBV-1 in Croatia.

Dynamics of most important fungal diseases of strawberries in Croatia and suggestions for integrated control

During trials (1998-2001) in the strawberry plantation of the cv. Marmolade we monitored the appearance of the most important fungal diseases, their biology and possibilities of integrated control. Monitoring the biology of the determined fungi we established the way of overwintering, appearance of fruiting bodies and discharging of spores as well as beginning of primary infection and first symptoms of the following fungi: Mycosphaerella fragariae, Diplocarpon earliana, Gnomonia comari, Phomopsis obscurans on leaves and Botrytis cinerea on fruits. While checking the efficacy of fungicides, those most effecting to leaf diseases were Folicur Multi and Quadris, and on B. cinerea Switch and Teldor. Testing the resistance on botryticides the fungus B. cinerea showed low level of resistance to pyrimethanil (anilinopyrimidines), dichlofluanid (phenilsulfamides) and high level to vinclozolin (dicarboximides).

First Report of Verticillium Wilt Caused by Verticillium dahliae on Olive Trees in Croatia

In April 2015, dieback of young olive trees (Olea europaea L., cvs. Istarska bjelica and Buza) occurred in a private orchard near Vodnjan in Istria, Croatia. From total of 50 trees, eight (16%) showed symptoms of dieback, with branches that had vascular browning in cross section and attached light brown, downwardly curled leaves. A fungus was consistently isolated from surface sterilized wood pieces of diseased branches, obtained in pure cultures on potato dextrose agar (PDA) by monohyphal isolation, and incubated at 25°C in darkness. Colonies grew slowly (1.9 mm/day), and the initial white color became gradually dark. Microscopic analysis showed hyaline, septate hypha with conidiophores bearing phialides in verticillate arrangement producing hyaline, single-celled, ovoid to ellipsoid conidia, 4.8 ± 0.7 µm × 2.4 ± 0.4 µm (n = 100). Dark parts of the colonies showed abundant dark-brown microsclerotia embedded in PDA. Solitary microsclerotia (rounded, elongated, or irregular in shape) and their aggregates were 22 to 117 µm and up to 404 µm in diameter, respectively. Based on morphology, the fungus was identified as Verticillium dahliae Kleb. (Inderbitzin et al. 2011). For molecular identification, the representative isolate MTH4 was used for PCR amplification and sequencing of its ITS region (GenBank accession no. KX061497) using the primer pair ITS5/ITS4, showing 99.8% homology to reference isolate of V. dahliae (PD322, HQ206718) (Inderbitzin et al. 2011) and 100% homology to isolate SDV1025 (KC834733) (Yu et al. 2016), confirming identification based on morphology. Pathogenicity of eight isolates obtained in this study (MTH1 to MTH8) was tested on 2-year-old olive plants (cv. Oblica), three per isolate. Their roots were gently washed with tap water to remove soil and immersed for 1 h in conidial suspension (105 spores/ml) obtained from 10-day-old colonies on PDA. Control plants were treated with sterile water only. All the plants were potted in steam-sterilized soil and kept in greenhouse at 24 ± 2°C, around 70% relative humidity, and 15 h of daylight while watered twice a week. Inoculated plants, but not controls, showed leaf chlorosis after 4 to 5 weeks, and dieback by week six. Reisolations of V. dahliae from inoculated plants were attempted as described above and were successful for all but control plants, thus satisfying Koch’s postulates. This is the first report of V. dahliae causing dieback of olive in Croatia. During this study, it was observed that approximately 15% of olive trees in neighboring orchards also showed symptoms of dieback ; therefore, V. dahliae can be suspected as more widespread in Istria than was previously known, and control measures are needed. Diplodia seriata De Not., previously found to cause similar symptoms on young olive in Croatia (Kaliterna et al. 2012), was not found in this study.

Identification of Fusarium Species Isolated From Stored Apple Fruit in Croatia / Identifikacija Vrsta Roda Fusarium Izoliranih S Plodova Jabuke Nakon Skladištenja

Abstract Several species of the genus Fusarium can cause apple fruit to rot while stored. Since Fusarium taxonomy is very complex and has constantly been revised and updated over the last years, the aim of this study was to identify Fusarium species from rotten apples, based on combined morphological characteristics and molecular data. We identified 32 Fusarium isolates from rotten apple fruit of cultivars Golden Delicious, Jonagold, Idared, and Pink Lady, stored in Ultra Low Oxygen (ULO) conditions. Fusarium rot was detected in 9.4 % to 33.2 % of naturally infected apples, depending on the cultivar. The symptoms were similar in all four cultivars: a soft circular brown necrosis of different extent, with or without visible sporulation. Fusarium species were identified by the morphology of cultures grown on potato-dextrose agar (PDA) and carnation leaf agar (CLA). Twenty one isolates were identified as Fusarium avenaceum and confirmed as such with polymerase chain reaction (PCR) using specific primer pair FA-ITSF and FA-ITSR. F. pseudograminearum,F. semitectum, F. crookwellense, and F. compactum were identified by morphological characteristics. F.avenaceum can produce several mycotoxins and its dominance in Fusarium rot points to the risk of mycotoxin contamination of apple fruit juices and other products for human consumption. Pathogenicity tests showed typical symptoms of Fusarium rot in most of the inoculated wounded apple fruits. In this respect Fusarium avenaceum, as the dominant cause of Fusarium rot in stored apple fruits is a typical wound parasite. Fuzarijsku trulež ploda jabuke može uzrokovati veći broj vrsta roda Fusarium. Budući da je taksonomija roda Fusarium vrlo kompleksna te je podložna neprestanim promjenama posljednjih godina, cilj ovog rada bio je identificirati vrste roda Fusarium, izolirane sa zaraženih plodova jabuke na temelju morfoloških karakteristika i molekularnih analiza. Skupljena su ukupno 32 izolata roda Fusarium sa zaraženih plodova jabuke kultivara Golden Delicious, Jonagold, Idared i Pink Lady, nakon skladištenja u ULO („Ultra Low Oxygen“) uvjetima. Fuzarijska je trulež zabilježena na 9,4 % do 33,2 % plodova zahvaćenih skladišnim bolestima. Simptomi fuzarijske truleži bili su slični kod sva četiri kultivara, a javljali su se u obliku smeđe, mekane nekroze koja se postepeno širi, uz pojavu ili bez pojave vidljive sporulacije na površini ploda. Vrste su identificirane na temelju morfoloških karakteristika na krumpirsko-dekstroznom agaru (KDA) i agaru s listićem karanfila (LKA). Ukupno je 21 izolat determiniran kao vrsta Fusarium avenaceum, što je potvrđeno i lančanom reakcijom polimerazom (PCR) uz uporabu specifičnog para početnica FA-ITSF i FA-ITSR. Vrste F.pseudograminearum, F. semitectum, F. crookwellense i F. compactum identificirane su na temelju morfologije. Istraživanjem je utvrđeno da je F. avenaceum, vrsta koja je potencijalni proizvođač nekoliko mikotoksina, dominantan uzročnik fuzarijske truleži ploda jabuke nakon skladištenja. Visok postotak zaraze fuzarijskom truleži može izazvati kontaminaciju sokova jabuke i ostalih proizvoda mikotoksinima, sekundarnim metabolitima vrsta Fusarium štetnih za ljudsko zdravlje. Testovi patogenosti pokazali su da većina izolata uzrokuje pojavu tipičnih simptoma fuzarijske truleži nakon inokulacije na oštećeni plod. Navedeno upućuje na činjenicu da je F. avenaceum, kao dominantan uzročnik fuzarijske truleži ploda jabuke tipičan parazit rana

Identifikacija vrsta roda Fusarium izoliranih s plodova jabuke nakon skladištenja

Abstract Several species of the genus Fusarium can cause apple fruit to rot while stored. Since Fusarium taxonomy is very complex and has constantly been revised and updated over the last years, the aim of this study was to identify Fusarium species from rotten apples, based on combined morphological characteristics and molecular data. We identified 32 Fusarium isolates from rotten apple fruit of cultivars Golden Delicious, Jonagold, Idared, and Pink Lady, stored in Ultra Low Oxygen (ULO) conditions. Fusarium rot was detected in 9.4 % to 33.2 % of naturally infected apples, depending on the cultivar. The symptoms were similar in all four cultivars: a soft circular brown necrosis of different extent, with or without visible sporulation. Fusarium species were identified by the morphology of cultures grown on potato-dextrose agar (PDA) and carnation leaf agar (CLA). Twenty one isolates were identified as Fusarium avenaceum and confirmed as such with polymerase chain reaction (PCR) using specific primer pair FA-ITSF and FA-ITSR. F. pseudograminearum,F. semitectum, F. crookwellense, and F. compactum were identified by morphological characteristics. F.avenaceum can produce several mycotoxins and its dominance in Fusarium rot points to the risk of mycotoxin contamination of apple fruit juices and other products for human consumption. Pathogenicity tests showed typical symptoms of Fusarium rot in most of the inoculated wounded apple fruits. In this respect Fusarium avenaceum, as the dominant cause of Fusarium rot in stored apple fruits is a typical wound parasite. Fuzarijsku trulež ploda jabuke može uzrokovati veći broj vrsta roda Fusarium. Budući da je taksonomija roda Fusarium vrlo kompleksna te je podložna neprestanim promjenama posljednjih godina, cilj ovog rada bio je identificirati vrste roda Fusarium, izolirane sa zaraženih plodova jabuke na temelju morfoloških karakteristika i molekularnih analiza. Skupljena su ukupno 32 izolata roda Fusarium sa zaraženih plodova jabuke kultivara Golden Delicious, Jonagold, Idared i Pink Lady, nakon skladištenja u ULO („Ultra Low Oxygen“) uvjetima. Fuzarijska je trulež zabilježena na 9,4 % do 33,2 % plodova zahvaćenih skladišnim bolestima. Simptomi fuzarijske truleži bili su slični kod sva četiri kultivara, a javljali su se u obliku smeđe, mekane nekroze koja se postepeno širi, uz pojavu ili bez pojave vidljive sporulacije na površini ploda. Vrste su identificirane na temelju morfoloških karakteristika na krumpirsko-dekstroznom agaru (KDA) i agaru s listićem karanfila (LKA). Ukupno je 21 izolat determiniran kao vrsta Fusarium avenaceum, što je potvrđeno i lančanom reakcijom polimerazom (PCR) uz uporabu specifičnog para početnica FA-ITSF i FA-ITSR. Vrste F.pseudograminearum, F. semitectum, F. crookwellense i F. compactum identificirane su na temelju morfologije. Istraživanjem je utvrđeno da je F. avenaceum, vrsta koja je potencijalni proizvođač nekoliko mikotoksina, dominantan uzročnik fuzarijske truleži ploda jabuke nakon skladištenja. Visok postotak zaraze fuzarijskom truleži može izazvati kontaminaciju sokova jabuke i ostalih proizvoda mikotoksinima, sekundarnim metabolitima vrsta Fusarium štetnih za ljudsko zdravlje. Testovi patogenosti pokazali su da većina izolata uzrokuje pojavu tipičnih simptoma fuzarijske truleži nakon inokulacije na oštećeni plod. Navedeno upućuje na činjenicu da je F. avenaceum, kao dominantan uzročnik fuzarijske truleži ploda jabuke tipičan parazit rana

Grapevine Trunk Diseases Associated with Fungi from the Diaporthaceae Family in Croatian Vineyards / Identifikacija Vrsta Roda Fusarium Izoliranih S Plodova Jabuke Nakon Skladištenja

Abstract Grapevine trunk diseases (GTD) have a variety of symptoms and causes. The latter include fungal species from the family Diaporthaceae. The aim of our study was to determine Diaporthaceae species present in the woody parts of grapevines sampled from 12 vine-growing coastal and continental areas of Croatia. The fungi were isolated from diseased wood, and cultures analysed for phenotype (morphology and pathogenicity) and DNA sequence (ITS1, 5.8S, ITS2). Most isolates were identified as Phomopsis viticola, followed by Diaporthe neotheicola and Diaporthe eres. This is the first report of Diaporthe eres as a pathogen on grapevine in the world, while for Diaporthe neotheicola this is the first report in Croatia. Pathogenicity trials confirmed Phomopsis viticola as a strong and Diaporthe neotheicola as a weak pathogen. Diaporthe eres turned out to be a moderate pathogen, which implies that the species could have a more important role in the aetiology of GTD. Bolesti vinove loze koje se danas u općeprihvaćenoj engleskoj fitopatološkoj terminologiji označavaju kao grapevine trunk disease (GTD) obuhvaćaju bolesti drva vinove loze s različitom simptomatologijom i etiologijom. Kao jedan od uzroka GTD-a navode se i fitopatogene gljive iz porodice Diaporthaceae. Iz ove porodice na vinovoj lozi do sada je u Hrvatskoj utvrđena samo vrsta Phomopsis viticola kao uzročnik bolesti crna pjegavost i Phomopsis cotoneastri. U svijetu se navodi još 15 vrsta koje spadaju u rod Diaporthe/Phomopsis s različitom patogenosti na vinovoj lozi pa se stoga neke vrste smatraju endofitima, a neke patogenima. Radi utvrđivanja etiologije bolesti drva vinove loze povezanih s gljivama iz porodice Diaporthaceae uzimani su uzorci bolesnog drva i rozgve vinove loze iz različitih vinogorja unutar svih 12 vinogradarskih podregija kontinentalne i primorske Hrvatske. Gljive su iz zaraženog drva izolirane u čistu kulturu na hranjivu podlogu. Taksonomski status izolata utvrđen je na temelju njihove fenotipske karakterizacije (karakteristike kolonija i spora) i analizom DNA-sekvencija molekularnog markera ITS (ITS1, 5.8S, ITS2). Najveći broj izoliranih gljiva identificiran je kao vrsta Phomopsis viticola, dok je manji dio izoliranih gljiva pripadao vrstama Diaporthe neotheicola i Diaporthe eres. Za vrstu Diaporthe eres ovo je prvi nalaz na vinovoj lozi u svijetu, a za vrstu Diaporthe neotheicola prvi nalaz u Hrvatskoj. Testovi patogenosti potvrdili su da je Phomopsis viticola izrazito patogena vrsta na vinovoj lozi, dok se vrsta Diaporthe neotheicola pokazala slabim patogenom, a vrsta Diaporthe eres utvrđena je kao srednje jak patogen pa bi mogla imati važniju ulogu u etiologiji GTD-a.