Diaporthe (anamorph = Phomopsis) species are plant pathogens and endophytes on a wide range of hosts including economically important crops. At least four Diaporthe taxa occur on soybean and they are responsible for serious diseases and significant yield losses. Although several studies have extensively described the culture and morphological characters of these pathogens, their taxonomy has not been fully resolved. Diaporthe and Phomopsis isolates were obtained from soybean and other plant hosts throughout Croatia. Phylogenetic relationships were determined through analyses of partial translation elongation factor 1-alpha (EF1-α) gene and ITS nrDNA sequence data. By combining morphological and molecular data, four species could be distinguished on soybeans in Croatia. Diaporthe phaseolorum is described in this study and its synonyms are discussed. Diaporthe phaseolorum var. caulivora is raised to species status and the name Diaporthe caulivora is introduced to accommodate it. A species previously known as Phomopsis sp. 9 from earlier studies on sunflower, grapevine, rooibos and hydrangea is reported for the first time on soybean, and is formally described as Diaporthe novem. The well-known soybean pathogen Phomopsis longicolla was also collected in the present study and was transferred to Diaporthe longicolla comb. nov. The presence of these species on herbaceous hosts raises once more the relevance of weeds as reservoirs for pathogens of economically important plants.
Weeds are alternative hosts of plant pathogens and when colonized may not exhibit disease symptoms. In 2008 and 2009, samples of weeds and plant debris were collected from 12 locations in eastern Croatia, and 300 Fusarium isolates colonizing them were identified. Strains were grouped and identified based on morphology and amplified fragment length polymorphism (AFLP) patterns. Portions of the b-tubulin and translocation elongation factor 1-a genes were sequenced from representative strains of each group to confirm the identifications. Fourteen
SUMMARYAsparagus is an interesting vegetable as a part of a healthy diet and has been widely studied due to the high nutritional value and potential positive effects on human health. Wild asparagus (Asparagus acutifolius L.) grows along the Adriatic coast and islands, while the cultivated asparagus (A. officinalis L.) is mostly grown in the eastern region of Croatia. The aim of this study was to determine total polyphenol content (measured spectrophotometrically) and antioxidant activity (DPPH) in wild and cultivated asparagus. No significant differences in total polyphenol content were found between wild and cultivated asparagus. The highest content of polyphenols in wild asparagus was 6. [mg GAE/g fresh weight] and 49.60 [mg GAE/g dry weight], and in cultivated asparagus it was 4.52 [mg GAE/g fresh weight] and 50.93 [mg GAE/g dry weight]. Antioxidant activity measured using DPPH (2,2-diphenyl-1-picrylhydrazyl) method showed statistically significant difference. Higher antioxidant activity was measured in cultivated than in wild asparagus. The results also indicate that increased polyphenol content leads to higher antioxidant activity, but more so in the cultivated asparagus. Based on the results, the cultivated asparagus is an excellent alternative to wild asparagus that should be protected from over-harvesting, thus preventing devastation of plant diversity of islands and coast.
A cane disease of blackberry {Rubus sp.) cv. Thomfree was observed in May and June 2010 in two growing regions in the eastem part of Slavonia in Croatia. Symptoms consisted of bleached areas between and around cane nodes with some canes showing wilt symptoms. Infected areas were covered with numerous, black pycnidia immersed in the epidermal tissue. Disease occurrence in orchards growing cv. Thornfree ranged between 1 and 15%. Thirty disease samples were collected, disinfected (1 min in 70% ethanol and 2 min in 1% NaOCl), and placed in a moist chamber for 4 days. Fungal sporulating structures were then picked off and placed on potato dextrose agar (PDA). Fungal isolates obtained were identified as a Phomopsis sp., tbe conidial state of Diaporthe (3), on the basis of cultural and morphological characteristics. Alpha conidia were unicellular, hyaline, fusiform, sometimes tapering toward one or both ends, biguttulate (sometimes with several guttules), and 5.2 to 9.7 x 1.4 to 2.7 |im (average 6.5 x 2.1 |jm). Beta conidia were hyaline, ascptate, filiform, hamate, and 16.6 to 28.2 X 0.5 to 1.5 um (average 24.0 x 1.1 |jm). The teleomorph was not observed. Biomolecular analysis was performed to identify the fungal species by sequencing the internal transcribed spacer (ITS) region spanning ITS 1, 5.8S rDNA, and ITS 2 of two isolates (Phkl and Phk2). The amplified product was sequenced (GenLab-Enea, Rome, Italy) and a BLAST search of the NCBI nucleotide database was performed. Sequences from Phkl and Phk2 (GenBank Accession Nos. HQ533144 and HQ533143, respectively) were identical to authentic and vouchered Diaporthe eres Nitschke (GenBank DQ4915I4, BPI 748435, and CBS 109767) ITS sequences in GenBank. Fungal isolates for pathogenicity tests were grown on PDA at 25°C for 7 days (12 h light/dark regimen). Inoculations were made on 30 to 40 cm long green shoots of potted plants of the blackberry cv. Thomfree. One-centimeter long wounds were made with a sterile scalpel and mycelia of D. eres were placed in the wounds. Inoculation sites were covered with a piece of wet cotton wool and aluminum foil to retain moisture. Three replications of 10 plants each were inoculated and these plus 10 control plants (inoculated with plugs of PDA only) were maintained in a growth chamber at 25°C. After 25 days, lesions developed on all 30 inoculated plants that averaged 15 mm long and control plants remained symptomless. D. eres was rcisolated from inoculated plants, thus completing Koch's postulates. Phomopsis spp. have previously been reported on blackberry canes in Serbia (1) and Yugoslavia (2,4), however, to our knowledge, this is the first report of the occurrence of D. eres (anamorph P. oblonga) on blackberry in Croatia.
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