THE FIRST REPORT OF TRUFFLES (TUBER AESTIVUM VITTAD.) IN ZADAR COUNTY

True truffles (Tuber spp.) are among the most appreciated mushroom in gastronomy. Due to their particular taste and limited natural availability the price of these underground mushrooms is high. In Croatia, the most important presence of Tuber spp. was reported in Istria. Since seven to eight decades ago there have been stories of Zadar’s truffles which are transmitted orally from generation to generation but, till now, no written document that could confirm it has been found. In this study, truffles were searched for on different locations in Zadar County, the presumed truffles were harvested and molecular analyses were performed to identify them. The results undoubtedly show that the summer truffle, Tuber aestivum Vittad. is present in Zadar County.

PCR primers comparisons for a successful Tuber spp. DNA region amplification in routine identifications / Primerjava PCR začetnih oligonukleotidov za uspešno pomnoževanje DNA regije Tuber spp. pri rutinski identifikaciji

Since late 20th century DNA sequencing became the method of choice method in precision species identification. The ITS region is one of the official fungal barcoding DNA markers, although in some cases sequencing of the ITS region may, due to misidentification, mislabeling or nomenclature errors in public databases, lead to incorrect or insufficient identification, as is currently a case in the genus Tuber. The aim of this study was to test, which ITS primer pairs are most appropriate and optimal for Tuber species DNA region amplification. Thereby we (1) compared amplification success for different Tuber species using fungal specific primer pair ITS1f and ITS4 and (2) compared amplification success using different ITS primer pair combinations in amplifying DNA region an example species Tuber aestivum. Based on results, Tuber aestivum was one of the most reluctant Tuber species in this study and in most cases failed to amplify with the above primer pair. After comparing different ITS primer pairs, we conclude that the primer pair ITS5 and ITS7 is the most appropriate primer pair for amplification DNA region of T. aestivum as it resulted in high amplification success from ectomycorrhizal root tips. Based on sequences, gained from public databases, we found that ITS1f and ITS6 primers have a mismatch in one base pair compared to the target sequence of Tuber aestivum, thus resulting in poor or no amplification success. Although primer pair ITS5 and ITS7 in our study was proven to be the most appropriate primer pair in amplifying DNA region Tuber aestivum species, further analysis about appropriateness of it for a general barcoding and identification of ectomycorrhiza in complex community samples is needed. Keywords: Tuber spp., ITS region, PCR amplification, ITS primers Izvleček Od konca 20. stoletja je določanje nukleotidnega zaporedja DNA postalo ena izmed pogosteje uporabljenih metod za določanje vrst. ITS regija je edna izmed uradnih glivnih DNA markerjev, čeprav lahko določanje nukleotidnega zaporedja le-te, v nekaterih primerih, predvsem zaradi napačne določitve, označevanja oziroma napak v nomenklaturi v javnih bazah podatkov, privede do napačne oziroma nenatančne določitve vrst, kar je trenutno težava pri določitvi vrst iz rodu Tuber. Namen te študije je bil testirati kateri pari ITS začetnih oligonukleotidov so najbolj primerni in optimalni za pomnoževanje DNA regij gliv iz rodu Tuber. S tem namenom smo v študiji (1) primerjali uspešnost pomnoževanja DNA regije različnih vrst iz rodu Tuber, z uporabo glivno specifičnih začetnih oligonukleotidov ITS1f in ITS4 ter hkrati (2) primerjali uspešnost pomnoževanja DNA regije vrste Tuber aestivum z uporabo različnih ITS začetnih oligonukleotidov. Na podlagi rezultatov ugotavljamo, da je vrsta T. aestivum izmed vseh analiziranih gliv iz rodu Tuber, bila najtežavnejša vrsta v naši študiji, saj je v večini primerov pomnoževanje DNA regije te vrste z uporabo glivno specifičnih začetnih oligonukleotidov ITS1f in ITS4 bilo neuspešno. Po primerjavi uspešnosti pomnoževanja z različnimi ITS začetnimi oligonukelotidi ugotavljamo, da sta bila v naši študiji ITS začetna oligonukleotida ITS5 in ITS7 najprimernejša za pomnoževanje DNA regije vrste T. aestivum, saj je bila uspešnost pomnoževanja iz ektomikoriznih vršičkov v tem primeru največja. Na podlagi T. aestivum nukleotidnih zaporedij pridobljenih iz javnih podatkovnih baz ugotavljamo, da je za začetna oligonukleotida ITS1f in ITS6 značilno neujemanje s tarčnim nukleotidnim zaporedjem (T. aestivum) v enem baznem paru, kar se lahko odraža bodisi v slabšem pomnoževalnem uspehu ali v nepomnoževanju na splošno. Kljub temu, da v naši študiji ugotavljamo, da sta začetna oligonukleotida ITS5 in ITS7 najprimernejša za pomnoževanje DNA regije glive T. aestivum, so potrebne nadaljnje analize, s katerimi bi potrdili splošno primernost omenjenega para ITS5/ITS7 za pomnoževanje DNA regije ne samo vrst iz rodu Tuber, temveč za določanje ektomikoriznih glivnih združb na splošno. Ključne besede: Tuber spp., ITS regija, PCR pomno­ževanje, ITS začetni oligonukleotidi

Mogućnosti uzgoja tartufa (Tuber spp.)

Cilj ovoga rada je opisati mogućnosti podizanja nasada tartufa te gospodarenje istim. U prvom dijelu rada opisan je tartuf kao gljiva, morfološka i biološka svojstva tartufa te pedoklimatski uvjeti rasta i razvoja. Prikazana su dosadašnja znanja o tartufima koja služe kao podloga razrade programa za njihov uzgoj. Detaljnije su opisani veliki bijeli tartuf te tamnosporni tartuf koji su najzanimljiviji za uzgoj. Također, prikazano je tartufarstvo u Hrvatskoj i svijetu kako bi se ukazalo na značaj te visokovrijedne gljive. Opisan je način obrade tla prije sadnje, plana sadnje te planirani način održavanja plantaže. Temeljem provedenih analiza i proučene literature može se zaključiti kako su određena područja u Hrvatskoj pogodna za uzgoj tartufa sa manjim odstupanjima potrebnih pedoloških uvjeta.

A Survey of the Knowledge of Truffles among Polish Foresters and Implications for Environmental Education

While the use of truffles in Poland has a long tradition, for historical reasons this knowledge was almost lost. Currently, truffles and truffle orchards are again receiving public attention. For example, the Polish State Forests supported the establishment of truffle orchards by the Forestry Research Institute. In recent years, knowledge concerning these unique hypogeous fungi has been disseminated systematically through scientific and popular publications, films, and electronic media. This study investigates the awareness of economically and culinary valued truffle fungi (Tuber spp.) among more than 1400 Polish foresters. The results show that 70% of interviewees were familiar with historical and contemporary information about growing and using truffles in Poland. Based on respondents’ age, education, type of work, and gender we attempted to identify whether these elements were associated with the state of knowledge about truffles. The results indicated that younger foresters were better informed about the presence of truffles in Poland and also about their use in the past in Polish cuisine. Environmental education was an important source of knowledge about truffle harvesting and the soils that are conducive to truffle development. Foresters who have provided forest ecology education and who are 36–65 years of age generally possessed better knowledge about truffles than other age cohorts. More than 30% of respondents expressed interest in educational courses to improve their knowledge of truffles. The results point to the need for forestry education concerning truffles and indicate the need for fostering sustainable agroforestry-centered initiatives disseminating this knowledge to the public.

ECOLOGICAL AND GENETIC ADVANCES IN THE CULTIVATION OF TUBER SPP.

Recent advances on the ecology and genetics of true truffles (Tuber spp.) are reported and their impact on truffle cultivation is discussed. New insights have been gained on truffle soil ecology and interrelationships of truffles with associated microorganisms in the soil. For instance, some bacteria seem to play a key role in truffle fruiting body formation and maturation. However, the most important advance in truffle genetics over the past 20 years has been the sequencing of the Tuber melanosporum genome and the discovery that truffles, like other Pezizalean fungi, are heterothallic. This finding has had a significant impact on research on truffles and many studies have been devoted to better understanding the distribution of the mating types in soil in natural and cultivated truffières. The characterization of the mating type idiomorphs of several Tuber species has led to the possibility of selecting mycelial strains for truffle cultivation in particular sites.