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Summary The composition of the endophytic fungal assemblages in Quercus cerris, Q. pubescens and Q. robur, three oak species most susceptible to decline, was investigated in Fagare ` (Padova), Ulignano (Pisa), and Radda in Chianti (Firenze). The endophytic assemblages were studied as a function of health status of the whole tree and of twigs and leaves. The effect of station altitude was tested on Q. cerris. Samples were gathered from healthy and declining trees. In April (bud opening time) 20 twigs and 10 leaves were collected from each tree. Fungi were isolated from twig segments and leaf portions. A total of 23 fungal genera with 27 different species were isolated, in addition to a few sterile mycelia. Eleven species were found to be common to all three oak species, while other species were found only on one oak species, suggesting host specificity. Among the fungal species isolated, some were known to be pathogenic (Apiognomonia quercina, Colpoma quercinum, Diplodia mutila, Phomopsis quercina). The colonization frequency (CF) of pathogenic species varied between 0.9% for A. quercina in Q. cerris and 60.2% for P. quercina in Q. robur. Analysis of variance showed a statistically significant difference among the oak species tested. The CF was higher in declining trees in comparison with healthy trees, and also in twigs vs. leaves. The CF was found to be higher for Q. cerris trees growing at sea level as compared with those situated at an elevation of 350‐400 m. In stands where, on account of particular ecological conditions, pathogenic behaviour is displayed simultaneously by more than one endophytic fungal species, trees accentuate their decline.

Summary Soil-borne species of Phytophthora were isolated from 19 of 30 examined oak forest areas in Italy. The frequency of isolated Phytophthora spp. (35.2%) was significantly correlated with soil pH and longitude of the sites. Eleven Phytophthora species were detected. Phytophthora cambivora, P. cinnamomi and P. cactorum were recovered from sites in central and southern Italy whereas P. quercina was isolated in the northern and central part of the country. Phytophthora citricola occurred all over Italy. Phytophthora quercina was the only species significantly associated with declining oak trees.

Summary Forty isolates of Diplodia mutila, a dieback agent on oaks were characterized by their capacity to produce heterokaryons, by their vegetative compatibility, and by their pathogenicity tests on Quercus cerris. The forty isolates were collected from Quercus cerris, Quercus frainetto, Quercus ilex, Quercus pubescens, Quercus robur and Quercus suber in different regions in Italy. Three nit mutants (nitt, nit and nitM) were identified with different nitrogen sources: sodium nitrite, sodium nitrate, uric acid, ammonium tartrate and hypoxanthine. On the two nutrient media employed, a potato dextrose agar (PDA) medium with chlorate (PDC), and a minimal agar medium with chlorate (MMC), isolates obtained from Q. cerris yielded the greatest number of mutants: 254 on PDC (out of a total of 583) and 190 on MMC (out of a total of 440). Nine vegetative compatibility groups (VCGs) were identified. There was a strong association between these VCGs and host species. All isolates tested except one from Q. cerris were pathogenic.

Summary Tests in vitro and in situ showed that both mycelial and conidial suspensions of Acremonium mucronatum were antagonistic for Diplodia mutila, a common pathogen on the apical twigs of declining Quercus cerris, Quercus frainetto, Quercus pubescens and Quercus robur. Antagonism was greater with the mycelium than with the conidial suspensions, and was strongest at a test temperature of 20°C. The results suggest that pathogenicity of D. mutila on declining trees may be significantly reduced by the presence of A. mucronatum.

Summary The callus-fungal method was employed to test the response to C. flaccidum of the highly susceptible P. nigra var. laricio and the resistant P. sylvestris, and to ascertain whether results obtained with this method matched in planta observations. Calli were inoculated with axenie cultures of C. flaccidum obtained by incubating basidiospores on modified Schenk and Hildebrandt medium. Several parameters were evaluated. Colony growth was more rapid on P. nigra var. laricio. Colonies were dense on P. nigra var. laricio, but sparse on P. sylvestris. Aerial hyphae growth was abundant on P. nigra var. laricio, but less frequent on P. sylvestris. Hyphal branching began after 18 h on P. nigra var. laricio and after 45 h on P. sylvestris. Necrosis of the host cells set in after 24 h on P. nigra var. laricio, and after 70 h on P. sylvestris. The number of cells with plasmolysis was much larger in P. nigra var. laricio than in P. sylvestris. These results were consistent with the known resistance of the two species on whole plants.

The penetration of Cronartium flaccidum into the cotyledons, primary needles and secondary needles of Pinus pinaster and P. nigra var, laricio was investigated. SEM examination showed that the fungus did not form appressoria and that the germ tube penetrated directly into the stomata.

The Authors observed, in Italy, over a number of years, isolated and grouped trees of the genus Quercus that declined over one or more vegetative seasons. The spread was very fast; in fact the trees declined in the course of 2–3 vegetative seasons. A series of symptoms was observed: production of epicormic shoots, yellowing of leaves, laceration of the bark, bleeding from the trunk. Many fungi were isolated: Armillaria spp., Cephalosporium spp., Cladosporium spp., Cylindrocarpon spp., Diplodia mutila, Hypoxylon mediterraneum, Phoma cavae, Pbomopsis quercina, Sporotrix spp. Future research will attempt to ascertain the possible causes of the decline.

An experiment on the effect of eight inoculum concentrations of basidiospores of Cronartium flaccidum, on development of symptoms on P. nigra spp. nigricans var. austriaca, P. nigra ssp. italica, P. nigra ssp. laricio var. calabrica (from Calabria and Corsica) showed that 60000 basidiospores/ml was the optimum concentration and P. nigra ssp. laricio was the most sensitive.

The effect of the fluorescent brightener Calcofluor (derivative of di-amine stilbene sulfonic acid) on the germination of basidiospores of Cronartium flaccidum (Alb. et Schw.) Wint. was tested by sowing spores on pine needles and then brightening them, and by labelling directly basidiospores on Calcofluor solution and subsequently sowing them on water agar. The first method enhanced germination most. It is recommended that 0.05% brightener in distilled water be used to maximise germination in pre-infection studies for resistance at the needle level.