First Observations of Mycosphaerella pini on Scots Pine in Finland.

Abstract

Red band needle blight of pines caused by Mycosphaerella pini (anamorph Dothistroma septosporum) has recently been recorded on Scots pine (Pinus sylvestris) at 14 rural districts in southern and central Finland. Scots pine is the most common and commercially most important tree species in Finland. Red bands with aggregations of conidial stromata on otherwise brown attached needles were frequently encountered on saplings and young trees in dense stands and sporadically on lower twigs of mature trees within 2 m of the ground. These symptoms and signs, typical for M. pini (1), were also observed on needles of P. contorta and P. cembra, which occur in Finland in low frequency. Symptoms of red band needle blight and abundant conidial stromata were found in March and April of 2008 after a mild and rainy winter. After this time, the frequency at which fresh acervuli were observed decreased. Conidia were isolated after squeezing conidial stromata into a drop of sterile water and rinsing out the drop onto water agar from where single conidia were picked up from under the microscope with a modified Pasteur pipette. The conidia were hyaline, smooth, thin walled, filiform, 2.0 to 2.7 (2.4) μm wide, and 15 to 37 (29.4) μm long. Germination of the conidia on water agar was 100%. The cultures grew slowly and reached a diameter of 4 to 10 mm within 3 weeks on modified orange serum agar (2) at 20°C and abundantly produced conidia. Complete sequences of the internal transcribed spacer (ITS) region including the 5.8S rRNA gene were obtained for three isolates from different rural districts: Hartola (61°34'N, 26°01'E), Kangasniemi (61°59'N, 26°39'E), and Suonenjoki (62°37'N, 27°07'E). These sequences are deposited in GenBank with Accession Nos. EU834294, EU834295, and EU834296 and are identical to each other and to more than 50 M. pini sequences in GenBank, including those of several Estonian and Austrian isolates. These isolates are deposited in the culture collection of the Finnish Forest Research Institute with identifiers Dot1, Dot2, and Dot8. Symptoms were reproduced after inoculation of 1-year-old Scots pine seedlings growing in seedling trays with 115 cm3 cavities. A conidial suspension (5 to 6·× 103 conidia ml-1) was prepared from two single-conidial cultures (Dot10 and Dot15), combined, and sprayed during July 2008 onto 192 seedlings until needles looked completely wet. Sixty-four seedlings were sprayed with distilled water as a control treatment. Seedlings were incubated outdoors in Suonenjoki and covered with a transparent plastic hood for the first 5 days after inoculation. The first symptoms (brown segments and red bands on needles) appeared on inoculated seedlings 1 month later, and conidial stromata appeared after another 2 to 4 weeks. M. pini was reisolated from the acervuli of 10 sample needles. Three months after inoculation, all inoculated seedlings showed symptoms while all noninoculated seedlings were healthy. It is possible that M. pini has spread recently from the south since it was identified for the first time in Estonia 2 years earlier (3). Although the Gulf of Finland separates Finnish pine forests from Estonian pine forests by approximately 50 to 100 km, spores may have been aerially disseminated over this distance. Alternatively, introduction of M. pini to Finland may have occurred on imported seedlings.