The diversity of endophytic fungi within single symptomless Norway spruce needles is described and their possible role as pioneer decomposers after needle detachment is investigated. The majority (90%) of all 182 isolates from green intact needles were identified as Lophodermium piceae. Up to 34 isolates were obtained from single needles. Generally, all isolates within single needles had distinct randomly amplified microsatellite (RAMS) patterns. Single trees may thus contain a higher number of L. piceae individuals than the number of their needles. To investigate the ability of needle endophytes to act as pioneer decomposers, surface-sterilized needles were incubated on sterile sand inoculated with autoclaved or live spruce forest humus layer. The dry weight loss of 13-17% found in needles after a 20-week incubation did not significantly differ between the sterilized and live treatments. Hence, fungi surviving the surface sterilization of needles can act as pioneer decomposers. A considerable portion of the needles remained green during the incubation. Brown and black needles, in which the weight loss had presumably taken place, were invaded throughout by single haplotypes different from L. piceae. Instead, Tiarasporella parca, a less common needle endophyte, occurred among these invaders of brown needles. Needle endophytes of Norway spruce seem thus to have different abilities to decompose host tissues after needle cast. L. piceae is obviously not an important pioneer decomposer of Norway spruce needles. The diversity of fungal individuals drops sharply when needles start to decompose. Thus, in single needles the decomposing mycota is considerably less diverse than the endophytic mycota.
Lophodermium piceae, the dominant needle endophyte of Norway spruce (Picea abies), is known to be extremely diverse. This work aimed to test the possible occurrence of cryptic species within the morphological species L. piceae. Genetic variation in 36 South Finnish L. piceae isolates originating from six localities was investigated by comparing DNA sequences of three genetic markers. One of the markers was the internal transcribed sequence (ITS) of the ribosomal DNA and the other two (LP1 and LP2) were based on sequence characterized amplified regions designed for L. piceae. The LP2 marker could be detected only from isolates of L. piceae but not from 20 other ascomycete species tested. This sequence, therefore, is considered as a species-specific marker for L. piceae. For comparison, ITS sequences of isolates representing two other Lophodermium species, L. pinastri and L. seditiosum, were also investigated. In a neighbor-joining analysis of ITS sequences all L. piceae isolates fell into one cluster, which was clearly separate from those of L. pinastri and L. seditiosum. Dendrograms of the three markers were incongruent indicating that the L. piceae population examined consisted of a single phylogenetic species. No geographical differentiation was observed. Our results confirm that L. piceae is a genetically highly diverse endophytic species.
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