Camarops species encompass members of wood-inhabi- ting ascomycetous fungi and represent an ecologically important fungal group with many members commonly occurring throughout temperate and tropical regions [1, 2]. The genus Camarops P. Karst. is characterized by applanate or flat-pulvinate stromata, with separate ostioles at the surface and very long tubular perithecia compo- sing a palisade layer of cream or brownish color [3]. C. hypoxyloides P. Karst was classified as the type species of the genus.
Since this genus was established in 1872, its classifica- tion has been changed. Camarops had been belonged to the Xylariaceae due to their stromatal affinities and brown ascospores as Hypoxylon and related genera [4, 5]. Alth- ough Doi [6] and Barr [7] have suggested it has affinities with the Sordariaceae, recent molecular data support that Camarops is placed in the Boliniaceae [8]. Currently, approximately 19 species have been reported worldwide [9]. One species, C. petersii (Berk. & M.A. Curt.) Nannf., was reported in South Korea by Japanese researcher Iwade in 1894 [10].
The national biological inventory organized by the National Institute of Biological Resources (NIBR, www. nibr.go.kr) has yielded numerous collections of fungi within Korea. While investigating fungal specimens of wood-inhabiting ascomyceteous fungi deposited at NIBR, we found an unreported Camarops species for Korea. To confirm its affinity with the genus Camarops, phyloge- netic analysis was carried out based on the sequence data of nuclear large subunit ribosomal DNA (nucLSU rDNA) region. Here we present a detailed description of a new species for Korea.
Macroscopic and microscopic characteristics were based on the deposited Camarops specimen (NIBRFG000010 6691) which was collected in 19 March, 2009 from Mt. Gaji in Ulsan-si, Korea. Measurements of micro-structures such as ascospores were made from slide preparations mounted in 3% KOH [11] using a NIKON 80i light microscope and the micro-structure of specimen was photographed using Digital Camera (Nikon D200).
The genomic DNA was extracted from the dry speci- men using AccuPrep Genomic DNA extraction kit (Bio- neer, Korea). The nucLSU region was amplified using LR 0R and LR5 primers [12]. The PCR conditions used were 95°C for 5 min, followed by 30 cycles of 1 min at 94°C, 1 min at 51°C, and 1 min at 72°C, and a final extension step at 72°C for 10 min. The PCR product was confirmed using electrophoresis on 1.2% agarose gel and purified using an AccuPrep PCR purification kit (Bioneer, Korea). DNA sequencing was performed at the DNA Synthesis and Sequencing Facility, Macrogen (Seoul, Korea), by using an ABI3700 automated DNA sequencer. For mole- cular identification, Camarops sequence was compared with the reference sequences in GenBank by using BLAST [13]. Sequences were edited and aligned with reference sequences retrieved from GenBank database using MEGA4 [14]. The phylogenetic tree was constructed with 1,000 bootstrap replicates using Neighbor-Joining method. The partial nucLSU region sequence was deposited in the GenBank database (KM042417).
Phylogenetic analysis based on the nucLSU rDNA reg- ion sequences showed that Camarops did not form mono- phyletic group (Fig. 1). This result was in agreement with previous result in which three genera (Camarops, Cornipulvina, and Camaropella) belonged to the family Bolini- aceae were intermingled in the nucLSU based phylogene- tic tree [2]. Although Camarops did not form a mono- phyletic clade with the closely related genera, Camarops species are well separated. Also the phylogenetic tree showed that C. ustulinoides and C. petersii were clearly separated. Our specimen (NIBRFG0000106691) placed in a clade comprising reference the C. ustulinoides AFTOL-ID 72 (DQ470941) with 95 % bootstrap values support and their sequences had 99 % similarity (842bp/845bp). The result indicated the specimen is C. ustulinoides.
Fig. 1. A phylogenetic tree of Camarops species inferred from Neighbor-Joining method based on nuclear large subunit (nucLSU) sequences. Bootstrap values more than 50% from a sample of 1,000 replicates are shown on each branch.
Camarops ustulinoides (Henn.) Nannf., Svensk Bot. 66: 370, 1972.
Basionym: Nummularia ustulinoides Henn., Hedwigia. 36: 227, 1897.
Stromata: peltate, button-shaped, or loaf-shaped, 10-40 mm × 5-10 mm × 2-4 mm high, with abrupt angular to rounded margins, bark adhering, upper surface convex to nearly plane, with punctuate ostioles with raised circu- lar rims, externally dull brown to blackish, internally tan to black, external stromatal layer 0.5-1 mm thick, very hard, completely encasing fragile perithecia and entostro- ma, entostroma cheesy to woody (Fig 2A and 2C).
Perithecia: 1-3 mm long × 0.3-0.5 mm diam, cylindri- cal to ellipsoidal, with periphysate neck, arranged in monostichous manner (Fig 2B and 2E).
Asci: 8-spored, cylindrical, the spores arranged in a partially biseriate manner, short- or long-stipitate, 23-35 μ m × 3-5 μ m [15].
Ascospores: grayish to brown to dark brown, ellipsoi- dal with one end somewhat acute, laterally compressed, smooth, 4-5 μ m × 2.5-3 μ m × 2-2.2 μ m, with a germ pore at the more pointed end (Fig 2D).
Habitat: on trunk of dead oak tree.
Specimen examined: KOREA, Ulsan, Mt. Gaji, on trunk of dead oak tree, 19 March 2009, Changmu Kim (NIBR FG0000106691; GenBank KM042417).
Geographical distribution: Costa Rica, French Guiana, Puerto Rico [2,3].
Camarops ustulinoides specimens represent brown to blackish stromata with bark adhering (Fig. 2A), elongated perithecia in section of stromata (Fig. 2B) and generally raised circular rims (Fig. 2C). This species is distinguished by elongated peritheica and raised circular rims from C. petersii. C. petersii has white outer layer and brown liquid exhibited at early stage. C. ustulinoides has not been com- monly reported from Asia and Europe [16].
