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ABSTRACTA new genus, Brevicollum, is established for two new species, B. hyalosporumon Syzygium samarangenseand B. versicoloron Volkameria inermis, in Japan. This genus is characterized by the presence of immersed ascomata with an excentric to central, short cylindrical ostiolar neck, thin ascomatal wall, clavate asci with a shallow ocular chamber, and broadly fusiform, hyaline to pale brown ascospores with a sheath. Molecular phylogenetic analyses based on a combined data set of sequences of 18S and 28S nuc rDNA genes and the second largest subunit of RNA polymerase II (RPB2) revealed that Brevicollumis a member of Neohendersoniaceae (Pleosporales, Dothideomycetes). The genera Brevicollum, Crassiparies, Medicopsis, and Neohendersoniaare accepted in Neohendersoniaceae. Revised descriptions of Crassipariesand its type species, C. quadrisporus, are provided based on the holotype and a newly obtained specimen and isolate. An asexual morph of C. quadrisporusis reported for the first time. The nuc rDNA internal transcribed spacer (ITS1-5.8S-ITS2 = ITS) phylogenetic tree of the newly collected species (B. hyalosporum, B. versicolor, and C. quadrisporus), and the unnamed or provisionally identified endophytic species, suggests a broader distribution of Brevicollumand Crassipariesspecies.

: Internal transcribed spacers (ITS) of the ribosomal RNA gene (rDNA) were sequenced for 236 isolates covering 25 Colletotrichum species collected in Japan. The Japanese isolates could be grouped into 20 ribosomal groups (RGs) based on the sequences of ITS1, correlating the species identified by the morphology. Colletotrichum gloeosporioides sensu lato separated into three RGs that were morphologically different. Colletotrichum destructivum, C. linicola and C. higginsianum were possibly conspecific. Colletotrichum dematium sensu lato including C. capsici and other species that produce falcate conidia except for graminicolous ones were separated into three RGs that were difficult to distinguish morphologically. In the phylogenetic study using ITS2 and the 28S rDNA domain 2 region, topologies compiled by neighbor-joining and maximum-parsimony methods were almost the same, reflecting the conidial morphology. The phylogenetic group 1 (PG1) produced conidia with acute ends, e.g., C. acutatum, C. destructivum and C. graminicola; PG2 produced those with obtuse ends, e.g., C. gloeosporioides, and C. orbiculare. Colletotrichum theae-sinensis, which produced the smallest conidia, was grouped as PG3, far from other species, indicating it should not belong to Colletotrichum. Grouping and phylogenetic analysis using ribosomal DNA was an effective tool to classify and identify Colletotrichum species without using morphology.

Abstract: RLGS is a technique to detect DNA polymorphism using restriction sites as landmarks. It identifies the landmarks through direct end-labeling, two-dimensional electrophoresis and autoradiography, giving a profile with many spots to allow the scanning of numerous DNA loci. We successfully performed the technique on fungi using isolates ofColletotrichum acutatum andC. gloeosporioides in anamorphic Ascomycotina,Rhizopus oryzae in Zygomycotina,Phytophthora nicotianae in Mastigomycotina (or Oomycota) andRhizoctonia solani in anamorphic basidiomycotina. RLGS of total genomic DNA digested with three restriction enzymes,Not, I,EcoR V andMbo I, reproducibly gave specific profiles of ca. 400 to 1.600 spots for each isolate. A polymorphic spot appearing to reflect a genetic difference between the twoColletotrichum species was found in the profiles of the isolates. No other common spots were found in any combination of isolates of the twoColletotrichum species, and thus the other spots on the profiles were regarded as unique to each isolate. These results indicated that RLGS could be applied, as a powerful fingerprinting technique based on genetic information from the whole genomic DNA, to search for useful DNA markers for taxonomic and genomic studies on many fungal species.

RLGS is a technique to detect DNA polymorphism using restriction sites as landmarks. It identifies the landmarks through direct end-labeling, two-dimensional electrophoresis and autoradiography, giving a profile with many spots to allow the scanning of numerous DNA loci. We successfully performed the technique on fungi using isolates of Colletotrichum acutatumand C. gloeosporioidesin anamorphic Ascomycotina, Rhizopus oryzaein Zygomycotina, Phytophthora nicotianaein Mastigomycotina (or Oomycota) and Rhizoctonia solaniin anamorphic Basidiomycotina. RLGS of total genomic DNA digested with three restriction enzymes, NotI, EcoRV and MboI, reproducibly gave specific profiles of ca. 400 to 1,600 spots for each isolate. A polymorphic spot appearing to reflect a genetic difference between the two Colletotrichumspecies was found in the profiles of the isolates. No other common spots were found in any combination of isolates of the two Colletotrichum species, and thus the other spots on the profiles were regarded as unique to each isolate. These results indicated that RLGS could be applied, as a powerful fingerprinting technique based on genetic information from the whole genomic DNA, to search for useful DNA markers for taxonomic and genomic studies on many fungal species.

: Severe wilt with spots and/or leaf and stem blight were found on a scrophulariaceous flowering plant, Nemesia strumosa, grown in Kagawa Prefecture, Japan, in February 1999. Wilted plants had numerous lesions and died early. A mitosporic fungus isolated repeatedly from the diseased plants was identified as Colletotrichum fuscum and was demonstrated to cause the disease. N. strumosa is a new host for C. fuscum, which has been known to attack foxglove (Digitalis spp.). The present disease was named “anthracnose of N. strumosa” as a new disease.

: In October 1998, a disease causing mainly foliar necrotic lesions was found on African marigold (Tagetes erecta) and French marigold (T. patula) grown in Miyagi Prefecture, Japan. Similar lesions also developed on stems and flowers, resulting in early blight of the affected organs. Plants with numerous lesions withered rapidly. A mitosporic fungus isolated repeatedly from the diseased plants was identified as Alternaria tagetica and demonstrated to cause the disease. The disease, as well as the fungus, is new to Japan. We propose the name “hanten-byô”, which means leaf spot in Japanese, for this disease.

: Severe rot of stem bases caused by Pythium aphanidermatum was found on ulluco (Ullucus tuberosus) grown in Kagawa Prefecture, Japan, in September 1999. The name “foot rot of ulluco” is proposed for this new disease.

: Severe blight of stems and leaves caused by Rhizoctonia solani AG-1 (IB) was found on yacon (Smallanthus sonchifolius), a Compositae tuber crop, grown in Ehime Prefecture, Japan, from August to September 1996 and 1999. We named it “Rhizoctonia blight of yacon” as a new disease.