Jouji Moriwaki

Grouping of Colletotrichum Species in Japan Based on rDNA Sequences

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.

A refined inoculation method to evaluate false smut resistance in rice

False smut, caused by Ustilaginoideavirens, is a serious disease of rice worldwide. To evaluate false smut resistance in rice, we developed a method combining the cultivation of the main culm of rice plants in the greenhouse and rapid preparation of a conidial suspension to inject into the leaf sheath. The method was used to evaluate false smut resistance in 18 varieties/lines of rice. For comparison, field trials were also carried out in 2007 and 2008. The results indicated that the greenhouse method was more reproducible than field trials: commercial varieties tested were resistant; almost all the forage varieties were highly susceptible; and blast-resistant varieties/lines were mostly resistant to false smut. Thus, this inoculation method will be useful for determining the level of false smut resistance in rice and for breeding resistant varieties.

Colletotrichum carthami comb. nov., an anthracnose pathogen of safflower, garland chrysanthemum and pot marigold, revived by molecular phylogeny with authentic herbarium specimens

Authentic herbarium specimens of Gloeosporium carthami and G. chrysanthemi as well as live Colletotrichum strains from Carthamus tinctorius, Chrysanthemum coronarium var. spatiosum, and Calendula officinalis were judged to be conspecific based on morphological and molecular phylogenetic analyses. Representative strains from three asteraceous plants were demonstrated by inoculation tests to possess specific pathogenicity to the host plants. We propose a new combination, Colletotrichum carthami, for the anthracnose pathogen of the asteraceous plants, designating our specimen of G. carthami as its epitype. We characterize the species, which differs from the closely related Colletotrichum simmondsii in its pathogenicity and molecular characteristics.

Anthracnose of sweet pepper caused by Colletotrichum scovillei in Japan

Severe fruit rot of sweet pepper was found in Shimane, Hyogo, Chiba, Toyama, and Nagano prefectures, Japan from 2005 to 2011. Dark, sunken spots with concentric rings of orange conidial masses appeared on fruits. Pathogenic isolates from diseased fruits in the prefectures were identified as Colletotrichum scovillei. This species was added to the pathogens of sweet pepper anthracnose in Japan. The representative isolate was pathogenic to sweet pepper, tomato and chili pepper fruits, kidney bean pod, azuki bean, pea and strawberry leaves, but a caused no symptoms on cucumber or carrot in inoculation tests.

Anthracnose of Japanese radish caused by Colletotrichum dematium

Anthracnose of Japanese radish found in Kagoshima and Miyazaki prefectures was demonstrated to be caused by Colletotrichum dematium based on inoculation experiments and morphological and molecular identification of the pathogenic fungus. Although symptoms of Japanese radish anthracnose caused by Colletotrichum higginsianum were similar to those caused by C. dematium, damage by the latter pathogen was more severe than that by C. higginsianum.

Stem necrosis of aster and Russell prairie gentian caused by Chrysanthemum stem necrosis virus

A new disease causing necrotic streaks on stems and necrosis on leaves of aster (Callistephus chinensis) and Russell prairie gentian (Eustoma grandiflorum) was observed in Toyama Prefecture, Japan, in September 2009. On the bases of host reactions, serological reactions, viral particle morphology, and the nucleotide sequence of the nucleocapsid protein gene, the causal pathogen was identified as Chrysanthemum stem necrosis virus (CSNV). This report is the first in the world of natural infection of aster and Russell prairie gentian by CSNV. We propose the name “stem necrosis of aster and Russell prairie gentian” for these new diseases.

Anthracnose of snapdragon caused by Colletotrichum destructivum

Severe spotting and blight of leaves caused by Colletotrichum destructivum were found on snapdragon (Antirrhinum majus L.), a scrophulariaceous ornamental, in open fields in Shizuoka Prefecture, Japan, from June through September 2004. The fungus is added to the group of the pathogens causing anthracnose of snapdragon.

A new combination for the causal agent of tea anthracnose: Discula theae-sinensis (I. Miyake) Moriwaki & Toy. Sato, comb. nov.

The causal agent of tea anthracnose, Gloeosporium theae-sinensis, was transferred to Discula based on morphological and molecular reidentification using living cultures.

Biodegradable Plastic-degrading Activity of Various Species of Paraphoma

The fungal strain B47-9, isolated from barley, was previously selected as an effective degrader of various biodegradable plastic (BP) films such as poly(butylene succinate-co-adipate) (PBSA) and poly(butylene succinate) (PBS). The strain has not been identified based on mycological methods because it does not form fruiting bodies, which are the key to morphological identification. Here, we performed molecular phylogenetic analyses of the nuclear ribosomal RNA gene regions and their internal transcribed spacer region of B47-9 and related fungi. The results suggest that B47-9 is closely related to the genus Paraphoma. Investigation of the abilities of six strains belonging to the genus Paraphoma to degrade BPs indicated that all strains could degrade PBSA and PBS films to varying degrees. Based on our approach, we conclude that strain B47-9 is a species belonging to the genus Paraphoma.

Anthracnose of bacopa caused by Colletotrichum destructivum

Severe spotting and blighting of leaves were found on bacopa (Sutera cordata), a scrophulariaceous ornamental, in greenhouses in Gunma Prefecture, Japan, from January through February 2007. After we isolated and identified the causal fungus as Colletotrichum destructivum and inoculated host plants with the isolate to confirm pathogenicity, we named this new disease anthracnose of bacopa.