First report of celery stunt anthracnose caused by <Emphasis Type="Italic">Colletotrichum simmondsii</Emphasis> in Japan

A new disease was found in Japan, on celery (Apium graveolens var. dulce) having severe chlorotic leaf spot, stunt, and dwarf with leaf curl. A spore suspension from the fungus isolated from affected plants induced identical symptoms 14 days after plants were sprayed. Identification and molecular characterization showed that the causal agent is Colletotrichum simmondsii. This report is the first of stunt anthracnose on celery caused by C. simmondsii. We propose the name “stunt anthracnose” for the new disease. Colletotrichum acutatum sensu lato, as reference pathogen of celery anthracnose, should be changed to C. fioriniae based on morphological 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 Madagascar periwinkle caused by species belonging to the Colletotrichum gloeosporioides species complex

Severe blight of stems was found on Madagascar periwinkle (Catharanthus roseus) in Shizuoka and Okinawa prefectures, Japan, in July 2003 and September 2004, respectively. Similar fungi were isolated from the diseased plants. The isolated fungi caused the disease after inoculation and subsequently were reisolated from diseased tissues. The fungi were identified as species closely related to C. siamense or C. tropicale according to sequences of the β-tubulin-2 gene, indicating that they belong to the Colletotrichum gloeosporioides species complex. This is the first report specifying the genetic relationships of the pathogens of Madagascar periwinkle anthracnose in the complex.     

Pathologic changes in pigs with prednisolone-induced recrudescence of herpesvirus infection

In pigs inoculated with Aujeszky's disease virus (ADV), recrudescence of herpesvirus infection was induced by daily administration of 1,000 mg of prednisolone for 5 days at 2 (group A) or 5 (group B) months after the primary infection. At necropsy in group A pigs, ADV was recovered from nasal secretions 3 to 9 days after prednisolone treatment initiation and from the brain cortex 10 days after treatment initiation; ADV was not recovered from group B pigs. In pigs of both groups killed 10 days after treatment initiation, 2 types of characteristic lesions were found. One type was a nonsuppurative encephalitis that consisted of neuronal necrosis, neuronophagia, and formation of eosinophilic intranuclear inclusion bodies. The 2nd type had basophilic intranuclear inclusion bodies in the enlarged endothelial cells of the kidneys, liver, lungs, adrenal glands, and lymph node sinusoids. Cells containing intranuclear inclusion bodies had immature and mature herpesvirus particles. Therefore, the brain lesions containing the eosinophilic inclusion bodies were considered to be due to ADV. Basophilic inclusion bodies in the endothelial cells were due to porcine cytomegalovirus. These observations indicated that prednisolone treatment resulted in recrudescence of ADV and porcine cytomegalovirus infections.     

Genotypes and allele frequencies of buried SNPs in a bovine single‐nucleotide polymorphism array in Japanese Black cattle

Single nucleotide polymorphism (SNP) arrays are widely used for genetic and genomic analyses in cattle breeding; thus, data derived from SNP arrays have accumulated on a large scale nationwide. Commercial SNP arrays contain a considerable number of unassigned SNPs on the chromosome/position on the genome; these SNPs are excluded in subsequent analyses. Notably, the position‐unassigned SNPs, or “buried SNPs” include some of the markers associated with genetic disease. In this study, we identified the position of buried SNPs using the Basic Local Alignment Search Tool against the surrounding sequences and characterized the relationship between SNPs and genetic diseases in Online Mendelian Inheritance in Animals based on the genomic position. We determined the position of 285 buried SNPs on the genome and surveyed the genotype and allele frequencies of these SNPs in 5,955 individual Japanese Black cattle. Eleven SNPs associated with genetic disease, which contained five buried SNPs, were found in the population with the risk allele frequency ranging from 0.00008396 to 0.46. These results indicate that buried SNPs in the bovine SNP array can be utilized to identify associations with genetic disorders from large scale accumulated SNP genotype data in Japanese Black cattle.     

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.     

Anthracnose of snapdragon caused by <Emphasis Type="Italic">Colletotrichum destructivum</Emphasis>

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.     

Anthracnose of <Emphasis Type="Italic">Polygonatum falcatum</Emphasis> caused by <Emphasis Type="Italic">Colletotrichum dematium</Emphasis>

Severe spotting, blight and drop of leaves caused by Colletotrichum dematium were found on potted plants of Polygonatum falcatum, a liliaceous ornamental, in open fields in Kagawa Prefecture, Japan, in May 2001. This new disease was named anthracnose of P. falcatum. Keisuke Tomioka, Jouji Moriwaki, Toyozo Sato contributed equally to this work. The fungal isolate and its nucleotide sequence data obtained in this study were deposited in Genebank, National Institute of Agrobiological Sciences and the DDBJ/EMBL/GenBank databases under accessions MAFF239500 and AB334523, respectively.     

Quantification of the rice false smut pathogen <Emphasis Type="Italic">Ustilaginoidea virens</Emphasis> from soil in Japan using real-time PCR

Ustilaginoidea virens is the causal agent of false smut disease of rice. In this study, we developed a real-time polymerase chain reaction (PCR) assay to clarify the relationship between false smut occurrence on rice and quantification of U. virens from soil in Japan. The method here described is sensitive, detecting less than 50 fg of pathogen DNA, and specific to the nuclear ribosomal DNA for U. virens when tested across 27 rice-pathogenic fungi and bacteria, 26 other fungi and bacteria and four plant species. As few as eight chlamydospores of U. virens per gram soil were detected when added to sterilized Gley and Ando soils. The real-time PCR assay for the soil samples was at least 100-fold more sensitive than the conventional and nested-PCR assays tested. By quantification of U. virens with real-time PCR using DNA extracted from naturally contaminated Gley soils and visual assessment of the disease in agricultural fields, a linear correlation between cycle threshold (C_T) values and the number of false smut balls was revealed. Therefore, this specific quantitative assay could be a useful tool for optimization of disease control strategies, and for studying the ecology of U. virens.