Powdery Mildew of Prairie Gentian: Characteristics,Molecular Phylogeny and Pathogenicity

In March 1999, we found prairie gentian (Eustoma grandiflorum) infected with powdery mildew in a greenhouse in Oita Prefecture, Japan. Morphological observation revealed that the causal fungus belongs to the mitosporic genus Oidium subgenus Pseudoidium [teleomorph: Erysiphe sensu Braun and Takamatsu (2000)]. Precise taxonomic position of the fungus, however, is uncertain due to lack of the perfect stage. We determined the nucleotide sequence of the rDNA ITS region of the fungus. Comparison of the sequence with those obtained from DNA databases of this fungal group revealed that the sequence is identical to those of powdery mildews from garden four-o'clock (Mirabilis jalapa) and broad bean (Vicia faba). Inoculation of an isolate from garden four-o'clock caused mildew on prairie gentian and broad bean, suggesting that the prairie gentian mildew originates from garden four-o'clock or broad bean. Molecular phylogenetic analysis indicated a close relationship of this fungus to Erysiphe baeumleri on Vicia spp. and E. trifolii on Trifolium pratense. From these results, we propose that prairie gentian mildew diverged from a Fabaceae-parasitic ancestor. Received 14 March 2002/ Accepted in revised form 28 May 2002     

Physiological Races and Vegetative Compatibility Groups of Fusarium oxysporum f. sp. lactucae Isolated from Crisphead Lettuce in Japan

One hundred and sixteen isolates of Fusarium oxysporum f. sp. lactucae obtained from 85 fields in three crisphead lettuce-producing areas in Nagano Prefecture, Japan were typed for races using differential cultivars Patriot, Banchu Red Fire and Costa Rica No. 4. They were also grouped into vegetative compatibility groups (VCGs) using complementation tests with nitrate non-utilizing (nit) mutants. Two California strains reported as F. oxysporum f. sp. lactucum, a type culture of F. oxysporum f. sp. lactucae, and 28 avirulent isolates of F. oxysporum obtained from crisphead lettuce were included for comparison. Among Nagano isolates, 66 isolates were identified as race 1, and 50 as race 2. Race 1 strains derived from Shiojiri and Komoro cities and race 2 from Kawakami village and Komoro city. All isolates of race 2 were biotin auxotrophs, and the race could be distinguished based on its requirement for biotin on minimal nitrate agar medium (MM). Pathogenic isolates were classified into two VCGs and three heterokaryon self-incompatible isolates. Strong correlations were found between race and VCG. All the race 1 strains were assigned to VCG 1 except self-incompatible isolates, and all the race 2 strains to VCG 2. The 28 avirulent isolates of F. oxysporum were incompatible with VCG 1 and VCG 2. California strains was vegetatively compatible with VCG 1, and they were assigned to race 1. Based on vegetative compatibility, these two races of F. oxysporum f. sp. lactucae may be genetically distinct, and F. oxysporum f. sp. lactucae race 1 is identical to F. oxysporum f. sp. lactucum. Received 7 May 2002/ Accepted in revised form 6 September 2002     

A Cucumber mosaic virus Isolated from Silene armeria

A Cucumber mosaic virus was newly isolated from Silene armeria and was characterized by biological, serological and molecular biological methods. Received 4 July 2001/ Accepted in revised form 28 August 2001     

Alternative Route of Infection for Bacterial Seedling Blight of Rice Caused by Burkholderia plantarii

Burkholderia plantarii , the pathogen of bacterial seedling blight of rice, was detected in paddy water. Its concentration rose in July and August. The bacterial concentration in the paddy water was always higher along levees than at distances more than 5 m from levees. Confirmed to be released into water when graminaceous weeds were immersed, B. plantarii survived for at least 4 days at 30°C. B. plantarii was splashed at least 30 cm upward by rain splash in the field. Harvested seeds, which had been sprayed with B. plantarii released from graminaceous weeds at the flowering stage, retained the bacteria. Bacterial seedling blight occurred when the seeds were then sown in nursery boxes. These results indicated that graminaceous weeds growing on levees of paddy fields are a source of infection of the disease and that rice seeds are infected through the paddy water. Received 23 May 2002/ Accepted in revised form 1 September 2002     

Whole-Cell Fatty Acid Composition to Characterize and Differentiate Isolates of Rhizoctonia Species Associated with Turfgrass Diseases in Japan

Cellular fatty acids were analyzed to characterize and differentiate 34 isolates of Rhizoctonia species representing binucleate Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB, AG 2-2 LP, R. circinata var. circinata and var. oryzae associated with turfgrass diseases in Japan. Myristic, pentadecanoic, palmitic, palmitoleic, stearic, oleic, linoleic and linolenic acids were consistently present in varying quantities in all isolates. Heptadecanoic and 9-heptadecenoic acids were present in isolates of Rhizoctonia AG-D (I), AG-D (II), R. solani AG 2-2 IIIB and AG 2-2 LP but not in isolates of R. circinata var. circinata and var. oryzae. Palmitic, oleic and linoleic acids were the major fatty acids found, constituting 88.30-98.37% of the whole-cell fatty acid content. The remaining fatty acids were present in smaller amounts. Isolates within a single group were closely clustered, whereas isolates from different groups were clearly distinguishable based on average linkage cluster analysis of cellular fatty acids. Principal component analysis, based on all fatty acids detected, confirmed the distinct separation of isolates representing the six groups of Rhizoctonia species obtained from turfgrasses. These results suggested that fatty acid analysis is useful for the characterization and differentiation of isolates of Rhizoctonia species associated with turfgrass diseases. Received 21 May 2001/ Accepted in revised form 28 September 2001     

Aromatic Substances Inhibiting Wheat Powdery Mildew Produced by a Fungus Detected with a New Screening Method for Phylloplane Fungi

Microorganisms isolated from wheat leaf surfaces were screened for inhibition of wheat powdery mildew. A new screening method, in which wheat leaves were inoculated with Blumeria graminis f. sp. tritici and incubated with the cultured microorganisms under non-contact conditions, was developed in the present study. Using this method, 10 phylloplane fungi that inhibited wheat powdery mildew were selected from 408 microorganisms isolated from wheat leaf surfaces. Among these 10 strains, a fungus designated as Kyu-W63 had an especially strong inhibitory effect. Kyu-W63 produced white colonies without spores when cultivated on PDA. Kyu-W63 had a strong aromatic odor when being cultured. Wheat powdery mildew was suppressed even though a membrane filter with a pore size of 0.45 μm was placed between the mycelial colony and wheat leaf segment. However, when activated charcoal was introduced, Kyu-W63 did not inhibit growth of B. graminis. It was presumed that volatile substances were involved in the inhibitory effect of Kyu-W63. GC-MS analysis was used to identify two substances produced by Kyu-W63 with molecular weights of 164 and 166. Kyu-W63 also inhibited the in vitro growth of four plant pathogenic fungi other than B. graminis. Received 19 September 2001/ Accepted in revised form 7 February 2002     

Ultrastructural Localization of Hydrogen Peroxide in Host Leaves Treated with AK-toxin I Produced by Alternaria alternata Japanese Pear Pathotype

The rapid generation of reactive oxygen species (ROS), called the oxidative burst, is one of the earliest host responses to pathogen infection or elicitor treatments. Therefore, we looked for the induction of ROS generation in Japanese pear leaves by the host-specific toxin, AK-toxin I using a cytochemical method for detecting H₂O₂. A small amount of non-specific generation of H₂O₂ was found in the cell walls in toxin- and water-treated susceptible and resistant leaves. Thus, the generation of H₂O₂ at cell walls appears to be caused by wounding stress during sampling. Specific generation of ROS, however, was found only in the membrane fragments and extended desmotubules characteristic of modified sites of the plasma membrane in the toxin-treated susceptible leaves. In addition, generation of H₂O₂ at plasma membranes was observed with higher frequency in toxin-treated susceptible leaves. This result indicates that the H₂O₂ generation was associated with damaged sites in the plasmalemma after toxin treatment and perhaps with the formation of membrane fragments from altered portions of the invaginated plasma membrane. Received 21 September 2001/ Accepted in revised form 25 October 2001