Hiromitsu Negishi

Culturable leaf-associated bacteria on tomato plants and their potential as biological control agents.

Culturable leaf-associated bacteria inhabiting a plant have been considered as promising biological control agent (BCA) candidates because they can survive on the plant. We investigated the relationship between bacterial groups of culturable leaf-associated bacteria on greenhouse- and field-grown tomato leaves and their antifungal activities against tomato diseases in vitro and in vivo. In addition, the isolated bacteria were analyzed for N-acyl-homoserine lactone (AHL) and indole-3-acetic acid (IAA) production, which have been reported to associate with bacterial colonization, and resistance to a tomato alkaloid (α-tomatine). Leaf washings and subsequent leaf macerates were used to estimate the population size of epiphytic and more internal bacteria. Bacterial population sizes on leaves at the same position increased as the leaves aged under both greenhouse and field conditions. Field-grown tomatoes had significantly larger population sizes than greenhouse-grown tomatoes. Analysis of 16S rRNA gene (rDNA) sequencing using 887 culturable leaf-associated bacteria revealed a predominance of the Bacillus and Pseudomonas culturable leaf-associated bacterial groups on greenhouse- and field-grown tomatoes, respectively. Curtobacterium and Sphingomonas were frequently recovered from both locations. From the 2138 bacterial strains tested, we selected several strains having in vitro antifungal activity against three fungal pathogens of tomato: Botrytis cinerea, Fulvia fulva, and Alternaria solani. Among bacterial strains with strong in vitro antifungal activities, Bacillus and Pantoea tended to show strong antifungal activities, whereas Curtobacterium and Sphingomonas were not effective. The results indicated the differences in antifungal activity among predominant bacterial groups. Analysis of α-tomatine resistance revealed that most bacterial strains in the dominant groups exhibited moderate or high resistance to α-tomatine in growth medium. Furthermore, some Sphingomonas and Pantoea strains showed AHL and IAA production activities. Strain 125NP12 (Pantoea ananatis) showed particular α-tomatine resistance, and AHL and IAA production had the highest protective value (91.7) against gray mold. Thus, the differences of these physiological properties among dominant bacteria may be associated with the disease suppression ability of BCAs on tomato plants.

Alternaria Leaf Spot on Mealycup Sage (Salvia farinacea Benth.) Caused by Alternaria alternata (Fr.) Keissler

In June 1995, a disease causing round to irregular-shaped, water-soaked, brown to blackish brown spots on mealycup sage (Salvia farinacea Benth.) was found in Atsugi-shi, Kanagawa Prefecture, Japan. The symptoms were seen only on leaves, not on neither flower petals or stems. The disease was also found in Setagaya-ku, Tokyo, Memambetsu-cho, Hokkaido and Shimoda-shi and Matsuzaki-cho, Shizuoka. An Alternaria sp. was frequently isolated from these diseased plants. The isolates were severely pathogenic to mealycup sage and caused lesions on the inoculated leaves. The isolates were also weakly pathogenic on scarlet sage (S. splendens Sellow ex Roem. and Schult.) but not on any other Labiatae plants tested. Based on morphological characteristics, such as size of conidia, chain number, and the short beak on conidia, the causal fungus was identified as Alternaria alternata (Fr.) Keissler. This report is the first on a mealycup sage disease caused by A. alternata. Because the symptom was restricted to the leaf, the common name of Alternaria leaf spot was proposed.