First Report of Tomato Powdery Mildew Caused by Leveillula taurica in Taiwan

Tomato (Solanum lycopersicum L.) is one of the common and important economic crops in Taiwan. In July 2018, tomato leaves with unusual yellowing and lesions were observed in Taichung, Changhua, Nantou, and Yunlin counties in Taiwan. Symptomatic leaves initially showed chlorotic, irregularly shaped patches on adaxial side of leaves; and days later, necrosis appeared in the patches center. The disease incidence was 4 to 25% and occurred mostly on large type tomato in the field and greenhouse while rarely on cherry tomato. Since the pathogen could not be cultured in potato dextrose agar, for initial pathogen observation, symptomatic leaves were examined directly under light microscope. Conidiophores were found emerged through abaxial stomata of the lesion, and were erect, single and sometimes branched, with single-celled and dimorphic conidia, suggesting the lesions were associated with a powdery mildew. The white to hyaline pathogens collecting directly from fresh signs were also examined. Primary conidia were lanceolate, tip point, 50 to 78 × 14 to 22 μm, ratio of 2.3 to 5.9; secondary conidia were cylindrical to clavate, 47 to 72× 14 to 22 μm, ratio of 2.5 to 4.6. Conidial germ tubes were mainly terminal to subterminal and sometimes lateral. Conidial appressoria were alobate to multilobed, and hyphal appressoria were nipple-shaped, lobed to multilobed or even coralloid. No chasmothecia were found in the field samples. One representative specimens of isolates TARI_PM-3 was deposited in the National Museum of Natural Science in Taiwan (accession number F0034683), and DNA extracted from the fresh conidia was amplified respectively with primers PMITS1 and PMITS2 (Cunnington et al. 2003) for the sequences of internal transcribed spacer region (ITS1+5.8S+ITS2, partial sequence). The segment of sequence (accession numbers MT370494 in GeneBank) showed 99.8% identity with the sequence of AB045000, which was identified as Leveillula taurica (Khodaparast et al., 2001). Based on morphology and molecular analysis, the fungus was identified as L. taurica (Braun and Cook 2012; Choi et al. 2019). To confirm the pathogenicity, conidial suspensions (4 × 104 conidia/ml) of L. taurica (isolate TARI_PM-3) were used to inoculate by dropping (10 μl/site) on the abaxial side of leaves of 4-week-old potted tomato (cv. Golden Lucky). The plants were covered with transparent plastic bags for 1 day and then maintained at 16 to 26°C in a greenhouse. Lesion symptoms of leaves similar to those in the field were observed 4 weeks after inoculation, while the controlled plants inoculated with ddH2O showed symptomless. The same fungus was observed on the necrotic patches of the inoculated leaves. In Taiwan, the recorded causing pathogens of tomato powdery mildew are Erysiphe orontii and E. cichoracearum (Tzen et al. 2019). To our knowledge, this is the first report of tomato powdery mildew caused by L. taurica. Typical lesions appeared in the beginning of the disease progress in the field. Through inoculation, the same pathogen could infect sweet peppers (Capsicum annuum L.), which has been also reported as a host of L. taurica (Tzen et al. 2019), suggesting these two crops could be as alternate hosts of L. taurica in the field. Cross-species infection should be taken into consideration while managing the disease.

Ampelomyces strains isolated from diverse powdery mildew hosts in Japan: Their phylogeny and mycoparasitic activity, including timing and quantifying mycoparasitism of Pseudoidium neolycopersici on tomato

A total of 26 Ampelomyces strains were isolated from mycelia of six different powdery mildew species that naturally infected their host plants in Japan. These were characterized based on morphological characteristics and sequences of ribosomal DNA internal transcribed spacer (rDNA-ITS) regions and actin gene (ACT) fragments. Collected strains represented six different genotypes and were accommodated in three different clades of the genus Ampelomyces. Morphology of the strains agreed with that of other Ampelomyces strains, but none of the examined characters were associated with any groups identified in the genetic analysis. Five powdery mildew species were inoculated with eight selected Ampelomyces strains to study their mycoparasitic activity. In the inoculation experiments, all Ampelomyces strains successfully infected all tested powdery mildew species, and showed no significant differences in their mycoparasitic activity as determined by the number of Ampelomyces pycnidia developed in powdery mildew colonies. The mycoparasitic interaction between the eight selected Ampelomyces strains and the tomato powdery mildew fungus (Pseudoidium neolycopersici strain KTP-03) was studied experimentally in the laboratory using digital microscopic technologies. It was documented that the spores of the mycoparasites germinated on tomato leaves and their hyphae penetrated the hyphae of Ps. neolycopersici. Ampelomyces hyphae continued their growth internally, which initiated the atrophy of the powdery mildew conidiophores 5 days post inoculation (dpi); caused atrophy 6 dpi; and complete collapse of the parasitized conidiphores 7 dpi. Ampelomyces strains produced new intracellular pycnidia in Ps. neolycopersici conidiophores ca. 8–10 dpi, when Ps. neolycopersici hyphae were successfully destroyed by the mycoparasitic strain. Mature pycnidia released spores ca. 10–14 dpi, which became the sources of subsequent infections of the intact powdery mildew hyphae. Mature pycnidia contained each ca. 200 to 1,500 spores depending on the mycohost species and Ampelomyces strain. This is the first detailed analysis of Ampelomyces strains isolated in Japan, and the first timing and quantification of mycoparasitism of Ps. neolycopersici on tomato by phylogenetically diverse Ampelomyces strains using digital microscopic technologies. The developed model system is useful for future biocontrol and ecological studies on Ampelomyces mycoparasites.