Identification, characterization and chemical management of Alternaria alternata causing blackcurrant leaf spot in China.

AIM: The aims of this study were to identify the pathogen causing blackcurrant leaf spot, assess the pathogenicity of different isolates, the host range, and the sensitivity to common fungicides, and test the effectiveness of field control for controlling A. alternata in blackcurrants in China, and potentially elsewhere. METHODS AND RESULTS: In 2020 and 2021, an uncommon leaf spot on blackcurrants was observed in Harbin (125°42'-130°10'E, 44°04'-46°40'N), Heilongjiang Province, China. Based on morphological, molecular characteristics, and phylogenetic analyses, 10 fungal isolates, identified as Alternaria alternata, were obtained from infected blackcurrant leaves of 10 infected plants in this study. To our knowledge, this is the first description of A. alternata as a causal agent of leaf spot on blackcurrants in China. A. alternata has a wide host range and infects eight of the 10 crop and ornamental plants evaluated, namely Sorbus pohuashanensis, Malus pumila, Rosa davurica, Padus racemosa, Hippophae rhamnoides, Crataegus pinnatifida, Pyrus ussuriensis, and Sambucus williamsii, but not Viburnum trilobum and Prunus tomentosa. Moreover, ten blackcurrant cultivars were screened and found to have contrasting levels of resistance to A. alternata. One was moderately resistant, four were resistant, four were susceptible, and one was highly susceptible. The A. alternata isolate was most sensitive to propiconazole-azoxystrobin, with EC50 values of 0.0038 µg ml-1 and efficacy ranging between 83.34% and 84.13% at 317 µg ml-1 in the field. CONCLUSIONS: The work reported that A. alternata is the pathogen that causes blackcurrant leaf spot in northern China. It can infect a variety of crops and ornamental plants. Considering the control cost and effect, propiconazole-azoxystrobin is more suitable for controlling leaf spot in the field.

Evaluation and control of Alternaria alternata causing leaf spot in soybean in Northeast China.

AIM: The aims of this study were to characterize the causal agent of soybean leaf spot and determine the pathogenicity of the pathogen to the main crops in Northeast China and detect the sensitivity of pathogens to the main chemical fungicides and the potted control effect. METHODS AND RESULTS: In 2020 and 2021, an uncommon leaf spot was observed in Harbin, Heilongjiang Province (125°42'-130°10'E, 44°04'-46°40'N), China. The pathogen can infect soybean leaves and cause leaf spot. We collected diseased soybean leaves and isolated four pathogen organisms, all of which were identified as Alternaria alternata through morphological and molecular identification. Koch's postulates were used to confirm pathogenicity. To the best of our knowledge, this is the first report on soybean leaf spot disease caused by A. alternata in northeast China. Moreover, A. alternata had a broad host range and caused leaf spot in most legumes. However, it did not infect medicated lentil (Dolicho Lablab L.) or tobacco (Nicotiana tabacum L.). Assessment of A. alternaria susceptibility to fungicides by spore germination method, isolates of A. alternata were most sensitive to flusilazole, with EC50 values of 0.0040-0.0053 µg ml-1. Through two pot experiments, the average control efficacy of 0.1 mg ml-1 flusilazole on soybean leaf spot caused by A. alternata was 80.7%. CONCLUSIONS: The work reported that A. alternata is the pathogen organism that causes soybean leaf spot in northern China. The pathogen organism can infect a variety of leguminous plants. Considering the control cost and effect, flusilazole is more suitable for controlling leaf spot disease in the field, and benzoxystrobin can be used as an alternative fungicide.

First report of Corn Ear Rot Caused by Fusarium asiaticum in China.

Corn (Zea mays L.) ear rot, caused by various pathogens, is one of the most significant diseases of corn worldwide. In September 2020, a survey was undertaken to identify pathogenic fungi associated with corn ear rot in Suihua city (46.63°N 126.98°E), Heilongjiang Province, China. The average disease incidence was 14.2% and 15.6% in each of two fields sampled (~5 ha) using a five-point method (100 plants/each point). Twenty tissue samples from 20 diseased ears, showing white or pink mold on the surface of corn ears, were surface disinfected in 0.5% NaOCl for 5 min, rinsed 3 times in autoclaved distilled water. After drying, four treated corn kernels (one kernel/each ear) were placed onto potato dextrose agar (PDA) amended with 50 µg/mL streptomycin. The plates were sealed with parafilm sealing film and cultured in the dark at 26℃ with 80% RH for 3 days in an incubator. A total of 12 morphologically similar fungal isolates were obtained and subcultured by transferring hyphal tips for 3-5 days. Single-conidium isolates were generated with methods reported previously (Leslie and Summerell 2006). Colonies on PDA, reaching 20.3-20.9 mm·d-1 at 26℃, consisted of white to pale yellow, locculent, lush and dense aerial mycelium with red to apricot color. Macroconidia of six isolates randomly selected on carnation leaf agar (CLA) were falciform with a foot cell, three- to six-septate, measuring 12.6-67.2 × 2.6-5.4 µm (n = 100) in the dark at 26℃ with 80% RH for 5 days. No microconidia were observed. Based on these characteristics, the isolates were preliminarily identified as Fusarium asiaticum (Chang et al. 2020; Leslie and Summerell 2006). Genomic DNA of three representative isolates YSF2, YSF4 and YSF7 were extracted and the translation elongation factor 1-α (TEF-1ɑ) gene was amplified and sequenced using the primers EF1-728F/EF1-986R (Carbone and Kohn 1999). The DNA sequences of YSF2, YSF4 and YSF7 were deposited in GenBank (OL631287.1, OP272129 and OP272130). Analysis of TEF-1ɑ sequences of YSF2, YSF4 and YSF7 showed that they were 100% identical to F. asiaticum isolates NRRL 26156 (AF212452.1) in NCBI and NRRL 13818 (AF212451.1) in Fusarium MLST. A pathogenicity test was performed on corn cv. Xinxin 1. Four days after silk emergence, 3 mL conidial suspension (106 macroconidia/ml) of each 12 isolates was individually injected into the center of the ear through the husk sideways, penetrating the kernels to a depth of about 5 mm (6 ears/each isolate) in the field (Guo et al. 2020). Six corn ears treated with sterile distilled water were used as the control. After inoculation, normal field management was carried out. All inoculated ears showed symptoms similar to those observed in the field 50 days after inoculation, while no symptoms were observed on the blank control ears. Five fungal isolates with the same colony morphology as the naturally occurring ear rot in the field were re-isolated from the inoculated corn kernels and confirmed to be F. asiaticum according to morphological characteristics and sequence analysis of five fungal isolates. F. asiaticum has previously been reported to cause corn ear rots in Japan (Kawakami et al. 2015). To our knowledge, this is the first report of F. asiaticum causing corn ear rot in Northeast China. Corn ear rot poses a threat to significantly reduce the quality of corn in a major production zone of maize in China. Therefore, its distribution needs to be investigated and effective disease management strategies developed.