First Report of Corynspora cassiicola Causing Leaf Spot on Jasminum sambac in China.

Jasminum sambac (Linnaeus) Aiton belonging to Oleaceae family, has been cultivated in China over 1500 years. It is an important tea crop in South China, and popular ornamental plant in North China. In June 2015, Corynespora leaf spot on Jasminum sambac was first observed in Caozhuang in Tianjin(E: 117°20', N: 39°13'). In a series of investigations, the disease incidence of Corynespora leaf spot on Jasminum sambac in Tianjin was found to be as high as 20%. The disease was also found in flower planting greenhouses in Beijing and Hebei region. The initial symptoms appeared as small circular spot with gray white in the center and dark brown in the edge. The lesions expanded rapidly to 5-10 mm diameter, surrounded by a yellow halo. The affected leaves became covered with the circular spots and defoliated eventually. Fifty-six leaf samples were collected from 20 greenhouses, then directly examined and disease tissues set aside for isolation. Conidiophores were either solitary or in clusters, straight or curved and un-branched, with 3-10 septate, pale to dark brown in color. Those were measured 80.6 to 234.8 μm long and 4.5 to 8.0 μm wide in size and were formed with 0-8 successive cylindrical proliferation (n = 50). Conidia borne from conidiophore, were single or in chains, obclavate to cylindrical, smooth. The average size of 50 conidia was 43.6 to 130.7 μm long×6.4 to 13.5 μm wide, with 4 to 8 pseudosepta (n = 50). The infected leaves were cut into small pieces, sterilized with 75% ethanol, rinsed in sterilized distilled water and then plated on potato dextrose agar (PDA). The isolated plates were incubated at 25℃ for 5 days. Thrity-five pure culture isolates were obtained from diseased leaves. The colonies were pale brown on the front and dark brown on the underneath, with thickly aerial mycelia. Morphological characteristics of the pathogen from the infected leaves and purified strains matched descriptions of Corynesora cassicola (Ellis and Holliday 1971). To confirm the pathogenicity, conidial suspensions (106 conidia mL-1 ) of 35 isolated strains were sprayed on the healthy plants at the seedling stage of Jasminum sambac respectively, 5 plants per strains. 10 healthy plants were sprayed only with sterilized water as control. Conidial suspensions were prepared by washing conidia from the pure C. cassiicola cultures. Conidial suspensions were adjusted to a concentration of 1 × 106 conidia/mL with the hemocytometer. All plants were incubated in artificial climate-room at 90% relative humidity, 25±2℃. After 7 days, small circular spots, similar to the field samples were observed on all plants inoculated with the pathogen. By comparison, control plants displayed no symptoms. The same pathogens were re-isolated from the symptomatic leaves fulfilling Koch's postulates. For molecular identification, rDNA-ITS and two housekeeping genes (TUB2 and EF-1α) gene were amplified and sequenced (Zhu et al. 2020). The sequences of three high pathogenicity isolats showed 99% homology to C.cassiicola (MK571399) with 464 bp for rDNA-ITS, 100% homology to C.cassiicola (MN512639) with 469 bp for TUB2 gene, 98.61% homology to C.cassiicola (MH263735) with 287 bp for EF-1α gene. And all 9 sequences were submitted to GenBank (GenBank accession No. MT334575-77 for rDNA-ITS, MT886387-89 for TUB2, and MT886390-92 for EF-1α).The molecular identification and morphological characteristics showed that the isolated strains from Jasminum sambac were identified as C.cassiicola. C. cassiicola has an extensive host range, covering 397 host plants (Farr and Rossman 2020). Recently, C.cassiicola has been reported on Jasminum mesnyi of Oleaceaein China (Zhang et al. 2018). To our knowledge, this is the first report of leaf spot caused by C.cassiicola on Jasminum sambac in China and also worlwide.