First Report of Leaf Spot Disease on Dictamnus dasycarpus Caused by Phoma dictamnicola in China.

Fraxinella, Dictamnus dasycarpus Turcz (Rutaceae), is a perennial herbal plant and mainly distributed in Eurasia and North America. It is often used to treat jaundice, cough, rheumatism, and other diseases and is extensively cultivated in the northeast and northwest of China (3). In June 2009, a severe foliar disease was observed on D. dasycarpus in medicinal plantations in Antu, China. The disease occurred on 100% of the plants and at least 25% of the surface was affected. In the early stages of disease development, symptoms were visible on the top and bottom of infected leaves as small brown spots. Subsequently, these spots became elliptical to irregularly shaped, with beige or grayish white centers and dark brown margins. Within the spots, numerous, dark brown or black, subglobose or ostiolate pycnidia measuring 152 to 367 μm in diameter were observed. Fungal isolates were obtained from the infected leaves on potato dextrose agar (PDA) medium, with conidia that were aseptate or one-septate and ellipsoidal or reniform, measuring approximately 4.7 to 12.6 × 2.1 to 4.5 μm. On the basis of these characteristics, the fungus was identified as a Phoma sp. Four well-sporulating isolates, designated as DdPh-1, DdPh-2, DdPh-3, and DdPh-4, were selected for further studies. The morphological and cultural characteristics of these four isolates were studied as described by Boerema et al. and the fungus was identified as Phoma dictamnicola Boerema et al. (1). The internal transcribed spacer (ITS) region of the nuclear rDNA was amplified and sequenced using primers ITS4/ITS5 (2). All four of the ITS sequences were identical (GenBank Accession No. FR681861) and were 99% identical to P. dictamnicola strains CBS507.91 (Accession No. GU237877) and KACC42445 (Accession No. EF600960). Pathogenicity tests were performed by spraying the leaves of healthy D. dasycarpus plants with a conidial suspension (1 × 10⁶ conidia/ml). Five plants were inoculated with each isolate (DdPh-1, DdPh-2, DdPh-3, and DdPh-4) and five plants were mock inoculated with sterile water. The plants were covered with plastic bags and kept in a greenhouse at 20 to 25° for 72 h. After 9 to 13 days, all inoculated plants showed characteristic symptoms as previously described, while the control plants remained healthy. The fungus was reisolated from the leaf spots of inoculated plants. Currently, the economic importance of this disease is limited, but it may become a more significant problem in production of D. dasycarpus with the cultivation area increasing. The fungus was found in the Netherlands and Korea, but to our knowledge, this is the first report of P. dictamnicola on D. dasycarpus in China. References: (1) G. H. Boerema et al. Phoma Identificatión Manual: Differentiation of Specific and Infra-Specific Taxa in Culture. CABI Publishing. Wallingford, U.K., 2004. (2) D. E. L. Cooke et al. Mycol. Res. 101:667, 1997. (3) S. S. Jiang et al. Biosci. Biotechnol. Biochem. 72:660, 2008.