Schisandra chinensis (Turcz.) Baill is a perennial liana belonging to the Schisandra genus of the family Magnoliaceae, which is cultivated in China as an important medicinal plant. In the summer of 2008, we observed a previously unknown foliar disease on the schisandras in Jingyu and Antu counties and the cities of Ji'an and Hunchun in Jilin Province. Symptoms appeared on the apex, margin, and center of leaves. The infection initially manifested as pale brown, small, necrotic spots on the leaves. Subsequently, these lesions became grayish brown in the center and dark brown with slight protuberances at the margins. Finally, these lesions developed concentric rings with a clear boundary separating them from the healthy tissue, were round to elliptical or irregular in shape, and had a diameter of 3 to 5 mm. In severely infected leaves, these spots eventually covered the entire leaf. Black spots (pycnidia) were produced on the infected leaf tissues in a humid environment. Fungus from infected leaf tissues was isolated on potato dextrose agar. The cultures were initially pale brown and turned dark green with age. Embedded pycnidia were generally formed after 5 days. The pycnidia were agglutinating, globose to subglobose, and measured 60.0 to 212.0 × 33.6 to 268.0 μm. Abundant conidia (4.06 to 7.2 × 1.65 to 3.53 μm) exhibiting zero to three oil droplets were produced by an 8-day-old colony; these conidia were ovoid or ellipsoidal, colorless, and aseptate; they were similar to conidia of Phoma glomerata. The internal transcribed spacer (ITS) sequence of rDNA of the isolated pathogenic strain (PG11; GenBank Accession No. GU724511) had 100% identity to P. glomerata (GenBank Accession No. HM769279). Therefore, the pathogen was identified as P. glomerata (Corda) Wollenw. & Hochapfel on the basis of morphology and ITS sequence data. To validate Koch's postulates, schisandra leaves were spray inoculated with a 2.5 × 105 conidia/ml suspension of the isolated pathogen. An equal number of healthy plants were inoculated with sterile water (control). After inoculation, 10 plants were covered with plastic bags for 3 days and maintained in a growth chamber at 25°C. After 8 days, all inoculated plants showed symptoms identical to those observed on the schisandra leaves infected in the field, whereas the controls did not show any symptoms. Reisolation of the fungi from lesions of inoculated leaves confirmed that the causal agent was P. glomerata. Diseases caused by P. glomerata have been reported on some plants (1,2). However, to our knowledge, this is the first report of leaf spot disease caused by P. glomerata on S. chinensis in China as well as in the world. References: (1) J. S. Chohan et al. Trans. Br. Mycol. Soc. 75:509, 1980. (2) T. Thomidis et al. Eur. J. Plant Pathol. 131:171,2011.
Can ITS sequence data identify fungal endophytes from cultures? A case study from Rhizophora apiculata