First Report of Medical Tree Peony Root Rot Caused by Fusarium solani in Tongling, China.

Abstract

Tree peony bark, a main component of Chinese traditional medicine used for alleviating fever and dissipating blood stasis, is mainly produced in Tongling, China. Recently, tree peony cultivation in this area was seriously affected by root rot, with approximately 20 to 30% disease incidence each year. The disease severely affects yield and quality of tree peony bark. During the past 2 years, we collected 56 diseased tree peony plants from Mudan and Fenghuang townships in Tongling. We found reddish brown to dark brown root rot in mature roots, especially on those with injuries. Plant samples collected were disinfected with 2% sodium hypochlorite and isolations were conducted on potato sucrose agar (PSA). Eleven isolates were obtained and all had white fluffy aerial hypha on PSA. Two types of conidia were produced; the larger, reaphook-shaped ones had three to five septa and the smaller, ellipse-shaped ones had one or no septum. The reaphook-shaped conidia were 20.15 to 37.21 × 3.98 to 5.27 μm and the ellipse-shaped conidia were 6.02 to 15.52 × 2.21 to 5.33 μm in size. Chlamydospores were produced, with two to five arranged together. Biological characteristics of the fungi indicated that the optimum temperature for the mycelial growth on PSA was 25 to 30°C and the optimum pH range was 5.5 to 7.0. The above morphological characteristics point the fungal isolates to be Fusarium solani. To confirm pathogenicity, 30 healthy 1-year-old tree peony seedling plants were grown in pots (25 cm in diameter) with sterilized soil and a conidial suspension from one isolate (FH-1, 5 × 105 conidia/ml) was used for soil inoculation. Inoculated seedlings were maintained at 28°C in a greenhouse with a 12-h photoperiod of fluorescent light. Seedlings inoculated with distilled water were used as controls. After 3 weeks, the roots were collected and rinsed with tap water. Dark brown lesions were observed in the inoculated mature roots but not in the control roots. To confirm the identity of the pathogen, F. solani strains were reisolated from the lesions and total genomic DNA was extracted with the cetyltriethylammnonium bromide method from the mycelia of the reisolated strains (1). PCR was performed using the fungal universal primers ITS4 (5'-TCCTCCGCTTATTGATATGC-3') and ITS5 (5'-GGAAGTAAAAGTCGTAACAAGG-3') to amplify a DNA fragment of approximately 590 bp. The purified PCR products were sequenced (Invitrogen Co., Shanghai, China) and shared 100% sequence identity with each other. A comparison of the sequence (JQ658429.1) by the Clustal_W program (2) with those uploaded in GenBank confirmed with the fungus F. solani (100% sequence similarity to isolate S-0900 from the Great Plains of the United States; EU029589.1). To our knowledge, this is the first report of F. solani causing medical tree peony root rot in China. The existence of this pathogen in China may need to be considered for developing effective control strategies.