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Molecular Identification of the Pathogenic Fungi of the Root Rot of Fritillaria thunbergii |
ZHANG Yu-Jia, LI Ji-Er, WEN Si-Si, XU Yun-Fei, ZHAO Wei-Chun* |
School of Life Sciences, Zhejiang Chinese Medical University, Hangzhou 310053, China |
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Abstract Fritillaria thunbergii is a genuine medicinal material which is one of the important cash crops in Jinhua city, Lishui city and other places in Zhejiang Province. In recent years, fungal diseases have occurred frequently, including root rot, which affected the yield and quality of F. thunbergii. In this study, pathogenic fungi of root rot disease of F. thunbergii were isolated and purified on potato dextrose agar (PDA) medium by diseased tissue isolation method. The pathogenic fungi were preliminarily identified by morphological observation of colony, mycelium and spore. Using the genomic DNA of fungi, the DNA fragments of internal transcribed spacer (ITS), glyceraldehyde 3-phosphate dehydrogenase (G3PDH), translation elongation factor 1 alpha (TEF-1α), RNA polymerase Ⅱ beta subunit (RPB2), heat shock protein (HSP60), and ITS-specific fragment of Fusarium solani (SOL) were amplified by PCR and sequenced, respectively. The homology of each DNA fragment was aligned in GenBank and the phylogenetic adjacent trees were constructed to identify the species of the fungi. The pathogenicity of the fungi was determined by inoculation of host plant with acupuncture and smear method. A total of 14 fungal isolates were isolated by PCR amplification and electrophoresis, and 4 fungi species causing the root rot disease of F. thunbergii were isolated and identified. By constructing a developmental tree and pathogenicity experiments, it was proved that the pathogenic bacteria were F. solani, F. oxysporum, F. incarnatum and F. equiseti, respectively. The partial sequence of the G3PDH gene of F. incarnatum (MN386064) and F. equiseti (MN386065) was obtained by PCR amplification for the first time. The results enrich the gene sequences of the 2 Fusarium species, and also provid a basis for subsequent related research.
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Received: 01 July 2022
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Corresponding Authors:
* weichunzhao@zcmu.edu.cn
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