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Background : The causative organism of Astragalus membranaceus (Hwanggi) root rot disease was known as Pythophthora drechsleri. In this study, a new species of root rot causative organism has been identified and reported.
Methods and Results : Hwanggi, which has root rot disease, was collected in Jungsun-gun, Gangwon-do. The diseased root was cut into 1 ㎝ size and disinfected with 1% sodium hypochlorite solution for 1 minute and washed with sterile distilled water. The dried samples were dipped in the PDA medium and cultured in at 25℃ incubator. The 21 isolates were cultured under the same conditions and genomic DNA was extracted. The genomic DNA of isolated strains was extracted using a kit of Biofact. The isolated strains were identified using internal transcribed spacer (ITS) region sequencing. The ITS region was amplied using the primer pair ITS1 (5’-TCCGTAGGTGAACCTGCGG-3’) and ITS4 (5’-TCCTCCGCTTATTGATATGC-3’). The PCR was performed in a final volume of 50 ㎕ containing 10 mM Tris-HCl (pH 8.3), 50 mM KCl, 1.5 mM MgCl2, each dNTP at a concentration of 0.2 mM, 1.25 IU of Taq polymerase, each primer at a concentration of 10 pmol, and 2ul of the DNA template. Sequencing was performed by asking Macrogen. The analyzed sequences were compared using the Blast program provided by NCBI. As a result of identification of 21 strains, 13 Fusarium spp., 1 Peronospora sp. (causative organism of downy mildew), 3 Acremonium spp., 3 Uncultured fungus, 1 Phyrtophthora sp.. The main pathogen of wilt disease was known as Fusarium sp., a soil-borne pathogen. After analyzing the systematic taxonomic relationships, the systematic diagram was created using the neighbor-joining method.
Conclusion : We have finally identified Phytophthora cryptogea as a causative organism of Hwanggi root rot disease and will check whether the disease has occurred on the inoculation through pot test.
