This document summarizes a study that identified viral diseases affecting the Cnidium officinale plant in South Korea in fall 2021. Samples showing virus-like symptoms were collected and total RNA was extracted. RT-PCR was performed using primers for known viruses that infect cnidium. All samples tested positive for Apple stem grooving virus, Cnidium Vein Yellow viruses 1 and 2, and Cnidium virus X, indicating mixed infections with one or more of these four viruses. The complete genomic sequence of a new betanucleorhabdovirus found in the plants, tentatively named Cnidium virus 1, is also described.

1. Identification of Viral Diseases Affecting Cnidium Officinal plant in fall season Bonghwa-
gun, Republic of Korea
Abstract:
Cnidium officinale (Korean name: Chunkug) is a perennial plant belonging to the family Umbelliferae, and is used as
a side dish and a medicinal plant to counteract a variety of diseases in Asia, particularly in Korea and China [8].
Several viruses have been reported infecting C. officinale, including cnidium vein yellowing virus (CnVYV-1 and
CnVYV-2; family Secoviridae) [9], cnidium virus X (family Alphaflexiviridae) [10], and cycas necrotic stunt virus
(family Secoviridae) [11]. In this study, the complete genomic sequence of a betanucleorhabdovirus, tentatively
named Cnidium virus 1 (CnV1), identified from a C. officinale plant, is described for the first time.
In Oct 15, 2021, C. officinale plants that showed virus like symptoms (leaf deformation, mild mottling, severe stunting,
leaf mosaic, and vein clearing) (Fig) were collected at the region of Bonghwa-gun, Gyeongsangbuk-do, South Korea.
The samples were put in explicitly labeled plastic bags and brought to Korean Research Institute of
Bioscience and Biotechnology(KRIBB) laboratory in building 6th
, 4th floor. For further studying, the
collected samples individually crushed to a fine powder and kept at -80 °C. The total RNA was extracted from each
sample using the Easy-spin™ Total RNA Extraction Kit (iNtRON Biotechnology, Sangdaewon-dong, South Korea).
To identify the infected samples, the RNA was extracted with the WizPrep™ Plant RNA Mini Kit and the cDNA was
synthesized with the N25 primer (25-mer) and RevertAid Reverse Transcriptase (Thermo Scientific, Waltham, MA,
USA). The cDNA was amplified with PCR using the AccuPower® ProFi Taq PCR PreMix (Bioneer, Daejeon, South
Korea) with 9 set of pairs of primers, which were designed from previously cnidium infected known reported viruses.
Reverse transcription and polymerase chain reaction (RT-PCR) was performed to confirm the presence of Cnidium
Vein Yellow virus-1 (CVYV-1), Cnidium Vein Yellow virus-2 (CVYV-2), Apple stem grooving virus (ASGV),
Cucumber mosaic virus-RNA1 (CMV-RNA1), Cucumber mosaic virus -RNA3 (CMV-RNA3), Cnidium virus X
(CnVX), Cnidium mottle virus (CnMoV) , Cycas Necrotic Stunt Virus-RNA1 (CNSV-RNA1) , Cycas Necrotic Stunt
Virus-RNA2 (CNSV-RNA2). All the primer were effectively amplified except Cycas Necrotic Stunt Virus (RNA1 and
RNA2) and Cucumber mosaic virus-RNA1. All samples were positive for ASGV, CVYV, CnVX in mixed infection
with one or more of the other four viruses.
Fig2. RT-PCR detecting result showed that the cnidium plant infection with one or more mixed viruses
(CVYV, ASGV, CMV, CnVX)
Reference
8. Choi, H. S., Kim, M. S. L., & Sawamura, M. (2002). Constituents of the essential oil of Cnidium officinale Makino,
a Korean medicinal plant. Flavour and Fragrance Journal, 17(1), 49–53.
9. Yoo, R. H., Zhao, F., Lim, S., Igori, D., Kim, S. M., An, T. J., Lee, S. H., & Moon, J. S. (2015). The complete
genome sequences of two isolates of cnidium vein yellowing virus, a tentative new member of the family
Secoviridae. Archives of Virology, 160(11), 2911–2914.
10. Honma, H., Tsushima, D., Kawakami, H., Fujihara, N., Tsusaka, T., Kawashimo, M., Nishimura, T., & Fuji, S.
(2019). Complete nucleotide sequence of a new potexvirus, ‘Cnidium virus X’, isolated from Cnidium officinale
in Japan. Archives of Virology, 164(7), 1931–1935.