Jeon, Chang-Wook
(Division of Applied Life Science (BK21 Plus), Gyeongsang National University)
,
Kim, Da-Ran
(Department of Plant Medicine and Institute of Agriculture & Life Sciences, Gyeongsang National University)
,
Gang, Geun Hye
(Korea National Park Institute for Biodiversity Conservation)
,
Kim, Byung-Bu
(Korea National Park Institute for Biodiversity Conservation)
,
Kim, Nam Ho
(Korea National Park Institute for Biodiversity Conservation)
,
Nam, Seong-Yeol
(Korea National Park Institute for Biodiversity Conservation)
,
Kwak, Youn-Sig
(Division of Applied Life Science (BK21 Plus), Gyeongsang National University)
Cypripedium japonicum is known to be the indigenous plant to Korea, Japan, and China. However, C. japonicum represents the most critically endangered plant species in South Korea. The plant is esthetically pleasing due to its flower, which is larger than any other orchidaceous species. Disease sympt...
Cypripedium japonicum is known to be the indigenous plant to Korea, Japan, and China. However, C. japonicum represents the most critically endangered plant species in South Korea. The plant is esthetically pleasing due to its flower, which is larger than any other orchidaceous species. Disease symptoms relating to gray mold were observed on C. japonicum in May 2019. The suspected pathogen was successfully isolated from the symptomatic leaf tissue and conducted a pure culture of the fungi. The conidia formed consisted of a colorless or light brown single cell, which was either egg or oval-shaped with a size of 7.1 to 13.4 × 5.2 to 8.6 ㎛. Molecular phylogenetic relationship analysis was also confirmed that the pathogen concerned belonging to the family of Botrytis cinerea. Therefore, the findings confirmed that the pathogen isolated from C. japonicum was consistent with the unique properties of B. cinerea.
Cypripedium japonicum is known to be the indigenous plant to Korea, Japan, and China. However, C. japonicum represents the most critically endangered plant species in South Korea. The plant is esthetically pleasing due to its flower, which is larger than any other orchidaceous species. Disease symptoms relating to gray mold were observed on C. japonicum in May 2019. The suspected pathogen was successfully isolated from the symptomatic leaf tissue and conducted a pure culture of the fungi. The conidia formed consisted of a colorless or light brown single cell, which was either egg or oval-shaped with a size of 7.1 to 13.4 × 5.2 to 8.6 ㎛. Molecular phylogenetic relationship analysis was also confirmed that the pathogen concerned belonging to the family of Botrytis cinerea. Therefore, the findings confirmed that the pathogen isolated from C. japonicum was consistent with the unique properties of B. cinerea.
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문제 정의
japonicum, which include the study of growing conditions of its natural habitat [5], the study of its seed germination [6], orchid mycorrhizal fungi’s properties [7] and its genetic diversity [8], studies in relation to its pathology is still non-existent, yet. Therefore, this study seeks to provide a fundamental set of data for long-term preservation and restoration of C. japonicum by investigating its habitats located in South Korea to elucidate the diseases prevalent in the plant.
제안 방법
DNA extraction was performed based on a technique involving the use of cetyltrimethylammonium bromide (CTAB) [10] and the genomic DNA was used to perform the sequencing processes. Identification task specifically looked at three nuclear DNA genes (G3PDH, HSP 60, and ITS) with the specific primer sets (Table 2) corresponding to each of the genes was used to amplify through polymerase chain reaction (PCR) [12]. The amplicons of the PCR were subjected to electrophoresis using 1% agarose gel.
To investigate the properties of the pathogen such as conidia, shapes, sizes, the fore-ends of hyphae, to the fungi were grown for 72 h on a potato dextrose agar medium (PDA: potato dextrose broth 24 g, agar 20 g/L) and incubated at 25 °C without exposure to light for 10 days.
대상 데이터
The sequencing analyses were carried out at Macrogen (Seoul, Korea). The sequences were assigned an accession number MN267696 (ITS), MT233447 (HSP 60), and MN267728 (G3PDH) by the National Center for Biotechnology Information (NCBI). The phylogenetic analyses were conducted using GenBank BLAST provided by the NCBI, which involved maximum likelihood and maximum parsimony method in MEGA 10 program.
이론/모형
The sequences were assigned an accession number MN267696 (ITS), MT233447 (HSP 60), and MN267728 (G3PDH) by the National Center for Biotechnology Information (NCBI). The phylogenetic analyses were conducted using GenBank BLAST provided by the NCBI, which involved maximum likelihood and maximum parsimony method in MEGA 10 program. As a result of conducting sequencing analyses based on the three genes ITS, HSP 60, and G3PDH the isolated pathogens were showed 100% similarity with previously registered B.
성능/효과
The phylogenetic analyses were conducted using GenBank BLAST provided by the NCBI, which involved maximum likelihood and maximum parsimony method in MEGA 10 program. As a result of conducting sequencing analyses based on the three genes ITS, HSP 60, and G3PDH the isolated pathogens were showed 100% similarity with previously registered B. cinerea stains in GenBank. The result of a phylogenetic relationship analysis also confirmed that the pathogen concerned belonged to the same family as that of B.
후속연구
The findings are anticipated to play a significant role in devising long-term strategies to preserve and restore C. japonicum and at the same time, the ongoing pathological investigation is necessary to promote the preservation of the plant in the Korean peninsula.
참고문헌 (15)
1 Barnes BV. The landscape ecosystem approach and conservation of endangered species . Ann Arbor (MI) : University of Michigan School of Natural Resource ; 1993 . Endangered Species Update.
2 Falk DA , Olwell P. Scientific and policy considerations in restoration and reintroduction of endangered species . Rhodora . 1992 ; 94 : 287 ？ 315 .
3 Wilson OE. The diversity of life . Harvard (MA) : Cambridge Univ. Press ; 1992 .
4 Lee BH , Han HK , Kwon HJ , et？al. Diversity of endophytic fungi isolated from roots of Cypripedium japonicum and C. macranthum in Korea . Korean J Mycol . 2015 ; 43 ( 1 ): 20 ？ 25 .
5 Cho YC , Kim HG , Koo BY , et？al. Dynamics and viability analysis of transplanted and natural lady’s slipper ( Cypripedium japonicum ) populations under habitat management in South Korea . Restor Ecol . 2019 ; 27 ( 1 ): 23 ？ 30 .
6 Yan X , Tian M , Liu F , et？al. Hormonal and morphological changes during seed development of Cypripedium japonicum . Protoplasma . 2017 ; 254 ( 6 ): 2315 ？ 2322 . 28550470
7 Gang GH , Cho G , Kwak YS , et？al. Distribution of rhizosphere and endosphere fungi on the first-class endangered plant Cypripedium japonicum . Mycobiology . 2017 ; 45 ( 2 ): 97 ？ 100 . 28781542
8 Tian HZ , Han LX , Zhang JL , et？al. Genetic diversity in the endangered terrestrial orchid Cypripedium japonicum in East Asia: Insights into population history and implications for conservation . Sci Rep . 2018 ; 8 ( 1 ): 6467 . 29691494
9 Kwon J-H , Cheon M-G , Choi O , et？al. First report of Botrytis cinerea as a Postharvest Pathogen of Blueberry in Korea . Mycobiology . 2011 ; 39 ( 1 ): 52 ？ 53 . 22783073
10 Graham J , Marshall B , Squire G. Genetic differentiation over a spatial environmental gradient in wild Rubus idaeus populations . New Phytol . 2003 ; 157 ( 3 ): 667 ？ 675 .
11 Staats M , van Baarlen P , van Kan JA. Molecular phylogeny of the plant pathogenic genus Botrytis and the evolution of host specificity . Mol Bio Evol . 2004 ; 22 ( 2 ): 333 ？ 346 . 15496556
12 White TJ , Bruns T , Lee S , et？al. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics In: Innis MA , Gelfand DH. , Sninsky JJ , et？al. editors. PCR protocols a guide to methods and applications . New York (NY) : Academic Press ; 1990 p. 315 ？ 322 .
13 KNA (Korea National Arboretum ). Rare plants data book in Korea . Pocheon (Korea) : Korea National Arboretum ; 2008 . (in Korean)
14 Lee JS , Choi BH. Distributions and red data of wild orchids in the Korean peninsula . Korean J Pl Taxon . 2006 ; 36 ( 4 ): 335 ？ 360 .
15 Rankou H. Cypripedium japonicum The IUCN red list of threatened species. 2014 . e.T46650A43315717.
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