토종 갓끈동부의 ITS1, 5.8S 및 ITS2의 염기서열을 이용한 계통 분석 Phylogenetic Analysis of Native Vigna sinensis in Korea Using DNA Sequence of Internal Transcribed spacer (ITS) Region원문보기
본 연구에서 밝혀진 갓끈동부의 ITS1, 5.8S 및 ITS2의 염기서열은 NCBI (National Center for Biotechnology Information)의 GenBank에 Vigna sinensis AY195581로 등록하였다. ITS1, 5.8S 및 ITS2의 총 염기서열 507 염기서열을 이용한 Vigna sinensis (AY195581)의 분자계통분석에서 Vigna unguiculata 및 그 아종들과 98~100% 범위의 염기서열 상동성을 보였다. Vigna unguiculata는 계통분석에 이용된 다른 종들로부터 독립된 하나의 cluster로 그룹핑(grouping)이 됨을 확인하였다. 본 계통분석은 Vigna unguiculata가 Vigna 속의 다른 종에 비해 비교적 최근에 분화되었으며, 현재 유전적인 변화가 많이 일어나고 있음 보이고 있다. 또한, Vigna 속, Vigna longifolia, Vigna vexillata, Vigna membranacea, Vigna friesiorum, Vigna monophylla, Vigna schimperi, Vigna nigritia, Vigna lasiocarpa, Vigna trichocarpa, Vigna diffusa의 다른 종들과 비교하여 유전적으로 독립적인 종임을 확인하였다. 본 연구의 Vigna sinensis의 ITS1, 5.8S 및 ITS2를 이용한 계통분석은 Vigna sinensis를 Vigna unguiculata로 분류하는 것이 타당한 것으로 보여진다. 본 종은 국내에서 멸종된 것으로 알려져 있었으나 최근 토착 식물로써 발견되었고 이 갓끈동부의 관련 식물 종들과의 분자계통학적 위치를 명확히 밝힘에 의의가 있다고 하겠다.
본 연구에서 밝혀진 갓끈동부의 ITS1, 5.8S 및 ITS2의 염기서열은 NCBI (National Center for Biotechnology Information)의 GenBank에 Vigna sinensis AY195581로 등록하였다. ITS1, 5.8S 및 ITS2의 총 염기서열 507 염기서열을 이용한 Vigna sinensis (AY195581)의 분자계통분석에서 Vigna unguiculata 및 그 아종들과 98~100% 범위의 염기서열 상동성을 보였다. Vigna unguiculata는 계통분석에 이용된 다른 종들로부터 독립된 하나의 cluster로 그룹핑(grouping)이 됨을 확인하였다. 본 계통분석은 Vigna unguiculata가 Vigna 속의 다른 종에 비해 비교적 최근에 분화되었으며, 현재 유전적인 변화가 많이 일어나고 있음 보이고 있다. 또한, Vigna 속, Vigna longifolia, Vigna vexillata, Vigna membranacea, Vigna friesiorum, Vigna monophylla, Vigna schimperi, Vigna nigritia, Vigna lasiocarpa, Vigna trichocarpa, Vigna diffusa의 다른 종들과 비교하여 유전적으로 독립적인 종임을 확인하였다. 본 연구의 Vigna sinensis의 ITS1, 5.8S 및 ITS2를 이용한 계통분석은 Vigna sinensis를 Vigna unguiculata로 분류하는 것이 타당한 것으로 보여진다. 본 종은 국내에서 멸종된 것으로 알려져 있었으나 최근 토착 식물로써 발견되었고 이 갓끈동부의 관련 식물 종들과의 분자계통학적 위치를 명확히 밝힘에 의의가 있다고 하겠다.
Cowpea (Vigna unguiculata (L.) Walp.) is recognized as a potential source of protein and other nutrients. The genus Vigna includes 100 wild species of plants. Especially, Vigna unguiculata includes annual cowpeas (ssp. unguiculata) and ten wild perennial subspecies. DNA sequence of internal transcri...
Cowpea (Vigna unguiculata (L.) Walp.) is recognized as a potential source of protein and other nutrients. The genus Vigna includes 100 wild species of plants. Especially, Vigna unguiculata includes annual cowpeas (ssp. unguiculata) and ten wild perennial subspecies. DNA sequence of internal transcribed spacer (ITS) region was determined for Vigna sinensis, one of native plant, which was found in recent but thought to have gone extinct in Korea. The seeds of Vigna sinensis used in this study were donated from Dong-Young Jo. The DNA sequence of ITS-5.8S-ITS2 for Vigna sinensis obtained from this study was deposited as Vigna sinensis AY195581 on GenBank of NCBI (National Center for Biotechnology Information). We investigated the sequence-based phylogenetic relationships of plants related and clarified its taxonomical position. DNA similarities among subspecies including Vigna unguiculata showed the range 98 to 100% in sequence-based phylogenetic analysis using total 507 base pairs of ITS1, 5.8S and ITS2. Vigna unguiculata and subspecies were grouped independently as one cluster from other Vigna species used in the phylogenetic analysis. In this study, based on the phylogenetic analysis using the ITS1-5.8S-ITS2 sequence of Vigna sinensis, it may be concluded to be classified to one of Vigna unguiculata substrains.
Cowpea (Vigna unguiculata (L.) Walp.) is recognized as a potential source of protein and other nutrients. The genus Vigna includes 100 wild species of plants. Especially, Vigna unguiculata includes annual cowpeas (ssp. unguiculata) and ten wild perennial subspecies. DNA sequence of internal transcribed spacer (ITS) region was determined for Vigna sinensis, one of native plant, which was found in recent but thought to have gone extinct in Korea. The seeds of Vigna sinensis used in this study were donated from Dong-Young Jo. The DNA sequence of ITS-5.8S-ITS2 for Vigna sinensis obtained from this study was deposited as Vigna sinensis AY195581 on GenBank of NCBI (National Center for Biotechnology Information). We investigated the sequence-based phylogenetic relationships of plants related and clarified its taxonomical position. DNA similarities among subspecies including Vigna unguiculata showed the range 98 to 100% in sequence-based phylogenetic analysis using total 507 base pairs of ITS1, 5.8S and ITS2. Vigna unguiculata and subspecies were grouped independently as one cluster from other Vigna species used in the phylogenetic analysis. In this study, based on the phylogenetic analysis using the ITS1-5.8S-ITS2 sequence of Vigna sinensis, it may be concluded to be classified to one of Vigna unguiculata substrains.
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제안 방법
The refined PCR products were directly sequenced with the use of a BigDye Terminator Cycle Sequencing Ready Reaction Kit (Applied Biosystems, USA). The cycle sequencing reaction was performed with a PCR Thermal CyclerMP (Takara, TP3080, Japan). The sequencing reaction was performed for 5 min at 96℃ at first and consisted of 25 cycles of the following: 10 sec at 96℃, 10 sec at 52℃, and 2 min at 60℃.
대상 데이터
The DNA sequence of ITS-5.8S-ITS2 for Vigna sinensis obtained from this study was deposited as Vigna sinensis AY 195581 on GenBank of NCBI (National Center for Biotechnology Information). DNA similarities among subspecies including Vigna unguiculata showed the range 98 to 100% in sequence-based phylogenetic analysis using total 507 base pairs of ITS1, 5.
PCR amplification was performed as described by Spiridonov and Moens [9]. Two sets of primers were used in the PCR reactions: (1) TW81 (5-GTTTCCGTAGGTGAACCTGC-3) and AB28 (5-ATATGCTTAAGTTCAGCGGGT-3) or (2) 18S (5-TT GATTACGTCCCTGCCCTTT-3) and 26S (5-TTTCACTCGCC GTTACTAAGG-3) [2, 12], respectively. PCR products were purified using QIAquick PCR or Gel Purification Kit (Qiagen Ltd, Crawley, UK).
이론/모형
1. Phylogenetic tree of ITS1-5.8S-ITS2 sequences for Vigna species using Neighbour-Joining method. A total of 507 bp aligned positions were used.
성능/효과
It is presumed that the cluster of Vigna unguiculata subspecies be branched comparatively lately to other species of genus Vigna, and now occurred much on genetic divergence. In addition, it is showed that the cluster of Vigna unguiculata subspecies is genetically independent, compared to other species of genus Vigna, Vigna longifolia, Vigna vexillata, Vigna membranacea, Vigna friesiorum, Vigna monophylla, Vigna schimperi, Vigna nigritia, Vigna lasiocarpa, Vigna trichocarpa and Vigna diffusa (Fig. 1).
In this study, DNA sequence of internal transcribed spacer (ITS) region was determined for Vigna sinensis, one of native plant, which was found in recent but was thought to have gone extinct in Korea. We investigated the Sequence-based phylogenetic relationships of plants related and clarified its taxonomical position.
In this study, based on the phylogenetic analysis using the ITS1-5.8S-ITS2 sequence of Vigna sinensis, it may be concluded to be classified to one of Vigna unguiculata subspecies. The fact of this study is that the phylogenetic position of Vigna sinensis, one of native plant, found in recent but thought to have gone extinct in Korea, is clearly identified from plants related of Vigna unguiculata through sequence-based phylogenetic analysis.
참고문헌 (13)
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