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2023 Vol.41, Issue 4 Preview Page

Research Article

Comparative Analysis between the ITS TaqMan SNP Genotyping Assay and CAPS for the Rapid Molecular Identification of Zoysia japonica and Zoysia sinica, Related Hybrid Lines, and the Habitat Distribution of Each Species

한국에 자생하는 들잔디와 갯잔디, 교잡종의 신속한 식별을 위한 ITS TaqMan SNP genotyping assay와 CAPS의 비교 분석 및 각 종의 서식환경별 분포

Dae-Hwa Yang1†
,
Ok-Cheol Jeong1†
,
Yu-Ryang Kim2
,
Mi-Jeong Kang2
,
Yang-Ji Kim2
,
Ji-Hi Son2
,
Seong-Seop Han2
,
Mi-Young Park2
,
Il-Doo Jin1
,
In-Ja Song1
,
Min-Ji Hong2
,
Hyeon-Jin Sun1
,
Hong-Gyu Kang1
,
Hyo-Yeon Lee12*
양 대화1†
,
정 옥철1†
,
김 유량2
,
강 미정2
,
김 양지2
,
손 지희2
,
한 승섭2
,
박 미영2
,
진 일두1
,
송 인자1
,
홍 민지2
,
선 현진1
,
강 홍규1
,
이 효연12*
1Subtropical Horticulture Research Institute, Jeju National University, Jeju 63243, Korea
2Faculty of Biotechnology, Jeju National University, Jeju 63243, Korea
1제주대학교 아열대원예산업연구소
2제주대학교 생명공학부
*Corresponding Author
†These authors contributed equally to this work.
31 August 2023. pp. 463-495
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Abstract

In this study, zoysiagrasses found in a major province of the Korean peninsula were collected and identified by molecular techniques. The zoysiagrasses used in this study were collected from mountainous national parks; ‘oreum’ areas; and from inland, coastal and island areas, including marine and coastal national parks on the west, east, and south coasts. Molecular identification was done by means of a cleaved amplified polymorphic sequence (CAPS) marker analysis based on the nrDNA-internal transcribed spacer (ITS) method established by our research team. Additionally, in this study the ITS-based TaqMan single-nucleotide polymorphism (SNP) genotyping assay was developed and used to perform molecular identification more rapidly and accurately. In total, 450 collected zoysiagrass plants were classified into three lines (234 Zoysia japonica plants, 62 Zoysia sinica plants, and 154 Z. japonica and Z. sinica hybrid plants) by molecular identification based on the ITS-based CAPS marker analysis and ITS-based TaqMan SNP genotyping assays. The results of the ITS-based TaqMan SNP genotyping assay were consistent with those of the ITS-based CAPS marker analysis. Both the ITS-based CAPS marker analysis and the ITS-based TaqMan SNP genotyping assay were found to be efficient molecular methods to identify the three species of zoysiagrasses. Specifically, the ITS-based TaqMan SNP genotyping assay method developed in this study produced results more rapidly compared to the ITS-based CAPS marker analysis method. The habitat distributions of each of the molecularly identified zoysiagrass plants investigated in this study were also assessed. The Z. japonica plants were mainly distributed in mountainous national parks and oreum areas as well as inland and coastal areas. The Z. sinica plants were distributed along the coast and on rocks and stones frequently submerged in seawater. The Z. japonica and Z. sinica hybrid plants were distributed more in the inland, coastal, and island areas than in the mountainous national parks and oreum areas. Findings related to the leaf blade width as a main external morphological classification characteristic of the molecularly identified 450 zoysiagrass plants are described below. The leaf blade width ranged from 4 to 5 mm for the Z. japonica plants, from 2 to 4 mm for the Z. sinica plants, and from 3 to 4 mm for the Z. japonica and Z. sinica hybrid plants. Additionally, some Z. japonica and Z. sinica plants and Z. japonica and Z. sinica hybrid plants with leaf blade widths of 2 to 3, 4 to 5, and from 2 to 3 and 4 to 5 mm, respectively, were also present. Because the ranges of the leaf blade width of each species overlapped, it was difficult to identify the hybrid lines using this metric. The feasibility of the developed molecular identification method for these three species of zoysiagrasses was confirmed in this study.

Keywords
DNA barcoding
main external morphological classification
molecular classification
leaf blade width
zoysiagrass

본 연구에서는 한국의 국립공원(산) 및 오름, 서해안 및 동해안, 남해안 부근의 해상국립공원을 포함한 해안 및 섬 등에서 한국잔디류 620점을 수집하였다. 이식, 보존 및 증식 중인 다량의 잔디 450개체는 분자생물학적 분류동정을 수행하였으며, 들잔디(Zj) 234개체, 갯잔디(Zs) 62개체, 들잔디와 갯잔디의 교잡종(H) 154개체로 식별되었다. 본 연구에서 종의 분류 동정 결과는 새로 개발된 ITS 기반 TaqMan SNP genotyping assay 분석과 기존 방법인 ITS 기반 CAPS 분석에서 그 결과가 일치하였으며, ITS 기반의 TaqMan SNP genotyping assay 분석법은 ITS 기반 CAPS 마커 분석법보다 더 신속하게 종 식별 가능함이 확인되었다. 분류 동정된 450개체의 들잔디, 갯잔디, 교잡종의 서식환경별 분포를 조사한 결과, 들잔디의 경우 주로 국립공원(산) 및 오름, 내륙, 해안가 등에 분포되었고, 갯잔디의 경우 바닷물에 자주 잠기는 바위나 돌, 해안가의 바닷물이 인접한 부근에서 분포되었으며, 들잔디와 갯잔디의 교잡종은 국립공원 산 및 오름보다 내륙 및 해안가, 섬지역에 걸쳐 다양한 장소에 분포되었다. 기존 외부형태학적으로 주요한 분류 동정의 형질인 잎의 너비(엽폭)가 들잔디와 갯잔디 교잡종의 분류동정에 이용할 수 있는지 분자생물학적으로 분류 동정된 450 개체의 한국잔디류의 주요 외부형태적 분류 특성인 엽폭을 조사한 결과, 엽폭의 크기는 대체로 들잔디 4–5mm, 갯잔디 2–4mm, 들잔디와 갯잔디의 교잡종 3–4mm로 분석되었다. 일부 엽폭 크기는 들잔디(2–3mm)와 갯잔디(4–5mm), 들잔디와 갯잔디의 교잡종(2–3mm과 4–5mm)으로 확인되어 엽폭 크기에 의하여 교잡종을 확실히 분류 동정하기에 쉽지 않았기 때문에 본 연구에서 개발된 신속정확한 분자생물학적 분류 동정법의 필요성을 재확인하였다.

키워드
DNA 바코드
주요 외부형태학적 분류
분자생물학적 분류
엽폭
한국잔디
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Information
  • Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
  • Publisher(Ko) :원예과학기술지
  • Journal Title :Horticultural Science and Technology
  • Journal Title(Ko) :원예과학기술지
  • Volume : 41
  • No :4
  • Pages :463-495
  • Received Date : 2022-05-30
  • Revised Date : 2022-11-14
  • Accepted Date : 2022-12-19
  • DOI :https://doi.org/10.7235/HORT.20230042
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