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nrDNA-ITS 염기서열 분석 및 CAPS marker를 이용한 Zoysia 속 들잔디와 갯잔디의 분자생물학적 동정

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Alternative Title
Molecular identification of Zoysia japonica and Zoysia sinica(Zoysia species) based on nrDNA-ITS sequences analyses and CAPS marker
Abstract
Zoysia plants are important turf grasses used for school playgrounds, parks, roadsides, golf courses, and athletic fields. Especially, Zoysia japonica and Zoysia sinica species growing near the beaches are not readily identifiable due to their similar morphology. It is thus desirable to have both morphological and molecular identifications. DNA barcoding using specific gene or non-coding DNA as barcode is one of the molecular identification methods for cryptic species. ITS (Internal Transcribed Spacer) is a non-coding DNA involved in nuclear ribosomal DNA, and is a DNA barcode popularly used because of more frequent evolutionary changes at the DNA level than other DNA barcode regions in plastid.
In this study, we collected 97 Zoysia turf grasses and classified into 66 Zoysia japonica and 31 Zoysia sinica species according to morphological standards for identification. To classify them based on the molecular method, the nrDNA-ITS regions including ITS1, 5.8S nrDNA, and ITS2 from four different grass species (Zoysia japonica, Zoysia sinica, Agrostis stolonifera, and Poa pratensis) were used for DNA barcoding. The substitutions and InDel (Insert/Deletion) sites in the nrDNA-ITS sequence of Z. japonica and Z. sinica were converted into the restriction endonuclease sites (BglI, StyI, and FauI) for CAPS (Cleaved Amplified Polymorphic Sequence) markers, and applied to collected the Zoysia grasses to verify whether the markers are presenting both seed and plant samples.
We successfully identified 43 Zoysia japonica, 25 Zoysia sinica, and 29 hybrids between Zoysia japonica and Zoysia sinica among the 97 collected grass samples. These results suggest that Zoysia japonica and Zoysia sinica can be identified at the species level using nrDNA-ITS barcode region and CAPS markers.
Author(s)
홍민지
Issued Date
2015
Awarded Date
2016. 2
Type
Dissertation
URI
http://dcoll.jejunu.ac.kr/jsp/common/DcLoOrgPer.jsp?sItemId=000000007583
Department
대학원 생명공학부
Advisor
이효연
Table Of Contents
목차 1
Abstract 3
Ⅰ. 서론 4
1장. DNA 바코드 nrDNA-ITS를 이용한 들잔디, 갯잔디, 크리핑 벤트그라스, 켄터키 블루그라스 잔디 동정 6
1. 서론 7
2. 재료 및 방법 - 9
1) 식물 재료 9
2) Genomic DNA 추출 - 9
3) nrDNA-ITS PCR 증폭 - 16
4) nrDNA-ITS cloning 16
5) nrDNA-ITS 염기서열 비교 분석 - 18
6) nrDNA-ITS 염기서열을 이용한 유연관계 분석 - 18
3. 결과 및 고찰 - 21
1) 잔디 nrDNA-ITS의 cloning 및 염기서열 비교 분석 21
2) 잔디 nrDNA-ITS 구간 내 ITS1, 5.8S rDNA, ITS2 염기서열의 비교분석 - 31
3) nrDNA-ITS 염기서열 기반의 잔디 간 유연관계 분석 - 34
2장. nrDNA-ITS 기반 CAPS marker를 이용한 들잔디와 갯잔디의 동정 36
1. 서론 - 37
2. 재료 및 방법 - 39
1) 식물 재료 - 39
2) CAPS marker 전환 및 분석 45
3) 잔디 형태적 분석 - 48
2. 결과 및 고찰 - 49
1) 잔디 nrDNA-ITS 염기서열 기반 CAPS marker 분석 - 49
2) 식별된 들잔디, 갯잔디, 교배종의 형태적 분석 - 61
Ⅲ. 종합고찰 64
Ⅳ. 참고문헌 67
Degree
Master
Publisher
제주대학교 대학원
Citation
홍민지. (2015). nrDNA-ITS 염기서열 분석 및 CAPS marker를 이용한 Zoysia 속 들잔디와 갯잔디의 분자생물학적 동정
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Faculty of Biotechnology > Molecular Biotechnology
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