기체크로마토그래피 질량분석기 (GC-MS)를 이용한 도라지의 뿌리 조직, 연령, 서식지에 따른 대사물질 비교 분석
Comparison on the metabolic profiles of Balloon flower (Platycodon grandiflorum) roots by layers, ages and habitats using gas chromatography mass spectrometry (GC-MS)
- 주제(키워드) 도라지 , 식물대사물질
- 발행기관 아주대학교
- 지도교수 박상규
- 발행년도 2015
- 학위수여년월 2015. 8
- 학위명 석사
- 학과 및 전공 일반대학원 생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000020618
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Plant are exposed to various abiotic and biotic stress factors. They produce diverse metabolic substances. To elucidate the change of metabolic profiles by layers, ages, seasons and habitats, Platocodon. grandiflorum sample were collected from November 2013 to September 2014 at five sites : Site 1, 2 and 3 were Balloon flower cultivation fields located in Hongcheon-gun, Kangwon-do. Site 4 is wild P. grandiflorum habitats located in Gangneung-si, Kangwon-do and site 5 is another wild P. grandiflorum habitat in Mt. Daebong. Humidity, electric conductivity, temperature and acidity were measured for soil environmental factors. For gas chromatography mass spectrometry (GC-MS) analysis, MeOH extraction was proceeded by using internal standards (13C-ribitol and 13C-sorbitol). For GC-MS analysis, the extracted samples were derivatizated using methoxiamination and MSTFA (N-Methyl-N-(trimethylsilyl) trifluoroacetamide). Data from GC-MS was analyzed by Automated Mass-Spectral Deconvolution and Identification System (AMDIS), National Institute of Standards and Technology (NIST) library, OpenChrom, MetAlign, MSClust and R program. There were significantly differences in GC-MS profiles of P. grandiflorum between above-ground, such as leaves and below-ground parts. Cortex and epidermis showed slightly different metabolic profiles. Such as epidermis, cortex and pith exhibited distinctive profile patterns by seasons. Between before and after, November 2013 and March 2014, epidermis samples showed very clear differences in profile. After winter, P. grandiflorum had higher sucrose concentration than in November. In the comparison among habitats, epidermis samples showed quiet different profiles from cultivation samples. However there is not clear patterns by ages because of habitat heterogeneity and genetic diversity. To study the effect of microbial communities or metabolic profiles, a growth experiment with different soils was conducted by using plants induced from same callus and stem. I made four type soil : soil of P. grandiflorum cultivation field (CS), autoclaved soil of P. grandiflorum cultivation field (ACS), soil of P. grandiflorum wild habitat (WS) and autoclaved soil of P. grandiflorum wild habitat (AWS). Metabolic profiles of P. grandiflorum were changed by soil condition. Also P. grandiflorum grown from different habitats have similar metabolic profiles after soil autoclaved indicating soil microbial communities may change metabolic profiles of plant below-ground as well as plant above-ground.
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Abstract ⅰ
Table of Contents ⅲ
List of Figures ⅳ
List of Tables ⅵ
I. Introduction 1
II. Materials and Methods
1. Study sites and sample collection 3
2. Extraction and derivatization for GC-MS analysis 7
3. GC-MS analysis 7
4. Plant embryogenic callus induction and regeneration 8
5. Growth experiment with soils 8
III. Results
1. Soil environmental factors 11
2. Metabolic profile change of P. grandiflorum by layers, seasons, ages and habitats 14
3. Metabolic profile change of P. grandiflorumtissue cultured plant by soil condition 24
IV. Discussion 30
V. References 33
VI. 국문요약 36
