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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 許輔 | zh_TW |
| dc.contributor.advisor | Fuu Sheu | en |
| dc.contributor.author | 吳映柔 | zh_TW |
| dc.contributor.author | Ying-Jou Wu | en |
| dc.date.accessioned | 2025-02-27T16:27:34Z | - |
| dc.date.available | 2025-02-28 | - |
| dc.date.copyright | 2025-02-27 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-02-13 | - |
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Frontiers in Plant Science, 10, Article 1638. | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/97158 | - |
| dc.description.abstract | 部分發酵之包種茶以其細膩香氣、獨特風味與健康效益聞名。製茶過程各步驟皆會影響包種茶最終的香氣與風味，特別是日光萎凋及室內萎凋與攪拌對香氣的形成至關重要。傳統製茶工藝透過長期經驗的積累與實踐逐步建立，但製茶過程關鍵步驟對香氣形成的科學意義與其分子機制大多仍不明確。因此，本研究鑑定與分析茶菁中揮發性化合物成分，並比較三種不同包種茶製程，以探討日光萎凋及攪拌對揮發性化合物組成與含量之影響，接著進一步分析標準製程中香氣成分與其生合成基因的關聯性，得到與揮發性化合物增加密切相關的基因，並比較未成熟與成熟鮮採茶菁揮發性化合物生合成基因的表現，以了解採收時間可能帶來的差異，最後分析並選殖 geranyllinalool 合成酶 TPS11。
本研究首先全面鑑定並分析茶菁中的揮發性有機化合物 (volatile organic compounds) 在三種不同製程中的含量變化，分別是包含日光萎凋與攪拌的標準製程 (normal procedure, NP)、無日光萎凋但包含攪拌的製程 (non-solar withering procedure, NSW) 和無攪拌但包含日光萎凋的製程 (non-shaking procedure, NSH)。總共鑑定出 80 種揮發性化合物，根據其生合成途徑可分為脂肪酸衍生揮發性化合物 (fatty acid derived volatile, FADV)、揮發性萜類 (volatile terpene, VT)、類胡蘿蔔素衍生揮發性化合物 (carotenoid derived volatile, CDV) 及胺基酸衍生揮發性化合物 (amino acid derived volatiles, AADV)。在標準製程 NP 中，揮發性化合物總含量波動性上升，最終 VT 與 AADV 顯著增加，而 FADV含量相對持平。在 NSW 製程中，揮發性化合物同樣波動性上升，但最終 FADV 含量高於 NP，VT 與 AADV 含量則低於 NP。NSH 製程揮發性化合物總含量波動，但上升幅度顯著低於 NP 與 NSW。此結果顯示日光萎凋與攪拌步驟對於茶菁中揮發性化合物組成的重要性，是包種茶製程中的關鍵步驟。在標準製程中，FADV 類別的茉莉內酯 (jasmine lactone)、VT 類別的橙花叔醇 (nerolidol)、β-羅勒烯 (β-ocimene)，與 AADV 類別的吲哚 (indole)、苯乙腈 (benzyl nitrile) 含量顯著提升，顯示這些揮發性化合物在包種茶香氣中具有重要角色。 進一步分析標準製程中，在日光萎凋與第一次攪拌期間，揮發性化合物含量與其生合成基因表現量之關聯性，以探討揮發性化合物生合成可能的分子機制。顯著相關的基因與揮發性化合物配對包括 FADV 類別的脂氧合酶8 (lipoxygenase 8, LOX8) 與 3-hexenyl iso-butyrate (r=0.99)；VT 類別的萜類合成酶2 (terpene synthase 2, TPS2) 與 β-羅勒烯 (r=0.94) 及芳樟醇 (linalool) (r=0.89)；及 AADV 類別的 tryptophan synthase β-subunit 2 (TSB2) 與吲哚 (r=0.98)。顯示不同類別的揮發性化合物生合成基因，皆會受到日光萎凋與攪拌過程的逆境誘導而表現量提升，進而促進相關揮發性化合物的生合成與累積，這些基因對於包種茶製程香氣的形成可能扮演關鍵角色。 此外，兩個 β-葡萄糖苷酶 (β-glucosidases)，β-GH3_1 和 β-GH1_1，在日光萎凋與攪拌期間基因表現量提升超過 30 倍，顯示其可能於包種茶製程中參與揮發性化合物的釋放。而 β-葡萄糖苷酶 GH5 家族之基因在未成熟茶菁中表現量較高，是否作用於未成熟或新鮮採收茶菁中揮發性化合物的釋放仍需未來深入研究。 最後，TPS11 為雙萜類 geranyllinalool 合成酶，其在日光萎凋期間表現量顯著增加，室內萎凋與攪拌階段波動性上升，顯示 TPS11 可能參與了包種茶製程 VT 的生合成，但其功能仍不明確。本研究進一步發現 TPS11 之表現量於室內萎凋後期達到高峰，並成功選殖及於大腸桿菌異體表現 TPS11。 綜合本研究結果，揭示了日光萎凋與室內萎凋過程中的逆境誘導之揮發性化合物生合成與相關分子調控機制，以及製茶步驟對於包種茶特徵香氣形成的關鍵作用。本研究的發現提供了傳統包種茶製程更深層的理解，以及包種茶香氣組成及其生合成的科學基礎，有助於進一步提升茶葉品質，並為茶葉生產技術的開發提供理論支持。 | zh_TW |
| dc.description.abstract | Semi-fermented oolong tea is renowned for its delicate aroma, unique flavor, and health benefits. Among its manufacturing processes, solar and indoor withering play critical roles in aroma formation. While the traditional practices of the processes have been established through accumulated experience and empirical knowledge, the scientific basis and underlying mechanisms remain largely unexplored. This study identified 80 volatile organic compounds (VOCs) in tea shoots and categorized them into fatty acid derived volatiles (FADVs), volatile terpenes (VTs), carotenoid derived volatiles (CDVs), and amino acid derived volatiles (AADVs) according to their biosynthetic pathways. Three manufacturing procedures were compared and revealed that indole, nerolidol, β-ocimene, benzyl nitrile, and jasmine lactone were accumulated predominantly only in the normal process, where both solar withering and shaking were incorporated. Key gene and VOC pairs with significant correlations and up-regulated during solar withering and the first shaking were identified, such as lipoxygenase 8 (LOX8) with 3-hexenyl iso-butyrate, terpene synthase 2 (TPS2) with β-ocimene and linalool, as well as tryptophan synthase β-subunit 2 (TSB2) with indole. Besides, two β-glucosidase, β-glucosidase3_1 (β-GH3_1) and β-glucosidase1_1 (β-GH1_1), were up-regulated by more than 30-fold during these stages. Additionally, three β-glucosidase5 (β-GH5), β-GH5_1, β-GH5_2, and β-GH5_3, were found to be highly expressed in immature tea shoots. Furthermore, TPS11, a geranyllinalool synthase, was cloned and expressed. TPS11 was up-regulated during solar withering, fluctuated during indoor withering and shaking, and peaked at late withering stage, suggesting the potential role to participate in VT biosynthesis during paochung tea manufacturing process. These findings provide insights into the regulation of VOC accumulation under stresses during withering, and emphasize the pivotal role of specific manufacturing processes in the formation of paochung tea characteristic aroma. This study offers a deeper understanding of the scientific basis behind traditional oolong tea manufacturing, and may contribute to the enhancement of tea quality. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-02-27T16:27:34Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-02-27T16:27:34Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 致謝 ii
摘要 iii ABSTRACT v CONTENT vii LIST OF TABLES x LIST OF FIGURES xi CHAPTER 1 INTRODUCTION 1 1.1 Introductory of Camellia sinensis and paochung tea 1 1.2 Aroma compounds in oolong tea 2 1.2.1 Fatty acid derived volatiles 2 1.2.2 Volatile terpenes and carotenoid derived volatiles 3 1.2.3 Amino acid derived volatiles 4 1.2.4 Glycosidically bound volatiles 4 1.2.5 Characteristic oolong tea aroma 5 1.3 Factors affecting aroma compounds in oolong tea 5 1.3.1 Tea cultivar and plantation position 5 1.3.2 Postharvest treatments 6 1.4 Recent studies of VOC biosynthesis during oolong tea processing 7 1.5 Scientific rationale of empirical-based oolong tea processing 7 1.6 The aim of this study 8 CHAPTER 2 ＭATERIALS AND METHODS 10 2.1 Chemicals and reagents 10 2.2 Plant materials and tea manufacturing process 10 2.3 VOC analysis 12 2.4 Transcriptome profiling 13 2.5 Hierarchical heatmap and correlation analysis of gene expression and VOC accumulation 14 2.6 Expression analysis with quantitative RT-PCR 14 2.7 Cloning and sequence analysis of TPS11 15 2.8 Heterologous expression of TPS11 15 2.9 Electrophoresis and western blotting 16 CHAPTER 3 VOLATILE PROFILE DURING ENZYME ACTIVE STAGE OF PAOCHUNG TEA PROCESSING 17 3.1 VOC composition and content in fresh tea shoots of TTES No.12 17 3.2 VOC profile of three paochung tea manufacturing procedures 18 3.3 Selected VOCs with specific profiles during paochung tea manufacturing procedures 19 CHAPTER 4 CORRELATION BETWEEN VOLATILE CONTENT AND GENE EXPRESSION DURING EARLY STAGE PAOCHUNG TEA PROCESSING 21 4.1 VOCs and related gene expression in FADV biosynthetic pathway 21 4.2 VOCs and related gene expression in VT biosynthetic pathway 22 4.3 VOCs and related gene expression in AADV biosynthetic pathway 24 4.4 GBV related gene expression and VOCs 25 4.5 Validation of transcriptome profiling with qPCR 26 CHAPTER 5 VOLATILE BIOSYNTHETIC GENE EXPRESSION IN FRESH TEA SHOOTS WITH DIFFERENT MATURITY 28 CHAPTER 6 MOLECULAR CLONING AND EXPRESSION ANALYSIS OF TEA GERANYLLINALOOL SYNTHASE 30 CHAPTER 7 DISCUSSION 33 7.1 Stress responses during solar withering trigger transcriptome reprogramming and volatile biosynthesis in tea shoots 33 7.2 Stress responses during indoor withering and shaking trigger transcriptome reprogramming and volatile biosynthesis in tea shoots 35 7.3 The potential role of GBV hydrolysis in releasing free VOCs during paochung tea processing 37 7.4 The biosynthesis of linalool, a representative VT during paochung tea postharvest manufacturing 39 CHAPTER 8 CONCLUSION 41 REFERENCES 43 TABLES 52 FIGURES 59 APPENDIX 98 | - |
| dc.language.iso | en | - |
| dc.subject | 機械性傷害逆境 | zh_TW |
| dc.subject | 萜類合成酶 | zh_TW |
| dc.subject | 萎凋 | zh_TW |
| dc.subject | 揮發性化合物 | zh_TW |
| dc.subject | 部分發酵茶 | zh_TW |
| dc.subject | terpene synthase | en |
| dc.subject | semi-fermented tea | en |
| dc.subject | volatile organic compound | en |
| dc.subject | withering | en |
| dc.subject | wounding stress | en |
| dc.title | 探討包種茶製程於香氣化合物組成及其生合成之角色 | zh_TW |
| dc.title | Investigating the Roles of Paochung Tea Manufacturing Processes in Aroma Compound Composition and Biosynthesis | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-1 | - |
| dc.description.degree | 博士 | - |
| dc.contributor.oralexamcommittee | 蘇南維;陳右人;謝淑貞;官彥州 | zh_TW |
| dc.contributor.oralexamcommittee | Nan-Wei Su;Iou-Zen Chen;Shu-Chen Hsieh;Yen-Chou Kuan | en |
| dc.subject.keyword | 部分發酵茶,揮發性化合物,萎凋,機械性傷害逆境,萜類合成酶, | zh_TW |
| dc.subject.keyword | semi-fermented tea,volatile organic compound,withering,wounding stress,terpene synthase, | en |
| dc.relation.page | 110 | - |
| dc.identifier.doi | 10.6342/NTU202500449 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2025-02-14 | - |
| dc.contributor.author-college | 生物資源暨農學院 | - |
| dc.contributor.author-dept | 園藝暨景觀學系 | - |
| dc.date.embargo-lift | N/A | - |
| 顯示於系所單位： | 園藝暨景觀學系 | |
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