Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/36777
DC FieldValueLanguage
dc.contributor林順福zh_TW
dc.contributor夏奇鈮zh_TW
dc.contributor賴宏亮zh_TW
dc.contributor陳宗禮zh_TW
dc.contributor葉茂生zh_TW
dc.contributor.advisor葉茂生zh_TW
dc.contributor.author林資哲zh_TW
dc.contributor.authorLin, Tzu-Cheen_US
dc.contributor.other中興大學zh_TW
dc.date2012zh_TW
dc.date.accessioned2014-06-06T07:57:49Z-
dc.date.available2014-06-06T07:57:49Z-
dc.identifierU0005-1508201115133300zh_TW
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dc.identifier.urihttp://hdl.handle.net/11455/36777-
dc.description.abstract為了探討台灣產藥用植物苦參(Sophora flavescens Ait.)之特性,收集自台灣及中國不同地區之苦參,進行農藝性狀調查、器官解剖、分子鑑定資料之建立,並以組織培養大量繁殖種苗,進而利用細胞懸浮培養生產有效性成分,最後檢測不同年生與不同海拔高度生育期之苦參根部有效性成分之含量。 根據15 項農藝性狀調查結果,以花蓮及玉里地區收集系生長較好。苦參器官解剖結果顯示,不同收集系之根、莖及葉部組織構造相似,可作為苦參基源鑑定之參考。進一步觀察不同收集系苦參染色體之數目,均為 2n=18 ,其核型公式為 2n=18 = 14m + 4sm。利用ITS (internal transcribed spacer)分子鑑定顯示,不同收集系苦參之間變異性相當小,具有高度的遺傳相似性,經鑑定結果屬於同一種。苦參真偽品藥材的5.8S rRNA-ITS2 序列分析呈現高度的序列相似性,但在 ITS2 序列中卻可以偵測到苦參與其偽品的鹼基對差異。再透過 5.8S rRNA-ITS2 之 PCR-RFLP技術,結合限制酵素剪切,可以有效正確的鑑別苦參藥材基源之真偽。 利用苦參無菌苗之莖段為培植體,在適量 BA (2.0 mg L-1) 處理下,可以建立苦參瓶苗大量繁殖系統,同時發現在較高濃度的 auxin 處理下有利於根的生長,但過高之生長素濃度則不利於後續植株之生長。本研究已成功大量繁殖瓶苗,配合高透氣性之 Grotex 透氣膜及藥包紙進行容器封口處理,可生產品質均一且無落葉現象的健康組培苗。同時藉由掃描式電子顯微鏡觀察移植存活植株及組培苗葉片,證實高透氣性封口置換處理會增加瓶內氣體的交換及氣孔功能的提昇。在苦參細胞懸浮培養方面,試驗結果發現培養 3週後,癒合組織會由生長遲滯期進入快速生長期,至第 7 週進入高原期。在不同初始的 pH條件下,檢測兩種生物鹼含量從培養初期緩慢增加,直到培養 4 週後才快速上升。總酚類化合物含量變化,亦有明顯之差異,以初始 pH 5.7培養 7 週含量最高。總黃酮類含量以初始 pH 5.2培養含量最高。 不同年生之苦參收集系,其苦參鹼及氧化苦參鹼含量不同,以 一 年生成分含量而言,甘肅收集系含量較高。總類黃酮及總酚類含量亦不同,以 一年生之丹大收集系含量較高。不同海拔高度生育期之氧化苦參鹼、苦參鹼、總類黃酮及總酚類含量亦有明顯之差異,以生長 12 個月所測得生物鹼及總酚類化合物含量最高,隨著發育時間的延長有上升之情形。總黃酮化合物含量在海拔 700 公尺及 1000 尺生長 6 個月時有最高之含量。zh_TW
dc.description.abstractIn order to understand the potential for untilization of Sophora flavescens Ait., a medical plant in Taiwan, seeds of Sophora flavescens Ait. were collected from Taiwan and china for planting. Investigation of agronomic characteristics, histological analysis, molecular markers for indentification were conducted for various collected Sophora flavescens lines. In addition, micropropagation and suspension cell culture were also conducted to produce seedlings and medical effective components, respectively. Influences of cultivation period and geographic altitude on root containing with effective components were compared. According to 15 agronomic characteristics investigated. The best plant growth were found from collected lines of hualien and yuli. Histological analysis of root, stem, and leaf structures were inspected and a result of higher similarities were observed among all collected lines. Examination of chromosome number showed that all collected lines with 2n=18=14m+4sm karyotype distribution. The high similarity and genomic conservation of ITS (internal transcribed spacer) molecular marker among all collected lines of Sophora flavescens Ait. revealed all collected lines are in same species. Although sequences of 5.8S rRNA-ITS2 were conserved among tested species, several base pairs variances were observed in the ITS2 sequences. Using ITS2 PCR-RFLP coupled with sacII restriction enzyme digestion was performed effectively to distinguish Sophora flavescens Ait. from some of its adulteration. The highest induction rate of callus, based on asepsis stem coupled with BA(2.0 mg L-1) treatment, were builded. The best condition with higher concentration of auxin treatment for rooting, the worse for growth of plantlet subsequently. There were less defoliation and higher quality of plantlet, covering Gortex membrane and dispense paper, to be obtained with higher survival rate. The observation of scanning electron microscope (SEM) revealed that acclimated plantlets with functional stoma were found from closure Gortex membrane and dispense paper container treatments. In addition, the suspension culturing system of compact callus from Sophora flavescens Ait. was established. Biomass of the suspension cells was found increasing dramatically starting from 3rd wks until 7th wks. Investigation of the effect of pH condition on 2 kinds of alkaloids contents in the suspension cells was conducted and a result showed that slowly increasing in the first 4 wks culturing before its accumulation dramatically. The highest total phenol content was found for 7 wks culturing in the pH 5.7 initiation medium. However, the highest flavonoid content was obtained from the culturing medium with pH 5.2 for initiation. Higher martine and oxymatrine contents were obtained in the one-year root of Gansu line among all tested lines with various root ages. Meanwhile a higher flavonoid and total phenolic compounds were in one-year old Danda line. Influences of cultivation period and geographic altitude for planting were investigated. Matrine, oxymatrine, alkaloids as well as total phenolic compounds were found significant different among various planting altitude. The content of alkaloids and total phenolic compounds were found increased with the growth period and the highest contents at 12-month cultivation period. the highest content of flavonoid was obtained from a 6-month-old plant growing either in 700 or 1000 m high altitudes.en_US
dc.description.tableofcontents目次 頁次 中文摘要………………………………………………………………...i 英文摘要……………………………………………………………….iii 表目次…………………………………………………………………..vi 圖目次…………………………………………………………………..iv 第一章 緒言…………………………………………………………1 第二章 苦參農藝性狀、器官解剖及核型分析之研究……………4 一、前言………………………………………………………………4 二、前人研究…………………………………………………………5 三、材料與方法………………………………………………………7 四、結果…………………………………………………………….11 五、討論…………………………………………………………….23 第三章 苦參分子鑑別與偽品物種5.8S rRNA序列及PCR-RFLP 之分子鑑定……………………………………………………27 一、前言………………………………………………………………27 二、前人研究…………………………………………………………28 三、材料與方法………………………………………………………31 四、結果………………………………………………………………35 五、討論………………………………………………………………51 第四章 苦參組織培養微體繁殖之研究……………………………55 一、前言……………………………………………………………..55 二、前人研究………………………………………………………..56 三、材料與方法………………………………………………………63 四、結果………………………………………………………………68 五、討論………………………………………………………………89 第五章 苦參癒合組織誘導及根部細胞懸浮培養有效性成分含量 之研究…………………………………………………………96 一、前言……………………………………………………………..96 二、前人研究…………………………………………………………97 三、材料與方法…………………………………………………….102 四、結果…………………………………………………………….107 五、討論…………………………………………………………….124 第六章 不同年生與不同海拔高度生育期之苦參根部有效性成分含量之研究……………………………………………………….......130 一、前言…………………………………………………………….130 二、前人研究……………………………………………………….131 三、材料與方法…………………………………………………….133 四、結果…………………………………………………………….136 五、討論…………………………………………………………….147 第七章 綜合討論………………………………………………….150 第八章 引用文獻…………………………………………………157zh_TW
dc.language.isoen_USzh_TW
dc.publisher農藝學系所zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1508201115133300en_US
dc.subjectSophora flavescens Ait.en_US
dc.subject苦參zh_TW
dc.subjectOrgan anatomyen_US
dc.subjectKaryotypeen_US
dc.subjectMolecular identificationen_US
dc.subjectMicropropagationen_US
dc.subjectsuspension cultureen_US
dc.subjectmatrineen_US
dc.subject器官解剖zh_TW
dc.subject核型分析zh_TW
dc.subject分子鑑定zh_TW
dc.subject微體繁殖zh_TW
dc.subject懸浮培養zh_TW
dc.subject苦參鹼zh_TW
dc.title台灣產苦參之鑑定、組織培養與活性成分分析之研究zh_TW
dc.titleStudies on Identification, In vitro culture, and Active Compound Analysis of Sophora flavescens Ait. in Taiwanen_US
dc.typeThesis and Dissertationzh_TW
item.languageiso639-1en_US-
item.openairetypeThesis and Dissertation-
item.cerifentitytypePublications-
item.grantfulltextnone-
item.fulltextno fulltext-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
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