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研究生:李佩珊
研究生(外文):Lee Pei-shan
論文名稱:地耳草(HypericumjaponicumTHUNBERGexMURRAY)組織培養之研究
論文名稱(外文):Studies on Tissue Culture of Hypericum japonicum THUNBERG ex MURRAY
指導教授:郭昭麟郭昭麟引用關係
學位類別:碩士
校院名稱:中國醫藥大學
系所名稱:中國藥學研究所碩士班
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:133
中文關鍵詞:地耳草組織培養金絲桃科癒傷組織大量繁殖
外文關鍵詞:Hypericum japonicumplant tissue cultureGuttiferaecallusmultiplication
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地耳草為金絲桃科 (Guttiferae/Clusiaceae) 植物Hypericum japonicum THUNBERG ex MURRAY之乾燥全草,始載於生草藥性備要,稱田基黃,植物名實圖考稱地耳草,是台灣民間常用的青草藥之一,現代研究顯示其還具抑菌、抗病毒、保肝、抑制癌細胞、增強免疫等多種藥理作用,是頗具開發價值的中草藥。
本次研究目的,是要探討地耳草各部位培殖體培養於不同植物生長素和細胞分裂素對地耳草植株再生及癒傷組織誘導的影響,進一步建立植株之大量繁殖的系統。將莖、葉、莖節等培殖體培養於含1 mg/L BA和TDZ之MS培養基中能誘導出癒傷組織;莖節培養於含1 mg/L kinetin之MS培養基能長出具完整根系之植珠;將培殖體培養於含1 mg/L ZT之培養基中能誘導最多芽體,且以莖節的誘導率最高,培養4週後再繼代至含1% 蔗糖之WPM培養基,能長出最多的不定芽。將不定芽繼代至含5% 蔗糖之 1/2 MS培養基中,並添加1 mg/L之IAA或kinetin,各能得到最健壯的地上部及最健全的根系,並能成功馴化至野外。
關於地耳草癒合組織培養的部份,以MS基礎鹽類含1 mg/L BA之固體培養基中,莖節培殖體可獲得最高誘導率,而最佳繼代培養條件為MS基礎鹽類添加1 mg/L BA、3% sucrose、0.3% gelrite及pH 5.70 ± 0.01之培養基,在25 ± 1 ℃之恆溫下進行暗培養,15天繼代一次為宜。降低 BA 濃度會誘導癒合組織長出不定芽,以0.1 mg/L BA 能使癒合組織再生最多芽體,在將之繼代至不含生長調節劑之1/2 MS 培養基並培養於光照環境,能使不定芽抽長並發根,亦達到了大量繁殖的目的。本研究建立地耳草植物快速大量繁殖途徑,期能經由本省地耳草之提供大量植株,對藥用植物的開發有所助益。
Material of Di-Er-Cao, locally called ‘Tian-Ji-Huang’, is prepared from the entire herbs of Hypericum japonicum THUNBERG ex MURRAY (Family: Guttiferae/Clusiaceae),. It is one of the popular traditional Chinese medicine (TCM).
H. japonicum has been used for the treatment of some infectious diseases, contagious hepatitis, acute and chronic hepatitis, gastrointestinal disorder, internal hemorrhage and tumors for a long time. The medicinal herb is short in supply recently due to the environmental deterioration and overuse of the weedicides.
The purpose of this study is to establish a tissue culture technique for the mass propagation of H. japonicum. In the study we found that the most efficient shoot formation was the node explants, cultured on MS medium supplemented with 1 mg/L ZT for 4 weeks, then transferred onto solid medium containing WPM basic salts with 1% sucrose. The best medium for rooting was half-strength MS basic salts with 5% sucrose and 0.4% gelrite, pH 5.70 ± 0.01. It was also found that the best explant for callus initiation was node and stem, cultured on MS basal medium supplemented with 1 mg/L BA. The best cultural condition for callus proliferation is subculturing callus every 15 days on MS basal medium supplemented with 1 mg/L BA and 0.4% gelrite, incubated in the dark at 25 ± 1°C.
A successful multiplication of shoots from a standardized explant makes it possible to obtain a great quantity of uniform plant material for biotechnological purposes. The results of our study hence could be helpful to expand an agrotechnology of H. japonicum production as for traditional Chinese medicine resources development.
目 錄 ii
圖目錄 ii
表目錄 v
中文摘要 vii
Abstract viii
略字解 ix
第一章 緒 言 1
第二章 總 論 2
第一節 地耳草之本草學考察 2
第二節 地耳草之藥用植物學考察 6
第三節 地耳草之生藥學考察 22
第四節 地耳草之成份 29
第五節 地耳草之藥理作用 32
第六節 植物組織培養學之考察 35
第三章 試驗部分 55
第一節 材料與方法 55
第二節 結果 67
第三節 討論 107
第四章 結 論 114
附 錄 116
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