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研究生:陳國峰
研究生(外文):Kao-Phone Chen
論文名稱:農桿根群菌對台灣紅豆杉插穗發根能力及生長之影響
論文名稱(外文):Effects of Agrobacterium rhizogenes infection on the rooting ability and growth of Taxus mairei cutting
指導教授:郭幸榮郭幸榮引用關係何政坤何政坤引用關係
指導教授(外文):Shing-Rong KuoCheng-Kuen Ho
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:森林學研究所
學門:農業科學學門
學類:林業學類
論文種類:學術論文
論文出版年:2000
畢業學年度:88
語文別:中文
論文頁數:66
中文關鍵詞:台灣紅豆杉扦插農桿根群菌紫杉醇10-去乙醯巴卡亭 Ⅲ生長
外文關鍵詞:Taxus maireicuttingAgrobacterium rhizogenestaxol10-deacetyl baccatin Ⅲgrowth
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台灣紅豆杉稀疏散生於台灣中高海拔山上,天然母樹多是胸徑100 cm的巨型大樹,木材是昂貴的雕刻用材,針葉又含有抗癌有效藥物紫杉醇,被譽為台灣的國寶級樹種。由於天然母樹樹齡頗高,許多具有高含量紫杉醇的母樹插穗,很難藉扦插發根保存此一珍貴種質。
本文探討三種農桿根群菌種包括R1600、R1601及R15834對不同胸徑級(18、62、78、91 cm,母樹代號J4、J5、J8與J10)台灣紅豆杉插穗發根的促進作用。經120天的扦插期後,經由混和模式統計分析,各母樹插穗的基本發根率(對照組)各為71%、58%、0%及10%,具極顯著差異。三種農桿根群菌處理對插穗發根率的促進都顯著優於對照與 IBA處理,其中又以R15834菌種促進發根最佳,其發根率分別為91%、94%、8%及38%,且經農桿根群菌接種插穗在120天時的發根數與發根總長分別為對照組與IBA處理的3倍與6倍。
為提高農桿根群菌的感染率,本文更進一步探討割傷插穗基部與在菌液中添加acetosyringone (AS)、proline與betaine對易發根 (J5)與難發根 (J8)插穗發根的影響,結果顯示接種農桿根群菌對易發根的J5插穗,在扦插90天與120天時都可提高發根率,發根數與發根總長,且均顯著高於對照組,但割傷與添加藥劑並無顯著的促進轉殖及發根作用。對於難發根的J8母樹,在扦插120天時割傷插穗以菌液加AS處理的發根率為21%顯著高於對照組。延長扦插期到150天時,未割傷插穗以菌液加AS及proline處理可提高發根率達73%,發根總長達17.1 cm,顯著高於插穗割傷及未割傷之對照組的11%及7.3 cm,顯示難發根母樹需要延長扦插期間。
本研究調查285天生J4農桿根群菌、IBA與對照組的扦插苗生長表現和紫杉醇及10-去乙醯巴卡亭Ⅲ(10-deacetyl baccatin Ⅲ,簡寫DB)的產量,在接種R1600與R15834扦插苗的扦插苗總乾重顯著高於未處理苗,但菌種與 IBA處理扦插苗間差異不顯著。每株苗木紫杉醇的產量在各種處理的扦插苗間差異不顯著,但DB的總產量皆比未處理苗高1倍以上。因此利用農桿根群菌處理扦插苗與 IBA處理苗一樣都可促進苗木的生長並提高DB的產量。
Taxus mairei trees are scarcely scattered in the high mountains of Taiwan. Most of them are very giant trees with more than 100 cm of DBH. Their wood is valuable for sculpture, while needles contain taxol, an effective anti-cancer drug. They are the national treasures of Taiwan.
Cuttage is a common way in traditional tree breeding to preserve these valuable germplasm with good wood quality and high taxol concentration. However, it is very difficult to root these old trees. To enhance the rooting ability of 4 trees with 18, 62, 78 and 91 cm of DBH (tree coded as J4, J5, J8, and J10), cuttings collected from these trees were treated with IBA at 0 (control) and 3,000 ppm and 3 strains of Agrobacterium rhizogenes. After cutting 120 days, the natural rooting percentages of control treatments among 4 trees were 71%, 58%, 0%, 10%, respectively, showing very significant different in rooting ability after statistical analysis of mix model. Rooting percentages of three strains bacterium treatments were significantly greater than that of control and IBA treatments. Agrobacterium R15834 had 91%, 94%, 8%, and 38% for 4 different-DBH trees, better than the other Agrobacterium strains. Rooting numbers of cuttings produced from bacterium treatments were 2-3 times greater than that from control and IBA treatments, showing significant differences by using t-test at P = 0.06.
To investigate the effects of wound treatment and transformation efficiency on rooting ability of easy- and hard-to-root trees (J5 and J8), cuttings treated with or without 1 cm wound near to the basal parts of cuttings and bacterium suspension supplemented with actosyringone (AS), proline or betaine were tried. For J5 tree, Agrobacterium treatments still improved rooting percentages, numbers, and length of cuttings after cutting for 90 and 120 days, significantly greater than did control. However, There were not significantly different among rooting ability of wounded and unwounded cuttings treated with bacterium adding efficient components. For J8 trees, wounded cuttings treated with bacterium suspension adding AS obtained 21% of rooting percentage after cutting for 120 days, significantly greater than the control. When increasing cutting period up to 150 days, unwounded cuttings treated with bacterium suspension adding AS plus proline obtained 73% of rooting percentage and 17.1 cm of root length, significantly greater than did control, in which 11% and 7.3 cm were obtained. This suggests that elongating rooting period for difficult to root is required.
The variation of growth performance and the concentrations of taxol and 10-deacetyl baccatin III (DB) in 285-day-old rooted cuttings of J4 trees treated with or without IBA and Agrobacterium was investigated. The dry weights of R1600 and R15834 treated plants were significantly greater than control ones. However, there is no significant difference among the dry weights of bacterium and IBA treated plants. The amount of taxol per plant were not significantly different among all the treatments of rooted cuttings. However, the amount of DB per plant treated with IBA and Agrobacterium were one time greater than the control plants. This indicates that the plants treated with Agrobacterium rhizogenes or IBA can both improve seedling’s growth and DB yield as well.
中文摘要………………………………………………..……………….1
英文摘要………………………………………………..….……………3
壹、前言……………………………………………………..………….5
貳、前人研究…………………………………………..……………….7
參、不同農桿根群菌對台灣紅豆杉插穗發根之影響……………….19
肆、割傷處理與菌液添加劑對插穗發根之影響…………….…...…...31
伍、台灣紅豆杉扦插苗之生長表現…………………………..….…...43
陸、台灣紅豆杉天然母樹及扦插苗之紫杉烷類分析………….….….51
柒、總結...................................................60
捌、引用文獻……………………………………………..…………….61
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