Thidiazuron促進赤桉優良營養系葉片癒合組織之植株再生

Thidiazuron Enhancement of Plant Regeneration from Leaf Calli of Superior Clones of Eucalyptus camaldulensis

張淑華(Shu-Hwa Chang) ； 何政坤(Cheng-Kuen Ho) ； 蔡錦瑩(Jin-Yin Tsai) ； 陳振榮(Zenn-Zong Chen)

《台灣林業科學》 15卷1期 (2000/03) Pp. 81-90

https://doi.org/10.7075/TJFS.200003.0081

摘要

由赤桉種原試驗林選取具優良表型的六個單株，取其芽體經試管微體繁殖培養後之試管苗作為試驗材料來源。為建立葉片再生體系統，先以48號營養系試管苗葉片，培養於添加cytokinins(TDZ、BA或kinetin)組合auxins(NAA或2,4-D)的B5培養基誘導癒合組織及芽體再生。發現葉片可於各種培養基中產生癒合組織，但只有TDZ可以誘導癒合組織再生芽體，且濃度限於0.1-0.5mg/L。TDZ濃度提高為1mg/L時，芽體生長無法完全，只觀察到擬芽體的產生，TDZ濃度再增加，則癒合組織大量增殖，而無法再生芽體。由BA與NAA培養基中所誘導的瘤狀癒合組織經移入TDZ培養基中可誘導芽體再生，其中以0.5mg/L TDZ效果最佳。培養基中加入100mg/L抗壞血酸可以抑制發生於繼代培養的癒合組織褐化，同時促進芽體分化。將分化完全的小芽移入低濃度BA或TDZ培養基，可誘導芽體抽長。來自BA抽長的芽體可順利發根，但培養於TDZ芽體，則需再移入不含任何植物賀爾蒙的培養基培養一個月，去除TDZ效應後，才可於發根培養基中順利發根。發根的小苗經移入溫室培養，可得95％以上的成活率。將此方法用於比較六種不同營養系間葉片的芽體再生能力，發現呈現相當大差異。

關鍵字

瘤狀癒合組織 ； 赤桉 ； 植株再生 ； 優良營養系 ； thidiazuron

並列摘要

Shoot subcultures via micropropagating shoot tips of Eucalyptus camaldulensis trees with superior phenotypes in the provenance plantations were used as materials in this study. To establish a regeneration protocol, leaf explants of clone
#48 cultures were cultured initially in E5 medium containing various combinations of cytokinins (TDZ, BA, and kinetin) and auxins (NAA and 2, 4-D) for inducing callus and shoot regeneration. All media produced calli from cultured explants, but only TDZ incorporation medium differentiated shoots from calli. However, its concentrations were limited to, the range of 0.1 to 0.5mg/L. With an increase of TDZ concentration to 1mg/L, calli formed a shoot-like body but could not induce shoots. At TDZ concentrations greater than 1mg/L, only abundant calli formed. All media containing BA or kinetin combined with different auxins failed to induce shoots. Nodular calli derived from BA and NAA incorporation medium could form shoots after they were transferred into TDZ medium, especially at 0.5mg/L. Callus browning occurring in subcultures could be controlled by adding ascorbic acid into medium, which increased shoot formation. These shoots elongated when shooting calli were cultured in media with lower BA or TDZ concentrations. Elongated shoots from BA-containing medium rooted well, while those in TDZ-containing medium did not form roots unless they were cultured in hormone-free medium for 1 mo to remove the TDZ residual effect. A 95% survival rate was obtained when these were transplanted in a greenhouse. Explants collected from 6 superior clones of E. camaldulensis were cultured following the protocol of shoot regeneration as previously established. Great variations of shoot regenerability among these clones were observed.

並列關鍵字

nodule callus ； Eucalyptus camaldulensis ； plant regeneration ； superior clones ； thidiazuron