Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/20612
標題: Benzothiadiazole,ABA及抗老化中藥處理對番木瓜組織培養苗抗低溫逆境能力之探討
The study of chilling tolerance of tissue-cultured plantlet of Carica papaya L. by treatments of benzothiadiazole,ABA and antiaging traditional Chinese medicine
作者: 陳冠吟
Chen, Kuan Yin
關鍵字: ABA
離層酸
chilling stress
SDS-PAGE
tissue-cultures plantlet of papaya
antioxidative enzymes
ATPase
低溫逆境
番木瓜組織培養苗
抗氧化酵素
出版社: 植物學系
摘要: 一、 摘要 以台農二號( Tai-nung No.2 )番木瓜( Carica papaya L. )組織培養苗為材料,分別添加ABA、BTZ及抗老化中藥何首烏及當歸於培養基中,進行不同溫度(25℃、15℃及10℃)處理,探討低溫對番木瓜組織培養苗葉片抗氧化酵素系統、原生質膜及液泡膜上ATPase活性之影響。另以SDS-PAGE分析可溶性蛋白質變化,期能暸解添加物是否能提高組織培養苗抗氧化能力,並增加對低溫之抗性。結果顯示,番木瓜組織培養苗處於15℃及10℃低溫環境下,脂質過氧化作用及離子滲漏程度均增加,顯示低溫已造成膜系之傷害,此時細胞內抗氧化酵素系統之酵素活性及原生質膜、液泡膜上ATPase之活性並未隨低溫處理而明顯增加,表示在低溫逆境中,活性氧無法充分藉由抗氧化酵素系統加以清除,導致活性氧的累積。綜合上述生理代謝變化結果,顯示出番木瓜組織培養苗不耐寒之特性。 木瓜組織培養苗先經ABA、BTZ及抗老化中藥何首烏及當歸進行前處理後,再進行低溫處理,結果顯示除脂質過氧化作用及離子滲漏程度降低外,培養苗抗氧化酵素活性增加。進一步分析添加BTZ、何首烏及當歸前處理對細胞原生質膜及液泡膜上ATPase活性之影響,結果亦顯示添加物可提昇培養苗ATPase之活性。本研究另以SDS-PAGE分析BTZ、何首烏及當歸處理之培養苗在低溫下,是否能誘導不同的蛋白質累積?結果顯示,經BTZ處理之培養苗,在25℃及10℃下,140.0、12.6、11.6、11.1及10.0 KDa蛋白質均有累積。15℃及10℃低溫下,何首烏處理可誘導特定128.6 KDa蛋白質累積,而當歸處理對番木瓜組織培養苗葉片可溶性蛋白質之影響在於可誘導小分子量(13.1、12.6、11.6及11.1 KDa)的蛋白質累積。 綜合上述各項生理機能測定得知,番木瓜組織培養苗不具耐寒之特性,經外加ABA、BTZ及何首烏、當歸之處理,可短暫提昇培養苗之耐寒能力。
Abstruct This study is focused on the effect of low temperature( 15℃and 10℃ )on the activity of antioxidative enzymes、plasma membrane ATPase and tonoplast ATPase of ABA、benzothiadiazole(BTZ)、Polygonum multiflorum thunb (hoshouwu )and Angelicae sinesis diel (danggui) treated tissue cultured plantlet of Carica papaya L.. Further more, the effect was made to investigate of the effects of low temperature on total soluble protein content in leaves of tissue-cultured plantlet of papaya. The goal of this study is to increase the oxidative ability and chilling tolerance by addition of various effectors to the culture media. The results showed the MDA and ions leakage of papaya were increased during chilling stress, at the same time, the antioxidative enzymes、plasma membrane and tonoplast ATPase activities were not followed. These results suggest that the antioxidative enzymes could not actively eliminate the activated oxygen generated by chilling treatments. The results revealed the chilling intolerance of papaya. The accumulation compounds were added to the culture media to enhance the ability of chilling tolerance.The effects of chilling on MDA、ions leakage were decreased , antioxidative enzymes activities were increased by ABA、BTZ、Polygonum multiflorum thunb (hoshouwu )and Angelicae sinesis diel (danggui) pretreatment. Further analyses on activities of the plasma membrane ATPase and tonoplast ATPase of BTZ、Polygonum multiflorum thunb (hoshouwu )and Angelicae sinesis diel (danggui) pretreatment of chilling revealed that all the additives could promote the ATPase activity , among which the plasma membrane ATPase increased most. SDS-PAGE was used to analyze the protein accumulation during chilling of BTZ、Polygonum multiflorum thunb (hoshouwu )and Angelicae sinesis diel (danggui) pretreatment of chilling. Several polypeptides with molecular mass of 140.0、12.6、11.6 and 11.1 KDa were all accumulation at 25℃、10℃ of BTZ pretreatment, A novel protein appeared, with molecular weight of 128.6 KDa under 15℃ and 10℃ of Polygonum multiflorum thunb (hoshouwu ) pretreatment. The accumulation of four low molecular mass polypeptides ( 13.1、12.6、11.6、11.1 KDa) was found in danggui-treated papaya plantlet. Based on the results,the pretreatments of ABA﹑BTZ、Polygonum multiflorum thunb (hoshouwu )and Angelicae sinesis diel (danggui) before chilling stress. could transiently increase the chilling tolerance of papaya.
URI: http://hdl.handle.net/11455/20612
Appears in Collections:生命科學系所

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