鳳梨EMS誘變及RAPD分子標誌分析

EMS Mutagenesis of Pineapple (Ananas comosus (L.) Merr.) and RAPD Analysis

陳昀皓(Yun-Hao Chen) ； 陳京城(Ching-Cheng Chen)

《植物種苗》 13卷2期 (2011/06) Pp. 37-51

https://doi.org/10.30077/SN.201106.0003

摘要

本研究以台農17號鳳梨冠芽為材料，探討EMS（甲基磺酸乙酯；ethyl methanesulphonate）誘變處理對冠芽上之休眠側芽再生率的影響，另外測試增殖培養之條件以利誘變處理後代之繼代繁殖，並以RAPD（Random Amplified Polymorphism DNA）分子標誌分析經誘變處理之植株，評估其輔助選拔誘變植株之可行性。增殖試驗結果顯示BA:NAA為2:0.2ppm之配方增殖效率最高，培養4週後平均增殖4.9倍，顯著高於未添加植物生長調節劑之對照組的1.4倍。經EMS 浸泡處理之冠芽上的休眠側芽再生率，隨處理濃度及處理時間增加而降低。而EMS溶液pH值調整為6.0之處理的休眠側芽再生率為36.5%，顯著高於對照組（pH 3.75）的9.5%，顯示EMS溶液pH值對芽體毒害反應有顯著的影響。RAPD分子標誌分析結果，10支引子共產生166個條帶，其中多型性條帶共73條，而不同EMS濃度處理與對照組植株之平均條帶相似度如下：0.02％為0.915，0.04％為0.891，0.06％為0.892，0.08％為0.874，0.1％為0.876，顯示當EMS處理濃度愈高時，所造成植株之變異度會隨之增加。

並列摘要

The present study was conducted to investigate the effects of using EMS (ethyl methanesulphonate) as a mutagen on the regeneration rate of dormant crown axillary buds of 'Tainung No.17' pineapple cultured in vitro and micropropagation of 'Tainung No.17' pineapple plantlet was also tested. RAPD (Random Amplified Polymorphism DNA) markers were employed to analyze the EMS treated plants and the probability of RAPD markers used for marker-assisted selection was assessed. Different concentrations of BA and NAA were supplemented in the cultured media and the results showed that the best proliferation rate was the treatment of 2:0.2 (ppm) BA:NAA which prolifered 4.9-fold plants over four weeks of incubation while the control only had a proliferation rate of 1.4. The results of mutation induction indicated that the regeneration rate decreased when the EMS concentration and treatment duration increased. The pH value of EMS solution had significant effect on the phytotoxity to cultured dormant buds. Using buffer to adjust 0.4% EMS solution pH value to 6.0 increased the regeneration rate to 36.5%, compared with 9.5% in the control (pH=3.75). Ten RAPD primers amplified a total of 166 fragments which contained 73 polymorphic bands. The average genetic similarity between the EMS treated plants and the control plants were 0.915 in the 0.02% treatment, 0.891 in 0.04% treatment, 0.892 in 0.06% treatment, 0.873 in 0.08% treatment, and 0.875 in 0.1% treatment, respectively. It seemed that the genetic dissimilarity of EMS treated plants gradually increased when the EMS concentration increased.