吳群英,李伯林,李景云
摘要:對羅漢果(Siraitia grosvenorii)組培苗采用熱處理和剝?nèi)∏o尖技術進行二次脫毒處理,脫毒效果采用間接酶聯(lián)免疫法和電鏡觀察法進行檢測。結果表明,適合材料熱處理的培養(yǎng)基為MS+0.3 mg/L 6-BA+NAA 0.1 mg/L+0.1 mg/L IBA,熱處理材料的頂芽呈錐形,利于剝?nèi)∏o尖;莖尖分化最佳培養(yǎng)基為MS+0.3 mg/L 6-BA +0.05 mg/L NAA+0.1 mg/L GA3;38.5 ℃熱處理10 d剝?nèi)¢L1.0~1.5 mm的莖尖進行二次脫毒培養(yǎng),存活率為80.00%,脫毒率為100.00%。
關鍵詞:羅漢果(Siraitia grosvenorii)花葉病毒;莖尖;熱處理;脫毒;酶聯(lián)免疫法
中圖分類號:S668.9 文獻標識碼:A 文章編號:0439-8114(2013)05-1185-03
Study on Duplicate Virus-free Techniques of Siraitia grosvenorii
Tissue Culture Seedling
WU Qun-ying1,LI Bo-lin2,LI Jing-yun3
(1. Biotechnology College, Guilin Medical University, Guilin 541004, Guangxi, China;
2. College of Life Science, Guangxi Normal University, Guilin 541004, Guangxi, China;
3. College of Basic Medicine, Guangxi University of Chinese Medicine, Nanning 530001,China)
Abstract: The tissue culture seedlings of Siraitia grosvenorii were treated by heat treatment and tip-stripping to obtain virus-free seedling. ELISA(enzyme-linked-immunosorbent serologic assay) method and electron microscope method were applied for virus detection. The results showed that the suitable medium for heat treatment was MS+0.3 mg/L 6-BA+0.1 mg/L NAA+0.1 mg/L IBA. Terminal bud after heat treatment was conical and easy for stripping stem tip. The optimum medium for inducement of tips was MS+0.3 mg/L 6-BA+0.05 mg/L NAA+0.1 mg/L GA3. Heat treatment for 10 d at 38.5 ℃ twice, stripping shoot-tip for 1.0~1.5 mm long, the survival rate was 80.00%; and the detoxication rate was 100.00%.
Key words: Siraitia grosvenorii virus (WMV-2-Luo); shoot-tip; heat treatment; virus-free; ELISA