羅漢果組培苗二次脫毒技術(shù)研究

吳群英，李伯林，李景云

摘要：對(duì)羅漢果（Siraitia grosvenorii）組培苗采用熱處理和剝?nèi)∏o尖技術(shù)進(jìn)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(jìn)行檢測(cè)。結(jié)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)￠L(zhǎng)1.0～1.5 mm的莖尖進(jìn)行二次脫毒培養(yǎng)，存活率為80.00%，脫毒率為100.00%。

關(guān)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(hào)：S668.9 文獻(xiàn)標(biāo)識(shí)碼：A 文章編號(hào)：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