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药蒲公英(Taraxacum officinale Weber)耐1.5% NaCl变异体的筛选及特性分析
引用本文:张新果,李银心,陈华,石武良.药蒲公英(Taraxacum officinale Weber)耐1.5% NaCl变异体的筛选及特性分析[J].生物工程学报,2008,24(2):262-271.
作者姓名:张新果  李银心  陈华  石武良
作者单位:1. 中国科学院植物研究所光合作用与环境分子生理学重点实验室,北京,100093;中国科学院研究生院,北京,100039
2. 中国科学院植物研究所光合作用与环境分子生理学重点实验室,北京,100093
3. 中国科学院遗传与发育研究所,北京,100101
基金项目:国家863计划课题(No. 2007AA091705)和中国科学院重要方向性项目(No. KSCX2-YW-N-003)资助。
摘    要:为提高药蒲公英的耐盐性, 用20~30 d大小的药蒲公英叶片诱导愈伤, 获得的愈伤以NaCl作为选择因子, 用直接筛选的方法, 每3周进行一次继代培养, 经3个月继代筛选获得了耐1.5% NaCl的药蒲公英愈伤组织, 将耐1.5% NaCl的药蒲公英愈伤组织接种在分化培养基上分化出芽, 之后将再生芽转接到生根培养基中进行生根培养, 经4个月得到了12株耐1.5% NaCl的药蒲公英再生植株。与野生型相比, 耐盐植株叶片宽大、叶柄粗短、叶表面覆盖白色细毛, 根粗壮较短, 花茎中部具2 cm左右的苞叶。RAPD(Random amplified polymorphic DNA , 随机扩增的多态性DNA)和SDS-PAGE检测表明, 耐盐植株与对照植株在DNA及蛋白水平上均存在明显差异。1.5% NaCl处理后, 与普通再生植株相比, 耐盐株系的抗氧化酶活性明显提高, 脯氨酸含量上升幅度更为显著, 而丙二醛(MDA)含量降低, 其主要药用成分黄酮的含量显著增加。这些结果说明耐盐植株的抗氧化防御能力明显增强。以上结果表明耐1.5% NaCl的药蒲公英再生植株为耐1.5% NaCl药蒲公英变异体, 这些耐盐变异体有望成为抗盐耐海水蔬菜家族的新成员。同时, 这些耐盐变异体植株比普通植株具有更高的医用商业价值。耐1.5% NaCl的药蒲公英再生变异体遗传稳定性的研究正在进行中。

关 键 词:活性氧清除剂    耐盐变异体    药蒲公英    叶片外植体
收稿时间:5/9/2007 12:00:00 AM
修稿时间:2007-06-14

Selection and Identification of Salt-tolerant Variants of Taraxacum officinale
Xinguo Zhang,Yinxin Li,Hua Chen and Wuliang Shi.Selection and Identification of Salt-tolerant Variants of Taraxacum officinale[J].Chinese Journal of Biotechnology,2008,24(2):262-271.
Authors:Xinguo Zhang  Yinxin Li  Hua Chen and Wuliang Shi
Institution:Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China; Graduate University of Chinese Academy of Sciences, Beijing 100039, China;Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China;Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China;Key Laboratory of Photosynthesis and Environmental Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
Abstract:In order to obtain salt-tolerant variant plants of Dandelion (Taraxacum officinale Weber), the leaf discs were excised from 20 to 30-day old seedlings to produce callus, then the induced calli were transferred to selection mediums containing 1.5% NaCl. After regenerating and rooting, these salt-tolerant calli finally developed into 12 variant plantlets. Compared with the wild-type, these regenerated plants produced more trichomes on their leaves, and had larger leaves and shorter petioles. Additionally, the dumpy roots and an approximately 2-cm bract in middle parts of the floricanes were clearly observed in these salt-tolerant plants. By RAPD( Random Amplified Polymorphic DNA) and SDS-PAGE analysis, these salt-tolerant plants showed differences from the control at DNA and protein levels. With 1.5% NaCl treatment, the antioxidant enzyme activity, proline content, and flavonoid concentration were higher in these salt-tolerant plants, whereas maloaldehyde concentration was significantly lower. Salt-tolerant lines of T. officinale showed stronger anti-oxidative activity and higher flavonoid contents.
Keywords:ROS scavenger  salt-tolerant variant  Taraxacum officinale  leaf explant
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