药蒲公英(Taraxacum officinale Weber)耐1.5% NaCl变异体的筛选及特性分析

Selection and Identification of Salt-tolerant Variants of Taraxacum officinale

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。

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.