卷叶贝母再生鳞茎生物碱高效积累相关基因资源发掘与激素调控机理研究

卷叶贝母是中药“川贝母”的珍贵来源植物，市场需求量大但产量低，供需矛盾突出，亟需寻找资源获得的新途径。甾体生物碱是卷叶贝母的主要药效成分。前期研究表明以卷叶贝母野生鳞茎为外植体经细胞分裂素和生长素诱导的再生鳞茎中生物碱含量明显高于野生鳞茎。基于RNA-seq的通路分析表明再生鳞茎与野生鳞茎之间在甾体生物碱生物合成途径的相关基因的表达量上存在差异。本研究拟在对卷叶贝母甾体生物碱合成关键基因克隆和功能鉴定的基础上，通过对培养过程中外源激素和代谢途径关键酶抑制剂的用量变化下相关代谢产物含量和基因表达量的变化进行分析，探讨激素在卷叶贝母甾体生物碱生物合成中的调控作用，为进一步阐释卷叶贝母甾体生物碱的合成规律，通过组织培养等途径获得卷叶贝母生物碱资源奠定基础。

Fritillaria cirrhosa D. Don is a precious source of traditional Chinese medicine of Chuan Bei Mu. Nowadays, F. cirrhosa is confronted with sharp conflictions between supply and demand. Effective ways to resolve this contradiction are needed. Steroidal alkaloids are the major effective components in F. cirrhosa. Previous research showed that the alkaloid content in regeneration bulbs induced by cytokinin and auxin was higher than that in the wild bulbs which are used as the explants. RNA-seq analysis showed differences between the regeneration and wild bulbs in expression levels of steroidal alkaloid biosynthesis pathway related genes. This project will clone the key genes in the steroidal alkaloid biosynthesis pathway and then verify their functions. We will also explore the role of hormones in the regulation of steroidal alkaloid biosynthesis by analyzing the content of metabolites and gene expression levels in the materials which cultured on the mediums with different dosage of hormones and/or inhibitors of some crucial enzymes in metabolic pathways. This will lay a foundation on obtaining alkaloids resources of F.cirrhosa by tissue culture, and explaining the synthesis mechanism of steroidal alkaloid in F.cirrhosa in further study.