长春花萜类吲哚生物碱含量测定及相关基因的表达分析

采用反相高效液相色谱法(RP-HPLC)对23个长春花品种中的重要萜类吲哚生物碱文多灵、长春质碱和长春碱含量进行了测定,发现这3种生物碱的含量在不同品种中存在较大差异。相关性分析表明,文多灵、长春质碱和长春碱这3种生物碱中,文多灵含量和长春碱含量有极显著的相关性(P0.01)。利用RT-PCR分析了香叶醇10-脱羧酶基因(g10h)和异胡豆苷合成酶基因(str)在萜类吲哚生物碱含量有显著差异的品种之间的表达水平差异,并结合生物碱含量数据结果,发现g10h和str的表达水平和文多灵和长春质碱的总含量有显著的正相关性(P0.05),说明g10h和str基因可以作为长春花中文多灵和长春质碱含量的参考基因标记。该研究对为选育高萜类吲哚生物碱含量长春花品种及长春花萜类吲哚生物碱代谢工程具有重要意义。

Measurement and Analysis of Important Alkaloids and Related Gene Expression in Catharanthus roseus

The contents of three important terpenoid indole alkaloids (TIAs) including vindoline,catharanthine and vin-blastine in 23 Catharanthus reseus varieties were measured by Reversed Phase High Performance Liquid Chromatography (RP-HPLC).It was found that the contents of the three alkaloids varied along with the different varieties.Correlation a-nalysis indicated that among the three alkaloids,vindoline and vinblastine contents were correlated at very significant lev-el (P < 0.01).RT-PCR was used to analyze the expression of geraniol 10-hydmxylase gene (g10h) and strictosidine synthase gene (str) in C.roseus varieties with distinct alkaloids contents, and the results, together with the alkaloids con-tent data analysis,showed that the expression of g10h and str was correlated with the combined contents of vindoline and vinblastine at significant level (P <0.01) ,indicating that g10h and str genes could be the reference gene markers for vindoline and vinblastine contents in C.roseus.This study is useful for developing high-TIAs content varieties and meta-bolic engineering of TIAs in C.roseus.