滇龙胆优质种质筛选及离体培养条件优选研究  

Study on the Screening of High Quality Germplasm and Optimization of in Vitro Culture Conditions for Gentiana rigescens

作　　者：赵志莲 徐立[1,2] 李海峰[3] 

机构地区：[1]大理农林职业技术学院,云南大理671003 [2]大理学院基础医学院,云南大理671000 [3]大理学院药物研究所,云南大理671000

出　　处：《湖北农业科学》2015年第17期4228-4231,共4页Hubei Agricultural Sciences

基　　金：云南省大理白族自治州科学研究计划项目(DL2012);大理学院2012应用开发基金项目(KY1219211210)

摘　　要：以大理州不同居群滇龙胆(Gentiana rigescens Franch.ex Hemsl.)、同一居群不同植株为试验材料,采用高效液相色谱法(HPLC)对滇龙胆根及根茎中龙胆苦苷的含量进行测定;再以龙胆苦苷含量≥7.0%的优质种质资源为外植体,采用正交试验设计方法研究培养基、蔗糖浓度、pH及培养温度对滇龙胆离体培养植株再生的影响。结果表明,大理州6个居群滇龙胆根及根茎中的龙胆苦苷含量较高,不同居群间龙胆苦苷含量具有显著差异(P<0.05),以鹤庆县居群的龙胆苦苷含量最高(80.70 mg/g),含量最低的巍山县居群也超过国家药典规定标准的2倍多;同一居群内不同植株滇龙胆根及根茎中龙胆苦苷含量差异较小,只有云龙县居群不同植株间具有显著差异(P<0.05),其他居群不同植株间无显著差异(P>0.05);滇龙胆生长点离体培养及植株再生最适培养基和培养条件是1/2MS+30 g/L蔗糖+pH 5.2+20℃/25℃变温培养,再生植株诱导率最高可达88.9%,试验初步建立了滇龙胆离体培养及植株再生体系。Gentiana rigescens Franch.ex Hemsl. plants from different populations and different individuals from the same population in Dali prefecture were selected as the experimental material. The gentiopicroside content of root and rhizome of G. rigescens was determined by High performance liquid chromatography （HPLC）. Then the high quality germplasm resources of which the gentiopicroside content was not lower than 7.0% were selected as the explants; and the effects of culture medium, sucrose concentration, pH value, and culture temperature on the in vitro culture and plant regeneration of G. rigescens were explored by orthogonal experiment design. The experimental results showed that the gentiopicroside content of root and rhizome of G. rigescens from 6 different populations of Dali prefecture was high, and there was significant difference among different populations in the gentiopicroside content （P﹤0.05）. Among them, the gentiopicroside content from Heqing county population was the highest （80.70 mg/g）; and from Weishan county population was the lowest, which was still more than twice of the standard gentiopicroside content of national pharmacopoeia. The difference of the gentiopicroside content of root and rhizome of G . rigescens from the same population was smaller. There was significant difference in the gentiopicroside content among different plants from Yunlong county population （P﹤0.05）, while no significant difference in other populations （P﹥0.05）. The optimal culture medium and culture conditions for the growing point of in vitro culture and plant regeneration of G. rigescens of Dali populations was 1/2 MS ＋ 30 g/L sucrose ＋ pH 5.2, and variable temperature culture at 20℃/25℃. Under the condition, the regenerated plant inductivity was the highest as the inductivity reached to 88.9%. This study preliminary construct the in vitro culture and plant regeneration system for G. rigescens from Dali population.

关 键 词：滇龙胆(Gentiana rigescens Franch.ex Hemsl.) 种质资源 龙胆苦苷 离体培养 植株再生 

分 类 号：S567.23[农业科学—中草药栽培]