摘要
目的:鉴别垂直生长(V类)和横向生长(L类)两类爵床并比较其地上部分及不同器官中化学成分含量的差异。方法:使用试剂盒提取两类爵床叶中的DNA,经聚合酶链式反应(PCR)扩增和双向测序得内部转录间隔区2(ITS2)序列,psb A-trn H序列;采用HPLC,测定两类爵床地上部分及花穗、叶和茎中爵床脂定B和金不换甲醚的含量;通过独立样本T检验和配对样本T检验,比较两类爵床地上部分及其不同器官中的爵床脂定B和金不换甲醚的含量差异。结果:将2个序列经过剪切后比对,V类和L类爵床的DNA序列完全一致;独立样本T检验结果表明,爵床脂定B的含量在两类爵床的地上部分中不存在显著差异,而金不换甲醚的含量为V类爵床显著高于L类(P <0. 05);配对样本T检验结果表明,爵床脂定B和金不换甲醚的含量在同类爵床的花穗、叶和茎中均存在显著性差异(P <0. 05),花穗和叶中的含量远大于茎;且金不换甲醚在V类爵床的花穗和叶中的含量要高于L类。结论:DNA条形码技术鉴别序列一致,表明两类不同生长形态的爵床为同种植物;不同生长形态的爵床植株中金不换甲醚的含量存在差异,V类爵床含量更高;同时,爵床茎中有效成分的含量极少,故在爵床的花穗和叶多的花、果期采收为佳。
Objective:To identify two different growth types vertical growth(V-class)and lateral growth(L-class))of Justicia procumbens,and compare the contents of chemical compounds between upper part and different organs.Method:DNA was extracted from the leaves of two growth forms of J.procumbens.Internal transcriptional spacer 2(ITS2)and psb A-trn H sequences were obtained by polymerase chain reaction(PCR)amplification and bidirectional sequencing.High performance liquid chromatography(HPLC)was used to determine the contents of justicidin B(JB)and chinensinaphthol methyl ether(CME)from the above-ground parts of V-class and L-class J.procumbens and their inflorescence,leaves,and stems.Independent sample T-test and Paired-sample T-test were used to compare the contents of JB and CME from two different growth forms of J.procumbens and their different organs.Result:The DNA sequences after shearing were shown to be identical between the V-class and L-class samples.Independent sample T-test showed no difference on the content of JB between the above-ground parts of V-class and L-class samples,but the content of CME was higher in V-class J.procumbens than L-class(P<0.05).Paired-sample T-test showed that the content of JB and CME were different in the inflorescence,leaves,and stems of J.procumbens(P<0.05),and the contents in inflorescence and leaves were much higher than those in stems.At the same time,we found the content of CME in inflorescence and leaves of V-class J.procumbens was higher than that in L-class.Conclusion:DNA barcoding and comparison of JB and CME content indicated that the two different growth forms of J.procumbens belong to the same species.The contents of CME were different in J.procumbens between different growth forms,and was higher in V-class.Meanwhile,due to the little content of JB and CME in stems,the best picking time of J.procumbens is the flowering and fruiting period with more inflorescence and leaves.
作者
姚云峰
熊羿屹
熊唯琛
甘娇娥
刘博
吴和珍
杨艳芳
YAO Yun-feng;XIONG Yi-yi;XIONG Wei-chen;GAN Jiao-e;LIU Bo;WU He-zhen;YANG Yan-fang(School of Pharmacy,Hubei University of Chinese Medicine,Wuhan 430065,China;Key Laboratory of Traditional Chinese Medicine(TCM)of Resources and Chemistry Hubei Province,Wuhan 430061,China;Collaborative Innovation Center of TCM of New Products for Geriatrics Hubei Province,Wuhan 430065,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第21期140-147,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(31570343)
关键词
爵床
垂直生长类
横向生长类
DNA条形码
爵床脂定B
金不换甲醚
Justicia procumbens
vertical growth
lateral growth
DNA barcoding
justicidin B
chinensinaphthol methyl ether
