| Object: To explore the correlation between interspecific genetic relationships and medicinal chemical constituents of Lindernia All., on the basis of the morphological, micromorphological and histological characteristics of the seven species distributed in Sichuan, China. To lay a good foundation for using the interspecific genetic relationships to seek resource plants rich in the same medicinal chemical components, and also for the development of their quality control standards. To provide reference for the further study of interspecific genetic relationships and phylogeny within Lindernia All.Method:The taxonomic status of the seven species of Lindernia All. distributed in Sichuan province, their resource distribution, alias and synonyms, chemical components, the clinical application and so on were determined by herbal textual researching, literature consulting and sorting. Morphological characteristics of the seven species were recorded by the classical morphological taxonomy and plant anatomy, along with the micromorphological features of their flowers and leaves, the histological characteristics of the transverse section of their leaves, stems and fruits.21typical quantitative indexes were chosed to measure. In order to screen out the indexes which were stable at intraspecific level, the SPSS statistical analysis software was used to calculate the mean value, standard deviation and coefficient of variation. Combined with20character indexes, the Q cluster analysis was conducted to get a reasonable dendrogram. The neighbor-joining (NJ) phylogenetic tree based on the psb-A sequences was constructed by DNA barcode technology. Qualitative and quantitative analysis were conducted by thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC). The content of oleanolic acid and chlorogenic acid in the seven species were determined and compared.Result: The seven species of Lindernia All. distributed in Sichuan province all had medicinal records. Lindernia crustacea and L. Antipoda were most commonly used. The results of cluster analysis based on micromorphological and histological characteristics indicated that L. angustifolia and L. anagallis had close genetic relationships, they were classified into a group at the first stage of development. L. nummularifolia and L. crustacea were grouped together at the second stage, and then L. procumbens and L. Antipoda were clustered into a branch. It was a little different from classical morphological taxonomy. The NJ phylogenetic tree based on the psb-A sequences divided L. angustifolia and L. anagallis into a group, L. antipoda and L. ruellioides into anther. Howerver, L. nummularifolia and L. crustacea were not clustered into a group, the result was consistent with the view proposed by morphological classification. Lindernia crustacea, L. antipoda and their analogues contained the common chemical components, beta-sitosterol and oleanolic acid. Moreover. L. crustacea, L. angustifolia and L. anagallis contained chlorogenic acid, while the other four species did not. Standard curve equation of oleanolic acid:Y=287367X-121845,r=0.9998. Standard curve equation of chlorogenic acid:Y=959457X-156644, r=0.9999. The linear range of oleanolic acid was0.84-5.04μg, while chlorogenic acid was0.62-3.72μg. The recovery rate of oleanolic acid was100.96%, while chlorogenic acid was99.601%. The content of oleanolic acid in L. anagallis was more than0.2%and the chlorogenic acid more than0.5%. There were large differences between the contents of oleanolic acid in L. ruellioides form different habitats.Conclusion:The seven species of medicinal Lindernia All. contained the common chemical components, beta-sitosterol and oleanolic acid. It accords with their efficacy of clearing heat and detoxicating, activating blood and removing stasis. Lindernia crustacean, L. angustifolia and L. anagallis contained chlorogenic acid; while L. antipoda and its analogues L. ruellioides did not. It suggests that there is a certain correlation between interspecific genetic relationships and medicinal chemical constituents with a genus, the classification and Chinese folklore selected medicinal plants based on their morphological features seems justified. The results of cluster analysis based on micromorphological and histological characteristics remind us to consider grouping L. nummularifolia into Sect. Torenioides (Benth.) Pennell. L. antipoda and L. ruellioides have similar morphological features, but their micromorphological and histological characteristics are quite different from each other, analysis suggests that the two species might be presenting two different evolution directions. The NJ phylogenetic tree based on the psb-A sequences supports the view proposed by classical morphological taxonomy that features of the androecium plays an important role in classification. With the high content of oleanolic acid and chlorogenic acid, L. anagallis could be a resource plants with the efficacy of reducing fever and causing diuresis for further study. As the large differences between the contents of oleanolic acid in L. ruellioides form different habitats, we inferred that the temperature, humidity and lighting may be the key factors which influence the accumulation of medicinal chemical constituents. |
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