DNA barcoding and TLC as tools to properly identify natural populations of the Mexican medicinal species Galphimia glauca Cav

Abstract

Galphimia glauca is a plant that is endemic to Mexico and has been commonly used since pre-Hispanic times to treat various illnesses, including central nervous system disorders and inflammation. The first studies investigating a natural population of G . glauca in Mexico showed that the plant has anxiolytic and sedative activities in mice and humans. The plant’s bioactive compounds were isolated and identified, and they belong to a family of nor -secofriedelanes called galphimines. The integration of DNA barcoding and thin-layer chromatography analysis was performed to clarify whether the botanical classification of the populations in the study, which were collected in different regions of Mexico, as G . glauca was correct or if the populations consist of more than one species of the genus Galphimia . We employed six DNA barcodes ( matK , rbcL , rpoC1 , psbA-trnH , ITS1 and ITS2 ) that were analyzed individually and in combination and then compared each other, to indicate differences among the studied populations. In the phylogenetic analysis, ITS1 and ITS2 markers as well as the combination of all DNA regions were the most efficient for discriminating the population studied. The thin-layer chromatography analysis exhibited four principal chemical profiles, one of which corresponded to the populations that produced galphimines. DNA barcoding was consistent and enabled us to differentiate the populations that produce galphimines from those that do not. The results of this investigation suggest that the studied populations belong to at least four different species of the genus Galphimia . The phylogenetic analysis and the thin-layer chromatography chemical profiles were convenient tools for establishing a strong relationship between the genotype and phenotype of the studied populations and could be used for quality control purposes to prepare herbal medicines from plants of the genus Galphimia .

Natural populations of Galphimia spp. attenuates peripheral and central inflammation

“…However, the inhibitory effect in the nitrite production increased with the highest concentration (100 µg/mL) of the methanolic extract, in comparison with the lowest one (25 µg/mL). The presence of galphimines in four (GM, HZ, QC and QJ) of the nine population here studied was demonstrated previously by means of TLC analysis (Gesto-Borroto et al 2019). Even though the anti-in ammatory effect of galphimines have been demonstrated before in the TPA-induced mouse ear in ammation test (González-Cortazar et al 2014), in the present work the methanolic extracts from the galphimines-producer (GM, HZ, QC, QJ) and non-galphimine-producer (MC, MM, MZ, SV and ZV) populations did not show signi cant differences in the inhibition of the NO synthesis.…”

“…in an in vitro experimental model. Four of these populations (GM, HZ, QC and QJ) produce galphimines, while ve (MC, MM, MS, SV and ZV) do not produce these metabolites (Gesto-Borroto et al 2019). All the studied populations showed a similar inhibitory effect in the nitrite production.…”

“…In order to clarify the identity of Galphimia species, we performed two molecular studies, using DNA barcoding analysis of natural populations botanically classi ed as G. glauca, including the populations studied in the present investigation. These studies suggest the presence of four species of the genus Galphimia among the collected populations (Sharma et al 2012b;Gesto-Borroto et al 2019). In consequence, it was considered to use the term Galphimia spp.…”

Implications of rbcL and rpoC1 DNA Barcoding in Phylogenetic Relationships of some Egyptian Medicago sativa L. Cultivars

“…Moreover, rpoC1 was proven to differentiate between different populations of the species Galphimia that produce the chemical compounds galphimines from others non-producing species. DNA barcoding investigation was confirmed by thin layer chromatography to identify the different chemical profiles (Gesto-Borroto et al, 2019). Both rbcL and rpoC1 regions were used individually and in combination in several plant studies (Hollingsworth et al, 2009;Singh et al, 2012;Inglis et al, 2018;Gao et al, 2019).…”

Verification of Thai ethnobotanical medicine “Kamlang Suea Khrong” driven by multiplex PCR and powerful TLC techniques

“…To alleviate these challenges, DNA barcoding and TLC were incorporated to identify commercial KSK crude drugs in this study. This integrated approach has also previously been applied to successfully identify and control the quality of ethnobotanical medicines, for example, Marsdenia tenacissima (traditional Chinese medicine) and Galphimia glauca (Mexican medicinal species) [34,35].…”