Interactions of a medicinal climber Tinospora cordifolia with supportive interspecific plants trigger the modulation in its secondary metabolic profiles

Abstract

Tinospora cordifolia (TC) is scientifically proven immunomodulatory drug being used for centuries. Ancient literature reported that inter-specific interactions change medicinal properties of TC. Thus, the current study is aimed to understand the influence of interspecific biotic interactions on chemo-profiles of TC. To explore it, TC samples collected from six co-occurring plants, i.e. Azarditchita indica, Acacia nilotica, Albezia lebbeck, Ficus benghalensis, Tamarandus indica and Acacia leucophloea were analyzed by HPLC-ESI-QTOF-MS. Mass data were subjected to multivariate analysis. Support vector machines (SVMs) was found to be best classifier (r² < 0.93). Data analysis showed the specific compounds in all TC due to inter-specific interactions. Data were further analyzed with SNK post-hoc test followed by permutative (n = 50) Bonferroni FDR multiple testing correction. The compound without any missing values reduced the number of variables to 133 (p < 0.01). Statistical analysis revealed that TC having interactions with A.lebbeck and A. nilotica formed the most distant groups. However, TC co-occurred with A. indica showed the highest number of up-regulated metabolites, including jatrorrhizine, chrysin, peonidin, 6-methylcoumarin and some terpenoids. Some metabolites, including jatrorrhizine and magnoflorine were quantified to confirm the accuracy of qualitative analysis. Results demonstrated the influence of inter-specific biotic interactions on TC chemo-profiles, hence its medicinal properties.

Figure 1

Base peak chromatograms of all seven groups extracted from total ion current chromatograms (TIC), showing visual differences among the chromatographic profiles of various groups. A, B, C, D, E, F and G represents the ALC, ANI, ALL, AIN, CON, FBG, TMI groups respectively.

Figure 2

(A) Box Whisker plots of various groups after filtration, significance and fold change analysis showing high variations across the groups. (B) Principle component analysis score plot representing % variations among the metabolites of all seven groups where variability along the x, y and z axis are 23.42%, 17.19% and 13.62% respectively. AIN, ALC, ALL and ANI represents the distant groups showing highest variations.

Figure 3

Venn diagram showing distribution of number of specific entities among seven different groups. The diagram shows overlapping and distinct metabolites indicated by the numbers in the intersections and circles, respectively.

Figure 4

Spearman correlation heatmap of the metabolites without missing values, with compounds marked on the side of the map. Each row represents a sample and each column represents a metabolite feature. Color key indicates the normalized abundance of each metabolite expression value across the samples. The red and blue color indicates the highest and lowest metabolite expression values respectively. The figure clearly depicts the distinctness of ALL and ANI having lowest levels of most of the metabolites.