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Abstract
Plant cell culture offers an alternative to whole plants for the production of biologically important specialised metabolites. In cultured plant cells, manipulation by auxin and cytokinin plant growth regulators (PGRs) may lead to in vitro organogenesis and metabolome changes. In this study, six different combination ratios of 2,4-dichlorophenoxyacetic acid (2,4-D) and benzylaminopurine (BAP) were investigated with the aim to induce indirect organogenesis from Bidens pilosa callus and to investigate the associated induced changes in the metabolomes of these calli. Phenotypic appearance of the calli and total phenolic contents of hydromethanolic extracts indicated underlying biochemical differences that were investigated using untargeted metabolomics, based on ultra-high-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UHPLC–qTOF–MS), combined with multivariate data analysis. The concentration and combination ratios of PGRs were shown to induce differential metabolic responses and, thus, distinct metabolomic profiles, dominated by chlorogenic acids consisting of caffeoyl- and feruloyl-derivatives of quinic acid. Although organogenesis was not achieved, the results demonstrate that exogenous application PGRs can be used to manipulate the metabolome of B. pilosa for in vitro production of specialised metabolites with purported pharmacological properties.
Highlights
Plant secondary metabolites are distinctive sources to pharmaceuticals, food additives, flavours, and medicines [1,2]
The white friable callus was sub-cultured onto solid medium with different combinations of auxins and cytokinins (2,4-D: BAP) as detailed in Table 1 to stimulate the undifferentiated cells towards root—or shoot—organogenesis
Plant cell cultures of B. pilosa were successfully initiated from stem and leaf explant material
Summary
Plant secondary (specialised) metabolites are distinctive sources to pharmaceuticals, food additives, flavours, and medicines [1,2]. The plant is noted for its medicinal value, containing a wide spectrum of natural products, which include aliphatics, aromatic compounds, terpenoids, flavonoids, hydroxycinnamic acids (HCAs), and HCA derivatives such as chlorogenic acids (CGAs) that may be synthesised as a number of distinct regio- or geometrical isomers [3,4,5]. In addition to differentiated stem and leaf tissues, undifferentiated cultured cells of Bidens pilosa were found to produce some of these bioactive specialised metabolites [5]. This plant contains a wide variety of important metabolites, the natural habitats of plants are being lost due to agricultural practices, urbanization, and other environmental disturbances such as global warming. These in vitro systems offer the ability to produce high-value
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