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. 2023 Dec 22;29(1):80.
doi: 10.3390/molecules29010080.

Evaluation of Salicylic Acid and Methyl Jasmonate as Elicitors in Phyllanthus acuminatus Hairy Roots by Non-Targeted Analysis Using High-Resolution Mass Spectrometry

Katherine Benavides  1 , Andrés Sánchez-Kopper  2 , Karol Jiménez-Quesada  1 , Raquel Perez  1 , Giovanni Garro-Monge  1
Affiliations

Evaluation of Salicylic Acid and Methyl Jasmonate as Elicitors in Phyllanthus acuminatus Hairy Roots by Non-Targeted Analysis Using High-Resolution Mass Spectrometry

Katherine Benavides et al. Molecules. .

Abstract

Phyllanthus acuminatus has been studied for its vast medical and industrial potential. Phytochemical investigations reveal that the genus is a rich source of lignans, flavonoids, phenolics, terpenoids, and other metabolites. However, the phytochemical profile elucidation of this species still needs further research. The use of eliciting compounds such as salicylic acid and methyl jasmonate has managed to increase the production of secondary metabolites in plant cell cultures. Hairy roots of Phyllanthus acuminatus were produced in 250 mL flasks with a 16 h light/8 h darkness photoperiod under diffused light with a culture time of four weeks. The elicitors salicylic acid and methyl jasmonate were tested in 50 μM and 200 μM concentrations. Non-targeted analysis was done for the different treatments using HR-MS. Identified metabolites were grouped in phenylpropanoids, phenols, and mucic acids, and statistical analysis of relative concentrations was achieved. A significant change in phenols' relative concentrations appeared in the elicitations with salicylic acid. Because of the elicitation treatment, specific compounds increased their concentrations, some of which have known pharmacological effects and are used in treating chronic diseases. The best elicitation treatment was salicylic acid 50 μM as it increased by more than 100% the general content of phenols and phenylpropanoid derivates and triplicates the concentration of mucic acid derivates in treated hairy root extracts. The application of non-targeted analysis showed interesting changes in phytochemical concentration due to elicitation in Phyllanthus acuminatus hairy roots.

Keywords: Phyllanthus acuminatus; elicitation; flavonoids; hairy roots; methyl jasmonate; phenols; phenylpropanoids; salicylic acid; terpenoids.

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Figures

Figure 1
Figure 1
Elicitors molecular structures.
Figure 2
Figure 2
Identified compounds in P. acuminatus hairy root extracts grouped by compound type.
Figure 3
Figure 3
Average total intensities for each compound class present in each elicitation in the positive ionization mode. Error bars stand for the standard deviation of triplicates. (A). Phenols and mucic acids derivatives. (B). Phenylpropanoids. (C). Terpenoids.
Figure 4
Figure 4
Average total intensities for each compound class present in each elicitation in the negative ionization mode. Error bars stand for the standard deviation of triplicates. (A). Phenols and mucic acids derivatives. (B). Phenylpropanoids. (C). Terpenoids. (D). Flavonoids.
Figure 5
Figure 5
Structures of interesting compounds found in Phyllanthus acuminatus presented an increase in concentration in elicitation experiments.
Figure 6
Figure 6
Specific compound variation with elicitation treatments, measurements in negative (A,C,E,G) and positive (B,D,F) ionization modes. Phenols and mucic acids derivates (A,B). Phenylpropanoids (C,D). Terpenoids (E,F). Flavonoids (G) Dendrograms grouped the compounds with similar behavior for the different applied treatments.
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