Methyl Jasmonate Effect on Betulinic Acid Content and Biological Properties of Extract from Senna obtusifolia Transgenic Hairy Roots.

Tomasz Kowalczyk,Michał Bijak,Przemysław Sitarek,Anna Merecz-Sadowska,Leslaw Gorniak,Aleksandra Spławska,Monika Szyposzyńska,Tomasz Śliwiński

doi:10.3390/molecules26206208

Abstract

It is known that Senna obtusifolia has been used in medicine since ancient times due to the content of many valuable compounds with a pro-health effect. One of them is betulinic acid, which is a pentacyclic triterpene with antimalarial, antiviral, anti-inflammatory and anticancer properties. In this work, a continuation of our previous research, an attempt was made to increase the level of betulinic acid accumulation by the cultivation of transgenic hairy roots that overexpress the squalene synthase gene in a 10 L sprinkle bioreactor with methyl jasmonate elicitation. We present that the applied strategy allowed us to increase the content of betulinic acid in hairy root cultures to the level of 48 mg/g dry weight. The obtained plant extracts showed a stronger cytotoxic effect on the U87MG glioblastoma cell line than the roots grown without elicitors. Additionally, the induction of apoptosis, reduction of mitochondrial membrane potential, chromosomal DNA fragmentation and activation of caspase cascades are demonstrated. Moreover, the tested extract showed inhibition of topoisomerase I activity.

Highlights

Plant in vitro cultures are a rich source of biologically active compounds of natural origin, gaining more and more importance in many areas of human life [1,2,3,4,5,6]
We present results that are a continuation of our previous research on an attempt to obtain transgenic Senna obtusifolia hairy roots with overexpression of the gene encoding squalene synthase, grown in a sprinkle bioreactor with elicitation, in order to increase the content of betulinic acid
The HPLC analysis showed that the analyzed extract from the transgenic hairy roots grown in a 10 L bioreactor with the use of methyl jasmonate (MeJA) contained 48 mg/g dry weight (DW) of betulinic acid. These results show an increase in the content of this metabolite, compared to transgenic hairy roots grown in a bioreactor without the use of an elicitor

Summary

Introduction

Plant in vitro cultures are a rich source of biologically active compounds of natural origin, gaining more and more importance in many areas of human life [1,2,3,4,5,6]. The results of our previous work have shown that a combination of many strategies, such as the genetic manipulation and cultivation of hairy roots in a bioreactor, can lead to a significant increase in valuable secondary metabolites in Senna obtusifolia hairy root cultures [17]. This valuable plant is widely used in traditional Chinese medicine to prevent and treat diabetes, hyperlipidemia, hypertension, photophobia, headache, dizziness, liver and Alzheimer’s disease [18,19]. Antimicrobial and moderate antiviral properties have been demonstrated for this extract [17]

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