Abstract

This study established the DNA barcoding sequences (matK and rbcL) of three plant species identified in the tribe Clauseneae, namely Clausena excavata, C. harmandiana and Murraya koenigii. The total phenolic and total flavonoid contents, together with the biological activities of the derived essential oils and methanol extracts, were also investigated. Herein, the success of obtaining sequences of these plant using two different barcode genes matK and rbcL were 62.5% and 100%, respectively. Both regions were discriminated by around 700 base pairs and these had resemblance with those of the Clausenae materials earlier deposited in Genbank at a 99–100% degree of identity. Additionally, the use of matK DNA sequences could positively confirm the identity as monophyletic. The highest total phenolic and total flavonoid content values (p < 0.05) were observed in the methanol extract of M. koenigii at 43.50 mg GAE/g extract and 66.13 mg QE/g extract, respectively. Furthermore, anethole was detected as the dominant compound in C. excavata (86.72%) and C. harmandiana (46.09%). Moreover, anethole (26.02%) and caryophyllene (21.15%) were identified as the major phytochemical compounds of M. koenigii. In terms of the biological properties, the M. koenigii methanol extract was found to display the greatest amount of antioxidant activity (DPPH; IC50 95.54 µg/mL, ABTS value 118.12 mg GAE/g extract, FRAP value 48.15 mg GAE/g extract), and also revealed the highest α-glucosidase and antihypertensive inhibitory activities with percent inhibition values of 84.55 and 84.95. Notably, no adverse effects on human peripheral blood mononuclear cells were observed with regard to all of the plant extracts. Furthermore, M. koenigii methanol extract exhibited promise against human lung cancer cells almost at 80% after 24 h and 90% over 48 h.

Highlights

  • Oxidative stress and autoxidation are known to be harmful to the human body

  • Anethole was the most dominant compound identified in essential oil, while C. excavata was found to contain the highest yield at 86.72% followed by C. harmandiana and M. koenigii at 46.09% and 26.02%, respectively

  • The total phenolic (TPC) and total flavonoid contents (TFC) in the extracts of the leaves of C. excavata, C. harmandiana and M. koenigii were expressed as mg gallic acid equivalent (GAE) and mg quercetin equivalent (QE) per gram extract

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Summary

Introduction

Oxidative stress and autoxidation are known to be harmful to the human body. The presence of these conditions can result from an imbalance between free radicals and antioxidants present in the body, which can be damaging to the human body’s cells, proteins and DNA. The tribe Clauseneae is comprised of five genera, namely Micromelum, Marillia, Glycosmis, Murraya and Clausena [5] The latter genus has been identified as a potent natural resource. Similar plant species have been utilized for numerous alternative purposes as ingredients in food or in pharmaceuticals This present study, has aimed to confirm the obscure major taxa of Clauseneae i.e., Clausena excavata, C. harmandiana and Murraya koenigii gathered from Chiang Mai, Thailand using the DNA barcoding sequences obtained from two candidates of barcoding loci (matK and rbcL). AAllll oobbttaaiinneedd sseeqquueenncceess wweerree vveerriiffiieedd bbyy tthheeiirr aacccceessssiioonn nnuummbbeerrss ((mmaattKK;; MMHH118877223399-- MMHH118877224433 aanndd rrbbccLL;; MMHH118877222222-MMHH118877222299)) aannddcocmompapraerdedto ttohethseequseeqnuceesnpceresvpioruevsliyoudselpyosdietepdoisniteGdenibnanGkenubsianngkBLuAsinSTg. BRLegAaSrTd.inRgemgaorldeicnuglamr iodleencutilfiacratidioenn,tgifeicnaotmioinc,DgNenAomwiacs DsuNccAeswsfausllysuecxcterasscftuedllyfroexmtreavcteerdy fsrpoemcimeveenr.yTswpeocimmaeinn. Based on matK DNA sequences, five samples of Clauseneae (PSRU_RUT001-PSRU_RUT002, PSRU_RUT003-PSRU_RUT004 and PSRU_RUT005) were found to be closely related to Murraya koenigii (L.) Spreng, Clausena weexrceafvoautanBdutromb.fe. To attain a maximum discrimination rate among closely related species, a combination of Internal Transcribed Spacers (ITS + matK + rbcL) has been suggested by Li et al [22]

Phytochemical Analysis of Essential Oils
Total Phenolic and Total Flavonoid Contents
Plant Materials and Chemicals
Collection of Plant Material for Genetic Variation Analysis
Alignment of Sequences and Phylogenetic Analysis
Determination of Total Flavonoid Contents
Determination of Total Phenolic Contents
Determination of Antihypertensive Activity
3.10. Antitumor Activity and Cell Toxicity Assay
Findings
Conclusions
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