Optimization of DNA extraction from seeds and leaf tissues of Chrysanthemum (Chrysanthemum indicum) for polymerase chain reaction.

Saba Hasan,Saumya Mishra,Faizuddin Sagar,Keshav Dwivedi,Saransh Shukla,Abhinav Vashishtha,Kuldeep Srivastava,Payal Jain,Jyoti Prakash,Agnivesh Sharma,Nausheen Khan,Swati Prakash Gupta

doi:10.6026/97320630008225

Abstract

Chrysanthemums constitute approximately 30 species of perennial flowering plants, belonging to the family Asteraceae, native to Asia and Northeastern Europe. Chrysanthemum is a natural cosmetic additive extracted from Chinese herb by modern biochemical technology. It has the properties of anti-bacterial, anti-viral, reducing (detoxification) and anti-inflammation. It possesses antioxidant characteristics, which could assist in minimizing free-radical induced damage. Therefore, it is widely used in skin and hair care products. Chemical composition of this herbal remedy includes kikkanols, sesquiterpenes, flavonoids, various essential oils containing camphor, cineole, sabinol, borneole and other elements that interfere with DNA, causing erroneous or no PCR products. In the present study, testing and modification of various standard protocols for isolation of high-quality DNA from leaf tissues and seeds of C. indicum was done. It was observed that the DNA obtained from seeds and leaf tissues with a modified cetyltrimethylammonium bromide buffer protocol was of good quality, with no colored pigments and contaminants. Also, DNA could be extracted from leaf tissues without using liquid nitrogen. Quality of DNA extracted from seeds was much better as compared to that extracted from leaf tissues. The extracted DNA was successfully amplified by PCR using arbitrary RAPD primers. The same protocol will probably be useful for extraction of high-molecular weight DNA from other plant materials containing large amounts of secondary metabolites and essential oils.

Highlights

Chrysanthemums, often called mums or chrysanths, are of the genus (Chrysanthemum) constituting approximately 30 species of perennial which is native to Asia and northeastern Europe
Plant material DNA was isolated from the leaves and seeds of C. indicum, which were collected upon 6-8 leaves stage
Three protocols were followed for DNA isolation from C. indicum: isoamylalcohol, cetyltrimethylammonium bromide (CTAB) and sodium dodecyl sulphate (SDS)

Summary

Introduction

Chrysanthemums, often called mums or chrysanths, are of the genus (Chrysanthemum) constituting approximately 30 species of perennial which is native to Asia and northeastern Europe. Chrysanthemum plants exhibit potential medicinal activities as shown by earlier studies, e.g., C. indicum showed inhibitory action against bacteria and viruses [1, 2]. C. indicum is a traditional herb, commonly used to treat various disorders, hypertension symptoms and several infectious diseases in Koreanand Chinese medicine [3, 4]. Organic extracts of C. coronarium fresh flowerheads have shown antibacterial [5], insecticidal [6] and antiparasitic activities [7, 8]. Pyrethrins, complex esters extracted from C. cinerariaefolium exhibited insecticidal effects [9] and in vitro activity against Herpes simplex virus [10]. The use of C. trifurcatum flowerheads has been reported in Tunisian traditional medicine to treat constipation [11]. Chrysanthemum plants have been shown to reduce indoor air pollution

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