Development and Application of RAPD-SCAR Marker for Identification of Phyllanthus emblica LINN.

Abstract

Phyllanthus emblica LINN. Syn: Emblica officinalis GAERTN. (Indian Gooseberry) family-Euphorbiaceae fruit is one of the top selling botanicals having diverse applications in healthcare, food and cosmetic industry. It has been well studied for immunomodulatory, 1) anticancer, 2) antioxidant 3) and antiulcer 4) activities. An official drug of Ayurvedic Pharmacopoeia 5) and Indian Herbal Pharmacopoeia, 6) it forms a main ingredient of various multi-component formulations. Correct genotype identification of the plant material, therefore, remains important for protection of both the public health and industry. Chemoprofiling and morphological evaluation are routinely used for identification of the botanical. Chemical complexity and lack of therapeutic marker(s) are some of the limitations associated with chemical approach while subjective bias in morphological evaluation limits its use.Molecular biology offers various techniques that can be applied for plant identification.7) Genetic polymorphism in medicinal plants has been widely studied which helps in distinguishing plants at inter-and/or intra-species level.8,9) PCRbased methods including Randomly Amplified Polymorphic DNA (RAPD) 10) can be effectively used for authentication of the medicinal plant material. Further, development of more specific, sensitive and reproducible markers like RAPD based Sequence Characterized Amplified Region (SCAR) can increase industrial application of the molecular techniques. These markers have been used for authentication of medicinal plant species of ginseng, 11) Artemisia 12) and other commercially important timbers like bamboo. 13)In the present study, we have chosen six Phyllanthus species (other than P. emblica) that are commercially and therapeutically important; and are widely used in Ayurvedic system of medicine for treating various ailments.14) SCAR marker for correct genotype identification of P. emblica from a species-specific RAPD amplicon is developed by comparative analysis. The marker is further used for authentication of commercial samples of P. emblica fruit powders (Amlachurna) as well as Triphalachurna, a multi-component formulation which contains fruit powders of P. emblica, Terminalia chebula ROTZ. and Terminalia belerica ROXB. MATERIALS AND METHODS Plant MaterialFresh leaf tissues of Phyllanthus distichus LINN., Phyllanthus reticulatus POIR., Phyllanthus urinaria LINN., Phyllanthus simplex RETZ., Phyllanthus niruri LINN. and Phyllanthus indofischeri BENNET. were collected from different parts of India (three samples of each species) and authenticated in Agharkar research Institute, Pune, India. Leaf tissues of eleven cultivars of P. emblica viz.; NA-06, NA-07, NA-10, Kanchan, Chakaiya, Francis, Banarasi, Hathizool, Dongri, Bansired and Anand-01 were collected from standard cultivated plots of Mahatma Phule Agricultural University, Rahuri, India. The collected fresh tissues were powdered in liquid nitrogen and stored in deep freezer (Ϫ70°C) till further processing.Three commercial P. emblica fruit powder sample...

Unani Murakkabat (Formulations): Need of Modification

“…The adulterant may be phyllogenetically close or distinct (eg. dried fruit pieces of pumpkin) from Phyllanthus emblica [5]. Pueraria tuberosa, commonly known as Vidarikand in India, is an important plant used in traditional medicine.…”

Development and significance of SCAR marker QG12-5 for Canarium album (Lour.) Raeusch by molecular cloning from improved RAPD amplification

“…The sequence characterized amplified region (SCAR) marker is another molecular marker that is more stable and is generally derived from RAPD, ISSR, or other markers (Dnyaneshwar et al, 2006;Su et al, 2007;Kumla et al, 2012;Fu et al, 2013;Rajesh et al, 2013;Mei et al, 2015;Zhang et al, 2015). Combining SCAR with RAPD simplifies the molecular analysis procedure, as the PCR primers used to develop SCAR markers are designed from the sequence of the RAPD amplicon (Kumla et al, 2012;Rajesh et al, 2013;Dutta et al, 2014;Fu et al, 2015).…”

Development of RAPD-SCAR markers for different Ganoderma species authentication by improved RAPD amplification and molecular cloning

“…The sequence characterized amplified region (SCAR) marker is another molecular marker, which is more stable and is generally derived from RAPD or intersimple sequence repeat (Dnyaneshwar et al, 2006;Su et al, 2007;Li et al, 2010;Kumla et al, 2012;Rajesh et al, 2013). When SCAR with RAPD are combined, molecular analysis is simplified, in which PCR primers are designed from the sequence of the RAPD amplicon to develop SCAR markers (Kumla et al, 2012;Rajesh et al, 2013).…”