Real time sequence characterized amplified region (RT-SCAR) marker: Development and its application for authentication and quantification of Catharanthus roseus L. Don.

Abstract

The objective of the present study was to develop a Real Time-Sequence Characterized Amplified Region (RT-SCAR) of Catharanthus roseus, an effective molecular marker for authentication and quantification. Here, we made a comparative study of SCAR and RT-SCAR marker. These newer RT-SCAR methods are useful for the correct identification and quantification of C. roseus. In particular, this technique is very attractive and can be used as a novel tool for quantification of C. roseus when the SCAR results cannot be quantified and the RAPD results are not consistent. RAPD analysis of collected samples from different geographical locations in India was carried out with 25 random primers. The RAPD polymorphic band was cloned, sequenced and from the sequence information, primers pair for SCAR was developed. The same primers was used for RT-SCAR marker by using SYBR green fluorescence-based detection in Real Time PCR. Utilizing this newly specific RT-SCAR marker, we detected the presence and quantity of C. roseus in all the analyzed several samples, by aiding 10% its substitute and adulterants at time of DNA isolation. No difference was found in pure sample and 10% adulterant sample with the help of SCAR but in the case of RT-SCAR difference was found in the DNA sample. This analytical strategy could be used further for large scale quantification of C. roseus's market samples.

Rapid and easy molecular authentication of medicinal plant Zingiber officinale Roscoe by loop-mediated isothermal amplification (LAMP)-based marker

“…Various methods, including macroscopy, microscopy and chemoprofiling have been reported for the quality control of crude ginger and its products. These methods are reported to have limitations in distinguishing Z. officinale from closely related species: molecular based identification is a great promise in resolving issues of controversial identity and quality control, when botanical identification, based on morphology becomes difficult, such as in the case of incomplete or damaged samples and in dried herbal products, but has not yet been capitalized by the traditional medicine sector (Chaudhary et al, 2012). DNA fingerprinting patterns provide the ultimate in individualization due to the stability of DNA in any plant part and also through variation in environment and also variation in phase of life cycle.…”

Analysis of Several Popular Cultivars of Madagascar Periwinkle (<i>Catharanthus roseus</i> (L.) G. Don.) using Biochemical Markers

“…According to the wide literature survey to date on the assessment of genetic diversity of Catharanthus species and cultivars were performed by means of secondary metabolites, AFLP, ISSR, RAPD and RT-SCAR etc. (Chaudhary et al, 2012;El-Domyati et al, 2012;Idrees et al, 2010;Sevestre-Rigouzzo et al, 1993;Shaw et al, 2009). Yet, there has been no previous report on the use of isozyme profiles to characterize the genetic diversity of highly medicinal plant Catharanthus roseus cultivars.…”