Development of RAPD, DAMD and ISSR markers for authentication of medicinal plant Cassia auriculata and its adulterant Cassia surattensis

Cassia auriculata is an important traditional medicinal plant commonly used in many Ayurvedic formulations, meant for diabetes, rheumatism, conjunctivitis, infertility and etc. But due to similar morphological characters and misidentifications, adulteration from Cassia surattensis has been observed. Since safety and efficacy of herbal products has become a major concern due to adulterations, DNA profiling which is an effective and cheap method can be used to solve this problem by discriminating the genuine material. However, no reports about the genetic identification for these species are available to our knowledge. Therefore, the present study was devoted to developing Random Amplified Polymorphic DNA (RAPD), Direct Amplification of Minisatellite-region DNA (DAMD) and Inter Simple Sequence Repeat (ISSR) DNA profiles to authenticate C. auriculata and C. surattensis. As the first step, fresh leaf samples from both plant species were randomly collected from Gampaha district, Sri Lanka and genomic DNA were extracted using modified Cetyltrimethylammonium Bromide (CTAB) protocols. Four short arbitrary primers, two core primers and two SSR primers were used respectively with three different Polymerase Chain Reactions (PCR) based molecular markers which were RAPD, DAMD and ISSR to develop DNA profiles. Out of eight primers, three arbitrary primers, OP-F03, OP-U10 and OP-U20 and one core primer, HBV(5) yielded clear and reproducible amplification products. These results clearly discriminated the medicinal plant C. auriculata and the adulterant C. surattensis providing a complementary tool for quality control of plant derived herbal medicinal products. However, both SSR primers couldn't authenticate two plants and further work is needed to develop ISSR DNA profiles for the authentication.