Genetic diversity analysis among commercially available brands of saffron through SSR markers and detection of adulteration in them through gene specific amplifications and HPLC
Saffron (Crocus sativus L.) being a vegetatively propagated triploid plant is believed to lack genetic variability. However, there are not many authentic reports to support this belief. Therefore, in the present investigation efforts were made to assess genetic diversity among commercially available brands of saffron through DNA based SSR markers. Owing to its high price, saffron has always been an item for adulteration. Therefore, efforts have also been made to detect adulteration among commercially available brands of saffron through EST markers and HPLC analysis. To achieve these objectives, DNA was extracted from dried stigmas of saffron marketed under seven different brand names. Integrity, quality and quantity of DNA was analyzed by using Bio-analyzer DNA chip 7500. Touchdown PCR was used for standardization of SSR markers for precise amplification. Subsequently, EST markers specific to immature and mature stigmas of saffron were amplified to detect adulteration by comparing with positive control i.e. with amplicons generated by DNA as template extracted from saffron corms collected from Kashmir valley. The same samples were also analyzed for their chemical constituents and compared with standards of crocin and safranal using HPLC. Simple sequence repeat markers amplified many polymorphic bands among seven samples which suggested presence of genetic diversity among the clones. Out of the total seven samples, only two were quite similar to positive control, the rest were polymorphic on the basis of similarity index. Similarly, the stigma specific EST markers also gave coherent banding profiles with positive control in only two of the seven tested samples. The current study points to conclusion that SSR markers can be successfully utilized to assess genetic diversity and to check clonal fidelity of various saffron cultivars, whereas, EST markers and HPLC may detect adulteration in commercially available brands of saffron.
Fougat, R.S., Subhash, N., Singh, A.S., Mistry, J., Sakure, A., Raj, S. and Modi, A. (2018). Genetic diversity analysis among commercially available brands of saffron through SSR markers and detection of adulteration in them through gene specific amplifications and HPLC. Acta Hortic. 1200, 7-16
Crocus sativus L., crocin, safranal, adulteration, SSR, genetic diversity