Molecular marker based assessment of genetic homogeneity within the in vitro regenerated plants of Crocus sativus L. – a globally important high value spice crop

Abstract

The global market for spice is steadily increasing and saffron is one of the costliest spices in the arena. However, there is a huge gap in the availability of quality planting materials (QPM) of saffron which can only be effectively redressed by appropriate application of plant tissue culture protocols. Nevertheless, one of the major embargoes in the sustainable utilization of in vitro technologies is occurrence of cryptic clonal variabilities defined as somaclonal variations. In the present research, we report a reproducible, cost effective and simple molecular marker mediated assessment of genetic fidelity within the in-vitro propagated shoots of saffron in order to ensure genetic uniformity during cormlet production. The in vitro propagated shoots of Crocus sativus were raised from field grown corms using an earlier reported method. The genetic stability of in vitro propagated shoots was then assessed using molecular markers namely, Start Codon Targeted Polymorphism (SCoT), Amplification of Minisatellite DNA (DAMD) and Inter Retro Transposon Amplified Polymorphism (IRAP). The isolated genomic DNA samples (mother and regenerated plants) were subjected to PCR amplification with 22 best reproducible primers (14 SCoT, 4 DAMD and 4 IRAP). The combined assay of molecular markers revealed a clonal variability of 7.81% thereby, indicating largely stable clonal lines. The present research documents a fast, reproducible and cost effective methodology for assessment of genetic variability amongst the regenerants of saffron and also ensures the true-to-type nature of the micropropagated plants with a target goal of sustainable propagation.