A robust workflow for indirect somatic embryogenesis and cormlet production in saffron ( L.) and its wild allies; and .

Saffron ( L.) and its wild relatives, and are of considerable significance in the pharmaceutical, nutraceutical, and ornamental bulbs industry. Towards the ultimate goal of the conservation of wild species and establishment of an efficient workflow for production of Crocuses, efficient protocols were developed for disinfection and production of cormlets in and its wild allies and . Moreover, the differential expression of the Somatic Embryogenesis Receptor-like Kinase () gene was evaluated as a potential molecular marker during embryogenesis between embryogenic and non-embryogenic calli. A highly efficient disinfection recipe and a low-cost TDZ-free protocol have been successfully developed for cormlet production in three species. MS medium containing 10.18 μM 2, 4-D + 4.44 μM BAP was most efficiently induced callus and somatic embryo formation. The highest conversion frequency and maximum cormlet weight were achieved in MS containing 5.37 μM NAA + 8.88 μM BAP. The expression was significantly much higher in embryogenic calli than non-embryogenic in all species. The current low-cost and easy-to-use recipe suggests a promising propagation workflow for mass production of uniform pathogen-free cormlets of species, as well as a platform to better conservation of wild species and effective gene and genome editing using CRISPR-Cas9 in future studies.