Research Projects

Crocus sativus (Family-Iridaceae) is renowned as source of world’s costliest spice known as saffron, primarily because it involves labor-intensive harvesting and needs particular climatic conditions for its cultivation. Saffron is traditionally used for a range of purposes including cooking, dairy products, confectionary, aroma, medicine, perfume ingredient etc. Although the global demand of saffron has increased greatly but conversely saffron cultivation areas and production in most of the traditionally producing regions of J&K have decreased drastically and is about to disappear because of lack of availability of quality corms, lack of proper R&D, urbanization, adulteration, climatic changes etc. These factors are making saffron production unprofitable, also the current saffron production in J&K is far lesser than the market demand i.e. 9.46 to 18.5 MT, which is further anticipated to increase by more than 13.0% in the next 10 years. Saffron plant being sterile & triploid in nature that grows vegetatively through corms, this complicates its breeding via traditional methods because of lack of sexual reproduction and that leads to slow its rate of multiplication and enhance in susceptibility towards corm rot (Fusarium species). A major limitation in saffron cultivation is because of prolonged dormancy period of around 4–5 months and this is a major reason for single annual flowering cycle. Occurrence of single time flowering in saffron plant impedes its productivity and profitability with lesser farmer income. In our early study we have found that 9-cis-epoxycarotenoid dioxygenase 3 (NCED3) gene is a key regulator for abscisic acid (ABA) biosynthesis, a hormone pivotal in inducing and maintaining saffron dormancy. Some attempts in previous studies using PGR’s have been carried out to induce flowering in saffron and no fruitful outcome was made to break long dormancy of saffron corms. Overcoming long saffron dormancy could lead biannual cultivation and significantly enhance yield under controlled conditions. The novel strategy to break dormancy in saffron through the mediation of plant growth regulators (PGRs) and CRISPR-Cas genome editing arises from the urgent need to enhance saffron production efficiency. By utilizing PGRs to manipulate growth cycles and employing PEG-mediated delivery of Cas9/sgRNA RNPs technology for precise genetic modifications by knockout of NCED3 gene, this thus project aims to develop a transgene-free method for biannual cultivation of saffron by establishing protoplast regeneration protocol . Recent advancements in new speed breeding techniques via transgene- free CRISPR-Cas genome editing technology has opened novel avenues for genetic improvement with desired traits in a short period of time. However, PEG-mediated delivery of Cas9/sgRNA RNPs into saffron protoplasts & regeneration of edited saffron plants has remains largely unexplored to date. By interfering with ABA buildup, targeted knockout of NCED3 gene shall break or shorten the dormant phase. This objective can be accomplished without adding transgenes (Selectable Antibiotic Marker Gene) by using CRISPR-Cas genome editing via Cas9/sgRNA RNPs into protoplasts, which is consistent with the global trend toward developing non-GMO crops. This project seeks to fill the gap of developing a robust transformation system that can facilitate genetic modifications of knocking out NCED3 gene without introduction of foreign DNA. The integration of PGRs will further optimize the growth conditions, ensuring that saffron can be cultivated more efficiently and sustainably. Thus, our proposed novel project aims to break saffron dormancy via PGR augmentation and by targeting key dormancy-regulating NCED3 gene using CRISPR-Cas RNPs delivery into saffron protoplasts, followed by regeneration into whole plants. Conclusively, project shall enable biannual saffron cultivation, enhance productivity and meet market demand sustainably for farmers benefit.