Canavalia gladiata (Jacq) DC., is a legume and considered as a good source for protein because of presence of quality protein similar to most edible food legumes and can be used as food and feed. The seeds of C. gladiata are known for several valuable phytochemicals, useful for food and health benefits properties. Still the plant is underutilized/ not consumed worldwide due to presence of some anti-nutritional factors (ANFs) such as Concavalin A, Protease inhibitor, tannins, flatulence factors, allergens and lysinoalanine phytates and L-canavine etc particularly in the seed. So, it is necessary to remove all these anti-nutritional compounds to eligible the seed of C. gladiata as food for human society to combat the nutritional deficiency. To increase the nutritional value as well to reduce anti-nutritional compounds from the seed different kind of processes such as sprouting, roasting, fermentation, de-hulling, cooking etc. have been adopted. But the above methods are not efficient enough to reduce anti-nutritional compounds. Therefore, there is a need to develop plantlets with null or reduced anti-nutritional compounds. Tissue culture method is a first and necessary strategy for any kind of further research regarding the reduction of anti-nutritional factors. Whether it is by gene editing or alteration in any regeneration influencing factors during micropropagation anything can be possible by the exploration of tissue culture technique. So, in this part of work, a simple, efficient and reliable micropropagation protocol for Canavalia gladiata (Jacq.) DC was developed by using in vivo nodal segment for the first time. The nodal segment of 3 rd -6 th position are found suitable for multiple shoot regeneration. The optimum shoot proliferation has been found in Murashige and Skoog's (MS) medium supplemented with BAP 2.0 mg/L showing 100% response with 5.66 average shoots/explant of average length 3.90 cm. For upscaling purpose, the in vitro nodes used as explant from primary shoots harvested from in vivo node. Again, MS medium with BAP 2.0 mg/L found as optimum medium for these in vitro nodes having 100% response with average shoot/ explant 2.00 of average length 4.96 cm. The seasonal influence on micropropagation has also studied where, Dec-Jan was observed as best season for explant collection showing 92% bud break followed by Feb-March. Half-MS supplemented with IBA 0.5 mg/L found as optimum medium (100% response) for rooting of in vitro regenerated shoots with average 4.0 roots/shoot and average root length 3.45 cm. The plantlet was acclimatized on a simple mixture of sand: soil (1: 1) and subsequently transferred to the garden under natural condition. Micropropagated plant developed through this protocol can be a step forward for further research work in reducing its anti-nutritional factor.
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