Efficient Micropropagation Protocol for Jatropha Curcas Using Liquid Culture Medium

Abstract

Although several studies have been made on the micropropagation of Jatropha curcas using agar base mediums, none of them have been by using liquid medium systems. The effects of explant type and temporary immersion system (test tube, jar with filter paper boat, and growtek bioreactor) on the micropropagation of J. curcas were studied. The explant type influenced shoot quality, multiplication coefficient (MC), and rooting. Leaf explant produced more and longer shoots than nodal explant. Use of filter paper (FB) boat prevented hyperhydricity and allowed proliferation of nodal explants cultured in liquid MS (Murashige and Skoog) medium supplemented 6-benzylaminopurine (BAP) and Kinetin (KN). The best shoot bud induction (92.1±3.1%) was achieved in liquid MS medium supplemented with 2.0 mg/L KN. Leaf regeneration efficiency was compared in growtek bioreactor and in jar containing liquid MS medium supplemented with 0.5 mg/L Thidiazuron (TDZ). The best shoot bud regeneration (78.7±2.1%) was obtained in growtek bioreactor. Shoot buds achieved from nodal segment and leaf were subcultured on filter paper boats in jar and bioreactor containing liquid MS medium supplemented with BAP, Indole butyric acid (IBA), Indole-3-acetic acid (IAA), and KN. Best shoot proliferation and elongation was obtained in filter paper boats containing liquid MS medium supplemented with 1.5 mg/L BAP, 0.5 mg/L IAA, and 0.2 mg/L KN. The number of multiple shoot buds was higher in leaf explants as compared to nodal explants and the highest number of multiple shoot buds was recorded from leaf explants. Up to 76.4% rooting efficiency was obtained when the shoots were ex vitro rooted. The generated plants well established in the nursery and grew normally in outdoor conditions. The protocol has good potential for application in large-scale propagation of J. curcas using liquid medium.