There are many problems related to seed germination and in vitro cultivation of Jatrophacurcas L, this research was to solve this obstacle. This study also aimed to use triphenyl tetrazoliumchloride (TTC) as a new substance for in vitro callus initiation and growth in plant tissue culture fieldsfor the first time. In addition, an efficient regeneration method via seed culture has been developed.For this purpose, different media (MS; B5; WPM with B5 vitamins; wetted cotton and water) were usedfor seed germination. High seed germination percentage for all treatments were achieved after 24days from incubation. WPM was effective medium in increasing the total length of seedlings. Themaximum dry weight (0.24 g) of Jatropha curcas shoots was recorded with B5 medium. In vitro shootformation, MS medium supplanted with 2.0 μM IBA +10 μM BA and MS medium+2.5 μM BA+2.5 μMNAA recorded the highest shoot formation percentage (100%) after one month. Concerning callusinduction, data implied that different growth regulators tested (TDZ, BA, KIN) and TTC as a newsubstance used in plant tissue culture could induce the formation of callus in cotyledons of J. curcasseedlings, furthermore, the combination of BA and Kin in various concentrations was an appropriatemedium for inducing the formation of callus and promoting its growth. Also, callus formation andgrowth were significantly affected by explant types. The balance between auxin and cytokinin is thelimited factor for callus induction. MS nutrient medium supplemented with 2,4-D or NAA or both helpedto form callus at high percentage. regarding antimicrobial activity, the antimicrobial potential ofdifferent plant extracts was screened against four pathogenic microorganisms and reference bacterialstrains, the ethanolic extract of the roots was found to show greater inhibition of B. cereus and E. coliwith zones of inhibition, 21.7 and 10.4 mm respectively, when comparing the extracts from in vivoplant and in vitro callus. Also, ethanol extract from the leaf of the mother plant showed higher inhibitoryzone than that of the leaf-derived callus against S. aureus, B. cereus and E. coli. On the other hand,butanol extract of hypocotyl-derived callus was achieved the highest fraction effect resulted fromdifferent fractionation of crude methanolic extract against antibacterial activity of E. coli, followed byethyl acetate- fraction (15.0 and 13.5 mm) respectively, in contrast, water- fraction came in the last.
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