Evaluation of In Vitro Germination and Callus Induction for Antimicrobial Activities of Jatropha curcas L. | ||
| Research Journal of Applied Biotechnology | ||
| Article 11, Volume 4, Issue 2, November 2018, Pages 104-124 PDF (928.12 K) | ||
| Document Type: Original Article | ||
| DOI: 10.21608/rjab.2018.106856 | ||
| Authors | ||
| Manal S. Soliman1; Khallil Elhalafawi2; Ibrahim A. Ibrahim1; Emad Mahrous1; Metwally Bekhit* 3 | ||
| 1Plant Biotechnology Dept. Genetic Engineering and Biotechnology Research Institute (GEBRI), University of Sadat City, Egypt. | ||
| 2Molecular Biology Dept. Genetic Engineering and Biotechnology Research Institute (GEBRI), University of Sadat City, Egypt. | ||
| 3Plant Biotechnology Dept. Genetic Engineering and Biotechnology Research Institute (GEBRI),University of Sadat City, Egypt. | ||
| Abstract | ||
| There are many problems related to seed germination and in vitro cultivation of Jatropha curcas L, this research was to solve this obstacle. This study also aimed to use triphenyl tetrazolium chloride (TTC) as a new substance for in vitro callus initiation and growth in plant tissue culture fields for 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 used for seed germination. High seed germination percentage for all treatments were achieved after 24 days from incubation. WPM was effective medium in increasing the total length of seedlings. The maximum dry weight (0.24 g) of Jatropha curcas shoots was recorded with B5 medium. In vitro shoot formation, MS medium supplanted with 2.0 μM IBA +10 μM BA and MS medium+2.5 μM BA+2.5 μM NAA recorded the highest shoot formation percentage (100%) after one month. Concerning callus induction, data implied that different growth regulators tested (TDZ, BA, KIN) and TTC as a new substance used in plant tissue culture could induce the formation of callus in cotyledons of J. curcas seedlings, furthermore, the combination of BA and Kin in various concentrations was an appropriate medium for inducing the formation of callus and promoting its growth. Also, callus formation and growth were significantly affected by explant types. The balance between auxin and cytokinin is the limited factor for callus induction. MS nutrient medium supplemented with 2,4-D or NAA or both helped to form callus at high percentage. regarding antimicrobial activity, the antimicrobial potential of different plant extracts was screened against four pathogenic microorganisms and reference bacterial strains, the ethanolic extract of the roots was found to show greater inhibition of B. cereus and E. coli with zones of inhibition, 21.7 and 10.4 mm respectively, when comparing the extracts from in vivo plant and in vitro callus. Also, ethanol extract from the leaf of the mother plant showed higher inhibitory zone 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 from different fractionation of crude methanolic extract against antibacterial activity of E. coli, followed by ethyl acetate- fraction (15.0 and 13.5 mm) respectively, in contrast, water- fraction came in the last. | ||
| Keywords | ||
| Jatropha curcas; Auxin; Cytokinin; germination; Tissue culture; Nutrient medium, thidiazuron; triphenyl tetrazolium chloride; antimicrobial activity | ||
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