Evaluation of In Vitro Germination and Callus Induction for Antimicrobial Activities of Jatropha curcas L.

Soliman, Manal S.; Elhalafawi, Khallil; Ibrahim, Ibrahim A.; Mahrous, Emad; Bekhit, Metwally

doi:10.21608/rjab.2018.106856

Evaluation of In Vitro Germination and Callus Induction for Antimicrobial Activities of Jatropha curcas L.

Document Type : Original Article

Authors

¹ Plant Biotechnology Dept. Genetic Engineering and Biotechnology Research Institute (GEBRI), University of Sadat City, Egypt.

² Molecular Biology Dept. Genetic Engineering and Biotechnology Research Institute (GEBRI), University of Sadat City, Egypt.

³ Plant Biotechnology Dept. Genetic Engineering and Biotechnology Research Institute (GEBRI),University of Sadat City, Egypt.

10.21608/rjab.2018.106856

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.

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