Grecian Foxglove, Digitalis lanata Ehrh. (Scrophulariaceae), is a perennial or biennial plant, which is a major source of digoxin and digitoxin. Its leaves are used for the preparation of these cardiac active glycosides for the treatment of cardio vascular disorders. This study was built on a suggestion that in Digitalis the explanted cells already contain all the genetic information that is required for the manufacture of digitoxin and digoxin. Accordingly, this genetic information will be present in callus from explant micro-culture. Activation by external factors such as light, the number of sub-cultures and type and age of the explants, should lead to stimulation, not only of callus size, but also the production of glycosides. Segments of series explants taken from different Digitalis plant organs (shoot tip, leaf, hypocotyl and root), 2 and 4 weeks old, obtained from plants grown in vitro and of 8, 12 and 20 weeks from plants cultivated in vivo culture, were cultured on aseptic MS basal solid medium containing 5.0 mg/L 2.4-D and 0.5 mg/L BA. On the other hand, callus derived from different explants was recultured (in initial amounts of 49-63 mg) on the same new sterilised MS medium in order to increase the mass of callus. The cultures were then maintained in a growth chamber at 26 o ± 2 C o and subjected to four different light photoperiods as follows: 10 hr. light /14 hr. dark cycle, 14 hr. light/10 hr. dark cycle, 16hr.light/ 8hr.dark cycle, and 18 hr. light/ 6 hr. dark cycle. The obtained results show that the greatest callus production, as well as the highest amount of glycosides, occurred in callus derived from 2 or 20 week old leaf explants among all the examined types and ages of explants. Repeating the subculturing three times decreased dramatically the dry weight of callus, but it significantly favoured digoxin and digitoxin content. The optimal callus growth was obtained at 16 hr. light/day but the best glycosidal content was achieved when callus was exposed to 18hr. light/day.
A b s t r a c t A r t i c l e I n f oSufficient and promising protocol for enhancement and production of hypericin in calli cultures of Hypericum sinaicum was established. The highest value of calli cultures was produced from nodal segment and root explants respectively. MS-medium supplemented with 3 mg/l of 2,4-D gave the best results of calli production under dark condition. However, the lowest calli frequency (28 and 20%) and fresh weights (1.4 and 1.0 g/Jar) were estimated with nodal and root explants cultured on MS medium supplemented with 0.25 mg/l 2,4-D + 0.25 mg/l TDZ under light condition. Calli growth dynamics were investigated during five weeks of cultivation. Total hypericin produced by calli cultures derived from nodal segment and root explant were evaluated using HPLC.
In this study, Egyptian Jatropha and Castor oils were obtained and transformed into their corresponding biofuels by catalytic cracking using heterogeneous catalysts (Alumina and Montmorillonite-HCL) with different ratios (0.2%, 0.4%, 0.6%, 0.8% & 1%). The conditions of biofuel production were studied including: catalyst type, catalyst ratio (%), conversion time and temperature. The specifications of the obtained products were comparable to American Society for Testing and Materials (ASTM) Standards. The suitable blends between the obtained biofuels and gas oil fuel were described. The specifications of the prepared biofuel blends were comparable to fuel properties of petroleum diesel according to American Society for Testing and Materials (ASTM) Standards.
Calli were induced from different explants of Jatropha curcas L. seedling on MS basal agar medium in presence of different plant growth regulators. at different levels for 4 weeks Random Amplified Polymorphic DNA (RAPD) markers were used for the assessment of genetic variation of callus grown in different hormonal combinations in comparison with the mother plant. A total of 8 arbitrary sequence primers were evaluated. Of 8 primers used for the RAPD analysis, primers showed consistent band patterns. In total, scorable bands were observed with the primers. The total number of amplicons produced per primer varied from 5 for OPA-07 and OPB-12 to as many as 8 bands for OPB-10, OPC-09 and OPR-17. The average number of bands per primer was 6.625. Out of 53 bands, 36 were polymorphic (67.92%). The average number of polymorphic RAPD bands was 4.5 per primer. the highest similarity (0.71) with mother plant was recorded in leaf-derived callus (S4) grown on MS solid medium with 2.5 µM NAA plus 2.5 µM IBA, while endosperm callus maintained on MS with hormonal combination 12.5 µM IBA + BAP-25.0 µM was found to show least similarity (0.54) with mother plant. In this concern, results have been suggested to be useful fast methods for comparing of genetic changes and variation in plants.
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.
Axillary buds Cultures of Digitalis lanata Ehrh were established from seedling, obtained under aseptic conditions. Shoot tip and single node explants were cultured on MS basal solid medium with various of BA concentrations. Shoots proliferation and growth were significantly influenced by BA levels and explant type. The highest amount of shoots was obtained on medium supplemented with 5.0 µM of BA, while digoxin and digitoxin contents affected by high level of BA (17.5 µM). A further subculture to medium containing high levels of BA promoted shoot growth and suppressed glycoside contents. Production of digoxin and digitoxin improved upon transferring shoots onto half strength on MS medium, vitrified shoots were appeared as well. Interestingly, addition progesterone as precursor, led to enhancement of digoxin and digitoxin contents, especially on rooted shoots.
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