In this study, in planta transformation of tomato (Solanum lycopersicum L.), using fruit injection and floral dip, is reported. Agrobacterium tumefaciens strain EHA 105 containing one of three constructs, i.e., pROKIIAP1GUSint (carrying the Apetala 1 [AP1] gene), pROKIILFYGUSint (carrying the LEAFY [LFY] gene), or p35SGUSint (carrying the β-glucuronidase [GUS] gene), was used for plant transformation. For fruit injection transformation, no significant effects (p>0.05) of the construct used were observed. The highest frequency of transformation was obtained following 48-h incubation of tomato fruit with bacterial cells harboring either one of the three constructs; transformation frequencies of 17%, 19%, and 21% for AP1, LFY, and GUS gene constructs, respectively, were obtained. When fruit maturity was evaluated in fruit injection experiments, mature red fruit resulted in higher frequency of transformants than immature green fruit with 40%, 35%, and 42% for AP1, LFY, and GUS gene constructs, respectively. For floral dip transformation, a higher number of transformants was obtained when the GUS gene construct was used instead of either the AP1 or LFY gene construct, thus suggesting a possible inhibitory effect of the flowering genes used. When flowers were transformed prior to rather than following pollination, they yielded a higher transformation frequency, 12% for the LFY construct and 23% for the GUS construct (p<0.05), although no transformant was obtained with the AP1 gene construct.All putative GUS-positive transformants were analyzed using polymerase chain reaction and confirmed for the presence of the transgene. Compared to control plants, transgenic plants carrying either the AP1 or LFY transgene flowered earlier and showed several different morphological characters.
BackgroundFour medicinal plants (Chrozophora hierosolymitana Spreng, Chrysanthemum leucanthemum L., Ephedra gerardiana Wall. ex Stapf, and Quercus dilatata L.) used by indigenous healers to treat various infectious diseases were selected for the present study. The major objective of the present study was isolation and characterization of antimicrobial components from the crude plant extracts using bioassay guided fractionation.MethodsSeven methanolic extracts of the four plants were screened to identify any antimicrobial agents present in them. The active crude plant extract was fractionated first by solvent partitioning and then by HPLC. Characterization of the active fractions was done by using spectrophotometer.ResultsAll the seven methanolic extracts showed low antifungal activity, however, when these extracts were tested for antibacterial activity, significant activity was exhibited by two extracts. The extract of aerial parts of Q. dilatata was most active and therefore, was selected for further analysis. Initially fractionation was done by solvent-solvent partitioning and out of six partitioned fractions, ethanol fraction was selected on the basis of results of antibacterial activity and phytochemical analysis. Further, fractionation was carried out by RP- HPLC and purified active subfractions were characterized by comparing their absorption spectra with that of the known natural products isolated from the plants of Quercus genus.Discussion and conclusionThe results suggest that this is the first report of the isolated antibacterial compounds from this genus.
In continuation to our efforts in finding potential therapeutic agents, a variety of biologically significant semicarbazones were synthesized by the reaction of different carbonyl compounds with phenyl semicarbazides through microwave irradiation. Initially, 18 semicarbazones were studied for their antimicrobial, antitumor, and antioxidant potential. None of the tested compounds showed any antibacterial activity; however, some compounds showed significant antifungal activity. Interestingly, all compounds showed antitumor activity when tested against tumors grown on potato discs. These compounds were also tested for their effect on OH radical-induced oxidative DNA damage. All the compounds showed DNA protection to varying extent. Based on the promising results of antitumor and antioxidant activities, another set of 24 semicarbazones was synthesized, and all of these semicarbazones were evaluated for their antioxidant potential. The results showed that the semicarbazones derived from 2-nitrobenzaldehyde and acetophenone were the most active 2,2-diphenyl-1-picrylhydrazyl 9 (DPPH) free radical scavengers. The overall results have led to the identification of some interesting compounds which seem to have great potential to be developed into effective anticancer drugs.
In this study, four plants [Chrozophora hierosolymitana Spreng (Euphorbiaceae), Ephedra gerardiana Wall. ex Stapf (Ephedraceae), Chrysanthemum leucanthemum L. (Astraceae), and Quercus dilatata L. (Fegaceae)] collected from different regions of Pakistan were screened to identify any chemotherapeutic agents present in them. Seven methanol extracts of these plants (leaf, stem, and root extracts of C. hierosolymitana; stem and root extracts of E. gerardiana; aerial parts of C. leucanthemum, and aerial parts of Q. dilatata) were examined for cytotoxicity using brine shrimp assay, antitumor activity using potato disc assay, and phytotoxicity activity using radish seed bioassay. Two methanol plant extracts, that is, leaf extract of C. hierosolymitana and root extract of E. gerardiana showed significant brine shrimp cytotoxicity activity ranging from 171.55 to 523.8 ppm. Six of the seven extracts exhibited tumor inhibition at all the three concentrations tested, ranging from 10 to 80%. All extracts showed growth and seed germination inhibition at high concentration against radish seeds, while two extracts (root extract of C. hierosolymitana and aerial parts of Q. dilatata) showed growth stimulating effects at lower concentrations. Phytochemical tests showed the presence of alkaloids, saponins, anthraquinones, terpenoids, flavonoids, flavones, tannins, phlobatannins, and cardiac glycosides in different concentrations in these extracts.
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