Authors:N. Bukhari, Iram Siddique, K. Perveen, I. Siddiqui and M. Alwahibi
Synthetic seed technology is an alternative to traditional micropropagation for production and delivery of cloned plantlets. Synthetic seeds were produced by encapsulating nodal segments of C. angustifolia in calcium alginate gel. 3% (w/v) sodium alginate and 100 mM CaCl2 · 2H2O were found most suitable for encapsulation of nodal segments. Synthetic seeds cultured on half strength Murashige and Skoog medium supplemented with thidiazuron (5.0 μM) + indole-3-acetic acid (1.0 μM) produced maximum number of shoots (10.9 ± 0.78) after 8 weeks of culture exhibiting (78%) in vitro conversion response. Encapsulated nodal segments demonstrated successful regeneration after different period (1–6 weeks) of cold storage at 4 °C. The synthetic seeds stored at 4 °C for a period of 4 weeks resulted in maximum conversion frequency (93%) after 8 weeks when placed back to regeneration medium. The isolated shoots when cultured on half strength Murashige and Skoog medium supplemented with 1.0 μM indole-3-butyric acid (IBA), produced healthy roots and plantlets with well-developed shoot and roots were successfully hardened off in plastic pots containing sterile soilrite inside the growth chamber and gradually transferred to greenhouse where they grew well with 85% survival rate. Growth performance of 2 months old in vitro-raised plant was compared with in vivo seedlings of the same age. Changes in the content of photosynthetic pigments, net photosynthetic rate (PN), superoxide dismutase and catalase activity in C. angustifolia indicated the adaptation of micropropagated plants to ex vitro conditions.
Authors:Md. Siddiqui, I. Chakraborty, P. Hazra and J. Ayala-Zavala
The investigations carried out so far on high pigment tomatoes are confined to their nutritional aspects only. We present the comparative results of the first study on the kinetics of changes in chemical and sensory quality attributes in puree prepared from two colour mutants [dark green (dg) and old gold crimson (ogc)] and seven normal tomato genotypes during storage. Puree of mutant tomatoes BCT-115 and BCT-119, carrying dg and ogc genes, showed the less significant changes in TSS (7.52 and 6.02 °Brix), acidity (3.16 and 3.05%), pH (4.04 and 4.03), total sugar (12.4 and 11.13%), ascorbic acid (20.74 and 19.69 mg/100 g), lycopene (7.78 and 542 mg/100 g), and β-carotene (3.08 and 2.26 mg/100 g) during two months storage at 25 °C. Nevertheless, puree prepared from Berika and BCT-115 (dg) had higher colour (7.63 and 7.13), taste (7.4 and 7.37) and flavour (7.3 and 7.37) sensory scores during two months of storage at 25°C. These results provide new data on the effect of genotypes on the stability of quality for storage of tomato puree and insist on the utilization of these genotypes for breeding new processing cultivars in the near future.
Authors:I. Siddique, N. Abdullwahab Bukhari, K. Perveen, I. Siddiqui and M. Anis
An in vitro propagation system for Cassia angustifolia Vahl. has been developed. Due to the presence of sennosides, the demand of this plant has increased manyfold in global market. Multiple shoots were induced by culturing nodal explants excised from mature plants on a liquid Murashige and Skoog  medium supplemented with 5–100 μM of thidiazuron (TDZ) for different treatment duration (4, 8, 12 and 16 d). The optimal level of TDZ supplemented to the culture medium was 75 μM for 12 d induction period followed by subculturing in MS medium devoid of TDZ as it produced maximum regeneration frequency (87%), mean number of shoots (9.6 ± 0.33) and shoot length (4.4 ± 0.46 cm) per explant. A culture period longer than 12 d with TDZ resulted in the formation of fasciated or distorted shoots. Ex vitro rooting was achieved when the basal cut end of regenerated shoots was dipped in 200 μM indole-3-butyric acid (IBA) for half an hour followed by their transplantation in plastic pots filled with sterile soilrite where 85% plantlets grew well and all exhibited normal development. The present findings describe an efficient and rapid plant regeneration protocol that can further be used for genetic transformation studies.
Authors:Omer A. Basudan, Perwez Alam, Nasir A. Siddiqui, Mohamed F. Alajmi, Adnan J. Alrehaily, Saleh I. Alqasoumi, Maged S. Abdel-Kader, Prawez Alam and Abd El Raheim M. Donia
A simple and sensitive high-performance thin-layer chromatographic (HPTLC) method was developed for the evaluation of biomarker β-amyrin in the leaves of fve different species of genus Ficus (Ficus carica, Ficus nitida, Ficus ingens, Ficus palmata, and Ficus vasta) grown in the Kingdom of Saudi Arabia. Chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates with solvents toluene–methanol (9:1, v/v) as the mobile phase. After development, the HPTLC plate was derivatized with p-anisalde-hydereagent to give well-resolved and compact spot of β-amyrin. Scanning and quantifcation were done at 550 nm. The system was found to give compact spot for β-amyrin at RF = 0.58. The linear regression analysis data for the calibration plots showed good linear relationship with r2 = 0.998 with respect to area in the concentration range of 100–900 ng. The regression equation for β-amyrin standard was found to be Y = 5.835X + 87. The precisions (n = 6) for β-amyrin were found to be 1.64–1.77% and 1.68–1.84%, respectively, for intra-day and inter-day batches, and the recovery values were found to be 97.6–98.3%. β-Amyrin was found to be present in three species, i.e., F. carica (0.29%, w/w), F. nitida (0. 5 4% w/w), and F. p almata (0.31%, w/w), while it was absent in F. vasta and F. ingens. The statistical analysis proves that the developed method for the quantifcation of β-amyrin is reproducible; hence, it can beemployed for the determination of β-amyrin in plasma and other biological fuids as well as in fnished products avai lable in the market.
Authors:Perwez Alam, Tawfeq A. Alhowiriny, Nasir A. Siddiqui, Saleh I. Alqasoumi, Omer A. Basudan, Azmat Ali Khan, Abdullah T. Alhowiriny and Nawazish Alam
Extensive research on Ficus species has shown their excellent cytotoxic potential which motivated the authors for further evaluation of its other species. In this article, the β-sitosterol content in the chloroform extract of the leaves of five Ficus species (Ficus carica [FCCE], Ficus nitida [FNCE], Ficus ingens [FICE], Ficus palmata [FPCE], and Ficus vasta [FVCE]) was estimated by a validated high-performance thin-layer chromatography (HPTLC) method along with cytotoxic activity. The chromatography was performed on glass-backed silica gel 60 F254 HPTLC plates with hexane and ethyl acetate (8:2, v/v) as the mobile phase. The developed plate was derivatized with p-anisaldehyde reagent, scanned, and quantified at λ = 550 nm. It furnished a compact and intense peak of β-sitosterol at RF = 0.17 ± 0.001. The contents of β-sitosterol (μg mg−1 of the dried weight of the extract) in the selected Ficus species were found as: FCCE (1.047 μg mg−1) > FVCE (0.771 μg mg−1) > FNCE (0.372 μg mg−1) > FPCE (0.309 μg mg−1), while it was absent in F. ingens. Methylthiazol tetrazolium (MTT) assay was used to compare the cytotoxic potential of all Ficus species against HepG2 (liver), HEK-293 (kidney), MCF-7 (breast), and MDA-MB 231 (breast) cell lines. The FCCE exhibited good cytotoxic property against HepG2, HEK-293, and MDA-MB-231 cells (IC50: 32.5, 41.4, and 47.3 μg mL−1, respectively), while FICE showed against HepG2 and MDA-MB-231 cells (IC50: 31.4 and 41.2 μg mL−1, respectively). The remaining Ficus extracts were found to be very less effective or insignificant. The cytotoxic property of FCCE is also supported by the HPTLC estimation of β-sitosterol which is reported to exhibit anticancer properties by interfering with multiple cell signaling pathways, including cell cycle, apoptosis, and proliferation. Our data suggest that the developed HPTLC method can be further employed in the analysis of marketed herbal formulations, and the active Ficus species can be further subjected to isolation of cytotoxic phytoconstituents.