Ghulam Mustafa

Synthesis and characterization of cupric oxide (CuO) nanoparticles and their application for the removal of dyes

In the present work, cupric oxide (CuO) nanoparticles (NPs) were prepared by adopting aqueous precipitation method using copper sulphate 5-hydrate as a precursor and NaOH as a stabilizing agent. This gives a large scale production of CuO-NPs which are utilized for the removal of methylene blue (MB) dye. The CuO NPs were characterized for the studying of their structure and composition from XRD which reveals the single phase monoclinic structure. The surface morphology of these NPs was carried out by using scanning electron microscopy (SEM). Moreover, the effect of optimization parameters such as time, concentration and temperature was also examined. Spectrophotometric technique was used to evaluate the removal of MB in aqueous solution by NPs. The equilibrium adsorption of cationic dye (MB) was carried out at various temperatures ranging from 303 to 318 K ± 2K. The adsorption isotherm equations like Langmuir, Freundlich and Dubinin-Radushkevich were applied and the values of their respective constants were evaluated by adopting graphical method. Thermodynamic and kinetic studies were also performed to determine the feasibility of the process. The maximum MB removal was observed to be 88.93%. The pH of point zero charge (pHPZC) of adsorbent was also estimated by pH drift method. The results indicate that aqueous precipitation method is a reliable and cheap method for the development of CuO-NPs which can be effectively used for the removal of dyes from effluents. This method is beneficial for the remediation of industrial waste.

Molecular cloning and comparative sequence analysis of fungal β-Xylosidases

Commercial scale degradation of hemicelluloses into easily accessible sugar residues is practically crucial in industrial as well as biochemical processes. Xylanolytic enzymes have a great number of possible applications in many biotechnological processes and therefore, these enzymes are continuously attracting the attention of scientists. Due to this fact, different β-Xylosidases have been isolated, purified and characterized from several bacteria and fungi. Microorganisms in this respect have gained much momentum for production of these significant biocatalysts with remarkable features. It is difficult to propagate microorganisms for efficient and cost-competitive production of β-Xylosidase from hemicelluloses due to expensive conditions of fermentation. The screening of new organisms with an enhanced production of β-Xylosidases has been made possible with the help of recombinant DNA technology. β-Xylosidase genes haven been cloned and expressed on large scale in both homologous and heterologous hosts with the advent of genetic engineering. Therefore, we have reviewed the literature regarding cloning of β-Xylosidase genes into various hosts for their heterologous production along with sequence similarities among different β-Xylosidases. The study provides insight into the current status of cloning, expression and sequence analysis of β-Xylosidases for industrial applications.

Reproducible in vitro regeneration system for purifying sugarcane clones

Genome purification of a selected clone of sugarcane is the key to developing homogenous lines. Generally,xa0 regenerated plants after transformation are heterogeneous at genome level, and several successive rounds of selection on antibiotic-containing medium and regeneration cycles are required to purify the genome to develop a homogenous population. Sugarcane is a vegetatively propagated plant and hence it requires to be grown in the field to harvest mature cane tops carrying meristematic tissues. In the present studies, stems of in vitro grown plants of four indigenous genotypes namely; HSF-242, US-778, HSF-243 and HSF-240, were subjected to regeneration. Five days post incubation at various levels of 2,4-D, the segments were placed on regeneration medium containing a combination of casein hydrolysate (500 mg/L), kinetin (0.5 mg/L) and benzylaminopurine (BAP, 0.5 mg/L). Response to regeneration was varied from basal to top sections. Nevertheless, more than 137 shoots were regenerated from basal segment, suggesting that the segment consisting of meristematicxa0 tissues responded well to in vitro conditions. This procedure may be considered as one of the best everxa0 published report on regeneration from in vitro grown plants to purify clones without subjecting the plants toxa0 field conditions and harvesting the mature cane. This technique was used to purify transgenic sugarcane plants carrying Bacillus thuringiensis gene. Key words : In vitro explant, basal stem segment, callus induction, proficient regeneration, Saccharum officinarum.

Comparative sequence analysis of citrate synthase and 18S ribosomal DNA from a wild and mutant strains of Aspergillus niger with various fungi

A mutation was induced in Aspergillus niger wild strain using ethidium bromide resulting in enhanced expression of citric acid by three folds and 112.42 mg/mL citric acid was produced under optimum conditions with 121.84 mg/mL of sugar utilization. Dendograms of 18S rDNA and citrate synthase from different fungi including sample strains were made to assess homology among different fungi and to study the correlation of citrate synthase gene with evolution of fungi. Subsequent comparative sequence analysis revealed strangeness between the citrate synthase and 18S rDNA phylogenetic trees. Furthermore, the citrate synthase movement suggests that the use of traditional marker molecule of 18S rDNA gives misleading information about the evolution of citrate synthase in different fungi as it has shown that citrate synthase gene transferred independently among different fungi having no evolutionary relationships. Random amplified polymorphic DNA (RAPD-PCR) analysis was also employed to study genetic variation between wild and mutant strains of A. niger and only 71.43% similarity was found between both the genomes. Keeping in view the importance of citric acid as a necessary constituent of various food preparations, synthetic biodegradable detergents and pharmaceuticals the enhanced production of citric acid by mutant derivative might provide significant boost in commercial scale viability of this useful product. Abbreviations CS - Citrate synthase, CA - Citric acid, RAPD - Random amplified polymorphic DNA, TAF - Total amplified fragments, PAF - Polymorphic amplified fragments, CAF - Common amplified fragments.

Differential Role of Indolebutyric Acid in Sugarcane Root Development

Auxin biosynthesis, metabolism, transport and signaling pathways are crucial for plant growth and development, including rhizogenesis. Sugarcane shoots regenerated under in vitro conditions develop roots without supplementary addition of exogenous plant growth regulators. However, robust rooting in engineered clones and in vitro multiplied shoots of sugarcane is of interest to biotechnologists for mass propagation and adaptation of plants in the environment. Here we report that Indolebutyric acid (IBA), being precursor of Indoleacetic acid, has differential role in regulating the development of roots. Shoots developed under in vitro conditions of an elite genotype US-127 were placed on rooting medium, containing different levels (0.0, 1.0, 2.0, 3.0, 4.0, 5.0 and 10.0xa0mgxa0L−1) of IBA. Under in vitro conditions, root length varied from maximum (6.99xa0±xa00.17xa0cm) to minimum (0.50xa0±xa00.07xa0cm) in plants supplemented with IBA from 0.0 to 10xa0mgxa0L−1 in the multiplication medium. A dramatic change in root growth was observed in these plants when shifted to peat moss-containing pots; the plants supplemented with higher levels (5.0xa0mgxa0L−1) of IBA showed maximum (25.62xa0±xa01.41xa0cm) root growth which was gradually decreased to minimum (9.81xa0±xa00.46xa0cm) with decreasing concentrations (from 5.0 to 0.0xa0mgxa0L−1) of supplementary IBA, confirming that the root development is inversely and directly proportional to increasing concentrations of IBA under in vitro and in vivo conditions, respectively. Hence, plants treated with IBA under in vitro conditions develop more established roots upon transplanting to open environment, suggesting that supplementary IBA is desirable for establishment of root system and for adaptation of in vitro grown and/or engineered Saccharum clones to open environment without compromising the mortality of transplants.

Fungal Biomass Protein Production from Trichoderma harzianum Using Rice Polishing

Industrially important enzymes and microbial biomass proteins have been produced from fungi for more than 50 years. High levels of crude protein as much as 45% are present in fungal biomass with balanced essential amino acids. The aim of this study was to access the potential of Trichoderma harzianum to produce fungal biomass protein from rice polishings. Maximum biomass yield was obtained at 5% (w/v) rice polishings after 72u2009h of incubation at 28°C at pH 4. Carbon and nitrogen ratio of 20u2009:u20091 gave significantly higher production of fungal biomass protein. The FBP in the 75 L fermenter contained 49.50% crude protein, 32.00% true protein, 19.45% crude fiber, 9.62% ash, 11.5% cellulose content, and 0.325% RNA content. The profile of amino acids of final FBP exhibited that all essential amino acids were present in great quantities. The FBP produced by this fungus has been shown to be of good nutritional value for supplementation to poultry. The results presented in this study have practical implications in that the fungus T. harzianum could be used successfully to produce fungal biomass protein using rice polishings.

Impact of Farm Households’ Adaptations to Climate Change on Food Security: Evidence from Different Agro-ecologies of Pakistan

Treatment Effects Model was applied to evaluate the impact of adaptations on household food security. A household Food Security Index (FSI) was constructed applying PCA. Adaptation strategies employed by the farmers in response to climate change were categorised into four groups namely: changes in sowing time (C1); input intensification (C2); water and soil conservation (C3); and changes in varieties (C4). Out of 15 mutually exclusive combinations constructed for evaluation, only 7 combinations were considered for estimating the treatment effects models because of limited number of observations in other cases. Results of only two of the 7 are discussed here, as the other 5 had very small number of adapters and the impact measures shown either insignificant results or had opposite signs. The first (C1234) combined all the four while the second (C234) combined the last three strategies. The results suggest that the households which adapted to climate changes were statistically significantly more food secure as compared to those who did not adapt. The results further show that education of the male and female heads, livestock ownership, the structure of house—both bricked and having electricity facility, crops diversification, and non-farm income are among the factors which raise the food security of farm households and their impacts are statistically significant. The variables which are significantly negatively associated with the food security levels include age of the head of household, food expenditure management, households having less than 12.5 acres of land— defined as marginal (cultivate 6.25 to 12.5 acres). Farmers of cotton-wheat, rice-wheat, and rain-fed cropping systems are found to be more food secure as compared to the farmers working in the mixed cropping systems where farm holdings are relatively small and high use of tubewell water adding to salinity of soils. It is crucial to invest in developing agricultural technologies to address issues of climate change relevant to different ecologies and farming systems; improve research-extension-farmer linkages; enhance farmers’ access to new technologies; improve rural infrastructure; develop weather information system linking meteorological department, extension and farmers; and establish targeted food safety nets as well as farm subsidy programs for marginal farm households

Expression profiling of bioactive genes from Moringa oleifera.

Plants are under constant assault by biotic and abiotic agents. When an elicitor is prologued, an immense reprogramming of plant gene expression and defense responses are initiated, which could be a natural source for potential drug development and insertional mutagenesis. In this regard, differential expression analysis of a medicinal plant Moringa oleifera was performed for bioactive genes at seedling stage, using differential display-RT-PCR technique. Infected seedlings with a fungus Fusarium solani collected at different time intervals, showed a massive change in their gene expression profile. The data analysis revealed that at least 150 pathogen-induced and about 60 suppressed genes were differentially expressed at 8-h postinoculation of the biotic stress. Fifty-five selective genes were disunited and reamplified. Sequence analysis of these potential genes illustrated that these genes had properties of some induced peroxidase mRNA, cell proliferation, others were mitogen activated protein kinases, ribosomal protein genes, defense regulating genes, and a few also had structural properties. Further studies about the utility of these genes in plant metabolism could assist to develop improved transgenic breeds with enhanced value of infection tolerance not only of M. oleifera but of other cultivars also.

Enzymes: Plant-based Production and their Applications

BACKGROUNDnEnzymes are biocatalysts that play key roles in the production of biomolecules. Transgenic plants can be valuable cost effective resource to produce enzymes with bona fide structure. Further, plants provide inexpensive production platforms for pharmaceuticals and nutraceuticals.nnnOBJECTIVEnThis review article summarizes the properties and importance of enzymes and describes how foreign proteins/enzymes accumulate in plant cells that can be used for commercial purposes.nnnCONCLUSIONnThe instances illustrated in this review evidently depict that plant enzymes involved in fundamental cellular activities are of great importance regarding plant growth and development. Investigating these enzymes and the metabolic pathways involved in their synthesis will certainly help to improve plant and human health. Furthermore, enzymes of industrial and pharmaceutical importance can be expressed in genetically modified plants to obtain enhanced expression. Considering easiness of obtaining desired expression, GM plants can offer a good alternate for large scale production of enzymes.

Isolation and Characteristics of Biotechnologically Important Antagonistic Thermophilic Bacteria from Rhizosphere of Haloxylon salicornicum

Rhizobacteria are an active part of microbial population in the rhizosphere of plants. In this study, twenty rhizobacteria were isolated from the rhizosphere of a perennial grass, Haloxylon salicornicum, found in Cholistan desert, an arid landmass near Bahawalpur Pakistan, in one set of experimental conditions. Colony characteristics, biochemical and molecular analyses of these isolates were performed. All isolates were bacilli, gram positive with off-white colonies and exhibited typical bacilli colony morphology. None of the isolates was gelatinase, urease, indole, H2S and catalase producer. Eleven isolates were amylase producers and 8 isolates were acid producers. All isolates fermented glucose, 3 fermented lactose and 19 fermented fructose. Molecular data revealed that out of twenty isolates, 14 isolates showed 91-99% identity with Brevibacillus borstelensis, 4 with Bacillus subtilis (97-98%) and 2 with Bacillus licheniformis (94-99%) through BLAST analysis. All identified bacterial isolates cladded with their respective groups in the phylogenetic tree. Many (11-15 out of 20) of the isolates were more effective in inhibiting growth of the tested bacterial strains as compared to the positive control (Ampicillin 50 μg/disc). We conclude that bacilli are the predominant form populating rhizosphere of this desert grass. Among the isolated bacteria Brevibacillus borstelensis, Bacillus subtilis and Bacillus licheniformis are the most predominant species.