Assessment of anticlastogenic activity of cinnamic acid: Anticlastogenic index (ACI) and model simulation

Cinnamic acid and its derivatives are an important member of the phenolic compound used in food supplements. They usually occur in various conjugated forms, more frequently esters and glycosides. Mice (Mus musculus, 2n = 40) were employed as an experimental mammalian system to assess the anticlastogenic activity of cinnamic acid (CA) induced by Endoxan. Four doses; 1/32, 1/16, 1/8 and 1/4 of the LD50 of CA (5, 10, 20 and 40 mg/kg, respectively) in combination with five administration times of CA were tested. Micronucleated polychromatic erythrocytes (PCE) and mitotic index (MI) were used as a sensitive short term genotoxic bioassays. The results obtained showed that low doses (1/32 LD50 and 1/16 LD50) decreased the percentage of PCE significantly compared with that of the positive control and closer to that of the negative control. The data of this study were used to calculate a new index called anticlastogenic index (ACI). The new index measures the anticlastogenic activity of a compound or an extract. The ACI beside the percentage of PCE can give a deeper look of the anticlastogenic activity of compounds. It also makes it easy to draw conclusions from the genotoxicity data. The maximum ACI of CA was achieved when both CA and Endoxan were given concurrently. On the other hand, the higher doses of CA (1/8 and 1/4 LD50) caused a significant increase in the percentage of PCE. This gives evidence that CA at high doses (20 and 40 mg/kg) would be considered as a positive clastogen itself. CA significantly decreased the mitotic index compared with the negative control. In addition, high doses showed sharp decrease in mitotic index. Direct significant correlation coefficient was found between the ACI and the mitotic index. The data also were used for the simulation of a model to predict the ACI of doses with different relative timing to a treatment of the clastogen that were not conducted experimentally in the limit of experiment data range.Key words: Anticlastogenic, cinnamic acid, anticlastogenic index, ACI, mitotic index, mice, model simulation of ACI

Optimization of chemiluminescent detection of mitochondrial RNA-protein interaction by nonradioactive mobility shift

RNA mobility shift is one among many procedures used to study RNA-protein interaction. Yet, there are some limitations for the radioactive RNA mobility shift including; 1) the risk of using radiolabeled nucleotides, 2) the long time to get the results; this could range from days to weeks, and 3) its high cost as compared to nonradioactive techniques. In this study, we optimized a nonradioactive procedure using dig-11-UTP nucleotide and chemiluminescent detection for mitochondrial RNA-protein interaction. The optimizations include the quality limiting steps such as using non-specific competitors of RNA probe, UVcross linking time, electrotransfer to membranes, application with various protein extracts, and the examination for false positive RNA-protein complexes using proteinase K digestion. The results show that the optimizations carried out in the study significantly enhanced the quality of the results obtained with this procedure.Keywords: RNA mobility shift, mitochondria, wheat, RNA-protein interaction, chemiluminescent detection, nonradioactiveAfrican Journal of Biotechnology Vol. 12(16), pp. 1915-192

Estimation of water pollution by genetic biomarkers in tilapia and catfish species shows species-site interaction

This study was aimed at the estimation of water pollution with heavy metals using four biomarkers as well as to study the species-site interaction. Two species of tilapia as well as two catfish species caught from four sites that represent differential environmental stresses were used for this purpose. Water samples and gills were analyzed for heavy metal contents. Three enzyme biomarkers (acid phosphatase (AP), glutamic oxaloacetic transaminase (GOT), glutamic pyruvic transaminase (GPT), metalothionein (MT) gene expression) as well as real time PCR quantification of metalothionein transcripts from liver were used to monitor fish response to water pollutants. Results showed various activities of the four biomarkers at the different studied sites. There were clear interaction between fish species and the level of heavy metals. Real time PCR evaluation of metalothionein gene expression revealed species variations at similar sites. It is concluded that there are various types of interaction of species at different sites. These types of interaction depend on the type of biomarker tested

Molecular and biological characterization of Trichogramma turkestanica

Parasitoids of Trichogramma (Hymenoptera: Trichogrammatidae) parasitize the eggs of many species of Lepidoptera and have been used for the biological control of numerous pest species. We collected this parasitoid from Taif governorate, KSA in summer of 2009. It is difficult to differentiate between Trichogramma species because of their small size and lack of differences in morphological characters. Therefore, different molecular markers were employed to characterize this species, including direct amplification of the internal transcribed spacer 2 (ITS2) of ribosomal DNA and by restriction fragment length polymorphism followed by sequencing. The results show that ITS2 region is 491 bp and indicated that this is a new stain of Trichogramma. We named this strain TaifKSA. From the tested restriction enzymes, only EcoRI and PstI cut the PCR product of ITS2 region. We compared the biological characteristics of the strain under investigation with other commercial strain (SQG) of the same species and no significant differences between them have been shown.Key words: Trichogramma turkestanica, TaifKSA, molecular identification, internal transcribed spacer 2 (ITS2), restriction enzymes, biological characteristics

Functional Analysis of SBPase Gene Promoter in Transgenic Wheat under Abiotic Stresses

Background and Objective: This study focused on evaluation of SBPase promoter in driving transgenes under drought, high light intensity and salinity as major abiotic stresses limiting plant growth and production in various areas of the world. Wheat production has remained constant despite the rapidly growing population around the world. Materials and Methods: This had made it necessary to develop other varieties with higher yield in order to satisfy the future demand. When elite wheat varieties’ photosynthesis is genetically manipulated, it gives a high possibility for yield increase. Lack of a properly defined molecular tool-box promoter for driving gene expression for manipulation of photosynthesis, hinders the progression of this field. Gus histochemical staining and quantitative PCR were used to examine the function of Brachypodium distachyon sedoheptulose-1, 7-bisphosphatase (SBPase) promoter to drive the expression of GUS in the stably transformed wheat plants under various stresses conditions. Results: Results revealed that B. distachyon SBPase promoter fully drive the GUS expression in the stable wheat transformants (cv. Cadenza) in the leaf tissues under normal and high light conditions. It also drove GUS gene expression under drought and salinity stress conditions. In addition, the promoter responds to light/dark/light alternation as light responsive genes. Conclusion: The Bd SBPase promoter provides innovative molecular tool to drive genes in wheat leaves under natural, drought, high light intensity and salinity conditions as well as to allow for multigene photosynthetic manipulation for yield increase under various abiotic conditions

Sustainable phosphorous management in two different soil series of Pakistan by evaluating dynamics of phosphatic fertilizer source

Phosphorous (P) plays the prominent role to promote the plants storage functions and structural roles, as it is recognized as a vital component of ADP, ATP, Cell wall as well as a part of DNA. Soils acts as the sink to supply P to plants because soil pH and its physical condition are the main factor which regulate the solubility and availability P element. Phosphorus is not deficient in Pakistani soils but its availability to plants is the serious matter of concern. A pot experiment was conducted to evaluate P dynamics in two different soil series of Pakistan (Bahawalpur and Lyallpur) using Maize as test crop. The treatments applied were T0: Control (without any fertilizer), T1: Recommended DAP @648 mg pot−1, T2: Half dose DAP @324 mg pot−1, T3: Recommended rate of TSP @900 mg pot−1, T4: Half dose TSP @450 mg pot−1. Soil analysis showed that Bahawalpur soil has sandy clay loam texture with 33% clay and Lyallpur series has sandy loam texture with 15.5% clay; furthermore, these soils contain 4.6 and 2.12% CaCO3, respectively. Results showed an increase in P concentration in roots (23 mg kg−1) with the application of half dose of TSP in Lyallpur series and lowest in Bahawalpur series (14.6 mg kg−1) at recommended dose of DAP. Concentration of P in shoots responded the same; increase at half dose of TSP (16.7 mg kg−1) and lowest at full dose of DAP in Bahawalpur series as (15.58 mg kg−1). Adsorbed P (17 mg kg−1) was recorded highest in Bahawalpur soil with more clay amount in pot with DAP application but lower in Lyallpur soil series (14 mg kg−1) with the application of applied TSP. The PUE was recorded highest in Lyallpur series with the application of half dose of TSP and it was 61% more than control and was Highest in Bahawalpur series was with the application of recommended dose of DAP is 72% more than control treatment. On estimation, results showed that applied sources made an increase in P availability than control, but TSP gave better P uptake than DAP unless of rates applied. Soil of Lyallpur series showed better uptake of P and response to applied fertilizers than Bahawalpur series which showed more adsorption of P by high clay and CaCO3 amount. Conclusively, the study suggested that soil series play a crucial role in choosing fertilizer source for field application

Quantitative response of wheat to sowing dates and irrigation regimes using ceres-wheat model

An experiment was conducted at Punjab Agricultural University, Ludhiana during 2014–15 and 2015–16, keeping four sowing dates {25th Oct (D1), 10th Nov (D2), 25th Nov (D3) and 10th Dec (D4)} in main plots and five irrigation schedules {irrigation at 15 (FC15), 25 (FC25), 35 (FC35) and 45 (FC45) % depletion of soil moisture from field capacity (FC) and a conventional practice} in sub plots. The objective of the study was to evaluate the performance of CERES-Wheat model for simulating yield and water use under varying planting and soil moisture regimes. The simulated and observed grain yield was higher in D1, with irrigation applied at FC15 as compared to all other sowing date and irrigation regime combinations. Simulated grain yield decreased by 19% with delay in sowing from 25th October to 10th December because of 8% reduction in simulated crop evapotranspiration. Simulated evapotranspiration decreased by 16%, wheat grain yield by 23% and water productivity by 15% in drip irrigation at 45% depletion from field capacity as compared to drip irrigation at 15% of field capacity. It was further revealed that the model performed well in simulating the phenology, water use and yield of wheat

Micronucleus test in fish genome: A sensitive monitor for aquatic pollution

The aquatic environment makes up the major part of our environment and resources, therefore its safety is directly related to the safety our health. In this study, three tilapia species (Oreochromisniloticus, Oreochromis aureus and Tilapia zilli) and Clarias gariepinus were employed to estimate water pollution using micronucleus (MN) test. The test has been used successfully as a mutagenic assay. It issimple, reliable, sensitive, and it does not depend on any karyotypic characteristics. Fish were collected from locations that display differential environmental stresses. Two main experiments were carried out. In the first one, blood samples were collected, fixed for 24 h and then were stained with Giemsa. In the second experiment, fish were acclimated for a week. Fish were fed and each specimen had received anIP injection of cyclophosphamide (2.6, 10, 40, mg/kg b.wt). After 24 h, blood samples were collected and MN frequencies were counted and statistically tested. Results from this study recommend the use ofthe micronucleus test in fish erythrocytes as a sensitive monitor for aquatic pollution. The results show also that the assay can be employed for the evaluation and the assessment of water pollution andaquatic mutagens

Defens in gene expression in some plant sources of Taif

Plant defensins are promising future strategy as antimicrobial agents; however the number of characterized plant defensins is very low. We isolated and studied the expression of plant defensins in six plants from Taif region. Using RT-PCR and two pairs of common as well as three pairs of specific primers, the defensin gene expression was analyzed. Plants showed differences in defensin gene expression in floral buds and leaves, though floral buds represented higher gene expression. The amino acid sequence of the six isolated cDNA sequences showed high similarity with other defensin accessions from the nucleotide database, especially in the cysteine rich motif. Phylogenetic analysis revealed that the isolated sequences share the common features of plant defensins, especially the 8 conserved cysteines. The results of this study help to detect some valuable Saudi plant sources for the development of natural peptides as a replacement for chemical antibiotics