Stability of RAPD markers for determining cultivar specific DNA profiles in rye (Secale cereale L.)

Summary Cultivar specific DNA profiles in rye were revealed by polymerase chain reaction (PCR) using randomly amplified polymorphic DNA (RAPD) sequences. Ten base primers were used for the amplification of genomic DNA of rye cultivars by PCR. RAPD analysis was found to be reproducible among samples between PCR runs. When amplification profiles of different rye cultivars were compared using various primers, the overall profiles were cultivar specific. However, not all primers revealed polymorphisms. These primers appear to amplify conserved sequences in all rye cultivars. Intracultivar studies were conducted on two of the cultivars. In the cultivar Imperial, no polymorphisms were observed among ten plants analyzed with five primers. In the cultivar Balboa, polymorphisms were observed among fifty plants with four of the ten primers analyzed. Despite the small amount of intracultivar variability, RAPD analysis has the potential to be a rapid and reliable method of cultivar identification in this outcrossing species.     

Nutritional significance of amino acids,vitamins and minerals as nutraceuticals in poultry production and health – a comprehensive review

Nutraceuticals have gained immense importance in poultry science recently considering the nutritional and beneficial health effects of their constituents. Besides providing nutritional requirements to birds, nutraceuticals have beneficial pharmacological effects, for example, they help in establishing normal physiological health status, prevent diseases and thereby improve production performance. Nutraceuticals include amino acids, vitamins, minerals, enzymes, etc. which are important for preventing oxidative stress, regulating the immune response and maintaining normal physiological, biochemical and homeostatic mechanisms. Nutraceuticals help in supplying nutrients in balanced amounts for supporting the optimal growth performance in modern poultry flocks, and as a dietary supplement can reduce the use of antibiotics. The application of antibiotic growth enhancers in poultry leads to the propagation of antibiotic-resistant microbes and drug residues; therefore, they have been restricted in many countries. Thus, there is a demand for natural feed additives that lead to the same growth enhancement without affecting the health. Nutraceuticals substances have an essential role in the development of the animals’ normal physiological functions and in protecting them against infectious diseases. In this review, the uses of amino acids, vitamins and minerals as well as their mode of action in growth promotion and elevation of immune system are discussed.     

Floristic composition and ecological gradient analyses of the Liakot Forests in the Kalam region of District Swat,Pakistan

Kalam Kohistan is a dry,temperate hilly region in the Hindu Raj Series of the Greater Hindu Kush Mountains with diverse forests.As plant distribution and composition is expression of a range of various environmental variables,ecological and floristic attributes of vegetation in the Laikot Forests were therefore evaluated via consecutive explorative trips.These forests have not been evaluated ecologically before due to harsh climatic conditions and issues of accessibility and the study in hand is one of the first in its nature.Quantitative ecological techniques were used to sample forest vegetation and identify species in quadrats of 2 m×2 m for herbs,5 m×5 m for shrubs and 10 m×10 m for trees.All the data of 195 plant species and environmental factors were analyzed via Two-way Cluster Analysis and Canonical Correspondence Analysis using PCORD and CANOCO software.Among 195 plant species from 63 families were 27 species of Asteraceae,the most-speciose family,followed by Poaceae(20 spp.).The dominant life form was therophyte(86 spp.;44%share),followed by phanerophyte(41 spp.;21%share).For the leaf size spectrum,most prevalent was nanophyll(73 spp.;44%)followed by microphyll(66 spp.,34%)and mesophyll(44 spp.;23%).Calcium carbonate,pH,potassium,sand,silt,organic matter,slope aspect and grazing significantly influenced species composition,distribution and habitat.The floristic variation in the region was diverse due to elevational and aspect gradients and thus devided into three zones.This vegetation zonation can be utilized for forest management,species and habitat conservation.Both in situ and ex situ conservation of threatened plant species may improve their conservation status in future if address properly.     

Genome-wide association analysis for stripe rust resistance in spring wheat(Triticum aestivum L.) germplasm

Stripe rust is a continuous threat to wheat crop all over the world. It causes considerable yield losses in wheat crop every year. Continuous deployment of adult plant resistance(APR) genes in newly developing wheat cultivars is the most judicious strategy to combat this disease. Herein, we dissected the genetics underpinning stripe rust resistance in Pakistani wheat germplasm. An association panel of 94 spring wheat genotypes was phenotyped for two years to score the infestation of stripe rust on each accession and was scanned with 203 polymorphic SSRs. Based on D' measure, linkage disequilibrium(LD) exhibited between loci distant up to 45 c M. Marker-trait associations(MTAs) were determined using mixed linear model(MLM). Total 31 quantitative trait loci(QTLs) were observed on all 21 wheat chromosomes. Twelve QTLs were newly discovered as well as 19 QTLs and 35 previously reported Yr genes were validated in Pakistani wheat germplasm. The major QTLs were QYr.uaf.2 AL and QYr.uaf.3 BS(PVE, 11.9%). Dissection of genes from the newly observed QTLs can provide new APR genes to improve genetic resources for APR resistance in wheat crop.     

Effects of sludge on germination and initial growth performance of Leucaena leucocephala seedlings in the nursery

A study was carried out to determine the influence of different types of sludges （municipal, industrial and residential） on field germination, growth and nodulation of L. leucocephala seedlings in the nursery. Before sowing of seeds, different combinations of sludges were incorporated with the nutrient deficient natural forest soils. Field germination, nodulation status and physical growth parameters of seedlings （shoot and root length, vigor index, collar diameter, leaf number, fresh and dry weight of shoot and root and total dry biomass increment） were recorded after three and six months of seed sowing. Field germination, nodulation status and growth parameters were varied significantly in the soil amended with sludges in comparison to control. The highest number of nodule was recorded from soil amended with residential sludge （ 1：1 ） and highest fresh and dry nodule weight was also found from the same combination in both three and six month old seedlings. In case of growth parameters, the highest growth was recorded from soil and residential sludge （ 1：1 ） combination compared to control. From the study, it can be recommended that soil amended with residential sludge （ 1：1 ） provide better field germination, growth and nodule formation of L. leucocephala in degraded soil.     

Molecular simulations of Taxawallin I inside classical taxol binding site of β-tubulin

A new taxoid Taxawallin I (1) along with two known taxoids (2-3) were isolated from methanolic bark extract of Taxus wallichiana Zucc. Structural characterization was confirmed by mass and NMR spectral techniques. Taxawallin I exhibited significant in-vitro anticancer activity against HepG2, A498, NCI-H226 and MDR 2780AD cancer lines. Tubulin binding assay was performed to assess its tubulin binding activity. Molecular docking analysis was performed to study the potential binding mode inside the taxol binding site of β-tubulin.     

Pharmacokinetics of thymoquinone in layer chickens following oral and intravenous administration

Thymoquinone (TQ) is the major constituent of Nigella sativa and known to possess a variety of pharmacological effects. This study was designed to evaluate the pharmacokinetic profile of TQ following oral (PO) and intravenous (IV) administration in layer chickens. The layer chickens were equally divided into two groups (six chickens in each group, total 12 chickens), and TQ was administered via PO and IV routes. For PO route, the dose was 20 mg/kg b.w. and for IV route, 5 mg/kg b.w. was administered, respectively. A sensitive and accurate High‐Performance Liquid Chromatography (HPLC) technique was validated for the quantification of TQ from plasma. The limit of detection (LOD) and limit of quantification (LOQ) were 0.02 µg/ml and 0.05 µg/ml, respectively with >80% recovery. Maximum plasma concentration (C_max) following PO and IV administration was 8.805 and 4.497 µg/ml, respectively, while time to reach at maximum concentration (T_max) was 1 and 0.1 hr, respectively. The elimination half‐lives were recorded as 1.02 and 0.978 hr, whereas the mean residence times were 1.79 and 1.036 hr following both PO and IV administration, respectively. The 85% PO bioavailability was indicative that TQ could be used for various therapeutic purposes in layer chickens.     

Physicochemical,including spectroscopic,and biological analyses during composting of green tea waste and rice bran

The aims of this study were to monitor the changes in physicochemical, including spectroscopic, and biological characteristics during composting of green tea waste–rice bran compost (GRC) and to define parameters suitable for evaluating the stability of GRC. Compost pile temperature reflected the initiation and stabilization of the composting process. The pH, electrical conductivity, NO₃ ⁻-N content, and carbon-to-nitrogen ratio were measured as chemical properties of the compost. The color (CIELAB variables), humification index (the absorption ratio Q _4/6 = A ₄₇₂ / A ₆₆₄ of 0.5 M NaOH extracts), absorption at 665 nm of acetone extracts, and Fourier-transform infrared (FT-IR) spectra were measured to evaluate the organic matter transformation; germination of komatsuna or tomato seeds was measured to assess the potential phytotoxicity of composting materials during composting. No single parameter was capable of giving substantial information on the composting process, the nutrient balance, phytotoxicity, and organic matter decomposition. The FT-IR spectra at 3,300, 2,930, 2,852, and 1,065 cm⁻¹ provided information on the molecular transformation of GRC during composting and they decreased over the composting. Most of the assayed parameters showed no further change after about 90 days of composting suggesting that GRC can be used for agricultural purposes after this period.     

Induced Degradation of Anthraquinone-Based Dye by Laccase Produced from <Emphasis Type="Italic">Pycnoporus sanguineus</Emphasis> (CS43)

In this study, in-house isolated laccase isoforms, i.e., Lac-I and Lac-II of the basidiomycete Pycnoporus sanguineus (CS43), were evaluated in relation to their Remazol Brilliant Blue R (RBBR) dye degradation capacity. A modified Dhouib medium additionally supplemented with 3% ethanol as a secondary inducer was used to propagate P. sanguineus CS43 for enhanced production of laccase under liquid state fermentation. The crude laccase extract was purified by passing through ion exchange diethylaminoethanol (DEAE)-Sepharose and gel filtration-based Sephadex G-200 column chromatography. The purified laccase fractions were subjected to the electrophoresis, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis revealed two laccase isoforms Lac-I and Lac-II with 66 and 68 kDa, respectively. To explore the industrial applicability, for RBBR dye, degradation efficiencies ranged from 82 to 88% after 3 h of incubation for both; Lac-I and Lac-II at both concentrations were recorded. However, with 8 U/mL, the degradation ranged between 70 to 80% during the first 5 min of incubation. Enhanced degradation of RBBR dye was obtained in the presence of violuric acid and N-hydroxypthalamide as laccase mediators. Finally, using RBBR as a substrate kinetic characterization of both Lac-I and Lac-II isoforms was performed that revealed K _m (0.243 and 0.117 mM for Lac-I and Lac-II) and V _max (1.233 and 1.012 mM/Sec for Lac-I and Lac-II) values, respectively.     

Resistance of beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) to endosulfan,organophosphorus and pyrethroid insecticides in Pakistan

Field populations of beet armyworm, Spodoptera exigua (Lepidoptera: Noctuidae), from Pakistan were assessed for their resistance to the chlorinated hydrocarbon endosulfan, the organophosphates chlorpyrifos and quinalphos, and the pyrethroids cypermethrin, deltamethrin, bifenthrin and fenpropathrin. Using a leaf-dip bioassay, resistance to endosulfan was high during 1998–2000 but declined to very low, to low levels during 2001–2007, following a reduced use of the insecticide. Organophosphates and pyrethroids were consistently used over the past three decades, and the resistance had been increasing to these insecticide classes. Generally, the resistance to chlorpyrifos and pyrethroids remained low from 1998 to 2002–2003, but resistance increased to moderate to high levels from 2003–2004 to 2006–2007. For deltamethrin, resistance was very high during 2004–2007. Quinalphos resistance remained low during 1998–2006. Correlation analysis of LC₅₀ and LC₉₀ values showed a positive correlation between organophosphates and pyrethroids, but no correlation between endosulfan and organophosphates or pyrethroids tested herein. These results suggest that the conventional chemistries should be replaced with new chemistries for the successful management of S. exigua.