Rapid virulence typing of Pasteurella multocida by multiplex PCR

The gram-negative bacterium Pasteurella multocida constitutes a heterogeneous species associated with wide range of disease in many animals. Isolates are classified into five groups based on capsular antigen (capA, B, D, E and F). Recently, a new valuable PCR-based method was introduced to determine the epidemiological correlation between P. multocida infection and existence of virulence genes including tbpA, pfhA, toxA and hgbB. However, this method is tedious and laborious. Thus, in the current study, we designed a reliable multiplex PCR method for rapid detection of virulence genes in P. multocida. Eighty seven strains of P. multocida isolated from various clinically healthy and infected hosts were examined by uniplex PCR method for each virulence associated genes. Based on our improved and simplified multiplex PCR method, rapid detection of four virulence genes was accomplished. It is proposed that its implementation may benefit the epidemiological investigations.     

Genetic diversity in populations of wild diploid wheat Triticum urartu Tum. ex. Gandil. revealed by isozyme markers

Genetic variation and its distribution within and among 23 populations of Triticum urartu collected from Syria, Lebanon, Turkey, Armenia, and Iran was estimated using isozyme markers at eight polymorphic loci. The number of alleles per locus (A= 1.21), percentage polymorphic loci (P= 20.1%), and mean gene diversity (H_e= 0.024) were relatively low. In a population from Lebanon, a high number of alleles per locus (A= 2.13) and percentage polymorphic loci (P= 87.5%) was found. On average, genetic variation among populations (G_ST= 0.407) was smaller than within-population variation (0.593). However, different patterns of genetic structure were found among various geographic regions. Interpopulation variation was highest for the Iranian populations (0.89) followed by the Turkish populations (0.66). A reverse pattern was observed for the Syrian (0.11) and for the Lebanese (0.13) populations. The Armenian populations exhibited similar interpopulation and within-population variation. Principal component and cluster analyses resulted in distinct grouping of the geographically proximal populations, with the exception of the two Iranian populations. The Turkish populations were different from the neighboring Armenian populations compared to other countries. The populations from southern Syria and those from Lebanon also exhibited a high degree of genetic diversity. The two most heterozygous loci, Mdh-2 and Pgi-2, separated the populations along the first and second principal components, respectively. Most of the rare alleles were scattered sporadically throughout the geographic regions. Rare alleles with high frequencies were found in the Turkish and Armenian populations. These results indicated that different geographic regions require specific sampling procedures in order to capture the range of genetic variation observed in T. urartu populations.     

Effect of tillage and fertilizer levels on wheat yield, nitrogen uptake and their correlation with carbon isotope discrimination under rainfed conditions in north-west Pakistan

Appropriate cultural practices need to be determined for enhancing crop yields with low inputs under rainfed conditions. A field experiment was conducted to study the effect of tillage practices and fertilizer levels on yield, nitrogen (N) uptake and carbon (C) isotope discrimination in wheat (Triticum aestivum L.) grown under semi-arid conditions at three sites in north-west Pakistan: NIFA, Urmar and Jalozai. Two fertilizer levels, 60 kg N ha⁻¹+30 kg P ha⁻¹ (L1) and 60 kg N ha⁻¹+60 kg P ha⁻¹ (L2), were applied to wheat grown under conventional tillage (T1) and no-tillage (T0) practices. Labeled urea having 1% ¹⁵N atom excess at 60 kg N ha⁻¹ was applied as aqueous solution in microplots within each treatment plot. A pre-sowing irrigation of 60 mm was applied and during the growing season, the crop relied entirely on rainfall (268 mm). Biomass yield, N uptake and stable C isotope composition (δ¹³C) of plants were determined at maturity. Yield of wheat was improved by tillage at two sites (Sites 1 and 2), while at the third site yield was reduced by tillage as compared with the no-tillage treatment. At Sites 1 and 2, nutrient addition (L2, 60 kg N ha⁻¹+60 kg P ha⁻¹) increased the yield of all plant parts (straw, grain and root) in contrast to Site 3 where only grain yield was increased significantly. Maximum grain yield of wheat was observed with tillage under nutrient level L2 at all sites. Generally, the tillage treatment did not affect the N content in plant parts compared with no-tillage (T0) treatment at all three sites. However, fertilizer N uptake by wheat was variable under different fertilizer levels and tillage practices. Nitrogen derived from fertilizer (Ndff) for grain at Site 2 was higher in tilled plots but was not affected by tillage practice at the other sites. The C isotope (δ¹³C) values varied from −28.96 to −26.03‰ under different treatments at the three sites. The δ¹³C values were less negative indicating more effective water use at Sites 2 and 3 compared to Site 1. The C isotope discrimination (Δ) values were positively correlated with yield of wheat straw (r=0.578^*), grain (r=0.951^**) and root (r=0.583^*). Further, the Δ in grain had significant negative relationship (r=0.912^**) with Ndff (%). The tillage practice exerted a positive effect on yield, N uptake and plant N derived from fertilizer by wheat compared to no-tillage. The positive correlation of Δ with grain, straw and root yields and negative correlation with the Ndff (%) by wheat suggest that this value (Δ) could be used to predict these parameters. However, further studies on different crops under varied environmental conditions are necessary.     

Biological nitrogen fixation associated with roots of field-grown barley (Hordeum vulgare L.)

Nitrogenase (C₂H₂) activity was measured in microbial media inoculated with barley root segments or corresponding rhizosphere soil. Three different media were used, Döbereiner's malate medium, a modified Ashby medium, and an acid nitrogen-free medium. Only Döbereiner's medium gave consistently positive results, and cultures inoculated with roots showed higher activity than cultures inoculated with corresponding rhizosphere soil. Similar experiments with roots and rhizosphere soil from wheat gave only negligible nitrogenase activity, whereas the tropical grass, Cynodon dactylon, gave higher activity than barley. Measurements on intact soil cores containing barley root systems showed an initial lag phase followed by a rather stable activity level over a period from 12 h to 48 h, and then the activity again decreased. The activity during the stable period corresponded to fixation of about 100 to 200 g N₂ ha^?1 24 h^?1. Measurements on isolated, washed barley roots showed only negligible nitrogenase activity.     

Allele mining of wheat ABA receptor at TaPYL4 suggests neo-functionalization among the wheat homoeologs

ABA receptors (PYR/PYL/RCAR) play a central role in the water loss control of plants. A previous report indicated that TaPYL4 is a critical gene in wheat that improves grain production under drought conditions and increases water use efficiency. In this study, we analyzed the sequence polymorphisms and genetic effects of TaPYL4s. Based on isolated TaPYL4 genes from chromosomes 2A, 2B and 2D, three haplotypes were detected in the promoter region of TaPYL4-2A, and two haplotypes were present in TaPYL4-2B and TaPYL4-2D, respectively. Marker/trait association analysis indicated that TaPYL4-2A was significantly associated with plant height in 262 Chinese wheat core collection accessions, as well as the drought tolerance coefficient (DTC) for plant height in 239 wheat varieties from Shanxi Province in multiple environments. However, the frequencies of favored drought-tolerant haplotype TaPYL4-2A-Hap2 were considerably low, accounting for only 10%, and lines with this certain Hap could be reserved in the breeding program. TaPYL4-2B was significantly associated with grain number, and the favored haplotype TaPYL4-2B-Hap1 was the dominant allele of above 90% in the collection. For TaPYL4-2D, there were no significant differences in these traits between the two haplotypes in either of the two panels. These results indicate that variation might lead to functional differentiation among the homoeologs and the haplotypes had undergone artificial selection during breeding. Two molecular markers developed to distinguish these haplotypes could be used for breeding in water-limited regions.     

Evaluation of the Reactivity of Organic Pollutants during O3/H2O2 Process

The reactivity of selected compounds in Lake Huron water was evaluated during ozone/hydrogen peroxide-based advanced oxidation process (AOP) and conventional treatment (coagulation?Csedimentation?Cfiltration). Elimination of these compounds via advanced oxidation and conventional treatment processes were strongly related to their molecular structures. Overall removal of target compounds was quite similar in effluents from both the AOP and the combined treatment process (AOP?+?conventional) with the exception of fluoxetine. Reaction rate constants for the decomposition of the target compounds were substantially higher during AOP compared to conventional treatment alone.     

Developing Pedotransfer Functions for Predicting FC and PWP

Using pedotransfer functions (PTF) is a useful way for field capacity (FC) and permanent wilting point (PWP) prediction. The aim of this study was to model PTF to estimate FC and PWP using regression tree (RT) and stepwise multiple linear regressions (SMLR). For this purpose, 165 and 45 soil samples from UNSODA and HYPRES datasets were used for development and validation of new PTFs, respectively. %Clay, geometric mean diameter (dg), and bulk density (BD) were selected as predictor variables due to the highest correlation and lowest multicollinearity. The results showed that clay percentage with W* = 0.89 and dg with W* = ?0.57 were the most effective variables to predict PWP and FC, respectively. The RT method had a better performance (R² = 0.80, ME = ?0.002 cm³cm^?3, RMSE = 0.05 cm³cm^?3 for FC and R² = 0.85, ME = 0.003 cm³cm^?3, RMSE = 0.03 cm³ cm^?3 for PWP) than SMLR in estimation of FC and PWP.     

Relationship of seed pericarp color with seed quality in sugar beet (Beta vulgaris L. var. altissima Döll)

Genetic Resources and Crop Evolution - Quantitative traits of seed pericarp color were used to evaluate the sugar beet (Beta vulgaris L.) seed quality. These traits were measured on 10 single cross...     

A simple and easy method to measure ammonia volatilization: Accuracy under field conditions

Field studies on soil ammonia(NH₃)volatilization are restricted in many countries owing to the high costs commonly demanded for accurate quantification.We assessed the accuracy of a simple,open chamber design to capture NH₃under field conditions,as affected by different chamber placement schemes.Urea-15 N was surface applied to lysimeters installed in the spaces between maize rows.Open chambers made from plastic bottles were installed on each lysimeter with variations in i)N rates(3,8,13,and 18 g m^-2),ii)the height of the chamber above the soil surface(0,5,and 10 mm),and iii)chamber relocation(static vs.dynamic).Reference lysimeters without chambers were used to measure NH₃losses by¹⁵N-balance.Losses of NH₃-N accounted for more than 50%of the applied N.Relocation of the chambers had no impact on their NH₃-trapping efficiencies,proving to be an unnecessary procedure.Variation in the height of the chambers above the soil surface affected the capture of NH₃,but the results still maintained high linearity with the NH₃losses quantified by the reference method(R²>0.98).When the same placement scheme used in the introductory study describing the chamber was utilized(static and touching the soil surface),we found a trapping efficiency of 60%,which was very similar to that(57%)obtained in the previous study.Our results show that this simple,open chamber design can be used with satisfactory accuracy under field conditions,provided that simple,standardized procedures are warranted.