Curr. Issues Mol. Biol., Volume 11, Issue 2 (July 2009) – 4 articles

The complexity of the wheat genome has delayed the development and application of molecular markers to this species and wheat now lies behind barley, maize and rice in marker development. However, improvements in marker detection systems and in the techniques used to identify markers linked to useful traits has allowed considerable advances to be made in recent years. To evaluate the genetic diversity 53 genotypes of Richard's selection were studied at National Agriculture Reseach Center (NARC) Islamabad. The present study found that RAPD analysis is a valuable diagnostic tool. Different sets of RAPD primers were used to study the polymorphism at molecular level. Highest number of amplifications was shown by primer OpG-2 in Richard's material. Coefficient of similarity as well as genetic distances among these three sets of materials was calculated by using Unweighted Pair Group of Arithamatic Means (UPGMA) function. The SHs derived genotypes of Richard's selection were highly polymorphic with a polymorphism percentage of 69.70 as compared to NUYT (rainfed) and elite Pakistani bread wheat varieties with a polymorphism of 44.44% and 61.11% respectively. Cluster analysis was done in which grouping of genotypes was done on the basis of genetic distances. Cluster analysis revealed that genotypes of Richard's genotypes are showing high level of among cultivar variation as compared to NUYT (Rainfed) and elite Pakistani drought tolerant bread wheat varieties. These genotypes were also phenotypically evaluated. Full article

How plant gene expression responds to grazing defoliation is critical for plant re-growth, survival, and composition in the natural and dairy farming grassland environments. Rice, with genome sequence available, was used as a model plant to study grazing-induced pathway selections. When seedlings were 18 cm in height, the top 12 cm was removed by simulated grazing. The gene expression activities within 2 to 24 hours in the remaining aboveground tissues were profiled using the Affymetrix Rice GeneChips and RT-qPCR. The seedlings responded to grazing by immediately adjusting their global gene expression, e.g., enhancing anaerobic respiration, starch-to-sugar conversion, sucrose synthesis, and sugar transport. The results suggest that (1) remaining aboveground tissues used anaerobic respiration as an emergency measure for energy/substrates supply; (2) Sink tissues reduced its demand after 2 h; (3) Sucrose synthesis enhancement around the 24th hour is likely driven by shoot re-growth. In total, the expression activity of 466 genes, involved in signal transduction, miRNA regulation, cell wall modification, metabolism, hormone synthesis, and molecule transporters, had been significantly changed at least twofold. These genes and their biochemical pathways identified provide insights into how plants respond to grazing at the molecular physiology level. Full article

Escherichia coli are commensal bacteria that can account for up to 1% of the bacterial population of the gut. Ruminant animals are reservoirs of the pathogenic bacteria E. coli strain O157:H7, and approximately 30% of feedlot cattle shed E. coli O157:H7. Feedlot and high-producing dairy cattle are fed high grain rations in order to increase feed efficiency. When cattle are fed high grain rations, some starch escapes ruminal microbial degradation and passes to the hindgut where it undergoes fermentation. Ten years ago researchers demonstrated that populations of total E. coli were higher in grain-fed than in forage-fed cattle, and when cattle were abruptly switched from a high grain diet to an all hay diet, total E. coli populations declined 1000-fold within 5 days and reduced the ability of the surviving E. coli to survive an acid shock mimicking passage through the human gastric stomach. This research provoked many questions about the effects of diet or E. coli O157:H7 populations that have not been conclusively answered to date. Subsequent research has shown that diet does affect E. coli O157:H7 populations, but the effects have varied in magnitude and impact. Further studies have demonstrated that the effects of forage feeding on E. coli O157:H7 populations may be due to concentrations of tannins and phenolic acids in forages. Other ration components such as rapidly ruminally fermented grains (e.g., barley) increase the shedding of E. coli O157:H7, and in some situations, feeding distillers grains can increase fecal shedding of E. coli O157:H7 due to VFA concentrations. Data from researchers across North America indicate that diet does impact STEC/EHEC populations in cattle prior to slaughter; however the economic, logistic and practical impacts of dietary changes must be examined and accounted for. Full article

Azoreductases reduce the azo bond (N=N) in azo dyes to produce colorless amine products. Crude cell extracts from Enterococcus faecalis have been shown to utilize both NADH and NADPH as electron donors for azo dye reduction. An azoreductase was purified from E. faecalis by hydrophobic, anion exchange and affinity chromatography. The azoreductase activity of the purified preparation was tested on a polyacrylamide gel after electrophoresis under native conditions and the protein that decolorized the azo dye (Methyl Red) with both NADH and NADPH was identified by mass spectrometry to be AzoA. Previously, the heterologously expressed and purified AzoA was shown to utilize NADH only for the reduction of Methyl Red. However, AzoA purified from the wild-type organism was shown to utilize both coenzymes but with more than 180-fold preference for NADH over NADPH as an electron donor to reduce Methyl Red. Also, its specific activity was more than 150-fold higher than the previous study on AzoA when NADH was used as the electron donor. The catalytic efficiency for Methyl Red reduction by AzoA from E. faecalis was several orders of magnitude higher than other azoreductases that were purified from a heterologous source. Full article