Effect of methylmercury chloride on the primary photosynthetic activity of higher plants

The effect of methylmercury chloride (MeHg) on the fluorescence characteristics of pea seedling leaves and thylakoids isolated from these leaves was studied by the pulse-amplitude-modulation (PAM) fluorometric method. In 3-4 days after the addition of MeHg (20 microM) to the nutritious solution, the maximal (Fv/Fm) and real (under steady state actinic light illumination) (deltaF/F'm) quantum photochemical yield of PS II decreased. The nonphotochemical fluorescence quenching coefficient in control (qN) decreased after its maximum value has been reached. In MeHg-treated samples, this decrease was not observed, possibly due to the disturbance of delta pH energy transducing processes in ATP synthase. This was confirmed by the results of experiments on isolated thylakoids. After MeHg (5 microM) treatment of thylakoids, the photophosphorylation rate and light-triggered Mg2+-dependent H+-ATPase activity were suppressed by 20-40%, depending on the duration of MeHg exposure. However, in experiments with isolated thylakoids, no decrease either in the electron transport rate or in the Fv/Fm ratio was observed. In total, the results obtained allow one to assume that MeHg at concentrations and time duration used directly damages the coupling complex. The PS II inactivation in leaves and algae cells may be a result of the oxidative stress processes.     

Non-significance of rhizosphere degradation during phytoremediation of MTBE

Methyl tertiary butyl ether (MTBE) is a gasoline additive associated with groundwater pollution at gas station sites. Previous research on poplar trees in hydroponic systems suggests that phytovolatilization is an effective mechanism for phytoremediation of MTBE (Rubin and Ramaswami, 2001), but the potential for microbial degradation of MTBE in the rhizosphere of trees had not been assessed. MTBE had largely been considered recalcitrant to microbial processes, but recent fieldwork suggests rapid biodegradation may occur in certain cases. This paper investigates the potential for rhizosphere degradation of MTBE at time frames relevant for phytoremediation. Three experiments were conducted at different levels of aggregation to examine possible degradation of MTBE by rhizosphere microorganisms that had been acclimated to low levels of MTBE for 6 weeks. MTBE soil die-away studies, conducted with both poplar trees and fescue grass, found no significant differences between MTBE concentration in vegetated and unvegetated soils over a two-week attenuation period. Closed chamber tests comparing hydroponic and rhizospheric poplar tree systems also showed essentially complete recovery of MTBE mass in both systems, suggesting an absence of degradation. Finally, rhizosphere microbes tested in aerated bioreactors were found to be thriving and metabolizing root materials, but did not show measurable degradation of MTBE. In all tests, the MTBE degradation product, Tert Butyl Alcohol (TBA), was not detected. The insignificance of MTBE degradation by rhizosphere microorganisms suggests that plant processes be the primary focus of further research on MTBE phytoremediation.     

Interfacial versus homogeneous enzymatic cleavage of mandelonitrile by hydroxynitrile lyase in a biphasic system

The question of an interfacial versus a homogeneous reaction is carefully addressed for the enzymatic biphasic cleavage of mandelonitrile to benzaldehyde by Prunus amygdalus hydroxynitrile lyase (pa-Hnl) (Hickel et al. [1999] Biotechnol Bioeng 36:425-436). Experimental evidence, including 1) the reaction ceases when the interface is populated by previously adsorbed denatured pa-Hnl, 2) the reaction continues even after washout of the bulk enzyme from the aqueous phase, 3) highly nonpolar organic solvents initially promote fast reaction kinetics that relatively quickly decay to zero product production, and 4) the reaction rate is nonlinear in the bulk enzyme concentration, provide robust grounds for an interfacial reaction. We also model enzymatic mandelonitrile cleavage assuming a homogeneous aqueous-phase reaction. The homogeneous reaction scheme does not simultaneously account for the experimental observations of a linear dependence of the reaction rate on organic/water interfacial area, no dependence on the aqueous-phase volume, and a nonlinear dependence on pa-Hnl aqueous concentration. Further, simple calculations demonstrate that the homogeneous reaction rate is at least three orders of magnitude slower than those observed by Hickel et al. (1999). We again conclude that enzyme adsorbed at the organic solvent/water interface primarily catalyzes the biphasic mandelonitrile cleavage reaction.     

Detailed glycan analysis of serum glycoproteins of patients with congenital disorders of glycosylation indicates the specific defective glycan processing step and provides an insight into pathogenesis

The fundamental importance of correct protein glycosylation is abundantly clear in a group of diseases known as congenital disorders of glycosylation (CDGs). In these diseases, many biological functions are compromised, giving rise to a wide range of severe clinical conditions. By performing detailed analyses of the total serum glycoproteins as well as isolated transferrin and IgG, we have directly correlated aberrant glycosylation with a faulty glycosylation processing step. In one patient the complete absence of complex type sugars was consistent with ablation of GlcNAcTase II activity. In another CDG type II patient, the identification of specific hybrid sugars suggested that the defective processing step was cell type-specific and involved the mannosidase III pathway. In each case, complementary serum proteome analyses revealed significant changes in some 31 glycoproteins, including components of the complement system. This biochemical approach to charting diseases that involve alterations in glycan processing provides a rapid indicator of the nature, severity, and cell type specificity of the suboptimal glycan processing steps; allows links to genetic mutations; indicates the expression levels of proteins; and gives insight into the pathways affected in the disease process.     

Complementarity of genes for resistance to greenbug [Schizaphis graminum (Rondani)], biotype E,in sorghum [Sorghum bicolor (L.) Moench]

Summary Gene complementarity among various sources of resistance to greenbug biotype E was assessed. Analysis of the F₂ generation of crosses between susceptible and resistant parents (mating 1) and among sources of resistance (mating 2) suggested that resistance in sorghum to greenbug biotype E was complexly inherited and, to some extent, dependent on the nature of both the resistant and susceptible parents. Positive transgressive segregation in the F₂ generations of both matings was found to be due to effective plus factors, contributed by both parents in a cross, which complemented each other. The number of plus factors ranged from one to two in the susceptible parents and from two to five in the resistant parents of mating 1, and from one to five in the parents of mating 2. The consistently significant reciprocal effects shown by Sarvasi and PI264453 indicated that these sources had major factors for resistance in their cytoplasms, which were expressed in all their crosses. The results from this study indicated that the sources of resistance complemented each other to give increased number of F₂ segregates with increased resistance. Thus, it should be possible to increase and diversify resistance of sorghum to greenbug biotype E by accumulating different, effective plus factors from various sources through recurrent selection.Contribution No. 90-106-J from the Kansas Agricultural Experiment Station, Kansas State University, Manhattan, KS 66506, USA     

The folate pathway is a target for resistance to the drug para-aminosalicylic acid (PAS) in mycobacteria

The increasing rate of multidrug-resistant tuberculosis has led to more use of second-line antibiotics such as para-aminosalicylic acid (PAS). The mode of action of PAS remains unclear, and mechanisms of resistance to this drug are undefined. We have isolated PAS-resistant transposon mutants of Mycobacterium bovis BCG with insertions in the thymidylate synthase (thyA) gene, a critical determinant of intracellular folate levels. BCG thyA mutants have reduced thymidylate synthase activity and are resistant to known inhibitors of the folate pathway. We also find that mutations in thyA are associated with clinical PAS resistance. We have identified PAS-resistant Mycobacterium tuberculosis isolates from infected patients, which harbour mutations in thyA and show reduced activity of the encoded enzyme. Thus, PAS acts in the folate pathway, and thyA mutations probably represent a mechanism of developing resistance not only to PAS but also to other drugs that target folate metabolism.     

Host Specificity of Salmonella typhimurium Deoxyribonucleic Acid Restriction and Modification

The restriction and modification genes of Salmonella typhimurium which lie near the thr locus were transferred to a restrictionless mutant of Escherichia coli. These genes were found to be allelic to the E. coli K, B, and A restriction and modification genes. E. coli recombinants with the restriction and modification host specificity of S. typhimurium restricted phage λ that had been modified by each of the seven known host specificities of E. coli at efficiency of plating levels of about 10⁻². Phage λ modified with the S. typhimurium host specificity was restricted by six of the seven E. coli host specificities but not by the RII (fi⁻ R-factor controlled) host specificity. It is proposed that the restriction and modification enzymes of this S. typhimurium host specificity have two substrates, one of which is a substrate for the RII host specificity enzymes.