QTL analysis of cleistogamy in soybean

Early-maturing cultivars of soybean [Glycine max (L.) Merr.] native to the shores of the Sea of Okhotsk (Sakhalin and Kuril Islands) and eastern Hokkaido (northern Japan) have a strong tendency to produce cleistogamous flowers throughout their blooming period. A previous study revealed that cleistogamy is controlled by a minimum of two genes with epistatic interaction, one of which is associated with a maturity gene responsible for insensitivity to incandescent long daylength (ILD). This study was conducted to determine the genetic basis of cleistogamy in more detail by QTL mapping. F(2) to F(4) progenies derived from a cross between a cleistogamous cv. Karafuto-1 and a chasmogamous cv. Toyosuzu were used. A molecular linkage map spanning 2,180 cM comprising 500 markers was constructed using 89 F(2) plants. The markers were distributed in 25 linkage groups. An interval mapping method to analyze categorical traits identified four QTLs for cleistogamy, cl1, cl2, cl3 and cl4, in molecular linkage groups (MLGs) C2, D1a, I and L, respectively. Alleles derived from Karafuto-1 had additive effects to increase probability of cleistogamy at cl3 and cl4, whereas the alleles had additive effects to decrease the probablity at cl1 and cl2. Progeny test confirmed the effects of cl3, which had the highest LOD score (5.20). Composite interval mapping revealed four QTLs for flowering date, fd5-fd8. Judging from relative location with markers and association with ILD responses, fd7 and fd8 may correspond to maturity genes E4 and E3, respectively. cl3 and cl4 were located at similar positions as fd7 and fd8, suggesting that the two maturity genes may control cleistogamy by either pleiotropy or close linkage.     

A Hierarchical Approach to Cooperativity in Macromolecular and Self-Assembling Binding Systems

The study of complex macromolecular binding systems reveals that a high number of states and processes are involved in their mechanism of action, as has become more apparent with the sophistication of the experimental techniques used. The resulting information is often difficult to interpret because of the complexity of the scheme (large size and profuse interactions, including cooperative and self-assembling interactions) and the lack of transparency that this complexity introduces into the interpretation of the indexes traditionally used to describe the binding properties. In particular, cooperative behaviour can be attributed to very different causes, such as direct chemical modification of the binding sites, conformational changes in the whole structure of the macromolecule, aggregation processes between different subunits, etc. In this paper, we propose a novel approach for the analysis of the binding properties of complex macromolecular and self-assembling systems. To quantify the binding behaviour, we use the global association quotient defined as K _c = [occupied sites]/([free sites] L), L being the free ligand concentration. K _c can be easily related to other measures of cooperativity (such as the Hill number or the Scatchard plot) and to the free energies involved in the binding processes at each ligand concentration. In a previous work, it was shown that K_c could be decomposed as an average of equilibrium constants in two ways: intrinsic constants for Adair binding systems and elementary constants for the general case. In this study, we show that these two decompositions are particular cases of a more general expression, where the average is over partial association quotients, associated with subsystems from which the system is composed. We also show that if the system is split into different subsystems according to a binding hierarchy that starts from the lower, microscopic level and ends at the higher, aggregation level, the global association quotient can be decomposed following the hierarchical levels of macromolecular organisation. In this process, the partial association quotients of one level are expressed, in a recursive way, as a function of the partial quotients of the level that is immediately below, until the microscopic level is reached. As a result, the binding properties of very complex macromolecular systems can be analysed in detail, making the mechanistic explanation of their behaviour transparent. In addition, our approach provides a model-independent interpretation of the intrinsic equilibrium constants in terms of the elementary ones.     

Short and long-term uptake of Hg in white lupin plants:Kinetics and stress indicators

The intrinsic characteristics of white lupin regarding biomass production and tolerance to abiotic stresses could make it a good candidate to be used in degraded mine soils containing mercury (Hg), but white lupin behaviour in response to Hg has to be previously evaluated. With this aim, kinetic parameters of Hg uptake in short and long-term experiments, and Hg resistance of white lupin plants using several stress indicators were studied. The plants were grown with increasing Hg doses in nutrient solutions (0, 5 and 10 μM). Hg uptake showed an active component in Hg influx, suggesting the existence of a low affinity root transporter that can be used for Hg uptake into white lupin root cells. K_m and V_max values obtained for the saturable component were 217.7 ± 27.6 μM and 3.78 ± 0.18 μmol Hg g FW^?1 h^?1. Hg accumulation was concentration–time-dependent, showing a saturable behaviour for the lower doses but a linear behaviour for the highest ones. A high ability of Hg absorption by white lupin was observed both in short and long-term uptake experiments. The highest Hg dose reduced biomass production especially in the shoots. Moreover, increases in chlorophylls, malondialdehyde, total thiols and phenols were observed in Hg-stressed plants. The enhancement of total thiols and phenols levels in roots reduced oxidative stress for the 5 μM dose, but not for higher Hg levels. The deleterious effect of Hg was less marked in root tissues, in spite of their accumulation of very high Hg amounts (99%) because of, at least in part, a combined increase in total thiols and phenolics able to minimize oxidative stress. Our results suggested that phenolic content in roots could be used as a new and easy-to-use indicator of Hg presence. On the whole, white lupin showed a certain ability to survive in Hg-contaminated media and it would be possible to include it in combined decontamination strategies.     

Microalgal adaptation to a stressful environment (acidic,metal-rich mine waters) could be due to selection of pre-selective mutants originating in non-extreme environments

At least six species of eukaryotic microalgae inhabit the acidic (pH 2.4–2.7), metal-rich mine waters from ponds in the copper mine district of Mynydd Parys (N Wales, UK). Consequently, these ponds constitute interesting natural laboratories for analysis of adaptation by microalgae to extremely stressful conditions. To distinguish between the pre-selective and post-selective origin of adaptation processes that allow the existence of microalgae in these ponds, a Luria-Delbrück fluctuation analysis was performed with the chlorophycean Dictyosphaerium chlorelloides isolated from non-acidic waters. In this analysis, natural Mynydd Parys pond water (MPW) was used as selective factor. Pre-selective, resistant D. chlorelloides cells appeared with a frequency of 1.6 × 10^?6 per cell per generation. MPW-resistant mutants, with a diminished Malthusian fitness, are maintained in non-extreme waters as the result of a balance between new MPW-resistant cells arising by mutation and MPW-resistant mutants eliminated by natural selection (equilibrium at ca. 19 MPW-resistant per 10⁷ wild-type cells). We propose that the microalgae inhabiting these stressful ponds could be the descendents of chance mutants that arrived in the past or are even arriving at the present.     

Changes in calcium-dependent membrane permeability properties in mitochondria of livers from arthritic rats

The involvement of the mitochondrial permeability transition pore (PTP) in the responses of mitochondria from adjuvant-induced arthritic rats to Ca(2+) addition was investigated. The respiratory activity, the Ca(2+)-induced osmotic swelling and the electrophoretic (45)Ca(2+) uptake were evaluated in the absence and in the presence of cyclosporin A (CsA), a well-known inhibitor of the mitochondrial PTP. The Ca(2+)-induced mitochondrial permeability transition (MPT) process occurred in mitochondria from arthritic rats even in the presence of a low Ca(2+) concentration. Whereas in the normal condition, the Ca(2+)-induced uncoupling of oxidative phosphorylation and osmotic swelling was observed in the presence of 10 or 20 microM Ca(2+) concentration, in the arthritic condition, these events occurred at 1.0 microM concentration. In addition, mitochondria from arthritic rats presented an impaired ability to accumulate (45)Ca(2+). All these effects were completely prevented by the administration of CsA. The results of the present study suggest that the higher sensitivity of mitochondria from arthritic rats to Ca(2+)-induced MPT may be an important factor in the pathogenesis of the arthritis disease.     

Positive feedback in cellular control systems

Feedback loops have been identified in a variety of regulatory systems and organisms. While feedback loops of the same type (negative or positive) tend to have properties in common, they can play distinctively diverse roles in different regulatory systems, where they can affect virulence in a pathogenic bacterium, maturation patterns of vertebrate oocytes and transitions through cell cycle phases in eukaryotic cells. This review focuses on the properties and functions of positive feedback in biological systems, including bistability, hysteresis and activation surges.     

A mortality survey of free range nutria (Myocastor coypus)

A review of the literature revealed little information on natural occurring diseases in wild nutria. In this report, a summary of necropsies performed on free-range animals from four different geographical areas, is presented. Fifty-two percent of the nutria had trauma (mostly by predation and road kill), 15% had poisoning by different toxics, and 11% had starvation. The rest died due to infectious diseases and miscellaneous causes, while 21 individuals had no significant lesions. The occurrence of infections seems sporadic with a far lower prevalence than in the farmed animals, while the incidence of poisoning is rather high. In addition, anthrax was diagnosed in two individuals. Thus, nutria are probably subject to mortality from a number of different human-induced causes rather than natural ones. Analysis of these records may provide insight into prevention of problem and better management practices.     

A new record and an evaluation of the phylogenetic relationships of Abrothrix olivaceus markhami (Rodentia: Sigmodontinae)

Recent phylogenetic studies of the subfamily of cricetid rodents (Sigmodntinae) have validated the taxonomic classification at the tribal level of the Andean Clade. It is possible that some endemic species from Patagonian South America are part of this new tribe, but previous studies have not evaluated this hypothesis due to the difficulty of obtaining samples. In this study, we evaluate the phylogenetic relationships of Akodon markhami (Pine, 1973), a species endemic to the Chilean Patagonia, using individuals recently captured at the type locality of this taxon. Our results indicate that this form of Akodon corresponds to a subspecies of Abrothrix olivaceus, and should be incorporated into the Andean Clade as a geographic race of this widely distributed species on the South American continent. Based on a molecular clock calibration, we dated the origin of this geographic race during the last glacial cycles of the Quaternary, as the result of a vicariant process.     

Metabolically engineered Escherichia coli for efficient production of glycosylated natural products

Significant achievements in polyketide gene expression have made Escherichia coli one of the most promising hosts for the heterologous production of pharmacologically important polyketides. However, attempts to produce glycosylated polyketides, by the expression of heterologous sugar pathways, have been hampered until now by the low levels of glycosylated compounds produced by the recombinant hosts. By carrying out metabolic engineering of three endogenous pathways that lead to the synthesis of TDP sugars in E. coli, we have greatly improved the intracellular levels of the common deoxysugar intermediate TDP‐4‐keto‐6‐deoxyglucose resulting in increased production of the heterologous sugars TDP‐L‐mycarose and TDP‐d ‐desosamine, both components of medically important polyketides. Bioconversion experiments carried out by feeding 6‐deoxyerythronolide B (6‐dEB) or 3‐α‐mycarosylerythronolide B (MEB) demonstrated that the genetically modified E. coli B strain was able to produce 60‐ and 25‐fold more erythromycin D (EryD) than the original strain K207‐3, respectively. Moreover, the additional knockout of the multidrug efflux pump AcrAB further improved the ability of the engineered strain to produce these glycosylated compounds. These results open the possibility of using E. coli as a generic host for the industrial scale production of glycosylated polyketides, and to combine the polyketide and deoxysugar combinatorial approaches with suitable glycosyltransferases to yield massive libraries of novel compounds with variations in both the aglycone and the tailoring sugars.