Characterization of salt stress-enhanced phosphoenolpyruvate carboxylase kinase activity in leaves of <Emphasis Type="Italic">Sorghum</Emphasis><Emphasis Type="Italic">vulgare</Emphasis>: independence from osmotic stress,involvement of ion toxicity and significance of dark phosphorylation

C(4) phosphoenolpyruvate carboxylase (PEPCase: EC 4.1.1.31) is subjected to in vivo regulatory phosphorylation by a light up-regulated, calcium-independent protein kinase. Salt stress greatly enhanced phosphoenolpyruvate carboxylase-kinase (PEPCase-k) activity in leaves of Sorghum. The increase in PEPCase-k anticipated the time course of proline accumulation thereby suggesting that water stress was not involved in the kinase response to salt. Moreover, osmotic stress seemed not to be the main factor implicated, as demonstrated by the lack of effect when water availability was restricted by mannitol. In contrast, LiCl (at a concentration of 10 mM in short-term treatment of both excised leaves and whole plants) mimicked the effects of 172 mM NaCl salt-acclimation, indicating that the rise in PEPCase-k activity resulted primarily from the ionic stress. Both NaCl and LiCl treatments increased the activity of a Ca(2+)-independent, 35 kDa kinase, as demonstrated by an in-gel phosphorylation experiment. Short-term treatment of excised leaves with NaCl or LiCl partially reproduces the effects of whole plant treatments. Finally, salinization also increased PEPCase-k activity and the phosphorylation state of PEPCase in darkened Sorghum leaves. This fact, together with increased malate production during the dark period, suggests a shift towards mixed C(4) and crassulacean acid metabolism types of photosynthesis in response to salt stress.     

Characterization of sterol uptake in leaf tissues of sugar beet

The uptake of cholesterol has been characterized in leaf discs from mature leaves of sugar beet (Beta vulgaris L.). This transport system exhibited a simple saturable phase with an apparent Michaelis constant ranging from 30 to 190 M depending on the sample. When present at 10 M excess, other sterols were able to inhibit cholesterol uptake. Moreover, binding assays demonstrated the presence of high-affinity binding sites for cholesterol in purified plasma membrane vesicles. In the range 1–60 M, cholesterol uptake showed an active component evidenced by action of the protonophore carbonyl cyanide m-chlorophenylhydrazone. Energy was required as shown by the inhibition of uptake induced by respiration inhibitors (NaN₃), darkness and photosynthesis inhibitors [3-(3,4-dichlorophenyl)-1,1-dimethylurea, methyl viologen]. Moreover, the process was strongly dependent on the experimental temperature. Uptake was optimal at acidic pH (4.0), sensitive to ATPase modulators, inhibited by thiol reagents (N-ethylmaleimide, p-chloromercuribenzenesulfonic acid, Mersalyl) and by the histidyl-group reagent diethyl pyrocarbonate. The addition of cholesterol did not modify H⁺ flux from tissues, indicating that H⁺-co-transport was unlikely to be involved. MgATP did not increase the uptake, arguing against involvement of an ABC cassette-type transporter. By contrast, cryptogein, a sterol carrier protein from the Oomycete Phytophtora cryptogea, greatly increased absorption. Taken together, the results reported in this work suggest that plant cells contain a specific plasma membrane transport system for sterols.Abbreviations CCCP carbonyl cyanide m-chlorophenylhydrazone - PCMBS p-chloromercuribenzenesulfonic acid - PMV plasma membrane vesicle - TLC thin-layer chromatography     

Inflammatory reaction and changes in expression of coagulation proteins on lung endothelial cells after total-body irradiation in mice

Inflammatory reaction is a classical feature of radiation exposure, and pneumonitis is a dose-limiting complication in the handling of hematological disorders treated with total-body irradiation. In the present study, we first evaluated the inflammatory response in C57BL6/J mice exposed to lethal doses of gamma rays treated with antibiotics or not. Both interleukin 6 and KC (also known as Gro1) were increased in the plasma 10 to 18 days after radiation exposure, independent of bacterial infection, whereas fibrinogen release was linked to a bacterial infection. Furthermore, both Il6 and KC were increased in the lungs of irradiated mice. Our second objective was to characterize the endothelial cell changes in the lungs of total-body-irradiated mice. For this purpose, a quantitative RT-PCR was used to determine the expression of genes involved in inflammatory and coagulation processes. We found that the adhesion molecules P-selectin and platelet endothelial cell adhesion molecule 1 were up-regulated, whereas E-selectin remained unchanged. Tissue factor expression was up-regulated as well, and thrombomodulin gene expression was down-regulated. The investigation by immunohistochemistry of adhesion molecules confirmed the increase in the basal expression of both P-selectin and platelet endothelial cell adhesion molecule 1 on pulmonary endothelial cells. All together, our results suggest the involvement of endothelial cells in the development of radiation-induced inflammatory and thrombotic processes.     

Anatomy and uses of the mature leaves of three species of Sabal (Arecaceae) of the Yucatan, México

This paper describes the leaf anatomy of Sabal mauritiiformis (Karst.) Griseb. & H. Wendl., Sabal mexicana Mart. and Sabal yapa Wright ex Becc., three of the four most representative species of the Yucatán Península, in Mexico. These species are locally used: in the roofing of traditional homes, as food (fruits and apical buds), and in the production of hats, brooms and handicrafts. Leaf samples were collected in secondary growth of lower montane rainforest in the state of Quintana Roo and in two home gardens in the state of Yucatán. Herbarium samples were obtained, and samples of blade and petiole were fixed in formaline-acetic acid-alcohol. Cross incisions were made on the blade and petiole, and were dyed with safranin and toluidine blue O. The results show that S. mauritiiformis and S. yapa are morphologically alike: both are tall, slim palm trees; the leaf in S. mauritiiformis is a shorter palm-like structure compared with the other two species. The shape of the main nerve, as seen in cross section, is rectangular in the three species. The hastula in the three species is acuminate and adaxial. The foliar anatomic structure is similar in the three species, although there are some differences. The adaxial an abaxial epidermis of the blade consist of one layer and, superficially, the anticlinal walls are straight; the stomata are intercostal, of the tetracytic type, present on both surfaces in S. mexicana and S. yapa and only on the abaxial surface on S. mauritiiformis. The hypodermis is one layer thick in S. yapa and in S. mexicana and two layers thick in S. mauritiiformis. In the three species the palisade parenchyma consists of several undefined strata as the cells are similar-in shape and size--to the cells in the spongy parenchyma, so there is no marked difference between these strata and the spongy parenchyma seems almost continuous. Both fibrous and vascular bundles are distributed between the hypodermis and the palisade parenchyma; the fiber bundles can be found towards the abaxial surface while the vascular and fiber bundles are located towards the adaxial surface. The fibers, in the three species, are elongated, with the pointed tips, undivided and unseptated. One to three wide vessels of metaxilem can be seen in the vascular bundles, those in S. yapa being the widest in diameter. The vascular bundles are surrounded by thick fiber sheaths which come in pairs. The anatomic structure of the petiole is similar to that of the blade, and is characterized by the many vascular and fiber bundles dispersed in the parenchymatous tissue, and which are very resistant. The histological structure of the blade and petiole reflects strength and flexibility, qualities which make these plants adequate in the construction of roofs for rural housing and other buildings.     

Genetic and morphological characteristics of the populations of Cyprinodon dearborni (Atherinomorpha: Cyprinodontidae) in Chacopata and Laguna de los Patos, Venezuela

Two populations of the fish Cyprinodon dearborni were compared, one from Laguna de Chacopata and the other from Laguna de los Patos, Cumaná, Venezuela. The morphometric method of the Box Truss was used; the organisms were photographed with a digital camera. In the images 18 measures were made to each organism, with the MOCHA 1.2 program. The tendencies of morphologic variation among populations were analyzed by main components using STATGRAPHICS 2.0 and SHEAR programs. The first four main components in males and females explained 88% of the total variation. The components two, three and four separate the two populations based on the body form of the animals. For the electrophoretic study, 13 enzymatic systems were analyzed in starch gels. The variation and genetic distance were calculated with the program GENES 2.0. Differences were observed in the systems G6PDH, GPI; ES and GLUD, that which can be used as isoenzimatic markers. Genetic diversity of Chacopata (H = 0.086, p = 0.079) was higher than in Laguna de los Patos (H = 0.029, p = 0.028), but both are within the average in populations of fish. The two populations presented a Nei distance of 0.058. The genetic and morphologic differentiation suggests an incipient process of separation whose causes could be the ecological conditions of their habitat, or processes of vicariant speciation.     

Mapping QTLs for resistance against <Emphasis Type="Italic">Globodera pallida</Emphasis> (Stone) Pa2/3 in a diploid potato progeny originating from <Emphasis Type="Italic">Solanum spegazzinii</Emphasis>

A "F1" diploid population between Solanum tuberosum 2 x and the wild Solanum spegazzinii was studied. It segregated for resistance against the potato cyst nematode Globodera pallida derived from the wild species. The inheritance had a quantitative nature. Linkage maps of AFLP and RFLP markers were constructed for both parents. Three QTLs were identified on the map of the resistant parent on chromosomes V, VI and XII, respectively. The first one had a major effect and explained more than 50% of the total variance of resistance. It is located in a cluster of resistance genes and may be the same locus as Gpa which has been described formerly. The two others explained about 20% of the total variance each. The QTL on chromosome XII is also in a cluster of resistance genes, and in an orthologous position with resistance genes against nematodes in tomato and pepper.     

Competing populations in flows with chaotic mixing

We investigate the effects of spatial heterogeneity on the coexistence of competing species in the case when the heterogeneity is dynamically generated by environmental flows with chaotic mixing properties. We show that one effect of chaotic advection on the passively advected species (such as phytoplankton, or self-replicating macro-molecules) is the possibility of coexistence of more species than that limited by the number of niches they occupy. We derive a novel set of dynamical equations for competing populations.