Leishmania infantum Nicolle, 1908 from southern Spain: Characterisation of the strains from human visceral and cutaneous leishmaniasis and from sandflies; with a numerical analysis of the isoenzymatic data

This study presents the results of the characterisation of 17 strains ofLeishmania by isoenzyme electrophoresis from a focus of leishmaniasis in southern Spain: two from human visceral leishmaniasis, four from human cutaneous leishmaniasis and 11 from sandflies. The 17 strains are grouped in 6 zymodemes characterised by their variability as regards to the electrophoretic mobility of the enzymes MDH, G6PD, NP and ME. Thus, we confirm the high intraspecific variability ofLeishmania (L.) infantum in a focus of southern Spain, as already suggested by previous studies. Zymodemes GR-15 and GR-17 are also described for the first time in Spain, and they characteristically possess the same relative electrophoretic mobility in the enzyme ME (93). Sixteen zymodemes of theL. infantum complex found in southern Spain were numerically analysed on the basis of the enzymatic profiles of 122Leishmania strains characterised from this area.     

Higher-plant chloroplast and cytosolic 3-phosphoglycerate kinases: a case of endosymbiotic gene replacement

Previous studies indicated that plant nuclear genes for chloroplast and cytosolic isoenzymes of 3-phosphoglycerate kinase (PGK) arose through recombination between a preexisting gene of the eukaryotic host nucleus for the cytosolic enzyme and an endosymbiont-derived gene for the chloroplast enzyme. We readdressed the evolution of eukaryotic pgk genes through isolation and characterisation of a pgk gene from the extreme halophilic, photosynthetic archaebacterium Haloarcula vallismortis and analysis of PGK sequences from the three urkingdoms. A very high calculated net negative charge of 63 for PGK from H. vallismortis was found which is suggested to result from selection for enzyme solubility in this extremely halophilic cytosol. We refute the recombination hypothesis proposed for the origin of plant PGK isoenzymes. The data indicate that the ancestral gene from which contemporary homologues for the Calvin cycle/glycolytic isoenzymes in higher plants derive was acquired by the nucleus from (endosymbiotic) eubacteria. Gene duplication subsequent to separation of Chlamydomonas and land plant lineages gave rise to the contemporary genes for chloroplast and cytosolic PGK isoenzymes in higher plants, and resulted in replacement of the preexisting gene for PGK of the eukaryotic cytosol. Evidence suggesting a eubacterial origin of plant genes for PGK via endosymbiotic gene replacement indicates that plant nuclear genomes are more highly chimaeric, i.e. contain more genes of eubacterial origin, than is generally assumed.Abbreviations PGK 3-phosphoglycerate kinase - FBA fructose-1,6-bisphosphate aldolase - GAPDH glyceraldehyde-3-phosphate dehydrogenase - TPI triosephosphate isomerase     

Early ordovician reefs from Argentina: Stromatoporoid vs stromatolite origin

Summary Late Arenigian biohermal reef mounds and biostromes within the shallow-marine platform facies of the upper San Juan Formation of the Precordillera (Western Argentina) represent a new Early Ordovician reef type. The meter-sized reefs are dominated byZondarella communis n.g. n. sp. The new taxon is characterized by domical, bulbous and laminar morphotypes exhibiting growth layers and thin horizontal and vertical as well as intermingled skeletal elements included within different sets. The fossil maybe compared with stromatolites and stromatoporoids but an interpretation as primitive stromatoporoids is favoured.     

Evaluation of three herbaceous index plant species for bioavailability of soil cadmium, chromium, nickel and vanadium

Uncultivated plants growing on disturbed sites may be useful for assessing the bioavailability of some metals in soils, and thus the potential for metal mobilization up the terrestrial food chain, an important element in ecological risk assessment. A planted chicory cultivar (Cichorium intybus L. var. foliosum Hegi.) and the uncultivated plants horseweed (Canada fleabane) (Erigeron canadensis L.) and dogfennel (Eupatorium capillifolium (Lam.) Small) were evaluated for their ability to act as index plant species for soil Cd, Cr, Ni, and V at two field sites where these metals had been applied five yr previously to two highly weathered sandy Ultisols. Soil Cd was available to all analyzed plant tissues of all three plant species at both sites, particularly on the sandier Blanton soil. Chicory was an effective index plant for Cd on the finer textured Orangeburg soil but functioned as an indicator plant (toxicity symptoms were observed) on the sandier Blanton soil. Horseweed and dogfennel were effective index plants for Cd in both contaminated soils. Soil Cr, Ni, and V were less bioavailable than soil Cd and plant metal uptake was more sensitive to residual soil Cr, Ni, and V than was soil extraction with double acid. Horseweed and chicory may have potential as index plants for soil Cr. Chicory may have potential as a Ni index plant. Chicory and dogfennel may have potential as V index plants.     

Immunochemical comparison of membrane-associated and secreted arabinogalactan-proteins in rice and carrot

Arabinogalactan-proteins (AGPs) occurring in suspension-cultured rice (Oryza saliva L.) cells, their conditioned medium and at the rice root apex were investigated using monoclonal antibodies and the AGP-binding -glucosyl Yariv reagent ( GlcY). A monoclonal antibody, LM2, was generated that recognized an acidic carbohydrate epitope common to two soluble AGPs occurring in the conditioned medium of proliferating rice cells, membrane-associated AGPs (rmAGP) in the cultured cells and two AGPs at the rice root apex. In addition, LM2 recognized AGPs secreted by suspensioncultured carrot (Daucus carota L.) cells. The two AGPs of the rice culture medium, srAGP1 and srAGP2, were discriminated by their mobilities during sodium dodecyl sulfate-polyacrylamide gel electrophoresis, reaction with GlcY, the presence of arabinogalactan epitopes and anion-exchange chromatography. The association of rmAGP with the plasma membrane was investigated by Triton-X-114/aqueous partitioning of both microsomal and plasma-membrane preparations and rmAGP was found to partition into the detergent phase, indicating that AGPs are hydrophobic plasma-membrane proteins in rice. This was in contrast to plasma-membrane AGPs of suspension-cultured carrot cells that partitioned into the aqueous phase. At the rice root apex most of the AGP was associated with the microsomal fraction and also partitioned into the detergent phase, although a distinct highmolecular-mass AGP entered the aqueous phase.Abbreviations AGP arabinogalactan-protein - GlcY -glucosyl Yariv reagent - ELISA enzyme-linked immunosorbent assay We gratefully acknowledge support from the Leverhulme Trust, the UK Biotechnology and Biological Sciences Research Council and the Royal Society.     

Development of PCR markers linked to resistance to wheat streak mosaic virus in wheat

Wheat streak mosaic virus (WSMV), vectored by the wheat curl mite (Acer tulipae), is an important disease of wheat (Triticum aestivum L.) in the North American Great Plains. Resistant varieties have not been developed for two primary reasons. First, useful sources of resistance have not been available, and second, field screening for virus resistance is laborious and beyond the scope of most breeding programs. The first problem may have been overcome by the development of resistance to both the mite and the virus by the introgression of resistance genes from wild relatives of wheat. To help address the second problem, we have developed polymerase chain reaction (PCR) markers linked to the WSMV resistance gene Wsm1. Wsm1 is contained on a translocated segment from Agropyron intermedium. One sequence-tagged-site (STS) primer set (WG232) and one RAPD marker were found to be linked to the translocation containing Wsm1. The diagnostic RAPD band was cloned and sequenced to allow the design of specific PCR primers. The PCR primers should be useful for transferring Wsm1 into locally adapted cultivars.This is Journal Series No. J-4041 of the Montana Agricultural Experiment Station     

A testcross procedure for selecting exotic strains to improve pure-line cultivars in predominantly self-fertilizing species

Methods for identifying germplasm carrying alleles with the potential to improve a particular single-cross hybrid have been proposed and discussed in recent years. There is a need for similar methods to be used in breeding crops for which pure-line cultivars, rather than hybrids, are the goal. The objective of this research was to develop a method to identify germplasm lines with the potential to contribute favorable alleles not present in a specified pure line or set of pure lines. Given a set of adapted pure lines (A ₁, A ₂ ..., A _m) to be improved and a set of germplasm lines (P ₁ P ₂ ..., P _f), the procedure consists of producing all f x m possible hybrids and evaluating them along with the parents. The testcross statistic T _ij is defined by T _ij=(F _ij–A _j)+(1–) (F _ij–P _i), where A _j, P _i, and F _ij represent the performance of thej ^th adapted line, the i ^th germplasm line, and their hybrid, respectively. The statistic is the mean value of T _ij over all adapted parents A _j. If =(1/2)(1+d), where d = the mean degree of dominance, then T _ij measures the potential for alleles from P _i to improve A _j and measures the potential for alleles from P _i to improve the set A ₁, A ₂ ..., A _m. Use of data on soybean and peanut hybrids published by other researchers suggests that the value assumed for d has little effect on the P _i chosen. The ability of the T _ij and statistics to identify germplasm strains carrying rare favorable alleles should be assessed in empirical studies.Joint contribution: OARDC (Journal Articale No. 161-94), USDAARS, Iowa Agriculture and Home Economics Expriment Station (Journal Paper No. J-16109; Project 2985), and Agreculture and Agri-Food Canada. Salaries and research support for S. K. St. Martin Provided by state and federal funds appropriated to the Ohio Agricultural Research and Development Center, Ohio State University     

Solubility of sickle hemoglobin measured by a kinetic micromethod.

We have developed a photolytic method to determine the concentration of reactive hemes in a solution in the presence of a trace amount of CO. By measurement of the bimolecular rate of CO binding, and by calibration of the rate constant under equivalent conditions, the concentration of the reactive hemes can be determined. In a solution of sickle hemoglobin, the molecules in the gel contribute negligibly to the recombination rate, allowing the concentration of the molecules in the solution phase to be determined. To optimize signal to noise, modulated excitation methods were employed, although the method could also be used with pulse techniques and suitable signal averaging. Because the optical method employs a microspectrophotometer, only a few microliters of concentrated Hb solution is required to reproduce the entire temperature dependence of the solubility previously determined by centrifugation using milliliter quantities of solutions of the same concentration. This should be especially useful for studies of site-directed mutants, and we present results obtained on one such HbS in which Leu 88 beta has been replaced by Ala. The free energy difference in the polymerization of the Leu 88 beta double mutant is consistent with known differences in the amino acid hydrophobicities. The calibration required for these experiments also provides an excellent determination of the activation energy for binding the first CO to deoxy Hb.