Partial purification and biochemical properties of acid and alkaline phosphatases from Myxococcus coralloides D

F. GONZÁLEZ, M.E. FÁREZ-VIDAL, J.M. ARIAS AND E. MONTOYA. 1994. Acid phosphatase and alkaline phosphatase from vegetative cells of Myxococcus coralloides D were purified by two chromatographic steps. The molecular weights were estimated by gel filtration and SDS-PAGE. Optimum pH, stability, optimum temperature and thermal inactivation studies were made for both enzymes. EDTA and other chelating agents inhibited alkaline but not acid activity. Mg²⁺ activated the alkaline phosphatase, while the acid phosphatase was inhibited by fluoride. Both enzymes degraded a number of phosphomonoesters, but were unable to hydrolyse either polyphosphates or cAMP. The K _m values of the acid and alkaline phosphatases for p -nitrophenylphosphate were 5.0 times 10^-3 mol ***l^-1 and 1.5 times 10^-3 mol l^-1, respectively.     

The intermediary cell: Minor-vein anatomy and raffinose oligosaccharide synthesis in the Scrophulariaceae

Minor-vein anatomy, sugar content, sugar synthesis, and translocation were studied in mature leaves of nine members of the Scrophulariaceae to determine if there is a correlation between companion-cell type and class of sugar translocated. Three types of companion cell were found: intermediary cells with extensive plasmodesmatal connections to the bundle sheath; transfer cells with wall ingrowths and few plasmodesmata; and ordinary companion cells with few plasmodesmata and no wall ingrowths. Alonsoa warscewiczii Regal., Verbascum chaixi Vill., and Mimulus cardinalis Dougl. ex. Benth. have intermediary cells and ordinary companion cells in the minor veins. These plants synthesize large amounts of raffinose and stachyose as well as sucrose. Nemesia strumosa Benth., and Rhodochiton atrosanguineum Zucc. have both intermediary cells and transfer cells and make proportionately less raffinose oligosaccharide than the species above. In N. strumosa, a single sieve element may abut both an intermediary cell and a transfer cell. The minor veins of Asarina scandens (Cav.) Penn. have transfer cells and what appear to be modified intermediary cells that have fewer plasmodesmata than other species, and occasional wall ingrowths. Asarina scandens synthesizes little raffinose or stachyose. Cymbalaria muralis P. Gaertn et al. and Linaria maroccana Hook.f. have only transfer cells and Digitalis grandiflora Mill. has only ordinary companion cells; these species make a trace of galactinol and raffinose, but no stachyose. Translocation experiments indicate that there is long-distance movement of raffinose oligosaccharide in these plants, even when it is synthesized in very small quantities in the leaves. We conclude that intermediary cells are as distinct a cell type as the transfer cell. In contrast to transfer cells, which are specialized for uptake of solute from the apoplast, intermediary cells are specialized for symplastic transfer of photoassimilate from the mesophyll and for synthesis of raffinose oligosaccharide. This supports our contention that raffinose oligosaccharide synthesis and symplastic phloem loading are mechanistically linked (Turgeon and Gowan 1990, Plant Physiol. 94, 1244–1249). Minor-vein anatomy and sugar synthesis may be useful characters in determining the phylogenetic relationships of plants in this family.We thank Andrea Wolfe and Wayne Elisens for helpful discussions on the taxonomy of the Scrophulariaceae. This research was supported by National Science Foundation grant DCB-9104159, U.S. Department of Agriculture Competetive Grant 92-37306-7819, and Hatch funds.     

Recognition of pathogenic Yersinia enterocolitica by crystal violet binding and polymerase chain reaction

Cefsulodin-Irgasan-Novobiocin (CIN) agar is used for the selective isolation and enumeration of Yersinia enterocolitica from clinical specimens and food. The medium contains crystal violet and about 1 mmol l^-1 calcium and can be used for the phenotypic characterization of strains that carry a virulence plasmid. At 32°C, irrespective of pathogenicity, colonies are translucent with a pale pink centre surrounded by a transparent border ('bullseye'), while at 37°C pathogenic strains grow as calcium-dependent microcolonies which, because of crystal violet binding, are intensely coloured. These results were confirmed by the polymerase chain reaction with primers directed at the vir F gene, which is present only in pathogenic strains of Y. enterocolitica. Pathogenic strains of Y. enterocolitica can be recognized by growth at 37°C on Yersinia selective agar.     

A chromosome-level genome assembly enables the identification of the follicule stimulating hormone receptor as the master sex-determining gene in the flatfish Solea senegalensis

Sex determination (SD) shows huge variation among fish and a high evolutionary rate, as illustrated by the Pleuronectiformes (flatfishes). This order is characterized by its adaptation to demersal life, compact genomes and diversity of SD mechanisms. Here, we assembled the Solea senegalensis genome, a flatfish of great commercial value, into 82 contigs (614 Mb) combining long- and short-read sequencing, which were next scaffolded using a highly dense genetic map (28,838 markers, 21 linkage groups), representing 98.9% of the assembly. Further, we established the correspondence between the assembly and the 21 chromosomes by using BAC-FISH. Whole genome resequencing of six males and six females enabled the identification of 41 single nucleotide polymorphism variants in the follicle stimulating hormone receptor (fshr) consistent with an XX/XY SD system. The observed sex association was validated in a broader independent sample, providing a novel molecular sexing tool. The fshr gene displayed differential expression between male and female gonads from 86 days post-fertilization, when the gonad is still an undifferentiated primordium, concomitant with the activation of amh and cyp19a1a, testis and ovary marker genes, respectively, in males and females. The Y-linked fshr allele, which included 24 nonsynonymous variants and showed a highly divergent 3D protein structure, was overexpressed in males compared to the X-linked allele at all stages of gonadal differentiation. We hypothesize a mechanism hampering the action of the follicle stimulating hormone driving the undifferentiated gonad toward testis.     

High-resolution mapping by YAC fragmentation of a 2.5-Mb Xp22 region containing the human RS, KFSD and CLS disease genes

The disease loci for X-linked Retinoschisis (RS), Keratosis follicularis spinulosa decalvans (KFSD), and Coffin-Lowry syndrome (CLS) have been localized to the same, small region in Xp22 on the human X Chromosome (Chr). To generate a high-resolution map of the available contig in this area, we have used the YAC fragmentation vectors pBP108/ADE2 and pBP109/ADE2 and generated fragmented YACs from a 2.5-Mb YAC (y939H7) spanning the mentioned disease gene candidate regions. Forty-seven fragmented YACs were generated and analyzed, ranging in size from 170 kb to over 2400 kb. The resulting YAC fragmentation panel was used to construct a detailed restriction map of the region and has been used to bin clones and markers. As a deletion panel, it will present a valuable resource for further mapping. Received: 31 December 1996 / Accepted: 22 February 1997     

Chiton integument: Metamorphic changes in Mopalia muscosa (Mollusca,Polyplacophora)

Summary The larval integument and juvenile girdle integument of Mopalia muscosa (Mollusca: Polyplacophora) were studied by light microscopy. Within 24 h of settlement, eight distinctive changes occur that characterize metamorphosis: loss of the functional prototroch and apical tuft, secretion of a cuticle over the mantle field followed by the secretion of calcareous shell plates and the extrusion of spicules into the cuticle, a 20% decrease in length, secretion of chitinous hairs and the incorporation of the lateral ciliated bands into the pallial grooves. Similar changes which were often not recognized as metamorphic have been reported for other species. Evidence for metamorphosis being a common developmental feature of chitons is presented.