Cuticular Membrane Fine Structure of Nicotiana tabacum L. Leaves

The cuticular membrane (CM) ofNicotiana tabacumL., includingthe cellin wall (CW), was examined to gain more informationabout the nature and chemical constitution of its fine structurefor possible inclusion in a model system, as recent literaturequestions its function as a major water permeability barrier.Different preparation techniques were used and the results evaluatedto select a method for future studies on tobacco leaf cuticles.Fixation with OsO₄included in the primary fixative, either asa vapour or in combination with other agents, followed by OsO₄aspost-fixative, gave good contrast of the CM. The lamellar structureof the tobacco cuticle proper (CP) was revealed by contrastingwith uranyl acetate and lead citrate. The fine lamellar structureof the CP was very clearly contrasted when KMnO₄was includedin the primary fixative. This was interpreted as indicatingthe tobacco CP to be polar. The reticulate fibrillar patternof the tobacco cuticular layer (CL) containing polysaccharideswas well contrasted when either OsO₄or paraformaldehyde wereincluded in the primary fixative. Cold fixation with glutaraldehydeand dimethyl sulphoxide and post-fixation with OsO₄revealedelectron-opaque material in the outer cutinized, irregularlyoutlined, region of the CW. These ultrahistochemical reactionsare discussed in relation to the known chemical compositionand possible water permeability of the CM. Cuticular fine structure; cuticular transpiration; Nicotiana tabacumL.     

Molecular and ultrastructural analysis of a nonchromosomal variegated mutant. Tomato mitochondrial mutants that cause abnormal leaf development.

Mutants were recovered in a population of cybrids formed following protoplast fusion between tomato (Lycopersicon esculentum Mill.) cv UC82 and Lycopersicon pennellii Corr. The cybrids were identified as individuals with recombinant cytoplasmic genomes but only tomato nuclear genomes. The mutants were identified based on two features, a variegated sectoring of light and dark green regions on their leaves, stems, and fruit, and reduced growth in the field. The mutants produced 50% of the shoot fresh weight and 20% of the fruit fresh weight of the parental type, UC82. The variegated sectoring was maternally inherited. The chloroplast genome in the mutants was indistinguishable from the chloroplast genome in UC82, when distribution of restriction endonuclease sites was used as an assay. The mitochondrial genome in the mutants, however, was recombinant, containing genes from UC82 and L. pennellii. Light microscopic analysis of the leaves of the mutants demonstrated an absence of the palisade layer in the variegated sectors. Electron microscopic analysis of these same regions demonstrated an absence of normal inner membranes in the mitochondria of these cells.     

Internal eliminated sequences interrupting the Oxytricha 81 locus: allelic divergence, conservation, conversions, and possible transposon origins

Internal eliminated sequences (IESs) often interrupt ciliate genes in the silent germline nucleus but are exactly excised and eliminated from the developing somatic nucleus from which genes are then expressed. Some long IESs are transposons, supporting the hypothesis that short IESs are ancient transposon relics. In light of that hypothesis and to explore the evolutionary history of a collection of IESs, we have compared various alleles of a particular locus (the 81 locus) of the ciliated protozoa Oxytricha trifallax and O. fallax. Three short IESs that interrupt two genes of the locus are found in alleles from both species, and thus must be relatively ancient, consistent with the hypothesis that short IESs are transposon relics. In contrast, TBE1 transposon interruptions of the locus are allele-specific and probably the results of recent transpositions. These IESs (and the TBE1s) are precisely excised from the DNA of the developing somatic macronucleus. Each IES interrupts a highly conserved sequence. A few nucleotides at the ends of each IES are also conserved, suggesting that they interact critically with IES excision machinery. However, most IES nucleotide positions have evolved at high rates, showing little or no selective constraint for function. Nonetheless, the length of each IES has been maintained (+/- 3 bp). While one IES is approximately 33 bp long, three other IESs have very similar sizes, approximately 70 bp long. Two IESs are surrounded by direct repeats of the sequence TTCTT. No other sequence similarities were found between any of the four IESs. However, the ends of one IES do match the inverted terminal repeat consensus sequence of the "TA" IESs of Paramecium. Three O. trifallax alleles appear to have been recipients in recent conversion events that could have been provoked by double-strand breaks associated with IES ends subsequent to IES transposition. Our findings support the hypothesis that short IESs evolved from ancient transposons that have lost most of their sequences, except those necessary for precise excision during macronuclear development.      

EDC3 phosphorylation regulates growth and invasion through controlling P‐body formation and dynamics

Regulation of mRNA stability and translation plays a critical role in determining protein abundance within cells. Processing bodies (P‐bodies) are critical regulators of these processes. Here, we report that the Pim1 and 3 protein kinases bind to the P‐body protein enhancer of mRNA decapping 3 (EDC3) and phosphorylate EDC3 on serine (S)161, thereby modifying P‐body assembly. EDC3 phosphorylation is highly elevated in many tumor types, is reduced upon treatment of cells with kinase inhibitors, and blocks the localization of EDC3 to P‐bodies. Prostate cancer cells harboring an EDC3 S161A mutation show markedly decreased growth, migration, and invasion in tissue culture and in xenograft models. Consistent with these phenotypic changes, the expression of integrin β1 and α6 mRNA and protein is reduced in these mutated cells. These results demonstrate that EDC3 phosphorylation regulates multiple cancer‐relevant functions and suggest that modulation of P‐body activity may represent a new paradigm for cancer treatment.     

Biodiversity and ecology of epilithic diatoms in the Agnéby River,Ivory Coast

The ecology and taxonomy of the epilithic diatom flora of the Agnéby River, Ivory Coast were studied in 2012. Ten sites were investigated and diatoms were sampled on glass slides immersed for a period of 30 days during the wet and dry seasons. Physico-chemical parameters were measured at each site while sampling diatoms. Five taxa were largely dominant: Planothidium comperei CE Wetzel, N’Guessan and Tison-Rosebery, Eolimna minima (Grunow) Lange-Bertalot, Planothidium piaficum (JR Carter and Denny) CE Wetzel and Ector, Cocconeis schroederi Foged and Cocconeis scutellum var. parva (Grunow in Van Heurck) Cleve. Electrical conductivity, temperature, dissolved oxygen, nitrite and phosphorus were found to influence the distribution of taxa.     

The biogeochemistry of basic cations in two forest catchments with contrasting lithology in the Czech Republic

The biogeochemistry of Ca, Mg, K, and Nawere investigated in two forested catchments in theCzech Republic, one underlain by leucogranite, theother by serpentinite. High weathering rates at theserpentinite site at Pluhv Bor resultedin Mg²⁺ as the dominant cation on the soilexchange complex and in drainage water. Other basiccations (Ca²⁺, K⁺, Na⁺) showedrelatively low concentrations and outflow instreamwater. The catchment exhibited high basesaturation in mineral soils (>70%), and nearneutral soil and stream pH, despite elevated inputsof acidic deposition. Slow growth of Norway spruceat Pluhv Bor may be caused by K deficiency, Mgoversupply and/or Ni toxicity. In contrast, thegranitic site at Lysina showed low concentrations ofbasic cations on the soil exchange complex and instreamwater. Soil and drainage water at Lysina werehighly impacted by acidic deposition. Soil pH wasextremely acidic (<4.5) throughout the soilprofile, and the base saturation of the mineral soilwas very low (<5%). Supplies of basic cationsfrom atmospheric deposition and soil processes wereless than inputs of SO^2- ₄ on anequivalence basis, resulting in low pH and highconcentrations of total Al in drainage water. Needle yellowing in Norway spruce was possibly theresult of Mg deficiency at Lysina. Because of theirextremely different lithologies, these catchmentsserve as valuable end-members of ecosystemsensitivity to elevated levels of acidicdeposition.     

Identification and mapping of a novel dominant resistance gene,TuRB07 to Turnip mosaic virus in Brassica rapa

Key message

A novel dominant resistance gene, TuRB07, was found to confer resistance to an isolate of TuMV strain C4 in B. rapa line VC1 and mapped on the top of chromosome A06.

Abstract

The inheritance of resistance to Turnip mosaic virus in Brassica rapa was investigated by crossing the resistant line, VC1 with the susceptible line, SR5, and genotyping and phenotyping diverse progenies derived from this cross. Both a doubled haploid population, VCS3M-DH, an F2 and two BC1 (F1 × VC1 and F1 × SR5) populations were created. Population tests revealed that the resistance to the TuMV C4 isolate in B. rapa is controlled by a single dominant gene. This resistance gene, TuRB07 was positioned on the top of linkage group A06 of the B. rapa genome through bulk segregation analysis and fine mapping recombinants in three doubled haploid- and one backcross population using microsatellite markers developed from BAC end sequences. Within the region between the two closely linked markers flanking TuRB07, H132A24-s1, and KS10960, in the Chiifu reference genome, two genes encoding nucleotide-binding site and leucine-rich repeat proteins with a coiled-coil motif (CC-NBS-LRR), Bra018862 and Bra018863 were identified as candidate resistance genes. The gene Bra018862 is truncated, but the gene Bra018863 has all the domains to function. Furthermore, the analysis of structural variation using resequencing data of VC1 and SR5 revealed that Bra018863 might be a functional gene because the gene has no structural variation in the resistant line VC1 when compared with Chiifu, whereas at the other NBS-LRR genes large deletions were identified in the resistant line. Allelic differences of Bra018863 were found between VC1 and SR5, supporting the notion that this gene is a putative candidate gene for the virus resistance.     

The tomato Orion locus comprises a unique class of <Emphasis Type="Italic">Hcr9</Emphasis> genes

Resistance against the tomato fungal pathogen Cladosporium fulvum is often conferred by Hcr9 genes (Homologues of the C. fulvum resistance gene Cf-9) that are located in the Milky Way cluster on the short arm of chromosome 1. These Hcr9 genes mediate recognition of fungal avirulence gene products. In contrast, the resistance gene Cf-Ecp2 mediates recognition of the virulence factor Ecp2 and is located in the Orion (OR) cluster on the short arm of chromosome 1. Here, we report the map- and homology-based cloning of the OR Hcr9 cluster. A method was optimised to generate clone-specific fingerprint data that were subsequently used in the efficient calculation of genomic DNA contigs. Three Hcr9s were identified as candidate Cf-Ecp2 genes. By PCR-based cloning using specific OR sequences, orthologous Hcr9 genes were identified from different Lycopersicon species and haplotypes. The OR Hcr9s are very homologous. However, based on the relative low sequence homology to other Hcr9s, the OR Hcr9s are classified as a new subgroup.Data deposition: The sequence of the Cf-Ecp2 Hcr9 gene cluster and the orthologous Hcr9 sequences have been deposited in the GenBank database (accession No. AY639600..AY639604)