In Situ Localization of Parental Genomes in a Wide Hybrid

In situ hybridization enabled DNA originating from the two parentalgenomes to be distinguished in plant hybrids. A probe of biotinylatedtotal genomic DNA from Secale africanum labelled the chromosomesof S. africanum origin but not those from Hordeum chilense inroot-tip chromosome spreads of the sexual hybrid between thetwo species. Hybridization of total genomic DNA from S. africanumto DNA on filters (dot blots) confirmed the distinction betweenDNA from Hordeum and Secale. The total genomic probe hybridizedto the whole length of the chromosomes from S. africanum remarkablyuniformly, labelling both euchromatin and heterochromatin, exceptat the centromeric region. The probe binding was visualizedas a yellow colour by the fluorescein-coupled detection systemwhich contrasted with the red fluorescing counterstain of theunlabelled chromatin. The chromosomes originating from bothparents could be seen and distinguished as red and yellow fluorescenceat all stages of the cell cycle. At interphase and prophase,the chromatin originating from the two parental genomes didnot mix. Chromosomes or groups of chromosomes occupied distinctdomains and also tended to be arranged in a Rabl configurationwith the centromeres clustered at one end of the nucleus. Wepropose calling the technique using total genomic DNA as a probe‘genomic in situ hybridization.’ Hordeum chilense, Secale africanum, hybrids, genomic in situ hybridization, DNA, repetitive sequences, chromosomes, chromosome disposition, nuclear order     

Size-Structures and Reproductive Characteristics in Diploid and Triploid Populations of Disporum sessile D. Don (Liliaceae)

Abstract Disporum sessile (Liliaceae), a perennial herb of temperate forests is composed of diploid (2n=16) and triploid (2n=24) populations. The size structure differed remarkably as triploid populations had few small plants and no seedlings. Triploid flowering plants were considerably larger than diploids. Triploids that flowered were 2.5 times larger than diploids that flowered and the size of vegetative ramets produced by triploids was twice as large. In triploids, fruiting rates were quite low only with inviable seeds and vegetative propagule size was greater than that of diploids. As regards growth parameters that help to increase plant size, triploids were superior to diploids. Differences in growth and reproductive parameters between diploids and triploids may contribute to forming different patch sizes.     

In Vitro Induction of Haploid, Diploid, and Triploid Androgenic Embryoids and Plantlets in Datura metal L.

Pollen plantlets of Datura metel L. have been successfully rearedin vitro through anther culture on a nutrient medium supplementedwith coconut milk (15 per cent v/v) The early segmentation patternof the proembryos and their subsequent differentiation intoheart, torpedo, and cotyledonary stages were traced. More than100 plantlets were grown to maturity on soil. They grew normallyand produced flowers. Root-tip and leaf-squash preparationsshowed variable numbers of chromosomes in their cells, and thisindicated the occurrence of ploidy plants among the population.Frequency determinations showed the predominant types to bediploid (70 per cent), triploid (24 per cent), and haploid (6per cent). Differences in the morphology of mature plants inregard to floral size, meiotic irregularities in PMCs, formationof giant pollen grains, differential fruitset, somatic mosaicismin respect of chromosome number, and aneuploidy of cells, wererecorded. The origin of diploid and triploid plants is discussed.     

Hybrid and Rogue Kinases Encoded in the Genomes of Model Eukaryotes

The highly modular nature of protein kinases generates diverse functional roles mediated by evolutionary events such as domain recombination, insertion and deletion of domains. Usually domain architecture of a kinase is related to the subfamily to which the kinase catalytic domain belongs. However outlier kinases with unusual domain architectures serve in the expansion of the functional space of the protein kinase family. For example, Src kinases are made-up of SH2 and SH3 domains in addition to the kinase catalytic domain. A kinase which lacks these two domains but retains sequence characteristics within the kinase catalytic domain is an outlier that is likely to have modes of regulation different from classical src kinases. This study defines two types of outlier kinases: hybrids and rogues depending on the nature of domain recombination. Hybrid kinases are those where the catalytic kinase domain belongs to a kinase subfamily but the domain architecture is typical of another kinase subfamily. Rogue kinases are those with kinase catalytic domain characteristic of a kinase subfamily but the domain architecture is typical of neither that subfamily nor any other kinase subfamily. This report provides a consolidated set of such hybrid and rogue kinases gleaned from six eukaryotic genomes–S.cerevisiae, D. melanogaster, C.elegans, M.musculus, T.rubripes and H.sapiens–and discusses their functions. The presence of such kinases necessitates a revisiting of the classification scheme of the protein kinase family using full length sequences apart from classical classification using solely the sequences of kinase catalytic domains. The study of these kinases provides a good insight in engineering signalling pathways for a desired output. Lastly, identification of hybrids and rogues in pathogenic protozoa such as P.falciparum sheds light on possible strategies in host-pathogen interactions.     

Characterization of Chromosome Stability in Diploid,Polyploid and Hybrid Yeast Cells

Chromosome instability is a key component of cancer progression and many heritable diseases. Understanding why some chromosomes are more unstable than others could provide insight into understanding genome integrity. Here we systematically investigate the spontaneous chromosome loss for all sixteen chromosomes in Saccharomyces cerevisiae in order to elucidate the mechanisms underlying chromosome instability. We observed that the stability of different chromosomes varied more than 100-fold. Consistent with previous studies on artificial chromosomes, chromosome loss frequency was negatively correlated to chromosome length in S. cerevisiae diploids, triploids and S. cerevisiae-S. bayanus hybrids. Chromosome III, an equivalent of sex chromosomes in budding yeast, was found to be the most unstable chromosome among all cases examined. Moreover, similar instability was observed in chromosome III of S. bayanus, a species that diverged from S. cerevisiae about 20 million years ago, suggesting that the instability is caused by a conserved mechanism. Chromosome III was found to have a highly relaxed spindle checkpoint response in the genome. Using a plasmid stability assay, we found that differences in the centromeric sequence may explain certain aspects of chromosome instability. Our results reveal that even under normal conditions, individual chromosomes in a genome are subject to different levels of pressure in chromosome loss (or gain).     

Complex Function and Expression of Delta during Drosophila Oogenesis

Delta (Dl) encodes a cell surface protein that mediates cell-cell interactions central to the specification of a variety of cell fates during embryonic and postembryonic development of Drosophila melanogaster. We find that the Delta protein is expressed intermittently in follicle cells and in germ-line cells during stages 1-10 of oogenesis. Furthermore, Delta expression during oogenesis can be correlated with a number of morphogenetic defects associated with sterility observed in Dl mutant females, including failure of stalk formation within the germarium and subsequent fusion of egg chambers, necrosis in germ-line cells, and multiphasic embryonic arrest of fertilized eggs. We have also identified a Dl mutation that leads to context-dependent defects in Dl function during oogenesis. Direct comparison of Delta protein expression with that of the Notch protein in the ovary reveals substantial, but incomplete, coincidence of expression patterns in space and time. We discuss possible roles for the Delta protein in cell-cell interactions required for cell fate specification processes during oogenesis in light of available developmental and histochemical data.     

Thermal Tolerance of Diploid Versus Triploid Rainbow Trout and Brook Trout Assessed by Time to Chronic Lethal Maximum

Synopsis An effect of ploidy on thermal tolerance in juvenile trout was assessed in a series of tests comparing time to chronic lethal maximum (CLMax). Diploid and triploid fish were produced from a common spawn for three different groups each of brook trout Salvelinus fontinalis and of rainbow trout Oncorhynchus mykiss. One or two CLMax tests were performed per group, on between 15 and 50 individuals per ploidy within groups. The tests involved exposure of fish to a progressive 2°C day⁻¹ water temperature increase and recording of the time at which each individual fish reached loss of equilibrium (LE). The time to LE data were rank transformed and analyzed as a randomized complete block design. Although relative performance varied among trials, the analysis indicated overall differences due to ploidy were small and nonsignificant among both brook trout and rainbow trout. Size proved to be significantly correlated with time to LE in the brook trout trials, but not in the rainbow trout trials. Two of the six groups included a large proportion of fish which had received a heat shock following fertilization, but were not successfully triploidized. In both cases, thermal tolerance of the heat-shocked diploids was similar to that of the non-heat shocked control diploids, indicating no persistent effect of the heat shock on thermal tolerance.     

Elimination of Viruses and Indicator Bacteria at Each Step of Treatment during Preparation of Drinking Water at Seven Water Treatment Plants

[This corrects the article on p. 1418 in vol. 49.].     

A Sponge-like Structure Involved in the Association and Transport of Maternal Products during Drosophila Oogenesis

Localization of maternally provided RNAs during oogenesis is required for formation of the antero–posterior axis of the Drosophila embryo. Here we describe a subcellular structure in nurse cells and oocytes which may function as an intracellular compartment for assembly and transport of maternal products involved in RNA localization. This structure, which we have termed “sponge body,” consists of ER-like cisternae, embedded in an amorphous electron-dense mass. It lacks a surrounding membrane and is frequently associated with mitochondria. The sponge bodies are not identical to the Golgi complexes. We suggest that the sponge bodies are homologous to the mitochondrial cloud in Xenopus oocytes, a granulo-fibrillar structure that contains RNAs involved in patterning of the embryo.     

小麦与高冰草体细胞杂种后代的核基因组和叶绿体基因组的遗传特征

以小麦(TriticumaestivumL.)与高冰草(Agropyronelongatum(Host)Nevski)体细胞杂种同一个克隆来源的F2-F6自交系Ⅱ-2、Ⅱ-Ⅰ-8以及由Ⅱ-Ⅰ-8F2分离形成的8-1(F3-F6)为材料,利用小麦叶绿体基因组的微卫星(Microsatellite)特异引物及随机扩增多态性DNA(RAPD)引物进行分析。结果表明,杂种株系的叶绿体基因组组成一致,均以小麦叶绿体基因组为主,仅在rpl14和rpl16基因的间隔序列中检测到双亲的特征带,表明有高冰草的叶绿体DNA在杂种中存在,并稳定遗传至第六代。RAPD分析表明,不同杂种株系中存在不同的高冰草核DNA片段,核基因组在传代中基本稳定。     

Parental Smoking in the Vicinity of Children and Tobacco Control Policies in the European Region

Objective

To ascertain patterns of parental smoking in the vicinity of children in Eastern and Western Europe and their relation to Tobacco Control Scale (TCS) scores.

Methods

Data on parental smoking patterns were obtained from the School Child Mental Health Europe (SCMHE), a 2010 cross-sectional survey of 5141 school children aged 6 to 11 years and their parents in six countries: Germany, Netherlands, Lithuania, Romania, Bulgaria and Turkey ranked by TCS into three level categories toward tobacco control policies.

Results

A slightly higher proportion of Eastern compared to Western European mothers (42.4 vs. 35.1%) were currently smoking in but the difference was not statistically significant after adjusting for maternal age and maternal educational attainment. About a fifth (19.3%) and a tenth (10.0%) of Eastern and Western European mothers, respectively, smoked in the vicinity of their children, and the difference was significant even after adjustment for potential confounders (p<0.001). Parents with the highest educational attainment were significantly less likely to smoke in the vicinity of their children than those with the lowest attainment. After control of these covariates lax tobacco control policies, compared to intermediate policies, were associated with a 50% increase in the likelihood of maternal smoking in the vicinity of children adjusted odds ratio (AOR) = 1.52 and 1.64. Among fathers, however, the relationship with paternal smoking and TCS seems more complex since strict policy increases the risk as well AOR = 1,40. Only one country, however belongs to the strict group.

Significance

Tobacco control policies seem to have influenced maternal smoking behaviors overall to a limited degree and smoking in the vicinity of children to a much greater degree. Children living in European countries with lax tobacco control policies are more likely to be exposed to second hand smoking from maternal and paternal smoking.