text:
Haploids and doubled haploid (DH) inbred lines have become an invaluable tool for maize genetic research and hybrid breeding, but the genetic basis of in vivo induction of maternal haploids is still unknown. This is the first study reporting comparative quantitative trait locus (QTL) analyses of this trait in maize. We determined haploid induction rates (HIR) in testcrosses of a total of 1061 progenies of four segregating populations involving two temperate haploid inducers, UH400 (HIR = 8%) and CAUHOI (HIR = 2%), one temperate and two tropical inbreds with HIR = 0%, and up to three generations per population. Mean HIR of the populations ranged from 0.6 to 5.2% and strongly deviated from the midparent values. One QTL (qhir1) explaining up to of the genetic variance was detected in bin 1.04 in the three populations involving a noninducer parent and the HIR-enhancing allele was contributed by UH400. Segregation ratios of loci in bin 1.04 were highly distorted against the UH400 allele in these three populations, suggesting that transmission failure of the inducer gamete and haploid induction ability are related phenomena. In the CAUHOI × UH400 population, seven QTL were identified on five chromosomes, with qhir8 on chromosome 9 having in three generations of this cross. The large-effect QTL qhir1 and qhir8 will likely become fixed quickly during inducer development due to strong selection pressure applied for high HIR. Hence, marker-based pyramiding of small-effect and/or modifier QTL influencing qhir1 and qhir8 may help to further increase HIR in maize. We propose a conceptual genetic framework for inheritance of haploid induction ability, which is also applicable to other dichotomous traits requiring progeny testing, and discuss the implications of our results for haploid inducer development.
IN VIVO induction of maternal haploids in maize has paved the way for large-scale production of doubled haploid (DH) inbred lines, which today form the backbone of the global hybrid maize industry. Traditionally, the maize plants' cross-breeding nature required recurrent self-pollinations for 6–10 generations to obtain sufficiently homozygous inbred lines (Hallauer et al. 2010). Application of DH technology reduces the time for inbred development by more than half compared to the traditional method and additionally provides several quantitative genetic, operational, logistical, and economic advantages (Nei 1963; Schmidt 2003; Melchinger et al. 2005; Seitz 2005; Smith et al. 2008; Chang and Coe 2009; Geiger 2009). By using haploid inducer genotypes as pollinators in crosses with source germplasm, ears obtained carry a proportion of seeds containing haploid embryos of maternal origin. Subsequent treatment of haploids with mitotic inhibitors facilitates chromosome duplication resulting in diploid and completely homozygous inbred lines (see Prigge and Melchinger 2012 for a detailed description of DH production).
Modern maize inducers have haploid induction rates (HIR) of about 8% on average (e.g., Röber et al. 2005; Prigge et al. 2011). The genetic mechanisms underlying in vivo induction of maternal haploids in maize are not yet fully understood. The two major hypotheses for possible mechanisms are: (i) failure of fertilization of the egg cell and subsequent parthenogenetic development of the reduced egg into a haploid embryo (Sarkar and Coe 1966; Chalyk et al. 2003; Barret et al. 2008), and (ii) normal fertilization followed by elimination of inducer chromosomes (Fischer 2004; Zhang et al. 2008; Li et al. 2009). Further, Kato (1997) suggested that the aberrant fertilization mechanisms leading to haploidy may be related to mechanisms leading to heterofertilization.
Continuous variation of segregating populations developed from inducer by noninducer crosses suggests that HIR is a quantitative trait (Lashermes and Beckert 1988). The first exploratory quantitative trait locus (QTL) mapping study, conducted with RFLP markers in an F3 population involving Stock6 (HIR = 2.3%; Coe 1959) as inducer parent, provided evidence for two QTL for HIR on chromosomes 1 and 2, together explaining 17.9% of the phenotypic variance (Deimling et al. 1997). Barret et al. (2008) used marker segregation ratio distortion analyses in small samples from the two phenotypic extremes of a segregating population developed from a cross between a noninducer and an inducer line and also identified a locus on chromosome 1 associated with HIR.
None of the most recently developed haploid inducers with high HIR have been subjected to genome-wide QTL analysis yet. Further, QTL must be examined for stable expression in different germplasm because high congruency of QTL in different genetic backgrounds is desirable to facilitate marker-assisted introgression approaches. Therefore, we conducted comparative QTL analyses for HIR in four populations involving two inducers, UH400 and CAUHOI. Our objectives were to (1) study the inheritance of in vivo haploid induction ability and its association with segregation distortion and embryo abortion rate (EAR), (2) estimate the number, genomic positions, and genetic effects of QTL associated with HIR and EAR, and (3) discuss possible mechanisms underlying in vivo haploid induction in monocots as well as implications of the results for fine mapping and improving HIR in maize.
Materials and Methods
Genetic materials
Four mapping populations involving haploid inducer inbred UH400 as pollinator with two temperate (CAUHOI, 1680) and two tropical (CML395, CML495) inbreds as female parents were generated (Table 1). UH400 was developed at the University of Hohenheim, Germany, and has a HIR of ∼8% averaged across several tropical (Prigge et al. 2011) and temperate (W. Schipprack, personal communication) environments. It carries the dominantly inherited marker gene R1-nj conferring a purple coloration of the scutellum and the aleurone of seeds (Nanda and Chase 1966; Neuffer et al. 1997), which can be used as embryo and endosperm marker, respectively, to identify putative haploid seeds. CAUHOI is a haploid inducer developed by China Agricultural University with high kernel oil content (78 g kg−1) and a HIR of ∼2% (Li et al. 2009). Inbreds 1680 (developed by China Agricultural University) as well as CML395 and CML495 (both developed by the International Maize and Wheat Improvement Center, CIMMYT) have no induction ability but excellent agronomic characteristics.
In all four populations, random F2 plants were selfed to produce F2:3 lines. In addition, for population CAUHOI × UH400, randomly chosen seeds from self-pollinated ears of F2:3 lines were bulked and grown as F2:4 lines. In summary, F2, F2:3, and F2:4 generations were evaluated for HIR in CAUHOI × UH400 (designated CAU-F2, CAU-F3, and CAU-F4, respectively), F2 and F2:3 generations in 1680 × UH400 (1680-F2 and 1680-F3), and the F2:3 generation was evaluated in CML395 × UH400 (CML395-F3) and in CML495 × UH400 (CML495-F3) (Table 1).
Marker assays and linkage map construction
Genotyping was performed differently for the two temperate (CAU-F2, 1680-F2) and the two tropical (CML395-F2, CML495-F2) populations following the protocols used at China Agricultural University and CIMMYT, respectively. For CAU-F2 and 1680-F2, leaf tissue from the parental lines and the F2 plants was harvested at the three-leaf stage. Genomic DNA was extracted following the procedures of Porebski et al. (1997). One-hundred eighty-six F2 individuals of populations CAU-F2 and 1680-F2 were genotyped with 90 and 113 polymorphic simple sequence repeat (SSR) markers, respectively, following standard protocols (Ninamango-Cárdenas et al. 2003). The SSRs were selected from the MaizeGDB database (http://www.maizegdb.org; Lawrence et al. 2004) to provide good coverage of the entire genome.
For CML395-F2 and CML495-F2, leaf tissue from the parental lines and the F2 plants was harvested at the 10-leaf stage, placed in glassine bags, and stored at −80° until lyophilization at CIMMYT's biotechnology facility. Genomic DNA was extracted following standard protocols (CIMMYT 2005). Parents of both populations were screened for polymorphism with the custom GoldenGate assay containing 1536 single nucleotide polymorphism (SNP) markers (Yan et al. 2010). SNP genotyping for the parental screening was performed using the Illumina BeadStation 500 G (Illumina, San Diego, CA) at the Cornell University Life Sciences Core Laboratories Center according to the protocol of Fan et al. (2006) and data were analyzed using the Illumina BeadStudio genotyping software. The genotyping of 220 (CML395-F2) and 233 (CML495-F2) F2 individuals with 324 and 297 polymorphic SNPs, respectively, was conducted by KBioSciences (United Kingdom) using simple-plex, automated high throughput KasPar chemistry assays. Genotypes for all populations are available as Supporting Information, File S1.
The four linkage maps were constructed by Haldane's mapping function with software JoinMap 3.0 (Van Ooijen and Voorrips 2001). Individuals and loci with >20% missing data were excluded from the analysis. A LOD (log10 of the likelihood odds ratio) value of 3.0 was used as critical threshold to declare linkage between two markers. The order of the mapped SSR markers on each linkage group was confirmed according to the IBM2 2008 Neighbors Map (http://www.maizegdb.org). SNP marker order was confirmed by their absolute physical location on the B73 reference maize sequence (http://www.maizesequence.org) and bin positions were identified from the sequence. As some markers were very close (<1.01 cM) to each other, they were combined into a "synthetic" marker resulting in a reduced number of loci used for QTL analysis (Table 1). Further, we tested expected allele frequencies of p = 0.5 of the UH400 allele with a standard χ2 goodness-of-fit test and declared segregation distortion when loci significantly (P < 0.05) deviated from the expectation.
Assessment of haploid induction and embryo abortion rates
The mapping populations were grown in completely randomized designs at the experimental stations of China Agricultural University near Beijing (China) in summer 2008 (CAU-F2 and 1680-F2) and 2009 (CAU-F4 and 1680-F3), and in Hainan province (China) in winter 2008/2009 (CAU-F3), as well as at CIMMYT's experimental station in Tlaltizapan (Mexico) in summer 2009 (CML395-F3 and CML495-F3). Field plots consisted of 11 plants in each experiment.
Determination of HIR was conducted on the basis of two haploid identification systems. For the CAU and 1680 mapping populations, the R1-nj color marker inherited from the inducer parent UH400 was scored on testcross seeds harvested from the nonpigmented Chinese hybrid ZD958 after pollination with the mapping populations. Three to five plants of ZD958 were pollinated with either pollen of individual F2 plants (CAU-F2 and 1680-F2) or bulked pollen of a minimum of five plants per family (CAU-F3, 1680-F3, and CAU-F4). Testcross seed with purple endosperm (indicating a regular triploid endosperm resulting from successful fertilization with inducer pollen) but colorless embryo (indicating a haploid embryo of ZD958 origin) were designated as putative haploid seed following Prigge and Melchinger (2012). To verify the effectiveness of the R1-nj color marker and to account for putative misclassifications (Röber et al. 2005; Prigge et al. 2011), all haploids detected by the above method were grown near Beijing in summer 2010 and visually scored for plant type before flowering; haploid plants had shorter statures, thin stalks, and erect and narrow leaves. For 1680-F2 and 1680-F3, we additionally determined the EAR as the proportion of completely collapsed testcross seeds exhibiting embryoless pericarps.
For the CML mapping populations, a single cross obtained by crossing two temperate inbred lines carrying the recessive mutation liguleless was used as tester (hereafter referred to as lglg tester). The specific genetic source (lg1 vs. lg2) is unknown for this germplasm. The lglg tester allows clear distinction between haploid and diploid seedlings because haploids are characterized by absence of the ligule and auricle combined with an upright-positioned leaf enveloping the shoot (Neuffer et al. 1997). All lglg tester plants were emasculated to avoid self-contamination. Bulked pollen of a minimum of five plants per F3 family was used to pollinate a minimum of eight lglg tester plants. All testcross seeds were grown in the greenhouse until the four-leaf stage and were visually rated for the liguleless phenotype, an indication of haploidy.
For all populations, each testcross ear was shelled separately. Three testcross ears per family were selected on the basis of the highest total number of testcross seeds to determine HIR (in all populations) and EAR (in 1680-F2 and 1680-F3). Repeatabilities (w2) for HIR and EAR were calculated with the statistical software PLABSTAT (Utz 2004) taking the values of the three individual ears as three replicates in a completely randomized design. Combining the three ears, HIR and EAR were computed for each family as the proportion of putative haploid and embryoless testcross seeds, respectively, detected in the total testcross progeny monitored. Parent–offspring regression was performed with CAU-F2, CAU-F3, CAU-F4, 1680-F2, and 1680-F3 and the regression coefficient was used as an estimator of the narrow-sense heritability (h2) for HIR and EAR of the corresponding populations.
QTL analyses
Analyses of QTL for HIR and EAR were performed with software PlabQTL (Utz and Melchinger 1996) using composite interval mapping (CIM; Jansen and Stam 1994; Zeng 1994) and a multiple regression procedure (Haley and Knott 1992). The appropriate number of marker cofactors and the appropriate genetic models were chosen on the basis of the smallest values of the modified Bayesian Information Criterion (mBIC, Baierl et al. 2006). Using the mBIC as significance criterion, we did not detect significant first-order epistatic interactions in any of the populations. Minimized mBIC were obtained for a model including additive and dominance effects for all populations except CAU-F2 and CML495-F3 for HIR as well as 1680-F2 and 1680-F3 for EAR, where no significant dominance effects were detected. Critical LOD thresholds were determined empirically with 1000 random permutations (Churchill and Doerge 1994) for each generation, population, and trait. LOD thresholds corresponding to genome-wide error rates of α = 0.25 were employed for CIM, which is common for explorative QTL analyses (Beavis 1998). The support intervals with a LOD fall-off of 1.0 were expressed as a position on the chromosome. The proportion of the genetic variance explained by the model was estimated as where is the adjusted proportion of phenotypic variance explained by the model and w2 is the repeatability of the trait. The proportion of explained by individual QTL was estimated as , where is the proportion of phenotypic variance explained by that QTL. All values of were normalized to sum to For those loci showing significant dominance effects, we calculated the dominance ratio (d/|a|) of the estimated dominance effect over the absolute value of the estimated additive effect.
Further, to account for the special features of the phenotypic distributions of HIR in QTL mapping, we used the two-part model developed by Broman (2003) implemented in the software R/QTL (Broman and Sen 2009) in addition to CIM. This model combines the analysis of the binary trait (i.e., zi = 1 vs. zi = 0, corresponding to HIR > 0% vs. HIR = 0%, respectively, of inducer F2 plant or F2:3/F2:4 family i) with standard interval mapping for those having zi = 1. For plants/families with haploid induction ability (i.e., zi = 1) and genotype g (AA, AB, or BB) at marker locus m in the parental F2 plant, the square root HIR is assumed to follow a normal distribution with mean μg and standard deviation σ (Broman 2003). As HIR is determined on the basis of the testcross progeny of i, the phenotype of each testcross seed ij can be coded by an indicator variable zij, with zij = 1 if seed ij is haploid or zij = 0 otherwise. Further, πg = Pr(zi = 1|mi = g) denotes the conditional probability of obtaining zi = 1, if the genotype of the parental F2 plant i at marker locus m is g, whereas μg = E(zij|mi = g, zi = 1) denotes the conditional expectation for testcross seed ij within the progeny of inducer i with HIR > 0%, if the genotype of the parental F2 plant i at marker locus m is g. Separate LOD scores were calculated to test whether the three genotype classes g have the same probability πg of inducing haploidy (i.e., zi = 1) and/or whether those families with zi = 1 have the same sample means μg (corresponding to HIR) according to three hypotheses (Broman 2003): (1) LOD(π) to test whether πAA = πAB = πBB; (2) LOD(μ) to test whether μAA = μAB = μBB; and (3) LOD(πμ) to test the combined hypothesis that both the πg and μg were equal. The corresponding critical LOD thresholds for the two-part model were determined empirically with 1000 random permutations.
Haploid induction and embryo abortion rates
The average number of testcross seeds used for determination of HIR was lowest in CML395-F3 (153) and highest in CAU-F4 (317; Table 1) and ranged from 50 to 640 for individual families. Genotypic variances for HIR were significant (P < 0.01) in all populations. Repeatabilities (w2) were intermediate for CML395-F3 and CAU-F3 and high for all other populations. Mean HIR were highest in CAU-F2 and lowest in the two CML populations and HIR ranged from 0 to 15.6% for individual families (Table 1 and Figure 1). Distributions of HIR were strongly skewed toward low values in populations where noninducers (1680, CML395, CML495) were used as parents. Population means deviated significantly (P < 0.001, Wilcoxon signed rank test) from the midparent values toward lower values in all but the CAU-F2 population. CML395-F3 and CML495-F3 harbored a larger number of families with HIR = 0% than the other populations. Heritability estimates for HIR obtained from parent–offspring regression (Figure 2) were lower for the CAU population (h2 = 0.32 and 0.41) than for the 1680 population (h2 = 0.80). Spearman's rank correlations were higher for HIR between F2 and F3 generations in both the CAU (rs = 0.51***) and the 1680 (rs = 0.71***) populations than for HIR between F3 and F4 generations of the CAU population (rs = 0.26**).
Significant and high w2 were estimated for EAR in 1680-F2 and 1680-F3 (Table 3). Mean EAR was higher in 1680-F3 than in 1680-F2 and the range of EAR was also wider in 1680-F3. Heritability of EAR was 0.77 on the basis of parent–offspring regression (Figure S1) and Spearman's rank correlation between EAR in F2 and F3 generations was 0.36 (P < 0.001). Moderate correlations existed between EAR and HIR of 1680-F2 (rs = 0.63***) and 1680-F3 (rs = 0.53***).
Linkage maps and segregation distortion
The length of the four genetic linkage maps ranged from 1353 to 1630 cM and the average marker intervals were 4.8 cM in CML395-F2, 5.2 cM in CML495-F2, 13.1 cM in 1680-F2, and 20.4 cM in CAU-F2 (data not shown). Significant deviations from expected allele frequencies were observed in all populations (Figure 3). Most notably, strong (P < 0.001) distortion occurred against the UH400 allele in bin 1.04 in 1680-F2, CML395-F2, and CML495-F2, and in a varying number of adjacent bins depending on the population. Chromosome 4 was affected in three of the four populations (although distortion happened against different alleles), while chromosome 7 was affected by segregation distortion only in CAU-F2 and 1680-F2 and chromosomes 5 and 10 were affected only in CML395-F2 and CML495-F2. Across the four populations, distortion was observed against the UH400 allele in 17 bins and against the non-UH400 allele in 11 bins.
QTL analyses
Five QTL for HIR were detected with CIM in CAU-F2 on chromosomes 3, 5, and 9 explaining together of the genetic variance (Table 2 and Figure 3). In CAU-F3 three QTL were detected on chromosomes 4, 7, and 9 with while in CAU-F4 one QTL was detected on chromosome 9 (qhir8; ). The HIR-enhancing allele was always contributed by UH400 except for qhir5 in bin 5.01 of CAU-F2.
In 1680-F2 and 1680-F3, a major QTL (qhir1; 63–66%) for HIR was detected on chromosome 1 (bin 1.04) and the favorable allele was contributed by UH400 in both generations. Furthermore, a minor QTL on chromosome 3 (qhir2) was detected in 1680-F2 resulting in for 1680-F2 and the favorable allele at qhir2 was contributed by the noninducer parent 1680. One QTL for EAR (qear1) was detected on chromosome 1 (bin 1.04) in 1680-F2 and 1680-F3 ( Table 3). qear1 shared one flanking marker each with qhir1 in 1680-F2 (umc1917) and 1680-F3 (umc1811). The EAR-enhancing allele was from UH400.
In CML395-F3 and CML495-F3, qhir1 was detected in bin 1.04 explaining and 9%, respectively. In both CML populations, SNP PHM5306 flanked qhir1 and the favorable allele was contributed by UH400. On the basis of the physical positions of flanking SNPs (Table 2), the delimited physical interval for qhir1 was about 40 Mbp (data not shown).
Significant dominance effects were revealed for qhir8 in CAU-F4 as well as for qhir1 in 1680-F2, 1680-F3, and CML395-F3 (Table 2). At these loci, the dominance ratio ranged from −0.37 to −1.71 indicating weak to strong recessive gene action adopting the nomenclature suggested by Stuber et al. (1987). Individuals homozygous for the UH400 allele at the nearest flanking marker (see Table 2) to qhir1 always showed HIR > 0% in 1680-F2, while about one-third of the individuals of this genotype class showed HIR = 0% in CML495-F3 (Figure 4).
Using the two-part model approach on HIR data, we confirmed qhir2, qhir6, and qhir8 for CAU-F2; qhir4, qhir7, and qhir8 for CAU-F3; and qhir8 for CAU-F4 (Table 2). Furthermore, qhir1 of 1680-F2, 1680-F3 and CML495-F3 was confirmed with this method. All QTL identified with the two-part model affected only the mean HIR of the nonnull phenotypes, except for qhir1 in 1680-F2 (Figure 5), which additionally affected the probability of occurrence of nonnull phenotypes.
Conceptual genetic framework for inheritance of haploid induction ability
Results of our study and others (Lashermes and Beckert 1988; Barret et al. 2008) showed that (1) the phenotypic distributions of HIR obtained with segregating populations are mostly skewed and (2) a large proportion of null phenotypes is observed. This suggests that the properties of haploid induction ability differ from those of most agronomic traits, which display continuous phenotypic variation that usually approximates a Gaussian distribution.
Haploid induction ability of maize inducer genotypes is considered to be a pollen characteristic, because pollen of inducers under evaluation is used to pollinate a tester and expression of haploidy is assessed on the testcross seed. Under this premise, the genotype of the inducer as such does not matter, but only its gametic array transmitted through the pollen. Hence, dominance effects should be absent for HIR because testcross genotypic values follow a simple additive model in the absence of epistasis (Bernardo 2002). However, segregation distortion as detected in our study at four QTL in three populations can potentially mimic dominance, because if gene frequencies in the gametic arrays produced by heterozygous F2 individuals and even more so of their selfing progenies in generations F3 and F4 deviate from 0.5, this causes a deviation from the additive genetic model for testcross progeny. Nevertheless, it cannot be ruled out entirely that haploid induction ability is not only a property of the pollen itself but also influenced by its parental genotype, similar to factors causing sporophytic self-incompatibility in plants (e.g., Hiscock and Tabah 2003), in which case our observation would reflect dominance between HIR-affecting alleles.
Genetic analysis of haploid induction ability is complicated further by the dichotomous nature of the trait. In the testcross progeny of inducer genotype i the phenotype of each testcross seed ij is coded by the indicator variable zij, with zij = 1 if ij is haploid or zij = 0 otherwise. For our conceptual framework, we assume that the phenotype zij is determined by the concentration yij of the unobservable genetic factor(s) X (e.g., expression level of gene(s) or concentration of metabolite(s)) in the pollen grain triggering the development of testcross seed ij. We suppose that yij displays a quantitative distribution and decompose it according to the model commonly employed in quantitative genetics: yij = Gi + εij, where Gi refers to the genotypic value of genotype i (i.e., Gi = E(yij|i)) for factor X and εij reflects the experimental noise as well as genetic segregation in case of a heterozygous inducer genotype. This situation is depicted schematically in Figure 6. If yij exceeds a certain threshold T, the phenotype of seed ij will be haploid. Consequently, , which takes zij = 1 if yij > T and zij = 0 if yij ≤ T, links the distribution of the unobservable variable yij with the phenotype zij. Thus, genotype i induces haploidy in testcross seed ij with a true probability of pi such that Pr(zij = 1) = pi, and it follows that Pr(zij = 0) = 1 – pi. Hence, the expectation of zij is E(zij) = pi and its variance is var(zij) = pi(1 – pi). The genetic factor(s) X affecting pi are unknown, but once identified (e.g., through gene expression studies with near-isogenic lines) they could potentially be measured on a continuous scale and the inheritance could be studied directly.
In summary, we propose that the trait haploid induction ability is a threshold character under polygenic control, yet leading to dichotomous expression (haploid vs. diploid) on the level of the individual testcross seed. Prominent examples of threshold characters include disease susceptibility (affected vs. not affected) in plants or litter size in large mammals (single vs. twin births; Falconer and Mackay 1996). Further, the two common assumptions for genetic analysis, i.e., normality of residuals and homoscedasticity, are not fully met for haploid induction ability. Composite interval mapping (Jansen and Stam 1994; Zeng 1994) and alternative approaches such as nonparametric tests (Kruglyak and Lander 1995) or two-part model tests (Broman 2003) are expected to work well for nonnormally distributed data as long as critical LOD scores are determined via permutation tests from the original data set. However, heteroscedasticity of error variances remains an unsolved problem and warrants further research.
Heritability of HIR
Between-family heritability estimates for HIR (represented by h2 estimates from parent–offspring regression) were lower than within-family heritability estimates (represented by repeatabilities w2 estimated from HIR of three ears per family). Since parents and offspring were not evaluated in the same environments, genotype-by-environment interactions for HIR may have reduced h2 estimates. Further, the discrepancy between h2 and w2 can be explained by natural selection disfavoring the haploidy-inducing gametes during selfing as suggested by the observed segregation distortion. From an evolutionary point of view, a higher proportion of haploids in the progeny caused by high HIR of the pollinator would result in fitness disadvantages, because haploid maize plants are less vigorous, often male sterile, and, therefore, generally less likely to produce offspring than diploid maize plants (Chase 1952; Chalyk 1994), which explains the counteracting force of natural selection. The latter may also explain the difficulties encountered during maintenance of inducers reported by maize breeders.
Surprisingly, h2 for HIR estimated from regression of the 1680 generations was twice as high as those estimated from regression of the CAU generations or the estimate Deimling et al. (1997) (h2 = 0.44) reported for HIR. Since h2 is a population parameter (Falconer and Mackay 1996), the different germplasm base explains this discrepancy. Further, from our conceptual genetic framework it follows that h2 for HIR estimated from a segregating population such as F2 or F3 depends on the genotypic variance for the probability parameter pi as well as on the sample size Ni of the testcross progeny used for determining HIR of inducer genotype i. Consequently, estimation of h2 remains difficult because (1) phenotyping of HIR in multilocation experiments on various testers is extremely laborious, and (2) heteroscedasticity as well as the associated segregation distortion complicate quantitative genetic analyses including estimation of h2, so novel biometric approaches are urgently needed.
Agreement between conceptual genetic framework and QTL results
To the best of our knowledge, this is the first study that used multiple populations and generations to identify QTL responsible for HIR. A common approach for comparison of QTL locations across populations is using joint linkage analysis as proposed by Blanc et al. (2006), but this was not possible because of a lack of common markers between the four populations. Yet, simple addition of LOD curves from CIM runs of individual populations revealed that only qhir1 was significant across both CML populations (see Figure S3 and Figure S4), while no joint QTL was significant for the two Chinese populations (data not shown). Further, a variable degree of segregation distortion in different populations complicates joint analysis. Therefore, we refrained from joint linkage analysis and compared QTL across populations using the "bin concept," which is well established in the literature (e.g., Gardiner et al. 1993; Schön et al. 2010).
The number of QTL and the magnitude of QTL effects obtained for HIR in the present study varied depending on populations and generations. There was a clear trend, however, for qhir1 in bin 1.04 having a major effect on HIR in populations involving noninducer genotypes as females (1680, CML395, and CML495). Previous studies have also identified this chromosomal region to segregate in populations involving only one inducer parent (Deimling et al. 1997; Barret et al. 2008). In contrast, no QTL was detected in bin 1.04 in CAU-F2, CAU-F3, and CAU-F4, which were derived from a cross of the two haploid inducers CAUHOI and UH400. Most likely, the CAU generations are not segregating for qhir1 because the favorable allele is fixed in the parents, which provides a simple biological explanation for the incongruency of QTL. In fact, further analysis of the bin 1.04 region with 16 markers that were all polymorphic between inducer line UH400 and noninducer line 1680 revealed monomorphism between inducers UH400 and CAUHOI (Figure S2). As large-effect QTL are subjected to strong selection pressure, qhir1 has likely become fixed quickly when selection for high HIR occurred during the development of UH400 and CAUHOI. Further, the two inducers may be identical by descent in this chromosomal region due to derivation from the same original inducer source, Stock6 (Coe 1959).
QTL detection frequencies obtained with 200 runs of fivefold cross-validation (Utz et al. 2000) were 95–100% for the large effect QTL in bins 1.04 (qhir1) for each of the populations except CAU, and 9.01 (qhir8) for the CAU populations in this study. This explains why these QTL were detected in more than one population and/or all generations of the respective population and underlines their significance for HIR. In contrast, QTL detection frequencies ranged from 23 to 80% for QTL with smaller effects and, accordingly, none of the minor QTL except qhir2 was detected in more than one population. The power of simultaneous detection of QTL in different experiments is obtained by multiplication and, therefore, low congruency of minor QTL across experiments meets expectations and has also been reported previously (Melchinger et al. 1998; Mihaljevic et al. 2004).
The use of cofactors in CIM has been shown to increase power of QTL detection (Jansen and Stam 1994; Zeng 1994). Analyses with the two-part model did not confirm all of the QTL identified with CIM. Higher critical LOD thresholds were employed for this model and the minor QTL identified with CIM (qhir3 and qhir5) fell short of the critical LOD during two-part analyses due to lower power for QTL detection (Broman 2003).
The two-part analyses also revealed that qhir1 in 1680-F2 significantly affected the chance of whether genotypes have HIR > 0%, while the remaining QTL affected only the genotype means once HIR > 0%. Hence, we speculate that qhir1 acts as a key modulator for HIR; i.e., genes located in bin 1.04 trigger genetic processes that enable passing of the threshold outlined in our conceptual genetic framework. In this case, introgression of qhir1 would equip any germplasm with haploid induction ability. However, in the two CML populations not all individuals homozygous for the inducer allele at the flanking markers of qhir1 showed HIR > 0%, a phenomenon known as incomplete penetrance (e.g., Barret et al. 2008; Eldar et al. 2009; Gaudet et al. 2010), while complete penetrance was observed in 1680-F2 and 1680-F3. Yet, genotypes at the actual QTL position are unknown and denser marker coverage would be necessary to more conclusively infer about penetrance.
The QTL qhir1 explained a much larger proportion of the genetic variance in 1680-F2 and 1680-F3 than in CML395-F2 and CML495-F2, possibly due to incomplete penetrance. This discrepancy can be attributed to different testers and the associated haploid identification systems employed for the different mapping populations (Kebede et al. 2011; Prigge et al. 2011). Further, the presence of different modifier genes (Allard 1999) acting as enhancers or suppressors of major genes in different germplasm sources as well as QTL by genetic background interactions (Pumphreys et al. 2007) may be responsible. Future studies should employ different testers (such as liguleless, glossy, R1-nj) and near-isogenic lines developed from crosses of inbreds with contrasting HIR to focus on modifier QTL regions in greater detail and investigate putative epistatic interactions. Our results suggest that populations developed from crossing two inducers will be particularly helpful in revealing QTL other than qhir1.
Effects of segregation distortion
Distorted allele frequencies were observed in all populations indicating the presence of segregation distortion loci (SDL) that cause linked markers to deviate from expected Mendelian segregation patterns due to gametic or zygotic selection (Xu 2008). It is expected that recombination frequencies and, consequently, marker positions during linkage map construction will not be affected by segregation distortion if only one SDL is present in a distorted region and codominant markers such as SSRs or SNPs are used (Lorieux et al. 1995; Lu et al. 2002). The total map lengths estimated for our F2 populations were comparable to those reported in previous studies with F2 populations (e.g., Melchinger et al. 1998; Mihaljevic et al. 2004). Further, both marker order as well as bin positions have been confirmed for all markers on the basis of published IBM2 2008 Neighbors Map or physical locations. Thus, our QTL results should be robust and reliable as far as the flanking markers and chromosomal bins are concerned.
In fact, had SDL been excluded from analysis, qhir1 would have been missed in our study. This confirms previous results indicating that genomic regions exhibiting strong segregation distortion are equally or even more likely to contain QTL (e.g., Wang et al. 2005). The strong distortion against the UH400 marker allele observed in the region of qhir1 in three populations of this study supports the putative association between HIR and transmission failure of the inducer gamete suggested by Barret et al. (2008). Perhaps the same or tightly linked loci are responsible for both causing segregation distortion as well as triggering induction of haploidy.
Under the assumption that qhir1 is responsible for the observed segregation distortion in bin 1.04, this relationship could potentially be exploited for fine mapping of qhir1. By saturating this QTL region with SNPs and identifying those SNPs with maximum segregation distortion, fine mapping of the QTL may be achieved without the need of elaborate designs in a simple large-size F2 population. Schneeberger and Weigel (2011) proposed a similar approach using next-generation sequencing technology for mutation mapping in Arabidopsis.
Putative candidate genes for haploid induction ability
Identification of candidate genes for haploid induction ability is premature because the underlying biological mechanisms are still unknown. Considering that limited pollen production capacities and protogyny have been observed for inducers (data not shown; W. Schipprack, personal communication), weak or partial male sterility may be involved in haploid induction. Particularly relevant are mutants producing pollen that does not always lead to complete fertilization of egg and central cells. Almost 50 male sterility genes (ms; Neuffer et al. 1997) are known. For example, mutants carrying ms12, which is located on chromosome 1, show partial male sterility, probably depending on the genetic background (Albertsen and Phillips 1981).
Further, defective kernel mutations (dek; Neuffer et al. 1997) map to similar chromosomal regions as QTL identified in our study in bins 1.04 (dek32), 3.02 (dek5, dek24), 3.06 (dek17), 4.03 (dek11), 5.01 (dek18), 5.04 (dek9, dek26, dek27, dek33), and 9.01 (dek12). These mutations are characterized by a variation of degenerated embryo and endosperm tissues. Several mutants such as carriers of dek12, dek32, and dek33 show completely collapsed kernels appearing as embryoless pericarps without any starch deposition (Sheridan and Neuffer 1980). This phenotype was employed to determine EAR in our study. Significant variation for EAR was detected and EAR estimates of >10% were observed, which is more than expected in normal crosses. The joint presence of qhir1 and qear1 in bin 1.04 as well as the highly significant rank correlations between HIR and EAR in 1680-F2 and 1680-F3, suggest a possible relationship with inducer properties. However, further genetic mapping or gene expression studies will be necessary to study the involvement of dek loci in HIR. Examining the genotypes of endosperm and embryo via comparative seed and leaf DNA-based analyses (Gao et al. 2011) may be helpful to study the fertilization process(es) inducing haploidy and to better understand the biological basis of in vivo haploid induction in maize.
In Arabidopsis thaliana, Ravi and Chan (2010) report the production of haploid plants by crossing the source germplasm with a mutant carrying a genetic alteration of the centromeric histone CENH3. In our study, no QTL for HIR was detected in bin 6.06, which harbors CENH3 of maize (Zhong et al. 2002). This indicates that in vivo induction of haploidy in maize is not governed by the mechanism reported by Ravi and Chan (2010), although such centromere-mediated genome elimination may also work in maize to produce haploids. Nonetheless, in vivo induction of haploidy may be fundamentally different in mono- and dicots and maize could serve as a model species for studying this phenomenon in monocots.
Implications for haploid inducer development
Transgressive segregants were identified in all populations except CML395-F3 and also noninducer parents contributed HIR-enhancing alleles, indicating that inducers with increased HIR can be generated through targeted parental recombination and selection. To reduce the laborious and time-consuming task of phenotypic selection for HIR, marker-based approaches such as genomic selection are suitable for increasing the frequency of favorable alleles for HIR in inducer development programs. This seems particularly relevant if haploid induction ability is indeed governed by one or few major genes influenced by several modifier genes (Allard 1999), as major genes will quickly become fixed, and gain from selection will then depend entirely on small-effect modifier genes, which will be extremely difficult to select for phenotypically.
In addition, targeted introgression of relevant QTL into adapted and agronomically superior noninducer germplasm will enable the development of custom-made inducers that can be employed for efficient DH line production in various breeding programs and agroecologies. qhir1 represents an interesting chromosomal region for map-based cloning and the required fine mapping can be achieved by developing near-isogenic lines. However, incomplete penetrance observed for this QTL in CML395-F2 and CML495-F2 suggests that the relationship between genotype and phenotype is not that reliable in all genetic backgrounds. Thus, phenotyping of marker-selected inducers still seems necessary as a final evaluation step.
We greatly appreciate the skilled technical assistance of Li Haochuan, Dong Xin, Guo Mingxin, Chen Liming, and co-workers at China Agricultural University as well as of Ciro Sánchez, Alberto Vergara, Ana Mely Islas Montes, and Germán Bastian de León, and co-workers at International Maize and Wheat Improvement Center (CIMMYT) during DNA extraction, testcross seed production, and HIR assessment. We are grateful to Jianbing Yan for facilitating the parental screening with the 1536 SNP chip and to Stefan Scholten and H. Friedrich Utz for valuable comments on an earlier version of this manuscript. This research was financially supported by National Projects nycytx-02 and 2009CB118400 at China Agricultural University, and by the Bill and Melinda Gates Foundation, Seattle, Washington, through the Drought Tolerant Maize for Africa (DTMA) project at CIMMYT. Vanessa Prigge is indebted to Eiselen Foundation, Ulm, Germany, for financial support of her research as a joint Ph.D. student between CIMMYT and the University of Hohenheim, Germany.
Supporting information is available online at http://www.genetics.org/content/suppl/2011/11/30/genetics.111.133066.DC1.
↵1 These authors contributed equally.
Communicating editor: A. Charcosset
- Received July 21, 2011.
- Accepted November 19, 2011.
- Copyright © 2012 by the Genetics Society of America


text:
Live blogging the Fashion Rocks
Someone actually named their daughter "Poppy"? And she's not even British?
I love me some skyscrapers. We all know that. But holy shit. Did you SEE how high Gwen Stefani's pumps were?? My lord, how the hell does she walk in those?
Rob Thomas, Helen was right - your voice is vocal orgasm. But baby, please don't wear your jeans so tight. This is supposed to be about FASHION. You look like something out of my freshman yearbook.
Granted, she sings in a foreign language, so I have no idea what she's saying. Maybe she's explaining herself. Because I do not get this herky-jerky dancing thing that Shakira does. It is not sexy and it's not attractive. It looks like someone showing off their double-jointed finger.
David Bowie usually dresses impeccably. And he looked pretty good tonight...if you couldn't see the lower half and the fact that his tailored suit pants came to just ABOVE HIS ANKLES! And...is that a black eye??
Gwen, you looked sooooo much better in the 1940s glamour dress than you do in that caftan-slash-nightgown. Why do you do that? Is that really worthy of introducing...
Duran Duran! The band so nice, they named it twice. Let me just say for the record, that we had a group of DD groupies in my high school in 1984. They wore lots of pins on their t-shirts that they bought at Spencer gifts with all the faces of the band members on them. They tried to circulate petitions to get the band to come play the Biloxi Coliseum (which, I'm assuming has been washed away now). I was NOT one of these girls. I did not color my hair Nick Rhodes Pink. But I liked their early music. "Rio" still ranks as one of my all-time favs (and I don't necessarily even think it's one of their best). Their video work was groundbreaking in the early days of Mtv. And I did think that John Taylor was H-O-T.
David Bowie & Arcade Fire?? How cool is THIS?? Oh please, please let this be cool and not suck.
OK, this is just getting boring now. It's the same performers coming out over & over in different pairings. I'm going to bed.

subset: commoncrawl


text:
Busy week! On Monday we went on a field trip to Devil's Hopyard in East Haddam, CT. The scenery was spectacular as the leaves are starting to peak now. We talked about the local geography of the area, the lore surrounding the name "Devil's Hopyard," and the importance of Chapman Falls to the local mill industry.
There were also lots of interesting rock formations that we saw and later read about, including glacial puckering, rock shelters, folds and mineral deposits (biotite and quartz crystals).
At home the Peeps wrote and illustrated the legend of Devil's Hopyard, which says that the Christian Devil, while passing through, got so mad when his tail got wet in the falls that he stomped the ground leaving the huge pothole formations the falls are known for. They're actually caused by rocks getting caught and spinning in the current. I tried to get pictures of them but unfortunately they didn't come out!
Meanwhile, Peep #1 continued with his Egyptian God/Goddesses pages:
He also made a canopic jar, which didn't turn out as well as his Anubis. He still liked it though. 🙂
Peep #2 continued to work on her Botany block. She's enjoying it so much that she wants to do one more week. I had planned to do two weeks in the fall, two in the spring, but since she's having so much fun with it we'll continue. Here are some examples of the things she did this week:
She also did some more Math activities from Land of Pome, as well as some illustrations to reinforce the "Time and Measurement" lesson from Oak Meadow Math 5:
In Grammar, they both learned about the Present Perfect, Past Perfect, and Future Perfect tenses. We will be exploring this more next week:
Finally, we finished our leaf garland and strung it above the kitchen chalk board! (Don't look too closely; you'll see our mistake in one of the leaves. ;))
We also made these really pretty window leaf transparencies using dried leaves and wax paper. We framed the first one but then decided we liked them better free floating, like leaves being blown about wildly against the windows…


text:
Table of Contents
- 1 Overview
- 2 The Women, Business and the Law
- 3 The FAO Gender and Land Rights Database
Since the end of communism in 1989, Poland's transition to a democratic state and free market economy has witnessed limited progress in gender equality for women. Wage discrepancies, traditional customs that stress the role of women in the private sphere, and inconsistent policies targeting gender discrimination and protecting women's rights must still be addressed before gender equality is achieved.
The collapse of the communism not only was accompanied by the deterioration of women's socio-economic status, but it also witnessed the growing political power of the Roman Catholic Church and its position to effectively impact the politics. In 1980s the Church used the opportunities created by democratic processes to intensify its campaign for a complete ban on abortion. As a result in 1989-1992 several attempts were made by fundamentalist forces in the Parliament to dramatically restrict the law on abortion, which was legal in Poland since 1956. In 1993 abortion was criminalised and a restrictive anti-abortion law was introduced, limiting the right to termination of pregnancy only to three conditions: 1) when the pregnancy threatens a woman's health or life; 2) when the fetus is severely and irreversibly damaged; and 3) when the pregnancy is a result of criminal offence (e.g. rape, incest).
Despite ongoing advocacy efforts of the community of women's rights advocates to liberalize abortion law, especially in the context of the growing phenomenon of the abortion underground in Poland, the restrictive legislation is still in force.
Poland ratified the CEDAW (CEDAW) in 1981, voluntary pledging its commitment and accountability to take all necessary actions to advance women's human rights. However, not enough effort is being undertaken by the Government to implement the provisions of the Convention in law and in practice. In Poland there is no comprehensive and consistent policy on women's rights and gender equality. Since 2005, the Government has not formulated nor implemented any National Programme of Action For Women. Furthermore, there is no sustainable institutional machinery for the advancement of women and each appointed Government establishes temporary structures to deal with the issue. Women's rights continue to be a highly politicized subject and the commitment to improve the situation of women is entirely dependent on the political party in force.
The Act on the Implementation of Some Regulations of the European Union Concerning Equal Treatment that entered into force in January 2011 established the office of the Government Plenipotentiary for Equal Treatment, which is now responsible for the Government's anti-discrimination and gender equality policies.
The above act is very limited in the scope of the protection it guarantees to various groups that are vulnerable to discrimination. For example, women are not protected in such critical areas of their lives such as education or access to healthcare. The law was adopted despite a severe critique of it by the human rights organisations in Poland. The Government introduced the law only in the context of the threat of financial penalties for not implementing the European Commission 's Directives.
Social Institutions
The Social Institutions and Gender Index (SIGI) measures gender-based discrimination in social norms, practices and laws across 160 countries. The SIGI comprises country profiles, a classification of countries and a database; it serves as a research, policy and advocacy tool for the development community and policy makers.
The SIGI covers five dimensions of discriminatory social institutions, spanning major socio-economic areas that affect women's lives: discriminatory family code, restricted physical integrity, son bias, restricted resources and assets, and restricted civil liberties. The SIGI's variables quantify discriminatory social institutions such as unequal inheritance rights, early marriage, violence against women, and unequal land and property rights.
In the Social Institutions and Gender Index 2014 Edition , Poland was not classified in the SIGI due to lack of full dataset. It has lower discrimination in restricted access to resources and assets and higher discrimination in restricted civil liberties. Read the full country profile and access the data here: http://www.genderindex.org/country/poland
Women are concentrated in the more poorly paid occupations and trades, especially in low paying and low status public sector occupations. In 2006, 52.9% of women aged 15 years and older were inactive in the labour market, among men – 37.0%. The greater labour market inactivity among women compared to men is related to traditional perceptions of women as being connected to the private sphere, and the belief that men are responsible to a greater degree for the material well being of a family. Only 5% of households with children up to the age of 12 take advantage of institutionalised Child care. Only about 3% children aged 0-3 attend childcare services; childcare remains unavailable to almost half of the children aged 3-5, the deficit being especially acute in rural areas. The present-day childcare crisis must be seen in a broader context of structural and cultural factors pertaining to the welfare system and the treatment of care-work by social policy in general. The trend in post-socialist countries after 1989 has been described by sociologists as the refamilisation of care. For Poland this process was additionally exacerbated by cultural factors, especially by pressure from the Catholic Church. During the 1990s the state largely withdrew from its former responsibilities in the realm of care (including childcare), while the market failed to fill the resulting gap. In effect, inequality between men and women deepened both at home and in the labour market.
One of the major problems for the labour market in contemporary Poland is the growing phenomenon of 'working poor'. The lowest earning are in the feminised professions and thus the phenomenon affects women to a larger extent than men. The Salary Reports in Poland 2009 by Sedlak & Sedlak Company shows that the lowest wages are earned by women in the garment industry. Cashiers (salespersons) – a strongly feminised occupation – were in the third position from the end. One in eight employed women do not fall into the category of 'working poor', with salaries that do not cover the cost of living, and thus that do not allow women to live in dignity. This has considerably contributed to creating social underclass (with a significant number of women) living in poverty.
The biggest gaps between men's and women's salary is among people with basic vocational and primary education (and also among people with tertiary education) – over 30%. This is particularly alarming for women with the lowest education, which means the lowest earnings.
The basic vocational education addressed to women neither corresponds to the current challenges in the labour market, nor leads to a salary which could contribute to reducing the huge gap in the income between women and men and guarantee a living wage. Among people with this level of education, the gender pay gap thus amounted to 33,3% in 2008.
The relationship between the activity rate and education is very apparent. In 2010, the activity rate of women with tertiary education was 79%, and that of women with primary or incomplete primary was only 13,4%. The data reveals an enormous polarisation of women in their economic activity, as well as a continuing trend of 'pushing' women with the lowest education outside the labour market and into the informal economy or emigration (often seasonal).
Legislation and Institutional Machinery
The formulation of equal rights of women and men found in the Constitution of the Republic of Poland guarantees the equality of women and men in employment, access to promotion, and equality of wages for work of equal value (art. 33). However, despite ongoing advocacy of the civil society organisations no gender equality or anti-discrimination act was adopted. The first version of the gender equality act was prepared in 1993 and since then it has been amended and debated in every four-year term. In December 2004, the Government adopted a bill on combating and preventing family violence.
In December 2010, Poland adopted the Act on the Implementation of Some Regulations of the European Union Concerning Equal Treatment. This law was only approved in the context of the obligation to implement the Directives of the European Commission on equal treatment and the threat of severe financial sanctions for not fulfilling this duty. Unfortunately, there was no political will to formulate comprehensive anti-discrimination legislation that would guarantee legal protection from discrimination for all groups in all areas of life. The newly adopted so called 'anti-discrimination' act establishes the Office of the Government Plenipotentiary for Equal Treatment (GPET), responsible for the coordination of implementation of the Government's anti-discrimination policies.
Until 2011, when the so called 'anti-discrimination' act was adopted there was no sustainable institutional machinery for the advancement of women in Poland. Each appointed Government established temporary structure and its competencies were highly dependent on the political will of the winning party to advance gender equality. For example, after winning the elections in 2005, the right wing, conservative political party – Law and Justice eliminated the Office of the Government's Plenipotentiary for the Equal Status of Women and Men, which had a severe consequences on the situation of women in Poland. Since 2005, the Government has not formulated nor implemented any National Programme of Action for Women.
Political Empowerment
Women in Poland gained the right to vote and run for office in 1918. Women attained their highest numbers in Sejm (Poland's Parliament) from 1980 to 1985, when they represented 23% of parliamentarians. After the 2005 elections, women's representation dropped. While the number of women parliamentarians remained at a similar level to the previous term's (94), the number of women in the Senate went from 23 to 13, resulting in Poland placing much lower, 49th
(out of 139) in the Inter-Parliamentary Union's worldwide ranking (IPU).
In Polish politics women are a minority. Their participation in Government, both at the national as well as the local level is below 30%. This is despite the fact that since 1989 the number of women parliamentarians has been constantly increasing. Until 2001, in each term of the Seym (the lower house of the Polish Parliament), women constituted between 10 and 13%. After the 2001 elections women constituted 20% of all parliamentarians. This sudden increase in the participation of women has to be linked to the voluntary adoption of quotas by three parties, the SLD (Democratic Left Alliance), UP (Labour Union) and UW (Freedom Union). These quotas required either gender to have at least 30% of the total number of candidates on electoral lists.
In 2010 women's rights advocates involved in a massive initiative called 'Women's Congress' drafted an act on parity on electoral tickets and launched the campaign for its adoption. They gathered over 100 thousand signatures in support of this initiative. In the legislative process the draft law was amended and quota system amounting 35% was introduced on electoral tickets to the Parliament. The results of the October 2011 elections show that the effectiveness of this measure is considerably limited. Although it led to the doubling of female candidates in the 2011 election compared to the 2007 election, women got only 24% of places in the lower House of Parliament. This is a record number of women in Seym, but still women remain to be significantly underrepresented in this decision-making body. Women's rights advocates continue to argue that there is a need to adopt parity with a zipper system (alternate order of women and men) on electoral tickets to effectively increase women's participation in political life.
In Poland, the Life expectancy at birth is 76 years for Gender differences in life expectancy, females have a higher life expectancy average rate with 80 years, whereas men can expect an average of 71 years. Thus, men and women in Poland have a higher average life expectancy compared to the global average of 66 years for men and 71 years for women. The adult mortality rate (per 1000 adults between 15-59 years) remains high with 137 in contrast to the global average of 176. The maternal mortality ratio is measured with 6 deaths per 100000 live births, which is comparably low when considering the global average of 260 deaths. The under-5 mortality rate has decreased significantly from 1990 till today, and is stated as 6 deaths per 1000 live births, which includes both sexes and is below the global average of 60 deaths. Prevalence of HIV occurs at one case per 1000 adults aged from 15-49 years, which is far less than the average of 8 cases globally. The prevalence of tuberculosis is also lower with 32 cases per 100000, than the global average with 201 cases. Tobacco smoking does not show a big contrast between the genders; 37,7% of 15+ years old men are regular smokers, and 29,6% of women smoke regularly. There is an equal proportion, 29,9% of both genders older than 20 years to suffer from obesityhttp://www.who.int/gho/countries/pol.pdf.
One of the major area of gender-based discrimination in Poland is sexual and Reproductive Category:Health and rights. Poland has one of the most restrictive anti-abortion legislation in Europe. What is more, the law is more restrictive de facto than de jure. Women face extreme difficulties with accessing lawful abortion, which resulted in a number of cases at the European Court of Human Rights in Strasbourg in the last couple of years.
- http://www.soros.org/initiatives/women/articles_publications/publications/equal_20050502
- UNDP, Gender Mainstreaming Report 2007: Poland [accessible on: http://www.undp.org.pl/gender/aktualnosci.php?news=1019] http://www.who.int/gho/countries/pol.pdf
- KARAT Coalition. Alternative Report on the Implementation of CEDAW in Poland 2012.
The Women, Business and the Law
Where are laws equal for men and women?
The Women, Business and the Law, 2012 presents indicators based on laws and regulations affecting women's prospects as entrepreneurs and employees. Several of these indicators draw on the Gender Law Library, a collection of over 2,000 legal provisions impacting women's economic status. This report does not seek to judge or rank countries, but to provide information to inform discussions about women's economic rights. Women, Business and the Law provides data covering 6 areas: accessing institutions,using property, getting a job, providing incentives to work, building credit, and going to court. Read more about the methodology.
For detailed information on Poland, please visit the Women, Business and
the Law Poland
The FAO Gender and Land Rights Database
The FAOGender and Land Rights Database contains country level information on social, economic, political and cultural issues related to the gender inequalities embedded in those rights. Disparity on land access is one of the major causes for social and gender inequalities in rural areas, and it jeopardizes, as a consequence, rural food security as well as the wellbeing of individuals and families.
Six categories
The Database offers information on the 6 following Categories:
- National legal frame
- International treaties and conventions
- Customary law
- Land tenure and related Institutions
- Civil society organizations
- Selected Land Related Statistics
For detailed information on Poland, please visit the report on Poland in the


text:
This year we exhibited at the Research & Results show in Munich
Ton Paans and Arti Shah manned the stand. Before the show, Arti commented "I'm looking forward to this opportunity to talk face to face with people from our industry about our comprehensive one-stop-shop for all sampling needs worldwide.".
After the show, we talked to Arti and Ton to find out how the show worked out
'Our impression was that R&R is more globally focused than the UK shows. It feels like the right place as we have a huge global reach and a base in Amsterdam. Research & Results is the leading focal point of B2B networking for the market research industry in Europe.' said Arti.
Arti also mentioned, 'R&R is well-known in the industry, where many people or businesses from across the globe exhibit or visit. We have a strong European element to Sample Answers and Ton has represented us here every year. As he is multilingual he connects with many individuals, speaking in German, Dutch etc. It's important that we attend for global brand awareness purposes and to expand our knowledge of other businesses/competitors in the industry" .
The show was very busy this year
'The famous Sample Answers pens remained popular on the stand' said Ton."There was definitely an increase of interest in online sample and we felt that we were reaching the right people within UK and Europe. Many countries and market research specialists were represented, as well as the big brands. We attended the Research Club party – great fun and a good way to socialise. We'll definitely be there next year." commented Arti.
"Only a relatively small share of the total attendance is the right target audience for Sample Answers, inevitable given our core business is telephone sampling.
Similar to last year, offline is a small pond to fish in, but there remains an interest amongst many visitors. Online is very dominant, but there is an increased awareness of and potential interest in hybrid solution" stated Ton.
"We gained an insight of other businesses' products and services and felt that R&R did a great job of expanding our know
ledge globally. We had some very interesting conversations with individuals who we've added as contacts
on LinkedIn now. All the people we met who enquired about our products and services have received an email follow up. It was a great show and experience' said Arti.
"One of the best things about R&R is that the agencies, including large ones, still attend and exhibit – not the case at many other industry events. Research and Results show remains, in my opinion, the best MR trade show by far. Interesting to have seen it develop from a parochial German event years ago to a truly international show. For me it is the pre-eminent trade show for the market research business in Europe, well liked and highly rated by everyone I have spoken to". continued Ton.
Ton Paans sums up the 2017 R&R experience
"We've already received some new enquiries, resulting in a worthwhile order. Overall, Research & Results was great for establishing new relationships and maintaining and reaffirming existing ones. In that respect, I have aways been happy with our attendance at R&R over the years".


text:
From The Garden
Soup & Sandwich Combo $11.00
Soup & Salad Combo
Specialty Platters
Homemade Burgers
Carriage House Creations
Croissant Sandwiches
Dinner Seasonal Features
Dinner Land Entrees
Dinner Fresh Seafood Entrees
Our Dinner Specialties
Children's Menu
Creamy Brie baked in flaky puff pastry and served with sliced apple, caramel, walnuts and toasted French baguette. For two or three to share.
Four jumbo shrimp coated in crispy panko and coconut. Served with raspberry Thai dipping sauce.
Button mushrooms stuffed with our crab imperial.
Herbed tortilla topped with gulf shrimp, lump crabmeat and assorted cheeses broiled for two to share.
Tender and sweet shrimp, chilled and served with cocktail sauce and lemon.
Jumbo lump & claw crab meats mixed with cream cheese & lightly seasoned. Served in a sourdough boule with toasted French baguette. Excellent choice for two.
Cup $4.25 - Bowl $5.25
Cup $3.75 - Bowl $4.75
Our House Dressing Tomato | Vinaigrette | Buttermilk Ranch | Bleu Cheese | Fat-free Honey Dijon | Honey Mustard | Thousand Island | Pepper Parmesan | Olive Oil & Balsamic Vinegar
Add your favorite topping:Crumbled Bleu Cheese, Crumbled Bacon or Shredded Cheddar - $1.00
Topped with fresh mushrooms, red onions, black olives, tomatoes, and crisp bacon. Served with warm Louisiana bacon dressing.
Topped with cajun chicken breast | $11Topped with sauteed gulf shrimp | $12Topped with marinated salmon | $14
Cooked and peeled in our kitchen.
Extra cheese, extra fruit, craisins, and toasted almonds.
With fat free honey dijon dressing.
Chicken, bacon, and bleu cheese.
Shrimp Salad, Crab Meat, Bacon, Avacado, Egg, Crumbled Bleu, Tomato, Cucumber and Carrot
All sandwiches served on soft potato roll with lettuce, tomato and mayo on the side. Soup and sandwich includes chips and pickle. No Substitutions Please!
Choose 1: Seafood chowder, cream of crab, soup du jour, or french onion.
Choose 1: White tuna salad, roasted turkey, baked ham, top round of beef, or chicken salad.
Cream of Crab or Seafood Chowder served in a toasted french bread bowl, served with a garden salad.
Choose 1: Garden $8.00 - Caesar $9.00 - Chef $10.00 - Spinach $10.00 - Cobb $11.00 - Shrimp $11.00
With a mushroom madeira sauce.
Our Signature Selection.
Market Price
Baked in lemon pepper butter.
Served over penne pasta with fresh vegetables topped with parmesan cream sauce. Topped with Herb Chicken $11.00 Topped with Lemon Sauteed Shrimp $12.00
Lettuce, tomato, onion, thousand island dressing, and american cheese.With bacon $11
Served with a touch of mushroom madeira sauce.
Cannonball BBQ sauce, flame grilled onions and peppers, topped with provolone.
Boneless Chicken Breast Topped with Chesapeake Crab Dip and Cheddar Cheese on a Brioche Roll.
Fresh spinach, melted provolone, and a side of thousand island dressing.
Lightly seasoned lump crab. Our Signature Selection.
Served with Lettuce, Tomato and Homemade Tarter on the side
Steak, melted provolone, sauteed mushrooms, and onions, on a toasted sub roll with lettuce, tomato, and mayonnaise.
With roasted peppers & onions.
Served with Lettuce and tomato on the side.
Roasted turkey on grilled rye with melted swiss, sauerkraut, and thousand island dressing.
Thinly sliced corned beef on grilled rye with melted swiss, sauerkraut, and thousand island dressing.
Grilled chicken breast topped with lettuce, tomato, mozzarella, and basil pesto.
Fresh sliced roast beef sauteed with carmelized onions on a toasted sub roll with melted provolone, served with au jus for dipping.
Roasted turkey breast, ham, muenster cheese, bacon, lettuce, tomato, and mayonnaise.
Homemade shrimp salad served with lettuce and tomato on the side.
Crab imperial with melted muenster cheese.
Served with lettuce and tomato on the side.
Horseradish Maple Glaze, Carmelized Oinions and Fennel Risotto with Aromatic Vegetbales
With Brandy Apples and Onions Served with Chef's Potato and Vegetable
Seared with Garlic, Spinach, Parmesan, Sun Dried Tomato Cream, Served with Garlic Redskins and Chef'f Vegetabled du jour
Served over Angel Hair Pasta
All entrees include fresh vegetable, potato du jour and homemade bread.
A side Garden, Caesar or Hearts of Lettuce Salad is available for $1.50
Add sauteed mushrooms to any entree for $5.00
(When Available) Certified Angus Beef prime rib slow roasted and carved to order. Served with creamy horseradish and au jus.
12oz. Certified Angus Beef NY strip, lightly seasoned and flame broiled. Topped with garlic and herb infused creamy boursin cheese.
6oz. Certified Angus Beef filet mignon, hand cut, lightly seasoned and flame broiled served over top poached asparagus and finished with jumbo lump crab tossed in bearnaise sauce.
Veal scallopini seared and topped with Romano and Parmesan cheeses finished with a garlic, white wine, and prosciutto pan sauce.
Tender veal scallopini seared alongwith poached asparagus and sauteed jumbo lump crab tossed with bearnaise sauce.
8oz Certified Angus Beef filet mignon, hand cut, lightly seasoned and flame broiled, your choice of bearnaise or mushroom madeira sauce.
The consumption of raw or undercooked shellfish, eggs, meats or poultry may increase the risk of food borne illness.
Jumbo and lump crabmeat in a classic mild sauce, seasoned lightly and broiled, served with homemade tartar sauce.
Jumbo and lump crabmeat in a lightly seasoned cream sauce and finished with flavored bread crumbs.
Jumbo gulf shrimp, sea scallops, and jumbo lump crab meat sauteed in a tarragon infused butter with grilled foccacia toast points.
Flavorful sea scallops broiled in a garlic and shallot butter. The slight hint of nutmeg makes this scallop dish stand apart.
Four jumbo gulf shrimp topped with our zesty crab imperial and finished with seasoned bread crumbs, served with clarified butter.
Marinated in a mixture of soy, Kentucky bourbon, garlic and brown sugar.
Gluten free menus are available from your server.
Carriage House Crab Cake and Filet Mignon with mushroom madeira.Substitute cold water lobster tail for crab cake - $40.00
Chicken breast seared and topped with provolone, sauteed spinach, mushrooms and caramelized onion tossed in a creamy chicken veloute.
Fresh mushrooms with sun dried tomatoes, roasted red peppers and garlic sauteed in pure olive oil served over fettuccini.
Stuffed with crab imperial and served with drawn butter.
Children Under 12! Prices includes your choice of one side
Broiled or Fried.


text:
Railroad work to detour traffic in New Prague
A detour will be going into effect at the intersection of New Prague's Main Street (Highway 19) and Union Pacific Railroad's crossings beginning next week. While the sign says it will be Monday, Oct. 1, a press release from the Minnesota Department of Transportation (MnDOT) says the detour begins Wednesday, Oct. 3, as crews will be working to replace two railroad crossings for the Union Pacific Railroad located between Highway 21 and Third Avenue Northwest. Traffic will be detoured to Highway 21, Scott County Road 2 and Highway 13 for three to seven days. Access will remain open to all downtown businesses via Highway 19 and local streets. The work is in conjunction with the planned reconstruction of Main Street in 2020. For more see an upcoming print issue of The Times. (Patrick Fisher Photo)


text:
Fundraise for Family Services of the North Shore
Fundraising at home, school, work, or in your community to support Family Services of the North Shore can be both fun and rewarding.
Ways to donate
Organizing a fundraising event with friends, classmates, family, or co-workers is a wonderful way to support the most vulnerable members of our North Shore community.
All types of events – such as golf tournaments, pub nights, garage sales, birthday parties, partial proceeds from product sales, or holiday giving – can result in meaningful fundraising success.
Here are some fundraising ideas you might consider:
- Host a game or movie night and suggest a donation as 'admission'.
- Organize a neighbourhood car wash, garage sale, bottle drive, coin drive, or bake sale.
- Ask for donations for services such as babysitting, yard work, car detailing, cooking, or cleaning.
- Organize a pancake breakfast, BBQ lunch, or fundraising dinner.
- Leave a collection box at reception where people can donate their extra change.
- Have your employees wear their favourite team jersey or casual clothes to the office in exchange for a donation to Family Services of the North Shore.
Inspired to throw a party or plan a creative event to raise funds for Family Services of the North Shore? We can help you get started and provide you with tips, tools, and resources to help make your event a success.


text:
Check back regularly here for Dehumidifier and home improvement tips and tricks to make sure you get the most out of your next Dehumidifier!
We are going to assist you to understand the plan
You have to be well informed on the subject of Dehumidifier in Alzada, MT. We will make sure that you know what can be expected. This is exactly why we try to make every effort to be sure that you are familiar with the plan and are not confronted by any kind of unexpected surprises. Begin by discussing your task with our customer service reps when you dial 800-239-9895. We will respond to all of your concerns and questions and schedule your preliminary appointment. We work together with you throughout the entire project, and our crew will appear promptly and ready.
Perfect Dehumidifier is there for all your Dehumidifier in Alzada, Wyoming requirements
You have got plenty of good reasons to rely on Perfect Dehumidifier to suit your needs when it comes to Dehumidifier in Alzada, MT. We have got the best customer care scores, the very best quality equipment, and the most helpful and successful money saving practices. We have got the expertise you will want to meet all your ambitions. Dial 800-239-9895 to reach Perfect Dehumidifier and consider your goals about Dehumidifier in Alzada.


text:
Dota IMBA/Buildings
All buildings grant multiplied gold bounties based on the game's global setting. Bonus gold is only granted to the hero that deals the last hit on a structure.
Towers now have a 400 base experience bounty, of which can be multiplied by the global setting.
Towers may gain random custom abilities when the appropriate game setting is enabled. All tower abilities fully stack with each other.
Towers may also improve their abilities if the appropriate game setting is enabled. When a tower is destroyed, a splash message visible to all players will appear and a sound is played, indicating that all towers on that team have been upgraded. Each tower that is destroyed increases the tower "level" by one, leveling up their current ability by one. When a destroyed tower causes the abilities to reach level 3, a new skill is learnt by all towers in addition to the first. These skills can level up as well, following the same rules as above. The last tower will have 3 level 3 abilities and 1 level 2 ability.
The first ability of all towers is mirrored by both teams. This means that Dire's Tier 1 Bottom Tower has the same first ability as Radiant's Tier 1 Top Tower. However, subsequently gained skills are not mirrored.
Tower Abilities[edit]
Reduces hero attributes on every hit
Attacks faster with each consecutive attack.
Damages enemies when attacked.
Return Damage: 30/45/60
Every inch of this tower is covered in angry thorns and vicious spikes.
Each attack causes grievous wounds, which increase damage taken from this tower.
Hits very fast, but not as hard.
Damage Reduction: 50
Bonus Attack Speed: 100/175/250
Always bullied by other towers for being a quick shooter.
Attacks its target multiple times.
Bonus Attacks: 1/2/3
Quarter circle forward x 2 + Heavy punch.
Berserker Tower
Hits harder and faster when on low health.
Attack Speed per Stack: 4/6/8
Damage per Stack: 2/3/4
Health Loss per Stack: 4%
I just want to live! STOP HITTING ME! RAAAWRGH!
Aegis Tower
This tower has increased armor and health.
Bonus Armor: 4/8/12
Bonus Health: 400/800/1200
Made of impregnable mirthil-duranium-vibranium-adamantium-orichalcum-unobtainium alloy.
Deals pure damage instead of physical.
Blinds targets at higher levels.
This tower is magic immune. Passively increases the magic resistance of all allies in range, and periodically purges their debuffs. On level 3, the purge also removes stuns.
Bonus Magic Resistance: 10%/20%/30%
Purge Interval: 5/4/3
Spellcasters feel sick just being near it.
Reduced damage, but hits every enemy in range.
Damage Reduction: 70/30/0
Bonus Range: 0/0/150
A tower with multiple personality disorder.
Melee Barracks have a 400 base experience bounty, of which can be multiplied by the global setting.
Ranged Barracks 200 have a base experience bounty, of which can be multiplied by the global setting.
To make achieving the goal of destroying the enemy Ancient harder, Ancients now receive multiple buffs and abilities that trigger when certain criteria are met. Ancients now gain a flat 15 armor bonus at all times, for a total of 30 armor. Ancient health has also been increased to 7000.
When the Ancient is first damaged, Poison Nova is cast. Subsequently, when its health falls;
- under 80% Overgrowth is cast;
- under 60% Eye of the Storm is cast and Poison Nova's cooldown is refreshed;
- under 40% Borrowed Time is cast and Eye of the Storm's cooldown is refreshed;
- under 30% Ravage is cast and Eye of the Storm's cooldown is refreshed;
- under 15%, all cooldowns are refreshed. This only occurs once.
Ancients use their abilities when there is an enemy within 600 range, or if they receive an instance of damage from any range.
Ancient Abilities[edit]
- Poison Nova is cast when the Ancient is first damaged, and subsequently whenever it is attacked while the ability is off cooldown.
- Triggers when the Ancient's health falls below 80%.
- Works on spell immune units, but if units gain spell immunity after the effect starts, they'll break free.
- Triggers when the Ancient's health falls below 60%.
- Refreshes the cooldown of Poison Nova when activated.
- Each blast targets the lowest health enemy in range.
- Triggers when the Ancient's health falls below 40%.
- Refreshes the cooldown of Eye of the Storm when activated.
- Triggers when the Ancient's health falls below 30%.
- Refreshes the cooldown of Eye of the Storm when activated.
- Ravage will hit invisible units.
- Ravage spreads at a speed of 1400.
To prevent fountain camping, each Fountain has a 1400 attack range and flying vision, +100 attack speed, passive Grevious Wounds (a skill similar to Fury Swipes), and every enemy unit in 1800 range is bashed whenever the Fountain attacks. A glowing marker on the floor indicates the range in which the fountain can attack enemy units. Mirana's Shooting Star is not able to enter the enemy fountain.
Fountains also now heal for 5% of max health and mana, an increase from the normal 4%. This would mean one can heal completely within 20 seconds in the Fountain.
Lua error: Error in "where" parameter: the string "#" cannot be used within #cargo_query..


text:
You Can't Take it with You
by Fayette County Community Theatre
Feb. 24 - Feb. 25 (2019)
The famous zany comedy about an extended family, from George S. Kaufmann and Moss Hart.
Directed by Melissa Weltner and Jenny French.
There are 19 parts so bring a friend!! Character descriptions for this zany, Pulitzer Prize-winning play are below:
Penelope Sycamore - Usually goes by Penny, Penelope is the mother of Essie and Alice, wife of Paul, and daughter of Martin. She writes plays and paints as hobbies because it makes her happy, but is terrible at both. Penny is a loving mother and wife who is constantly concerned with the welfare of her family. Her main goal is to make sure everyone is happy, particularly her daughter Alice. She is a main character.
Essie Carmichael - Wife of Ed, daughter of Penny and Paul Sycamore, Granddaughter of Martin, sister of Alice. She is childish. As a hobby she makes candy that Ed sells. Essie dreams of being a ballerina. She has spent 8 years studying with Boris Kolenkhov, but is a terrible dancer.
Rheba - The African-American maid and cook to the Sycamore family. She is treated almost like a part of the family. She is dating Donald. In the words of Mrs. Sycamore, "The two of them are really cute together, something like Porgy and Bess."
Paul Sycamore - Father of Essie and Alice, husband of Penny, Son-in-law of Martin. He is a tinkerer who manufactures fireworks in the basement with the help of his assistant Mr. De Pinna. His hobby is playing with erector sets.
Mr. De Pinna - The ice man who came inside to speak to Paul eight years before, and never left. He helps Mr. Sycamore build fireworks, and moonlights as a model in Mrs. Sycamore's paintings.
Ed Carmichael - Husband of Essie, son-in-law of Paul and Penny. He is a xylophone player, and distributes Essie's candies. Ed is an amateur printer who prints anything that sounds good to him. He prints up dinner menus for his family and little quotes that he places in the boxes of Essie's candy. He also likes to make masks.
Donald - The African-American boyfriend of Rheba, who seems to serve as volunteer handyman for the Sycamores.
Martin Vanderhof - Referred to mostly as Grandpa in the play. Father-in-law to Paul, father of Penny, grandfather of Alice and Essie. He is an eccentric happy old man who has never paid his income tax because he doesn't believe in it, as he feels that the government wouldn't know what to do with the money if he paid it. Once a very successful businessman, he left his job 35 years prior for no reason other than to just relax. He lives his life by the philosophy 'don't do anything that you're not going to enjoy doing'. He goes to circuses, commencements, throws darts, and collects stamps.
Alice Sycamore - Fiancée of Tony Kirby, daughter of Paul and Penny, Granddaughter of Martin, sister of Essie. She is the only "normal" family member. She has an office job, and is rather embarrassed by the eccentricities of her family when she has Tony and his parents at her house, yet she still loves them. She tends to be a pessimist.
Wilbur C. Henderson - An employee of the IRS. He comes to collect the tax money owed by Grandpa, and can't understand why the latter won't pay income tax.
Tony Kirby - Fiancé of Alice, Son of Mr. and Mrs. Kirby. He sees how, even though the Sycamores appear odd, they are really the perfect family because they love and care about each other. His own family is very proper and has many issues none of them will admit. He is vice president of Kirby and Co.
Boris Kolenkhov - A Russian who escaped to America shortly before the Russian Revolution. He is very concerned with world politics, and the deterioration of Russia. He is the ballet instructor of Essie, aware that she is untalented at dancing, but knows that she enjoys dancing so he keeps working with her. He likes the Greeks and the Romans, questions society, and is interested in world affairs.
Gay Wellington - An actress whom Mrs. Sycamore meets on a bus and invites home to read one of her plays. She is an alcoholic, gets very drunk and passes out shortly after arriving at the Sycamore's home.
Anthony W. Kirby - Husband of Mrs. Kirby, father of Tony. He is a very proper man who is president of Kirby and Co. and secretly despises his job. His hobby is raising expensive orchids. He is also a member of the Harvard Society, the Union Club, the National Geographic Society, and the Racquet Club.
Miriam Kirby - Wife of Mr. Kirby, mother of Tony. She is an extremely prim and proper woman and is horrified by the goings-on in the Sycamore household. Her hobby is spiritualism.
G-Man 1, 2, 3 - Three agents who come to investigate Ed because of the communist quotes he prints up and places in Essie's candy boxes, such as "God is the State – the State is God".
The Grand Duchess Olga Katrina - She was one of the Grand Duchesses of Russia before the Revolution, another being her sister, the Grand Duchess Natasha. Since then she has been forced to flee to America where she has found work as a waitress in Childs Restaurant. The rest of her family has had a similar fate, such as her Uncle Sergei, the Grand Duke, who is now an elevator man. She loves to cook as a hobby.


text:
August is Peach & Watermelon Month
Blog, Save the Date
August is Peach & Watermelon Month
Blog, Save the Date
Peaches and Watermelons are some of Executive Chef Demetrio Zavala's favorite summer ingredients – see how we've incorporated them into our special dishes!
Perhaps one of the most creative preparation of peaches can be found in our Roasted Peaches dish, made with blue cheese funnel cake (yes, funnel cake), aged balsamic, and corn powder.
Our famous Brussels Sprouts will now be made with pistachio honey vinaigrette, roasted peaches and basil crema. A peachy entree is our Lavender Spiced Duck, made with pickled peaches, coffee corn puree, barley, and mole jus. And for Dessert enjoy our Peach "Mel"ba made with Spiced Pound Cake, Peach Compote, Pistachio Meringue, and Raspberry Sorbet!
Among our watermelon-themed dishes we have Charred Octopus with sofrito lentils, salsa verde compressed watermelon, and jalapeno vinaigrette, our Watermelon & Cucumber with bacon oil, balsamic onions, tomato water, and blue cheese vinaigrette.
Entrees include our NY Strip with feta cheese funnel cake, watermelon romesco and pickled shallots. Finish it off with our Watermelon Consomme Dessert!
This is a wonderful opportunity to savor summer's beloved fruits! Each dish will be available for Lunch and Dinner, from the 1st through the 31st. Hang on to that warm weather and dine al fresco on our patio.
For reservations, give us a call at (202) 872-8700, or reserve your spot online!


text:
Introduction: Rail and Stile With Stick & Cope Profiles
Rail and Stile is a common construct for Frame & Panels and Doors. While you could create the edge profiles manually, using Catalog Features to create the Stick and Cope profiles allows for quick creation of parts, as well as easy replacement of profiles for another project.
This tutorial will show how to create a part with a Stick profile on the edges of the part, as well as Cope profiles on the Ends. Profiles will be added to both edges and ends so as to allow the base part to be copied and used for all Rail & Stile parts. You only need to suppress the profiles not needed for the current part, thereby saving time by re-using the original part as much as possible.
You will want to read the Instructable on how to do an Extrude Cut Catalog Feature and be able to create your own Catalog Features before tackling this project. Also, you may want to be able to create your own raised panel to insert inside your new Rail and Stiles. The Raised Panel using Sketch Catalog Feature instructable will help with that as well
Teacher Notes
Teachers! Did you use this instructable in your classroom?
Add a Teacher Note to share how you incorporated it into your lesson.
Step 1: CREATE a BOARD
Start by creating a rectangular board (see Modeling Woodworking Board or Sheet Part) in an Alibre Part file. For this Instructable we will start with a board that is 3/4" thick, 2" wide, and 12" long. This is about the minimum thickness and width you will want for your Stile in order to drill it for a European hinge. The length is arbitrary, and you will size the completed part to suit the desired length of your Stiles and Rails.
Step 2: APPLY STICK PROFILES
While you can start with either the Stick or Cope profile, I prefer to start with the Stick profile and shape the long sides of my part. While the Extrude Cut Catalog Feature is the easiest, you can also just use a Sketch Catalog Feature and do an Extrude Cut once the sketch is constrained.
Inset your Catalog Feature of the Stick profile on the reference plane along the midpoint of the length of your part. If constructed according to our suggestions in the Board Instructable, this should be the YZ reference plane. It may help to orient to the Right End view to make positioning the double ended stick profile. Insert two copies of the Stick profile by dragging the preview first to near the final position on one edge and pressing APPLY, and doing the same again for the opposite edge. It helps with being able to constrain the Catalog Feature by making sure you have inserted the profile near to the final position. Yet, keep the profile just off the final position so you can visually verify your constraints when applied.
It is important to insert the Catalog Features on the reference planes and not the ends of the board, as those ends will be profiled by the Cope. While you could apply the Stick profile to the ends (since the Cope will be applied later) it keeps everything tidy by inserting the Stick profile on the reference planes. You will have to insert the Cope on the reference planes after the edge is profiled by the Stick cut.
Close your Catalog Feature window once you have applied both Stick Profiles.
Step 3: CONSTRAIN STICK PROFILES
Now you need to edit the Sketch for both of your Stick Profiles and locate them on the edges of your board.
Constrain the profile to the face of the board using the Colinear constraint. This will keep the Profile cut in the proper location. If the profile is such that you can not use a constraint to locate it, then dimension the profile off the face of the board and label the dimension of the constraint in the Equation Editor. Try not to use the Coincident constraint (why will be explained next).
While you can also constrain the location of the profile across the width of the board, I have found with experience that a constraint may come loose with drastic changes in a boards width. It is instead safer to dimension the Stick profile location off one of the axes. Dimensioning off an edge may also result in the dimension coming loose with drastic changes in the boards width. It is a simple matter to dimension the location of the profile by using the Width parameter set up when you constructed your board (following the Board creation Instructable). Simply dimension the offset from the axis to profile on each size as Width/2.
End your edit of each Sketch after constraining and dimensioning the location of your Catalog Feature. I prefer using F5 as that will Regenerate All features. If you have used and Extrude Cut Catalog Feature, you now have profiles on both sides of your board. It you used a Sketch Catalog Feature, you just need to Extrude Cut each of your sketches.
We will use the Suppress command later to remove individual Cut Features to create all the different Stiles and Rails for a Frame and Panel from copies of this single part file.
Step 4: COPE CATALOG FEATURES
Similar to the insertion of the Stick profile Catalog Features, Inset Catalog Feature for the end Cope profiles. Select the reference plane about which the width is constructed, which should be the ZX plane. While is may seem easier to work with parts constructed in their final orientation, it is good to become used to working with parts always constructed where the width is in the Y direction, the Length in the X direction, and the thickness in the Z direction. This convention will hold you in good stead when finally working with Cutlist programs and doing nesting of Sheet Goods.
Just as I was loosening up on my construction orientation, along came the need to have these parts manufactured using CNC equipment. Back into the rigid part orientation conventions as the software to process these parts required consistent orientation in order to define the grain direction. If you think you will never work with CNC, keep in mind that there are many small shops offering CNC services. Just visit the ShopBot forum and see how active this group of woodworkers are who have built their own more affordable CNC equipment. There is probably someone in your area who can do work for you at an reasonable price.
Here is where double ended Catalog Features of your Cope profile really help. While you can insert the Stick profile on opposite faces to orient the Catalog Feature, you probably no longer have flat faces on which to insert your Cope Catalog Features. You will not be able to inset a Catalog Feature on the back side of a reference plane, and will be stuck with a single direction and only being able to rotate about the axis of the profile.
Insert two copies of the Cope Catalog Feature, dragging them into approximate position on each end of the board.
Step 5: CONSTRAIN COPE CATALOG FEATURES
Constrain the Cope profile to the Face of the board as before using the Colinear constraint or dimension. Avoid constraining the profile along the length of the board as this is where the constraint is most often broken. Radical changes in the length of the board when using this part to make Stile and Rails most often breaks the locating constraint. Instead locate the Cope profile using a dimension, and if constructed using standard parameter labels, that would be the dimension of Length/2. Here is where it really helps to constrain your board parts using the Symmetric constraint. If your parts are not symmetrically constrained, the dimension to locate the profiles may contain a little more complicated formula.
Extrude Cut the same way your did for the Sticks.
Step 6: CREATE STILES AND RAILS
Now that we have a Part file with all the profiles required to create all of your Stile and Rails, Save As the current part file to every required part for your project. It may be useful to keep this master part as a Template file, from which to create additional parts in the future. Simply name it indicating the shape of the profiles used.
While you can just save a single Stile, it can be useful to have both a Left Stile and a Right Stile. There are occasions where the Stile is longer in length below the reference plane than above, as the reference planes are a useful tool to constrain the panel often inserted into the center of a Stick and Cope construction. And then there is the Mid Stile for multiple panel assemblies.
Similarly, create a Top Rail, Bottom Rail, and Mid Rail as required. Top and Bottom Rails are often different widths, with the Bottom Rail wider for a more stable looking appearance.
Step 7: SUPPRESS UNWANTED STICK AND COPE PROFILE FEATURES
Open each new part in turn and Suppress the Stick or Cope Profile features that are not desired for the current part.
For the Left and Right Stile, Suppress the Cope profile on both ends and the Stick profile on one side. You can simply delete the unwanted features, but Suppress allows greater flexibility to resurrect the profile should it be required in the future.
For the Top and Bottom Rails, Suppress the Stick profile on one side only. Since you most likely require a separate Top and Bottom Rail, you will Suppress the top Stick profile for the Top Rail, and the bottom Stick profile for the Bottom Rail.
Adjust the length of each part to suit your requirements, and keep in mind the added length required because of the Cope profiles on the ends of the Parts.
Step 8: CREATE ASSEMBLY AND CONSTRAIN YOUR PARTS
Create a new Assembly and insert all the parts required for your Stick and Cope door, Frame and Panel, or other construct.
I have found it best to turn on the Reference Geometry for each of the parts, and constrain them where possible by the Reference Geometry. This allows the greatest flexibility in re-use of the assembly, and better use of parameters to offset the assembly about the Reference Planes. By offsetting the Stiles vertically , you can construct a Frame and Panel door where you can use the Reference Geometry of the assembly to constrain it in the proper location on a Cabinet.
I prefer to constrain the Stiles vertically by their Reference Planes, Rails horizontally by their Reference Planes, and all parts along the Z axis by their Reference Planes.
After doing this, you can constrain the Rails vertically to the ends of the Stile, and the Stiles horizontally by some matching stick and cope geometry.
Add in the Panel, which is hopefully constructed to match about the Reference Planes, and constrain if where possible by the Reference Geometry.
Step 9: CONFIGURATIONS
Those Alibre users with the Professional or higher version, can use Configurations to create each of these Stile and Rail parts within the same part file. From the original configuration, a new configuration is created for each Stile and Rail. All the configurations are locked, and the appropriate Catalog Features suppressed in each configuration, so all that remains are the appropriate profiles to suit that part.
Another benefit of using Configurations is that the dimensions of all the different parts can be linked together. You can add parameters for the Overall panel Height and Width, and have the Width and Length for each configuration and its associated Stile or Rail conform to the specified overall dimensions. Once you set up the formula for each configuration while it is locked, you can then Unlock All, and when you change the Overall dimensions, the individual part dimensions will change to follow suit. No need to keep all configurations locked, as the setup made while configurations are locked will remain intact. As long as changes are only made to parameters that are not unique to a configuration, the formulas setup within each configuration will remain and follow suit to changes in the overall dimensions contained in those formulas.
We have created a Frame and Panel assembly that can consist of a single or multiple panels, and that will change the dimensions of every individual component based on the overall size of the Panel Assembly and the number of panels specified.