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Many highly beneficial traits (e.g. disease or abiotic stress resistance) have been transferred into crops through crosses with their wild relatives. The 13 recognized species of tomato (Solanum section Lycopersicon) are closely related to each other and wild species genes have been extensively used for improvement of the crop, Solanum lycopersicum L. In addition, the lack of geographical barriers has permitted natural hybridization between S. lycopersicum and its closest wild relative Solanum pimpinellifolium in Ecuador, Peru and northern Chile. In order to better understand patterns of S. lycopersicum diversity, we sequenced 47 markers ranging in length from 130 to 1200 bp (total of 24 kb) in genotypes of S. lycopersicum and wild tomato species S. pimpinellifolium, Solanum arcanum, Solanum peruvianum, Solanum pennellii and Solanum habrochaites. Between six and twelve genotypes were comparatively analyzed per marker. Several of the markers had previously been hypothesized as carrying wild species alleles within S. lycopersicum, i.e., cryptic introgressions.
Each marker was mapped with high confidence (e<1 x 10-30) to a single genomic location using BLASTN against tomato whole genome shotgun chromosomes (SL2.40) database. Neighbor-joining trees showed high mean bootstrap support (86.8 ± 2.34%) for distinguishing red-fruited from green-fruited taxa for 38 of the markers. Hybridization and parsimony splits networks, genomic map positions of markers relative to documented introgressions, and historical origins of accessions were used to interpret evolutionary patterns at nine markers with putatively introgressed alleles.
Of the 47 genetic markers surveyed in this study, four were involved in linkage drag on chromosome 9 during introgression breeding, while alleles at five markers apparently originated from natural hybridization with S. pimpinellifolium and were associated with primitive genotypes of S. lycopersicum. The positive identification of introgressed genes within crop species such as S. lycopersicum will help inform conservation and utilization of crop germplasm diversity, for example, facilitating the purging of undesirable linkage drag or the exploitation of novel, favorable alleles.
Cryptic introgression; Linkage drag; Breeding; DNA sequence; Solanum speciesBackground
Introgression is the transfer of genes of one species into the gene pool of another via hybridization. As a phenomenon, it has been an important topic in animal and plant genetics research for many different reasons. For example, introgression has been implicated in the adaptation of modern humans [1] and is of concern to conservation biologists due to loss of integrity of wild bird and mammal populations [2,3]. In plants, introgression is a key concept in studies of the risks of contamination of natural populations by genetically modified (GM) crops. More commonly in crops, favorable genes from wild relatives are intentionally transferred into breeding lines for cultivar development. This has been particularly valuable in crop species that are relatively low in genetic diversity.
According to a review of crop introgression breeding [4] the major functional categories of beneficial traits transferred from wild species are resistance or tolerance to abiotic stress or disease, yield, cytoplasmic male sterility or fertility restorers for hybrid production, and quality traits. Among the pioneering uses of wild crop relatives during the late 19th to early 20th centuries were the transfer of disease resistances into grape (Vitis vinifera) [5] and sugarcane (Saccarum officinarum) [6]. A 1986 review of 23 crops estimated that 6% of total annual economic value in the US was contributed by crop wild relatives [7]. For 13 major crops of global importance, it was estimated that 46 wild species have been used in released cultivars, and that furthermore, the introgression breeding approach is increasing [4]. Lack of information on pedigrees, unpublished activities within the private sector and changes in taxonomy are some of the factors that contribute to the uncertainty of the collective impacts of crop introgression breeding [4].
Some of the earliest tomato introgression breeding in the US may have been done indirectly and unwittingly via the French variety Merville des Marchés. Recent phenotypic data collected for Merville des Marchés PI 109834 showed it to be variable in fruit size and smoothness ( http://www.ars-grin.gov/cgi-bin/npgs/acc/display.pl?1129442 webcite); its genotype was segregating, showed population admixture, and was an outlier based on genetic distance relative to many other S. lycopersicum accessions [8,9]. We postulated that these were indications of S. pimpinellifolium in its ancestry (this idea was examined in the current study). The Fusarium wilt-resistant processing variety Marvel [10] was selected from Merville des Marchés in the early 1900s, and Marvel was a parent of Marglobe released in 1925 [11], which in turn can be found in the pedigree of many important varieties from the 1930s through the late 1950s (H.M. Munger's tomato pedigree chart provided by E.D. Cobb, Cornell University, 2012). Direct introgression of tomato with wild species in the US commenced in the 1930s concurrent with collection expeditions to geographic centers of origin. The first released cultivar, developed from Marglobe x S. pimpinellifolium, was aptly named Pan American [12]. Introgression breeding efforts of tomato increased globally post World War II, involving the screening of a wide range of traits and all wild tomato species [13]. Such efforts continue to be of utmost priority today using sophisticated tools such as introgression libraries for gene discovery [14,15].
Compellingly, of 96 introgressed traits tallied in released crop cultivars for 11 species (cassava, wheat, millet, rice, maize, sunflower, lettuce, banana, potato, groundnut, tomato), 55 of them were in tomato (Solanum lycopersicum L.); the next highest numbers were found in rice and potato with 12 traits each [4]. The emphasis and success of introgression breeding in tomato encompasses several factors including its intrinsically narrow genetic base, relative ease of crossing with several wild taxa, production demands based on growing conditions and market niche, its susceptibility to pests and pathogens, and its sensitivity to abiotic factors. In addition to resistance or tolerance to dozens of bacterial, viral, fungal, insect, and nematode pathogens, hundreds of favorable genes or quantitative trait loci (QTL) for abiotic stress resistance, flower and fruit traits, yield, and plant architecture have been mapped in wild tomato species [16] and thus hold the potential to be exploited.
Introgression breeding carries a cost, namely, genetic linkage of non-targeted loci that are eliminated through repeated backcrossing. Linkage drag can persist within a genome despite backcrossing, especially if recombination is suppressed. Several examples of linkage drag in tomato and other crops have been quantified using molecular markers [17-21]. Linkage drag can denote favorable, deleterious or neutral alleles that become inadvertently incorporated into breeding lines or cultivars.
In this study we apply the term 'cryptic introgression' [22] to describe latent genetic variation in S. lycopersicum that originated from wild tomato species. Various scenarios can be evoked for its origins ranging from linkage drag, hybridization between feral S. lycopersicum and wild relatives, to crossing in open-pollinated populations by wind or insect vectors with pollen of introgressed cultivars [23]. Cryptic introgression is of interest in germplasm collections such as those conserved at United States Department of Agriculture, Agricultural Research Service (USDA, ARS) Plant Genetic Resources Unit (PGRU) because it can indicate novel genetic variation for exploitation by end-users, or conversely, reveal unfavorable and hence undesirable alleles with respect to crop improvement.
In previous reports we hypothesized the detection of cryptic introgression in 5% to 10% of DNA markers that were resequenced in tomato germplasm panels [9,24,25]. The aim of the current study was to gather additional evidence on these alleles by resequencing and analyzing the same markers in several accessions of wild tomato species and one accession of weedy S. lycopersicum (Table 1). Although variation within wild species gene pools made it impracticable to attempt to discover the 100% identical homologous allele, the assumption was that introgressed alleles would be more closely related to the alleles of a particular wild species than to their S. lycopersicum homologs. To identify introgressed alleles we also used evidence from mapped locations of markers, phenotypic descriptions, and historical origins of lines and accessions.
Table 1. Tomato samples analyzed in this study
Results and discussion
Markers and in silico mapping
For the 47 markers used in this study (Additional file 1: Table S1), nucleotide primers for PCR and sequencing were originally designed against S. lycopersicum sequences except for the Conserved Ortholog Set II (COSII or C2) and unigene (U) markers, which were designed against Euasterids [27,28]. Molecular markers have shown good success rates of transferability among distantly related wild tomato species such as S. lycopersicum and S. pennellii (for examples see [29-31]). In the current study, some primer pairs did not amplify or give clean or homologous reads in every wild tomato sample. The COSII and U markers did not outperform the other markers in terms of successfully generating high quality sequence data because most of them contained introns, which sometimes carried small indels in a heterozyogus condition. Such heterozygous indels were major contributors to poor quality reads and hence missing data.
Additional file 1. Table S1.Virtual mapping of 47 sequence-based markers. BLASTN results of 47 markers against tomato whole genome shotgun chromosomes from version 2.40 of the WUR assembly, current as of 10 Jan, 2012. Table S2.Evolutionary relationships among taxa at 47 markers. Bootstrap support and evolutionary divergence of 47 tomato markers sequenced in wild and cultivated tomato accessions. Marker names in bold were previously hypothesized as carrying introgressions.
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In our current germplasm panel (Table 1), S. lycopersicum and S. pimpinellifolium had no missing markers; sequences were available for at least two of the four green-fruited taxa for the final set of 47 markers. S. peruvianum LA1537, S. peruvianum G32592, S. arcanum, S. pennellii and S. habrochaites gave data for 32, 42, 40, 35 and 38 markers, respectively. The latter two species are usually self-incompatible (SI) and carried greater numbers of polymorphic markers within accessions than the inbred accessions of S. peruvianum and S. arcanum that were sampled. Mean SNP frequency across the polymorphic markers was 0.0127 (n = 17) for S. habrochaites, 0.0082 (n = 17) for S. pennellii, 0.0126 (n = 5) for S. peruvianum LA1537, 0.0058 (n = 2) for S. peruvianum G32592 and 0.0078 (n = 7) for Peru Wild. TA496, Tomate and S. arcanum had no polymorphic sites in any of the markers.
S. pimpinellifolium showed unusually high polymorphism of 0.0055 (n = 17) relative to the other self-compatible taxa. As one explanation, this accession was categorized as an admixture population in a simple sequence repeat (SSR) genotyping study of S. pimpinellifolium population structure [32], so naturally represents two dissimilar S. pimpinellifolium genomes. Because the seed source traces back to Vaughan Seed Co. in USA and Horticultural Experiment Station, Ontario, Canada [33], another possibility is that S. lycopersicum was incorporated into its pedigree by one of those entities. This may be evidenced by comparing the previously estimated D, number of mutations per kb, [34] between the two species [35] which was approximately four-fold greater than our estimate reported below.
The majority of markers were mapped with high confidence to a single map location within the genome (Figure 1, Additional file 1: Table S1). This is the first report in which a whole-genome sequence [36] and web based tools were available with which to do this for the expressed sequence tag (EST) -based markers [24]. Accordingly, 12 of the EST-based markers were newly mapped and markers 437_2, 2189_1 and 2819_5 were revised with respect to previously predicted chromosomal location based on identities to restriction fragment length polymorphism (RFLP) markers [25]. All chromosomes were represented by the 47 markers; numbers of markers per chromosome ranged from one on chromosome 12 to eight on chromosome 6. Small gaps were observed in the alignments but close examination revealed that these were in masked regions rich in runs of poly A or poly T. Only marker 1675_1 did not initially provide any hits. A lowered stringency of 1 x 10-30 subsequently found the probable position.
Figure 1. Chromosomal map locations of 47 markers sequenced in this study. Nine markers with dashed outlines showed cryptic introgressions. Also shown on the map (color) are documented introgressions used in tomato breeding that were mentioned in this report, S. habrochaites: blue, S, lycopersicum Peru Wild: red, S. peruvianum: purple, S. pimpinellifolium: orange, S. pennellii: green.
Four markers had two BLASTN hits each. These were 1260_2 for two tightly linked (2,849 nt apart) sequences on chromosome 6, 2325_3 on chromosomes 1 and 3, 2582_1 on chromosomes 4 and 5, and 2819_5 for two linked (14,114 nt apart) regions on chromosome 6. A check of the primer regions found multiple mismatches in predicted primer binding sites of the secondary hit for three of the markers; these were assumed to have amplified as single-copy. For marker 2819_5 the forward primer had a single mismatch and the reverse had no mismatches. The predicted amplicon in the mismatch region was 86% identical to the reference TA496 sequence. When the mismatch sequence was included in MEGA cluster analysis, it separated from all other (wild and cultivated) alleles with 87% bootstrap support, and the mean D among sequences increased 8-fold (D = 1.5 for n = 10 alleles versus D = 12 for n = 11 alleles). Therefore, it is unlikely that this paralog amplified and confounded the results. All markers had previously been screened in our lab for amplification of single bands and highly homozygous sequences within S. lycopersicum[24,25]. Results of in silico mapping confirmed that this set of markers provides robust results in sampling single-copy S. lycopersicum genes. All sequences were deposited into the European Molecular Biology Laboratory (EMBL), European Nucleotide Archive (ENA) data base as accession numbers HE977919-HE978211.
Clustering patterns and divergence estimates among taxa
PI 99782 Tomate was chosen for the current study to represent a 'pre-introgression breeding' genotype. It bears small, slightly ribbed and unimproved fruit with scarring and cracking ( http://www.ars-grin.gov/cgi-bin/npgs/acc/display.pl?1127604 webcite) and was homozygous for the common S. lycopersicum haplotype at 48 of 50 markers for which it had been sequenced [25]. For 38 of the markers there was high bootstrap support for the red-fruited clade that consisted of S. lycopersicum including Tomate and S. pimpinellifolium alleles (Additional file 1: Table S2). Bootstrap values ranged from 54% to 100% (mean ± standard error = 86.8 ± 2.34%). These bootstrap values subtracted from 100% can be interpreted as estimates of the probability of Type I error, i.e., falsely accepting a cluster that is not signified by the data (see [37] for discussion).
Results underscored the close relationship of S. lycopersicum to S. pimpinellifolium because their alleles were frequently identical or highly similar. The placement of green-fruited species with respect to the red-fruited clade and each other varied among loci. This can result from incomplete lineage sorting or introgression [38]. An example was the placement of S. habrochaites near S. pimpinellifolium for marker TG11 (Additional file 2: Figure S1); this example was also reported by Nesbitt and Tanksley [35].
Additional file 2. Clustering and network analyses of tomato genotypes. Neighbor joining trees for 47 markers and hybridization networks for nine markers sampled from wild and cultivated tomato.
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Of the nine markers that did not support the red-fruited clade, seven had previously been hypothesized as carrying introgressions (Additional file 1: Table S2). These are discussed below. The other two markers, 1287_1 and 2280_1, showed patterns that contained S. peruvianum within the red-fruited clade (Additional file 2: Figure S1) seemingly due to lack of resolution. At marker 1287_1 S. peruvianum haplotypes were only one or two mutational steps from Tomate. These mutations were not shared with any other taxa. At marker 2280_1 only four unique haplotypes were observed. These were S. habrochaitesS. pennelliiS. arcanum and {TA496, Tomate, S. pimpinellifolium, Peru Wild, G32592, LA1537}. For the 38 markers that supported the red-fruited clade, accepted taxonomic relationships among tomato species were generally supported [39,40].
Divergence estimates (D) among loci are associated with a high variance over evolutionary time due to differences in mutation and recombination rates, selective constraints, and influences of various factors such as random sampling of gametes and demography. Average D ranged from 5 for marker 4301_3 to 52 mutations per kb for marker C2_At1g44575 (mean ± standard error = 16.5 ± 1.29). At the aggregate scale the mean D ± standard error from PI 99782 Tomate was as follows: TA496 D = 0.001 ± 0.0005, Peru Wild D = 0.002 ± 0.0006, S. pimpinellifolium D = 0.002 ± 0.0004, S. arcanum D = 0.020 ± 0.0022, S. peruvianum LA1537 D = 0.020 ± 0.0025, S. peruvianum G32592 D = 0.022 ± 0.0024, S. habrochaites D = 0.021 ± 0.0024 and S. pennellii D = 0.022 ± 0.033.
The lack of precise resolution in distinguishing taxa by clustering, or by average divergence from Tomate in the case of the green-fruited taxa was a function of shared polymorphisms in many instances, e.g., at 24 markers at least one single nucleotide polymorphism (SNP) within a species was also segregating between other species pairs. This has been observed in other studies of crop species and their closely related wild relatives ( [41] and references therein). In addition, random sampling contributed to low resolution, e.g., S. peruvianum haplotypes appeared to be derived from Tomate at marker 1287_1 based on a small number of noninformative SNPs.
Evidence for introgression
In previous studies we reported nine markers (Table 2) with highly diverged alleles within S. lycopersicum and hypothesized that this was due to introgression from wild species. These were 220_1, 437_2, 2325_3, 2534_1R (redesigned into 2534_1b), 2486_1 [24], 2819_5, C2_At1g73180 [25], U146140 and C2_At1g44575 [9]. Of the 47 markers in the current study, seven of these nine showed patterns in cladograms that did not cluster together members of the red-fruited clade (Additional file 2: Figure S1). Hybridization networks (Additional file 2: Figure S1), descriptive information of the accessions, map positions of markers, and species origins of documented introgressed disease resistance alleles (e.g. [16,26,42-44]) were used as total evidence to categorize the divergent markers into putative linkage drag during introgression breeding versus natural out crossing with S. pimpinellifolium (Table 2). This categorization did not constitute proof of natural hybridization versus introgression breeding. However, it was a useful concept from which to synthesize independent lines of evidence and can serve as a basis for future hypothesis testing of the two scenarios.
Table 2. Tomato markers tested for cryptic introgression
Linkage drag was inferred for at least three of the four markers at which TA496 carried an allele that was highly divergent from all other members of the red-fruited clade. All four mapped to chromosome 9 and spanned from 5.77 MB to 54.71 MB (Table 2). Introgressed disease resistance loci documented on chromosome 9 [42] include Ve1 (0.06 MB) and Ve2 (0.05 MB) both from Peru Wild, Frl (physical map position not annotated), Tm-2a (13.62 MB) and Sw-5 (67.30 MB), all three from S. peruvianum, and Ph-3 (66.71-66.78 MB) from S. pimpinellifolium (Figure 1, Table 2). LA1537 was the most closely related allele to TA496 in cladograms for the three markers spanning 5.77 – 17.00 MB on chromosome 9, which encompasses the Tm-2a locus at position 13.62 MB. LA1537 is an inbred accession that was derived from PI 128650, the original source of Tm-2a (Table 1).
Hybridization networks placed TA496 in various positions for each of the three markers surrounding Tm-2a: near the red-fruited alleles with reticulation back towards S. peruvianum for marker 2486_1 (Figure 2a and Additional file 2: Figure S1), between the red-fruited alleles and S. peruvianum with reticulation back towards both for marker 2534_1b (Figure 2b and Additional file 2: Figure S1), and within the green-fruited wild species alleles between the two S. peruvianum accessions, with no reticulation for marker 220_1 (Figure 2c and Additional file 2: Figure S1). The size of the Tm-2a introgressed region was estimated by mapping in segregating F2 populations [19]. RFLP marker TG101 at chromosome 9 location 50.41 MB was tightly linked (≤ 1 cM) to Tm-2a. The chromosomal position of Tm-2a has been characterized as very near the centromere with extremely repressed recombination [47]. It is therefore probable that markers 220_1 (5.77 MB), 2486_1 (5.90 MB) and 2534_1b (17.00 MB) were part of the introgressed segment in TA496.
Figure 2. Examples of splits networks at markers with cryptic introgressions inS. lycopersicum. Dashed boxes indicate accessions with introgressed alleles (a-d: hybridization networks using marker U146437 as a control, e: parsimony splits network); a) TA496 showed reticulation with S. peruvianum LA1537 and other green-fruited species, b) TA496 showed reticulation with red-fruited and green-fruited species, c) TA496 nested within green-fruited species, d) Merville des Marchés PI 109834 was closely related to S. pimpinellifolium, which showed reticulation to Tomate PI 99782, e) parsimony splits network used only conservative, global signal within the marker to illustrate the hybrid origin of Merville des Marchés PI 109834.
The TA496 allele at marker 437_2 (54.71 MB) was not as definitively related to S. peruvianum and its origin was more difficult to interpret. The cladogram placed TA496 in an intermediate position between two clades, namely, {S. pimpinellifolium, Tomate, S. arcanum, Peru Wild, S. peruvianum} versus {S. habrochaitesS. pennellii} (Additional file 2: Figure S1). The hybridization network showed reticulation with S. habrochaites and the red-fruited alleles (Additional file 2: Figure S1). Several genes commonly found in tomato varieties have originated from S. habrochaites (Cf-4Tm-1Ol-1Cyc-B and Del) or S. pennellii (I1 and I3) but none of these map to chromosome 9 [43,45]. The chromosome 9 introgressions listed in Table 2 were judged to be commonly used in cultivars based on an informal survey of descriptions from online seed catalogs (unpublished observations) as well as comprehensive reviews of tomato breeding (e.g. [16,26,42-44]).
Linkage drag of 437_2 in TA496 with Ph-3 (66.71 MB) or Sw-5 (67.30) was rejected because they originated from S. pimpinellifolium and S. peruvianum, respectively. Based on a BLASTN search of TA496 marker 437_2 against the Solanaceae PlantGDB-assembled Unique Transcipts (PUTs) in the Solanaceae Genomics Resource database ( http://solanaceae.plantbiology.msu.edu/ webcite), TA496 and S. habrochaites shared a sympleisiomorphy at nucleotide position 46 that was absent from all other alleles that we resequenced. This provided tentative but inconclusive evidence of a direct relationship between the introgression and S. habrochaites. As alternative evidence, FM6203, a progenitor of TA496, purportedly carries Asc resistance (pers comm. S. Loewen, University of Guelph, 2005) for which S. pennellii has served as one of the original sources on chromosome 3 in tomato [48].
The remaining five markers with divergent alleles showed patterns consistent with introgression from S. pimpinellifolium in natural populations. At four markers (2325_3, C2_At1g44575, 2819_5 and U146140) the divergent alleles were more closely related to red-fruited rather than green-fruited taxa with bootstrap values ranging from 82% – 100% (Additional file 1: Table S2), although markers 2819_5 and U146140 did not support monophyly of red-fruited alleles (Additional file 2: Figure S1). Hybridization networks showed red-fruited species alleles to be distinct from green-fruited species alleles for 2325_3 and C2_At1g44575, while 2819_5 and U146140 showed connections between red-fruited and green-fruited species (Additional file 2: Figure S1). Marker U146140 nicely illustrated the S. lycopersicum x S. pimpinellifolium hybrid origin of Merville des Marchés PI 109834 (Figures 2d 2e). Marker C2_At1g73180 showed an unusual pattern in that PI 196297 (and three additional accessions with the identical allele, Table 2) and PI 390510 were divergent from all other alleles in the cluster analysis with 99% bootstrap support (Figure 3a and Additional file 2: Figure S1). This suggested a potential paralog. However, no heterozygotes were observed, the marker did not map to more than one genomic location, and COSII markers were designed to amplify highly conserved single copy genes [27].
Figure 3. Extreme divergence of marker C2_At1g73180.a) Clustering at marker C2_At1g73180 showed the distinctiveness of PI 129026 and PI 196297 with 99% bootstrap support, b) the splits network showed a combination of introgression of PI 129026 and PI 196297 (among others, see Table 2) with S. pimpinellifolium and retention of ancient polymorphisms that reticulated down to the root; the latter supported genetic hitchhiking and rejection of selective neutrality.
The divergent C2_At1g73180 alleles were unlikely to have originated from a green-fruited species because chromosome 8 does not carry any introgressed disease resistance alleles [16,42-44]. The hybridization network depicted PI 196297 and PI 390510 as branching from S. pimpinellifolium, with complex reticulations at the base of the red-fruited clade that extended through the green-fruited taxa, down to S. pennellii at the root (Figure 3b). Among the seven unique polymorphisms carried by PI 196297 and PI 390510, two were non-conservative amino acid substitutions, two were synonymous and three were intronic. One possibility is that selection at or near this locus has caused ancient polymorphism to have been retained. A significant HKA test [49] (χ2 = 7.36, P = 0.007) strengthened this interpretation. Therefore, diversity at this marker showed patterns of both natural selection and introgression.
Additional evidence that these five markers represent S. pimpinellifolium introgressions in natural populations includes geographical origins of three accessions from Ecuador where the two species hybridize extensively [50], original collection of three accessions dating to 1935 and 1938 (likely precluding the influence of direct introgression breeding), the primitive fruit phenotype of fasciation (four accessions) or wild cherry tomato (one accession, n.b., cherry tomato was described as an admixture of S. lycopersicum and S. pimpinellifolium by [35]), and previously reported introgression of PI 196297 with S. pimpinellifolium by Rick [23]. In estimates of population structure [51] PI 109834, PI 129128, PI 258474 and PI 258478 all showed high probability of membership in the second of two populations inferred for S. lycopersicum genotypes, consistent with interspecific hybridization [9].
It was useful to delineate cryptic introgression within S. lycopersicum into linkage drag stemming from breeding versus natural hybridization with S. pimpinellifolium, although this categorization was not definitive and should be subjected to further scrutiny. Sequences of the wild tomato species markers in the context of their physical map locations have strengthened our previous interpretations of detection of introgressed alleles in domesticated tomato [24,25]. Genomic tools for fine resolution of introgressed regions in crop species are increasingly available. The strengths and weaknesses of comparative genotyping to verify introgression have been illustrated here using Solanum section Lycopersicon taxa.
In the current study the implications of linkage drag of introgressed alleles on phenotype remain unknown. At least one marker (2534_1b) codes for an enzyme involved in fruit ripening (Table 2). In cultivars or breeding lines it will be more useful to estimate the proportion of a genome that harbors linkage drag. This should be feasible for TA496 using bioinformatics given the vast amount of public EST sequence data available for this particular line and wild tomato species ( http://solgenomics.net/tools/blast/dbinfo.pl webcite as of June 2012 reported 323,465 Lycopersicon mRNAs). Importantly, an understanding of linkage drag will help to distinguish it from selection during crop improvement. Four of six markers that previously rejected neutrality tests (437_2, 2486_1, 2534_1b and 2819_5) (Table 2 in [25]) were found to be introgressed rather than selected. It is anticipated that next-generation sequencing will be utilized to more rapidly eliminate linkage drag in crops [52].
Natural hybrids will carry high proportions of wild alleles making them somewhat easy to detect using large numbers of molecular markers. The intrinsic value of naturally introgressed germplasm was recognized in common bean (Phaseolus vulgaris) as a source of new alleles for traits such as disease resistance [53]. In S. lycopersicum, if horticultural effects of introgression are subtle then accessions such as Merville des Marchés may be prime sources to screen for new alleles. A search of the literature for accessions that were part of the current study (Table 2) found that PI 129128 showed high lycopene content, similar to lines containing pigment mutations such as ogc, hp, and dg[54].
Finally, it is worth noting that Rick [23] reported the potential of natural hybridization of S. lycopersicum with Solanum chilenseS. habrochaites (formerly Lycopersicon hirsutum f. glabratum), and Lycopersicon peruvianum (now revised into four species, [55]) in regions of Chile, Ecuador and Peru where sympatric populations grow in close contact, although he found no evidence of this based on phenotypes and severe postzygotic barriers are well known. Genotyping of populations sampled from these regions would provide evidence to reexamine whether introgression from these wild tomato species into S. lycopersicum has played a role in the crop's evolutionary history.
Plant material
Marker genotypes used in this study were previously reported for Solanum arcanum and S. lycopersicum[9,25] or were newly collected from each of five wild tomato accessions and one weedy S. lycopersicum accession Peru Wild. The specific accessions of S. habrochaitesS. pennelliiS. pimpinellifolium and Peru Wild have served historically as sources of important disease resistance alleles for tomato cultivars (Table 1). The two S. peruvianum accessions were chosen because they were known to be inbred and predicted to be highly homozygous. One of these, S. peruvianum LA1537, originated from accession PI 128650 which was the original source of Tm-2a[56,57]. Two plants per each of the six accessions were sampled as seedlings for genomic DNA isolation and sequencing of markers. Three accessions included in this study had previously published sequences for the markers [25] (Table 1). These were − a breeding line with documented multiple introgressions in its pedigree including Tm-2a (TA496) [57], an accession that predated tomato introgression breeding (Tomate, PI 99782), and S. arcanum (G 32591) a naturally self-fertilizing accession formerly classified as Lycopersicon peruvianum. For a few markers, published sequences from additional S. lycopersicum accessions (PI 109834 Merville des Marchés, PI 129026, PI 129128, PI 196297, PI 258474, PI 258478, PI 390510, Table 1) were included in analyses because they were previously reported as carrying highly divergent alleles, i.e. putative introgressions, at those loci [9,25].
DNA sequences
Genomic DNA extraction from seedlings, PCR amplification and two-pass sequencing were as described in Labate et al. [28]. Initially, 49 of 50 markers from Labate et al. [9] were sequenced. These represent random loci including expressed genes (expressed sequence tag, EST-based), highly conserved genes (COSII and U) and arbitrary loci. Marker 1523_4 was excluded without testing because it tended to give poor quality sequence within S. lycopersicum. Markers 175_1 and 1909_2 were dropped during this study because they did not consistently amplify S. lycopersicum homologs in wild tomato species, leaving 47 markers representing approximately 24 kb in total (Additional file 1: Table S1). Software packages phred, phrap and Consed [58,59] and Staden [60] were used for assembly and base calling of reads. Pregap4 (ver. 1.5) of the Staden package was configured to apply a base-calling algorithm "Estimate Base Accuracies" that is different from phred in order to independently verify the data. Sequence data were trimmed to remove primer binding sites and low quality ends (phred<40), and manually aligned in BioEdit [61]. All SNPs and heterozygous positions were confirmed by visual examination of trace files by two people. If the two plants from one accession had different sequences they were kept distinct; if they were identical they were treated as a single representative sequence of that accession. Heterozygous sites were manually edited to use IUPAC nucleotide ambiguity codes. GeneSeqer (ver. 08 Oct. 2008) [62] was used to compare exon and intron prediction for all markers against previous annotations [28].
All sequences were mapped using BLASTN [63] against tomato whole genome shotgun chromosomes (SL2.40) database with an e-value threshold of 1 x 10-40 on the SGN web site [64]. Gene models within markers and adjacent regions were identified in the SGN genome browser using ITAG2.3 Release: genomic annotations. For marker 437_2, tomato sequences were compared to transcribed sequences from an evolutionary out group (potato, Solanum tuberosum) by BLASTN searches of the Solanaceae Genomics Resource database at Michigan State University ( http://solanaceae.plantbiology.msu.edu/ webcite).
Statistical analyses
For each of the 47 markers, relationships among genotypes (also referred to as taxa) were first examined by applying the neighbor-joining (NJ) clustering method [65] as implemented in MEGA 4.0.2 [66] with 1,000 bootstrap replicates. Genetic distance and average evolutionary divergence (D) were estimated using the Jukes-Cantor method [34]; positions with alignment gaps or missing data were eliminated in pairwise sequence comparisons. For each of 11 markers that did not support the red-fruited clade based on MEGA results, consensus trees were generated by Phylip ver. 3.69 using Seqboot to produce 100 datasets by bootstrap resampling, Dnadist to estimate genetic distances using the Jukes-Cantor method, Neighbor to produce unrooted NJ trees and Consense to compute a consensus tree by the majority-rule consensus tree method [67]. SplitsTree4 ver. 4.12.3 [68] was used to create splits networks from DNA sequences or NJ trees [69]. Hybridization splits networks were created using the consensus tree of marker U146437 as a control that highly supported the red-fruited clade (99%) plus the consensus tree of the marker being tested for an introgression, with balanced sets of taxa (same taxa in each tree). A neutrality test [49] of marker C2_At1g73180 was carried out in DnaSP v. 5.10 [70] using marker TG11 as a control and S. arcanum as the out group.
ARS, Agricultural research service; COSII, Conserved Ortholog Set II; EMBL, European Molecular Biology Laboratory; ENA, European Nucleotide Archive; EST, Expressed sequence tag; NJ, Neighbor joining; GM, Genetically modified; PGRU, Plant Genetic Resources Unit; PUT, PlantGDB-assembled Unique Transcipts; QTL, Quantitative trait loci; RFLP, Restriction fragment length polymorphism; SGN, Sol genomics network; SI, Self-incompatible; SNP, Single nucleotide polymorphism; U, Unigene; USDA, United States Department of Agriculture.
Competing interests
The authors declare that they have no competing interests.
Authors' contributions
JAL and LDR conceived and designed the study. JAL acquired, analyzed and interpreted the data, and drafted the manuscript. LDR gave final approval of the version to be published. Both authors read and approved the final manuscript.
We thank S. Sheffer and P. Kisly for excellent technical support and E. Cobb for stimulating discussions. This work was funded by CRIS project 1910-21000-019-00D. USDA is an equal opportunity provider and employer.
The use of trade, firm, or corporation names in this publication is for the information and convenience of the reader. Such use does not constitute an official endorsement or approval by the United States Department of Agriculture or the Agricultural Research Service of any product or service to the exclusion of others that may be suitable.
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They had been called many names before; Demons. Angels. Gods and Seraphs. Yes, under many aliases and titles they lived and operated in their daily lives. However, despite all of their titles; no matter the acrimony or praise, they had always known themselves to be one thing:
The Children of the Ex Machina.
The Children of the Ex Machina possessed blood passed down from generations; linking them to a bond closer than siblings. They possessed abilities that linked them to a higher existence; their very lineage setting a course for transcended existence. They were strong, capable, superhuman...
Yet, they were weak.
For you see, the Children of the Ex Machina were just as your average human; albeit, much more capable. They did not do battle themselves. No, these great beings relied on much more powerful; extensively more mysterious entities to deal with their bouts.
The Deus; creatures of the Forbidden Realm with powers that transcended all understanding, whose background varied in number as the stars in space. The Deus are sworn to follow their master's orders to the very end. Many of the Deus are loyal and kind; following this ingrained need to fulfill their master's wishes. Others were not as willing. Their personalities, abilities, levels of power, and all else completely varied.
The Children of the Ex Machina all lived among one another as siblings; peaceful and blissfully unaware of the suffering of mortal coils. Each held different views, but lived happily under the same sky.
It was only when they grew aware of their conflicting views that warfare broke out. For you see, not even beings transcending man themselves could deny the temptation of absolute power and dominance. They fought for the top, never stopping or faltering.
For over 5,000 years; this had been the cycle.
Now, the power of the Ex Machina has become a sport. Those with the power of the Ex Machina compete against one another in underground contests. These contests were formed and funded by hundreds of shadowy organizations; unknown even the the highest of participants.
Now, as of today; those who have the power of the Children of the Ex Machina were known by a much more colloquial term.
There have been multiple attempts to sabotage the game of Ex Machina, all failing. However, there is a new disturbance in the game that will threaten not only the stability of the sport; but the lives of the Summoners. All marked with the brand of the Ex Machina are threatened by this mysterious force.
How will you deal with this threat? Where do you stand in the world of Summoners?
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Managing eutrophication is a key element in maintaining the earth's biodiversity....
Managing eutrophication is a key element in maintaining the earth's biodiversity. Eutrophication is a process mostly associated with human activity whereby ecosystems accumulate minerals. This unit explains how this process occurs, what its effects on different types of habitat are, and how it might be managed.
By the end of this unit you should be able to:
- Describe the principal differences between a eutrophic and an oligotrophic ecosystem.
- Explain the mechanisms by which species diversity is reduced as a result of eutrophication. (Questions 2.1 and 2.2)
- Contrast the anthropogenic sources that supply nitrogen and phosphorus to the wider environment, and describe how these sources can be controlled. (Question 3.1)
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- 1 Introduction
- 2 Effects of eutrophication
- 3 Causes and mechanisms of eutrophication
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Eutrophication describes the biological effects of an increase in the concentration of nutrients. The collective term 'nutrients' refers to those elements that are essential for primary production by plants or other photosynthetic organisms. Eutrophication is most often caused by increases in the availability of nitrogen and phosphorus, commonly present in soil and water in the form of nitrate and phosphate, respectively. However, altered concentrations of any plant nutrient may have a recognizable biological effect. Eutrophication can occur in any aquatic system (freshwater or marine), and the term is also used to describe the process whereby terrestrial vegetation is affected by nutrient-enriched soil water.
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Long time disinfonauts will surely remember one of the legends of modern day conspiracy investigations, Danny Casolaro, and his revelations of 'the Octopus." Casolaro met an untimely end and his investigation petered out. He's not forgotten, however, and his life and work are about to be immortalized in a stage play, reports American Theatre:
… Read the restA journalist is hot on the trail of a conspiracy whose scope seems ever-expanding. Nicknamed "The Octopus," the plot connects shadowy private-security contractors with an international criminal bank, government piracy of law-enforcement database software, a corrupt Native American reservation where experimental weapons are developed, and even Reagan's October Surprise.
The journalist hopes to break the whole story all at once and find fame and fortune. According to his book proposal (quoted in Jonathan Vankin and John Whalen's The World's Greatest Conspiracies), he's after a "web of thugs and thieves who roam the earth with their weapons and their murders, trading dope and dirty money for the secrets of the temple."
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Las Isletas. Estado Anzoategui
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Oregon schools started the year with an almost $1 billion uptick in state education funding, yet not much has changed.
State Treasurer Ted Wheeler is proposing a first-of-its-kind college scholarship program considered risky by investment professionals.
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Oct. 25 (Bloomberg) -- BankInvest, a Danish asset manager which owns a 0.25 percent stake in G4S Plc, will vote against the security provider's $8.2 billion takeover of ISS A/S at next week's shareholder meeting, its stock portfolio manager said.
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G4S Chief Executive Officer Nick Buckles needs at least 75 percent of shareholders to approve the purchase of ISS, the world's largest cleaning service provider, at a Nov. 2 shareholder meeting.
Crawley, England-based G4S plans a 2 billion-pound ($3.2 billion) rights offer to fund the purchase. G4S is buying Copenhagen-based ISS from EQT Partners and Goldman Sachs Capital Partners for a total value of 5.2 billion pounds, of which 3.7 billion pounds is assumed debt.
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PORT OF SPAIN, TRINIDAD - The Trinidad and Tobago Under-15 squad will be looking to mirror the success of the T&T Under-17s as they leave for Jamaica today for the West Indies Cricket Board Under-15 competition which bowls off on Friday.
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MIDDLESEX, ENGLAND - Middlesex suffered more bowling injury scares and several other self-inflicted wounds on day one of the LV County Championship clash with Warwickshire at Uxbridge yesterday.
Both Toby Roland-Jones and Corey Collymore sustained ankle injuries, the...
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KARACHI: Spearheading Pakistan's pace attack at the ICC Champions Trophy, Junaid Khan's failure to make an impact has given him the drive to live up to expectations in the Caribbean.
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Here's a story I filed Monday evening:
About the only thing the rushing attacks for Oregon and Stanford have in common is how many yards they accumulate.
When the No. 7 Ducks meet the Cardinal at 12:30 p.m. Saturday in Stanford Stadium, the two most prolific rushing offenses in the Pac-10 will square off. But they'll go about running the ball in vastly different ways.
"We're obviously a shotgun, zone- running operation, and they're an I-formation, pound-it-at-you deal," UO coach Chip Kelly said. "And they're really good."
The Ducks lead the conference and are eighth nationally with 233.2 rushing yards per game. Stanford is second in the Pac-10 and 16th in the country at 205.6 yards per game on the ground.
The Cardinal are getting 124.2 yards per game from the conference's leading rusher, Toby Gerhart. He's a bruising runner who has sublime speed but spends most of his Saturday afternoons between the tackles.
Contrast that with Oregon's option look, which spreads a defense out and attacks the edges. Less than half the Ducks' rushing production comes from their leading rusher, LaMichael James, who is third in the Pac-10 at 114.8 yards per game.
Quarterback Jeremiah Masoli is ninth in the conference with 62.3 yards per game, after exploding for 164 yards in a 47-20 victory over USC last week. His relationship with James in the option was something to behold against the Trojans, particularly in the third quarter.
"It's always been pretty good, but you can never get too much experience with the reps (in practice)," Masoli said. "It definitely helps. It's way better than it was at the beginning." Click here for the complete story.


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It was late last night when I was writing this, so I didn't go into as much backstory detail as I would have liked. I just added to it (copied below). The initial thought was to remove the electronics of the TV, insulate nearly the whole interior and replace it with a flat screen TV. To do this, we would have had to reduce the size of our current TV, so that wasn't going to happen. We have a few ideas on what to do with the interior now though, so I won't spoil the end (unless we find another idea that was really unique but won't work).
can't wait to see the end product
subscribed this is totally effing awesome. Even more awesome would be if your final plan is a touch screen jukebox (that's what I would want done with it!)
My Beer Review Blog: Beer Review Dude My Craft Beer Social Network: Craft Beer Network
Yeah... I gotta see it, too. Subscribed.
Did you save the magnifying screen for solar brewing? Google that.
HAHA THIS sir is madness!! I love it
The Best Homebrew and Beer Review VIDEOS on the internets:http://www.youtube.com/user/BeerGeekNation Buy Beer Geek Nation shirts - http://www.beergeeknation.com/store


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The room was clean... The bed was a little hard, and the room could use updating. Did have a flat screen tv. Shower was very good. Your basic room. No frills but fair. Family run. The staff was friendly, and extremely helpful. Continental breakfast was pretty basic.
- Reservation Options:
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- Also Known As:
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I create interactive experiences
based on game design principles
using app & internet technology.
I have collaborated with
innovators, artists and entrepreneurs
to create interactive
guided meditations,
television shows,
music albums,
and more.
Check out strenghts, weaknesses & skills and earlier work to see if I can offer you are looking for. Read what others say, browse source code (if you're technically inclined) or view Mark's resume.
Appjenou!? by Tin Men and the Telephone is an exploration of improvised music incorporating unparalleled opportunities for interaction. It opens up a whole new gamut of ways to listen and watch and join in. Using dazzling visualistions, you can manipulate the mood of the music, choose a solo, mute instruments, manipulate effects and more. Watch the video below:
As a member of dreamyourweb, Made by Mark developed a data-collection app to facilitate research on clean water facilities in Mozambique. The app features offline data collection, GPS-tagging, complex routing and photos. Results and photo's can be downloaded online for further processing in SPSS. Read the article here.
Check out the example data-collection app:
Showrush offers innovative online events. While viewers are watching the event on the internet, they can participate using various interactive options, or even join the show using their webcam!
Julien de Waal, CEO of Showrush, writes:
Mark is a driven individual who took on the innovative Showrush software project with enthusiasm & professionalism. I was impressed with his approach to structuring the project every step of the way as well as his skills & vision to develop a solid & customer friendly software. Mark works well within a team, is patient & is considered a hard worker! It's a pleasure working with him.
As lead developer, Made by Mark is responsible for all web technology, facing challanges such as realtime synchronization, webcam video streaming and scalability.
Self-financed project born from a passion for meditation. The app features an interactive guided meditation. Interactive, because the app modifies the guidance in reponse to user input. More details at www.marknis.com.
These are my strenghts
I'm always learning
(well, not anymore...)
I'm always excited to use the newest technology
I know my basics
...and I even have some other tricks up my sleeve
Download my resume for more information on my education and work experience.
Hello, my name Mark Marijnissen,
I'm 25 years old and I live in Nijmegen, the Netherlands.
I have been programming since I was a little kid. It all started in Logo. Then I moved on to Visual Basic. By the time I was twelve years old, I created a professional website for a friend - it was still HTML and animated GIFs those days :) In high-school I programmed things such as Blackjack on my graphical calculator, and after that I picked up Object Oriented programming in Delphi.
Now, I have a bachelor in Artificial Intelligence and I am working as freelance app developer.
But my life is more than app development. I like to create music (using keyboard, guitar, ableton push). I used to paint and draw, too. Because I am so fond of creative experiences, I enjoy collaborating with performers and artists.
I love the clarity and calm that mindfulness and meditation give me. I'm creating an interactive meditation app, and sometimes I guide a meditation at a local meditation group.
I like to practice non-violent communication to love and connect with the people around me - I am hosting a practice group in Nijmegen once a week.
Wolfstraat 180 | |
6531LR Wolfstraat | |
The Netherlands | |
KvK: | 50210041 |


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Hedge Fund Tech Compliance Update
On Tuesday, we began our webinar recap by looking at Form PF requirements and recommendations and other essentials for maintaining an effective compliance program. The second half of our webinar focused on technology compliance, specifically around message archiving, email security and mobile device management. Let's take a closer look at some of the content that was covered. If video is more your style, you can watch a replay of the webinar here.
Record Retention & Message Archiving
The SEC currently requires investment advisers to retain all internal and external electronic business communications. Rule 204-2 mentions the following specific measures:
In order to meet the requirements of the SEC, firms must retain and archive more than just email. Instant messages, Bloomberg and Thomson Reuters messages and other electronic communications are also considered required archival material.
Categorized under: Hedge Fund Regulation Security Communications Trends We're Seeing
Hedge Fund Form PF: Getting the compliance basics right
Is your firm registered with the SEC? Do you manage one or more private funds with assets of at least $150 million? If you said yes to these questions, then you have some homework to do. Under SEC regulations, your firm is required to file Form PF.
During a recent webinar, we asked ACA Compliance Group to talk us through the requirements and recommendations for filing Form PF as well as some additional compliance program recommendations. Below is a short recap of ACA's presentation. To listen to the full replay of our event, click here.
Form PF: Requirements & Recommendations
Depending on your firm's fund type and assets under management (AUM), the deadline for your Form PF filing may be sooner rather than later. Larger funds - including hedge fund managers, liquidity managers and private equity managers - will need to file sooner, while the majority of registered investment advisers won't need to file until early next year.
Categorized under: Hedge Fund Regulation Hedge Fund Due Diligence Trends We're Seeing
What Makes a Hedge Fund IT Provider Proud?
Anytime I write a self-promotional (i.e. Eze Castle Integration-focused) article on Hedge IT I feel the need to add the caveat that we try really hard to make all our articles educational. However, sometimes pride gets in the way and we have to share with you our proudest accomplishments.
To that end, today's post is going to be focused on blowing our own horn about the award-winning streak the company has been on. These awards are a direct reflection of our amazing employees.
Proud moment #1: Winning the Help Desk Institute Team Excellence Award for External Support. This award recognized our world-class help desk organization for their outstanding client support. In winning this award, we joined a very elite group that includes Dell Perot Systems, EDS and Novell as previous winners. Anyone who has dialed into our help desk knows our team is truly deserving of this award.
Categorized under: Eze Castle Milestones
Disaster Recovery Game Changer: From SAN-to-SAN to SAN-to-Cloud
Happy 5th of July!
We interrupt this holiday week to talk about how cloud services are expanding disaster recovery options for users of Storage Area Networks (SAN that is).
A traditional SAN-to-SAN disaster recovery scenario may rely on host-based replication technology transmitting data and applications between two mirror SAN environments. While this works for some firms, cloud technology presents a viable, cost-effective alternative where data and applications from a firm's production SAN are replicated to a cloud environment for disaster recovery.
Categorized under: Disaster Recovery Business Continuity Planning Cloud Computing
Happy Fourth of July from Eze Castle Integration!
On behalf of the entire team here at Eze Castle Integration, we would like to wish all of our readers, clients, partners and friends a very happy and safe Independence Day holiday. Enjoy!
Categorized under: Trends We're Seeing
MPLS Networks Simplified
What is an MPLS Network?
MPLS (Multi-Protocol Label Switching) is a mechanism in high-performance telcom networks that directs data from one place on the network to another based on short path labels rather than long network addresses. MPLS is highly scalable and protocol agnostic.
In an MPLS network, packets of data are assigned labels, and all packet-forwarding decisions are made solely on the contents of these labels, eliminating the need to examine the packets themselves. As a result, end-to-end circuits can be created across any type of transport medium, using any protocol.
At Eze Castle, we like to boast that our private cloud services are delivered via an MPLS network which connects our data centers. That sounds good, but what are the real benefits of this type of network infrastructure? We asked our vice president of networking services, Mike Abbey, for some insights. Here's what we learned.
Categorized under: Business Continuity Planning
Data Jurisdiction in the Cloud: The Patriot Act Fear Factor
Cloud computing has gained popularity over the years and is now fast approaching a global scale as hedge funds around the world leverage this innovative technology to help improve efficiency and cut costs. However, cloud computing raises unique data regulation and jurisdiction considerations as cloud environments span multiple geographic locations and data is not tied to one physical location. In today's article we will look at data regulation and jurisdiction considerations for UK companies utilising US headquartered cloud providers.
Many cloud service providers are increasingly serving customers outside their home markets and using service delivery models that require the transmission of data across borders, which has led to a great deal of fear about the rights of access under the USA PATRIOT Act and the geographical extension of those.
Beyond the US, in December 2011 the European Commission published results of its cloud computing consultation, which showed a lack of understanding about the EU legal framework that cloud computing should be implemented within. It also signaled that there is still a widespread need for clarification on rights, responsibilities, data protection and liability in the cloud, especially in cross-border situations.
Categorized under: Hedge Fund Regulation Cloud Computing
Cloud Adoption in the Investment Industry: Survey Results Part 2
Last week, we revealed Part 1 of our cloud adoption trends survey results and detailed how hedge funds and investment firms are currently leveraging the cloud, as well as what kinds of cloud deployment models they are using (private clouds take the cake).
Some additional data points we learned as a result of this survey include the driving factors influencing firms' decisions to use the cloud, potential barriers to cloud adoption and the key evaluation criteria for cloud services providers. Let's take a closer look at what survey respondents had to say relative to these categories.
Factors Influencing the Decision to Use the Cloud
There are a multitude of factors that alternative investment firms need to take into consideration as they evaluate cloud offerings. Survey respondents were asked to rank the importance of several factors related to their cloud decision-making, including cost, flexibility, functionality and speed.
Categorized under: Cloud Computing Security Trends We're Seeing
Cloud Adoption Survey Results Revealed: Part 1
Survey Says: Cloud computing is no passing trend!
If you missed it yesterday, we officially announced the results of our "Cloud Adoption Trends within the Investment Management Industry" survey during a live webinar. You can listen to the webinar recording here.
Our cloud survey set out to investigate how hedge funds and investment management firms are using cloud services today, as well as to provide insight into the factors influencing this trend and also the barriers to adoption. Here's a look at some of the findings:
Respondent Profile
Our online survey, which was conducted between March and May of 2012, surveyed 125 financial services firms in the United States. Of those 125 firms, 65 percent identified as investment management firms. Additional firm types represented included hedge funds (16%), private equity firms (16%) and fund-of-funds (4%).
Categorized under: Cloud Computing Trends We're Seeing
World IPv6 Launch Day is Here. What Does that Mean?
It's alive! It's alive!
Well actually tomorrow, June 6 2012, it (aka IPv6) will officially be alive, but that doesn't work with our Hedge IT blog calendar so today we look at IPv6.
As a refresher, since the inception of the Internet, we have been using IPv4, which totals about 4.3 billion Internet addresses. But with the increasing number of wireless technologies that support the Internet (smart phones, tablet, etc.), these addresses are depleting.
Enter IPv6. The new IPv6 protocol uses 128-bit addresses and allows for substantially more IP addresses – trillions upon trillions of new addresses. The World IPv6 Launch Day marks a key milestone as companies shift their infrastructures to the new protocol, which will eventually completely replace IPv4.
Categorized under: Infrastructure Cloud Computing Communications Security Trends We're Seeing


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23rd Annual Holiday Concert
December 13 at 7:30 pm and December 14 at 2:30 pm
Tickets may be purchased at the lobby box office prior to each performance.
Welcome to Champlin Park Vocal Music!We are the 300 voices of Champlin Park High School, a comprehensive four-year high school serving the northern suburbs of Minnesota's Twin Cities since 1992. Our five principal ensembles--Concert Choir, Treble Choir, Chamber Choir, Bel Canto, and Rebel Men--present four concerts each year and are joined by a number of extracurricular ensembles like Rebelution (our amazing vocal jazz group), plus Viva Voce and Carolers. Now in our 23rd season, we perform unique and challenging repertoire under the direction of Ms. Laura Tempel and Ms. Margaret Sabin. We've had the opportunity to work with Dr. Galen Darrough, Dr. Edith Copley, and Phil Mattson, among others. We have performed at the ACDA Minnesota Fall Convention and the Choral Arts Finale at Orchestra Hall. Every other year, we travel to sing somewhere other than Minnesota. In 2013 we toured Colorado & The Rockies and in 2011, New Orleans! This year, we're heading to Texas & The Gulf.
We'd love to have you in the audience for an upcoming concert and showing your support for the performing arts at Champlin Park. Get involved with the CPHS Vocal Music Association and like us on Facebook, follow us on Twitter, watch on YouTube, and listen on SoundCloud!
Showing posts 1 - 0 of 28.
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Lancs groundsman takes top gong
The England and Wales Cricket Board today announced the 2009 Groundsman of the Year Awards.
Matt Merchant from Old Trafford is the winner of the four-day category with Mick Hunt of Lord's awarded the runner-up prize.
In the one-day category, the winner is Neil Godrich from Derby with the runner-up prize shared between Sean Williams of Bristol and Phil Frost of Taunton.
A commendation went to Andy Peirson (Canterbury) in the four-day pitch category, while Bill Gordon (Brit Oval), Paul Marshall (Northampton), Mick Hunt (Lord's), and Nigel Gray (Rose Bowl) were commended in the one-day category.
John Moden at Fenners is the winner for pitches at MCC Universities with Richard Sula at The Parks runner-up.
Ross Spry at Cheltenham was awarded the Outgrounds prize with Micky Stewart and Christian Dunkerley at Scarborough earning the runner-up award.
Alan Fordham, head of operations (first-class cricket), commented: "The work of all head groundsmen is highly valued and we are delighted to make these annual awards, which recognise the head groundsmen who have prepared the best pitches this season."
Every pitch is rated at the end of the match by each of the umpires. The winners of each category are the groundsmen whose pitches have achieved the highest average rating across the course of the season.


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OK, so I'm stuck really early on and am getting frustrated.
I assume that I have to make Wallace look like the picture on the magazine but I don't have a clue on how to do that, and trying everything on everything doesn't seem to accomplish anything. (I worked out what to do with the nut butter, but that doesn't seem to be getting me anywhere, either.)
EDIT: Don't worry, I worked it out in the end - it wasn't Wallace that I had to alter.


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If sin(theta) < 0 and tan(theta) > 0 then: Answer choices: 90degrees < theta <180degrees 180degrees<theta<270degrees 270degrees<theta<360degrees 0degrees<theta<90degrees This...
If sin(theta) < 0 and tan(theta) > 0 then: Answer choices: 90degrees < theta <180degrees 180degrees<theta<270degrees 270degrees<theta<360degrees 0degrees<theta<90degrees This...
the value of x and the measure of the angles
A "Sparky" is a new measure for angles invented by Pre-Calculus students at Arizona State University. An angle corresponding to one full rotation around a circle measures exactly 170 Sparkies...
1) in triangle ABC <B=53 degrees, b=6.2 cm, and <C=47 degrees. Find the length of side a 2) A kayak leaves a dock on Lake Athatbasca, and heads due north for 2.8 km. At the same...
I'm not sure how to solve this, or what a ratio means.
Find m<1 using the given information. m<1=5x, m<4=2x+90 X= <1=
A tetrahedron has edges of 5cm each. What is the angle between any two faces? Please explain how to answer this question. Thanks :)
If supplement of an angle has a measure 78 less than the measure of the ange, what are the measures of the angles?
Find the length of side BD.
Determine the interior angles of triangle ABC for A(5,1), B(4, -7), and C (-1,-8)
sinP= cosP= tanP= cscP= secP= cotP=
what is the degrees of a hexagon
A rectangle has EI = 3x+8 FI=5x-6 and m<IGF=29 Find EG and m<IFE
Bisectors of Triangles As you well know by now, being able to deduce key information from a limited set of facts is the basis of geometry. An important type of segment, ray, or line that can help us prove congruence is called an angle bisector. Understanding what angle bisectors are and how they affect triangle relationships is crucial as we continue our study of geometry. Let's investigate... read more
Angle Properties of Triangles Now that we are acquainted with the classifications of triangles, we can begin our extensive study of the angles of triangles. In many cases, we will have to utilize the angle theorems we've seen to help us solve problems and proofs. However, there are some triangle theorems that will be just as essential to know. This first theorem tells us that if... read more
Pairs of Angles In geometry, certain pairs of angles can have special relationships. Using our knowledge of acute, right, and obtuse angles, along with properties of parallel lines, we will begin to study the relations between pairs of angles. Complementary Angles Two angles are complementary angles if their degree measurements add up to 90°. That is, if we attach both angles... read more
Introduction to Angles We begin our study of angles by learning what they are, how to name them, and ways in which angles can be classified. These concepts are explained below. Angle An angle is formed when two rays meet at a common endpoint, or vertex. The two sides of the angle are the rays, and the point that unites them is called the vertex. The vertices are shown... read more
Lines and Angles Comprehension of lines and angles is critical in the study of geometry. Whether we know it or not, we see all sorts of angles on an everyday basis. The corners of a room, the way a ladder rests against a house, and the inclination of a hill all have angles that can be measured. In order to continue our study of geometry, it will be necessary to learn about the different... read more
Angle Properties, Postulates, and Theorems In order to study geometry in a logical way, it will be important to understand key mathematical properties and to know how to apply useful postulates and theorems. A postulate is a proposition that has not been proven true, but is considered to be true on the basis for mathematical reasoning. Theorems, on the other hand, are statements... read more


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Cotton canvas ankle sandals from A.P.C with contrast elasticated ankle strap. Features a reinforced heel cap, ridged rubber sole, and easy slip-on styling. • Cotton canvas ankle sandal • Elasticated ankle strap • Reinforced back • Wedged platform View Size Chart
1.25" heel 2.5" ankle height View Size Chart


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Before babies became Hollywood's hot new fashion accessory, most stars kept their parental endeavors limited to their pets. And even throughout Hollywood's recent birth boom, celebs still manage to find the time to get themselves photographed while walking their dogs. While some famous pooch-lovers have fetching doggystyle (UPGRADE), others look like they should be taken to the pound (DOWNGRADE). Tell us which of these stars are improved by their puppies, and which ones just look like a dogface.
BWE: Paul Scheer Hijacks The Airwaves
From Product Shop NYC:
I thought that our "exclusive interview" with "Jeff Mangum" would immediately be exposed as a fake, especially considering that the interview was done by comedian Aziz Ansari, and that "Jeff" talked about releasing a soloflex machine, starting a dating show for VH1, killing the singer of Beirut, and the fact that he sounded exactly like Best Week Ever's Paul Scheer. Do I need to be any clearer about this?
Apparently. Paul Scheer just so happened to post the interview with the Neutral Milk Hotel frontman on his site, which I think means that he was the "Jeff Mangum" all along! No! Could it be? Well, head on over and listen to the hilarious interview and find out for yourself.
ICYMI: Rocky 6(0)
Pop Quiz: What's the funniest part of the official Rocky Balboa trailer? Is it:
a) That a video game may be intricate to the plot of the film
b) The antagonist's name is Mason "The Line" Dixon
c) Paulie
d) Despite everything, it still looks better than Rocky V
Watch it now, then choose your answer. Good luck.
While You Were Prepping For Little Man
- Ben Affleck talked Kevin Smith into giving him a cameo in Clerks II. Unfortuantely Smith chose not to enforce the Jersey Girl clause that's supposed to keep the two men 1000 yards away from one another.
- Weezer has called it quits. Fans are encouraged to trash their last three albums pretend this announcement actually came in 1997.
- The reverend at Ken Lay's funeral compared the former Enron CEO to Jesus and Martin Luther King Jr. Okay, so let's see; they're all dead, and… you know what, that's the only similarity I can think of.
- During a show in Hollywood, Bon Jovi told guitarist Richie Sambora to "sign a prenup" next time. Yeah, the poor guy needs to stop attaching himself to poor, struggling women like Heather Locklear and Denise Richards.
- Zinedine Zidane headbutted Italian defender Marco Materazzi during the World Cup final because Marco insulted his mother and sister. Coincidentally, this is the same reason Zidane was never able to cut it on Yo Mamma.
Pat Or No Pat
The banker on Deal Or No Deal is a shady character. You never see him, never hear him, never know what's going on in his little banker head. He just lurks in the shadows and occasionally blogs… but that's it.
Well, the folks over at Collegehumor may have discovered the true identity of the banker– it's Pat O'Brien! Now we finally know what's actually going on during those drawn out phone calls to Howie Mandel. It all kind of makes sense now…
Watch it here! (audio NSFW)
Best Night Ever: Wenesday, July 12th
It's Best Night Ever for Wednesday, July 12th! Shea Hess is here to walk you through the best of Wednesday night tv, including Rock Star: Supernova, America's Got Talent, and Project Runway!
Featuring music by The Vitamen and The Station Myth.
- UNINVITING VIEW: Shannen Doherty, who will temporarily fill Star Jones' vacant chair on The View, shows off a new face that isn't exactly what I want to see in the morning. (Hot Online News)
- "AWESOME" HUSBAND MOVE: K-Fed is planning to record a duet with his first baby momma, Shar Jackson. (The Bosh)
- IMPRESSIVE RESUME: Johnny Depp has been engaged to five women, all of which are hotter than most women you'll ever get to see in person. Well, almost all of which. (Us Weekly)
- WANNABE HOMEWRECKER: Cameron Diaz had better watch her back, because Rachel Bilson might try making Justin to "Bye Bye Bye". (Popsugar)
- MASTURBATORY COMIC BOOK TRAILER: I don't know that this trailer is the direction Sony wants to take their prized Spiderman franchise into, especially after the lukewarm reaction to "Gay Superman". (Junkiness, slightly NSFW)
SIZZLER: 'Idol' Reject Made R. Kelly-esque Sex Video
It's one thing for an aspiring hip hop artist to emulate R. Kelly's career, but DJ Boyd seems to have taken that admiration just a bit too far. You probably don't even remember who DJ Boyd is, and you shouldn't, so here's a quick refresher – after making it to the "Top 70″ on the 5th season of American Idol, Boyd was dismissed and joined the ranks of rejects deemed unworthy of competing against the likes of Clay Aiken. But back home in Utah, Boyd held onto his dreams of musical stardom – he signed to "Big Fedi" records, released an album you've probably never heard, and constantly reminded people that he was once on TV for a few moments. Nothing too impressive, unless of course you're two teenage girls who he has just filled with booze, in which case you might have found him attractive enough to have sex with, even allowing him to videotape the encounter. But when that Smirnoff Ice buzz finally wore off and you woke up hungover with the realization that you'd just slept with some J-list reality show failure, you might have regretted your decision so much that you called the police and reported to what this 27 year-old sleazebag did to you and your friend (who happens to be 15, a year older than yourself), resulting in his arrest and a possible judgement much harsher than even the nastiest thing Simon Cowell could have said to him. As the local court justices he's now facing probably won't be nearly as drunk as Paula Abdul, this wannabe Idol could be in some serious trouble.
LISTEN UP: BWE's Daily Dose of the Best Music Ever
- Scenestars serves up a few classic tracks from indie pop fuzz-freak OG-hipsters Stereolab, from their forthcoming "best of" album.
- Stereogum says The Slip is a band to watch, which means that in about 6 minutes every blog-reading indie-kid on the planet will be doing just that – so hurry over and grab yourselves a good seat!
- If you still haven't discovered the sweet sounds of Bishop Allen, you're just not listening. So stop what you're doing, check out Indie Mix Tapes, and hear the hotness.
- The early birds over at The Pelican's Perch get the
wormword out on the awesome new album from Regina Spektor by posting three of the latest tracks. - MOKB shakes up the sounds of Snowglobe, a great indie rock band from Memphis who happen to be friends of mine. Do yourselves a favor and give them a listen!
BWE: Dare To Air Promos
A while back we asked some filmmakers to put together interesting Best Week Ever promos. Anything goes. Here's what the folks over at Hungryman came up with: Timothy- a man who truly is having the Best Week Ever.
Check out Timothy At The Bus Stop and At The Office as well. What a guy!


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Lil Herb x Earl Sweatshirt – Knucklehead (Prod. by Randomblackdude)
For RedBull's 20before15 event, Earl Sweatshirt and Chicago rapper Lil Herb dropped a song together called "Knucklehead." The song can
For RedBull's 20before15 event, Earl Sweatshirt and Chicago rapper Lil Herb dropped a song together called "Knucklehead." The song can
Earl Sweatshirt has just released two new songs, "SONG FROM 2012 HIGH 6AM RAMBLING AT THE 5TH" (a slowed, screwed
Cleveland, OH rapper just released the newest single off his upcoming CleveLAfornia mixtape, Cherish The Day, and it features a hot verse
To top off a great year of music from The Internet (which included their full-length Feel Good, touring with Mac Miller,
Domo Genesis just dropped a hot verse on a standout track from Curren$y affiliate Trademark Da Skydiver's newest project, Flamingo Barnes
Mike G just released the newest addition to his ongoing music giveaway, Mike G Mondays. The track "Nesta", produced by
I had gone to the liquor store after a hard hungover blaze and purchased myself a bottle of Wild Turkey
Some new music from Mike G today! After sitting down for a conversation on the "Kinda Neat" podcast, Mike let
Frank was in Munich, Germany tonight for the first show of his "You're Not Dead" world tour. The set list
Some new music from Domo Genesis today! Table Tennis sees Domo going in over a smooth Thelonious Martin produced beat.
Another new Chopped & Screwed track from Mike G. This time it's the new Wale remix of his track Bad
Another new Chopped & Screwed track today from Mike G! This time he stays within the Odd Future camp and


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American Standard Version (ASV) | New International Version (NIV) |
1 And there was a famine in the days of David three years, year after year; and David sought the face of Jehovah. And Jehovah said, It is for Saul, and for his bloody house, because he put to death the Gibeonites. | 1 During the reign of David, there was a famine for three successive years; so David sought the face of the LORD. The LORD said, "It is on account of Saul and his blood-stained house; it is because he put the Gibeonites to death." |
The American Standard Version is in the public domain. (The American Standard Version Bible Online) | Scripture quoted by permission. Quotations designated (NIV) are from THE HOLY BIBLE: NEW INTERNATIONAL VERSION®. NIV®. Copyright © 1973, 1978, 1984 by Biblica. All rights reserved worldwide. (New International Version Bible Online) |


text: Not Remotely ImportantThe old man still looked up at the sky. Every night, when he had the chance, he would wrap up warm and take a flask of hot tea out to the seat beside his telescope. It was an act of tribute now. He had long since let go of the idle fancy that he would ever see that man, that most wonderful man again. Sometimes it was a truly bitter memory, another friend lost to war, and how he hoped that the poor creature, that ancient blazing angel had been wrong. Perhaps he was out there somewhere, fully recovered, too embarrassed to return. Too afraid?Wilfred knew it was still dangerous. His granddaughter visited often, particularly when his bungalow was situated just around the corner from her comfortable family home. It was a miracle to see her so content since those miserable Christmases past, but she had found happiness. Her husband treated her well and the world was just bursting with opportunities for her now she had the freedom and time to consider. The distant expressions she showed