You've got a knot in your compound curve. Specifically, the Z-shape's path is reversed. How to unravel: select the compound path 'Zep' and break it apart (Ctrl-K). The 'e' and the 'p' will look like they've lost their counters (the inside 'holes') but don't worry, they're there. Deselect and then click on the Z shape only to select it. Hit Ctrl-10 to bring up the node editor tool, right click (or use icon on Command bar) to get to 'Reverse SubPath'.
Click off the object to deselect and then select everything including the background oval and use 'Ctrl-L' to Combine the objects into what AI users call a combined path. The letters will magically appear as transparent holes.
You can also use the Shaping tools to do the cut-out, but since nothing overlaps the oval, this is faster. First of all thanks to Broacher on your 'reverse subpath' solution. I've had this problem many times when I intersect or weld items and some pieces appear to go missing but the paths are there. This happens almost all the time when I'm doing script with tail designs and I weld the tail to the script that has been converted to curves and all welded together. My previous solution was to not weld it, export it as a high resolution black and white bitmap, import the bitmap and auto-trace it (but then it loses quality).
Now replying to JourneyGrafix's last post, after you set the outline to 1pt and then convert the outline to an object you then need to select the fill and the new outline object and weld them together. If you go into View and Wireframe View you can see what's going on. After you convert the outline to an object the original object is still there only it's outline has been removed and turned into a new object. The easiest way to see what's going on is select 'Zep', set your outline, press Ctrl+Shift+Q (to 'Convert Outline to Object') and then click on another color while that new object is still selected.
You'll see the outline change to a different color than the fill. Just click the outline and the fill and choose 'Weld' so the 'Zep' part will slightly overlap the oval in the background. Thanks Risket for the follow-through. The way I often handle this situation is by using a tool that is unique to CorelDraw-- in fact, it's one of those rare cases where I will actually bring in art from an AI file to do this trick.
Corel's Contour tool is great at quickly adding an expanded 'outline' to not only individual vector shapes, but to whole groups. Ideal for t-shirt stuff like type overshooting an oval, for example. Basically, you just set the contour to outside, step of 1, and enter your offset width. Once the contour is complete, you then convert it (break, in Corel's parlance) to a single curve and go from there. Mind you, there's a slight price for all this -- especially if we're talking knife cutters. Corel is kinda 'easy' when it comes to calculating the points and often a tight corner or a complex area gets more complex than desired. But that's Corel.
In a previous post, I promised that we we would run a future post “synthesizing some of the more recent court orders to ask if the orders draw an optimal balance between the need to protect IP owners against infringement, whilst at the same time safeguarding the interests of a raided party.” I am still in the. Warez download corel draw 12. Com a reliable community where all people can download free including downloads categories Rasmussen university online degrees platforms. We provide high-secure harry potter prisoner of azkaban game file 32caf5b1eb mastodon skye rarest dvd audio extractor 6. InformationWeek 2.
Lots of stunt driving, just get to know where the potholes are.
Anguillonema amolensis n. Is described and illustrated based on its morphological, morphometric, and molecular characters. The new species is characterized by its 575 to 820 μm long and wide body (body width at vulva = 30 to 59 μm), irregularly ventrally curved after fixation, five to six lines in lateral fields, 6.0 to 7.5 μm long stylet with small rounded knobs, pharynx lacking a median bulb, pharyngo-intestinal junction anterior to nerve ring and excretory pore, females with monodelphic-prodelphic reproductive system, 15 to 19 μm long conical tail with broad rounded tip, and males absent. The new species is compared with two known species of the genus, Anguillonema poligraphi and A.
Molecular phylogenetic studies of the new species using partial sequences of small subunit (SSU) rDNA revealed that it forms a clade with an unidentified nematode species and two species of the genus Howardula. In phylogenetic analyses using partial sequences of the 28S rDNA (D2-D3 segment), the new species formed a monophyletic group with species belonging to two genera Howardula and Parasitylenchus. The genus Anguillonema belongs to suborder Hexatylina, and there was not a consensus on its taxonomic position until 2000 (a genus dubium in )., in his second volume of book series on free-living nematodes of Hungary, proposed a resolved taxonomic position for the genus under the same suborder, Hexatylina, the family Neotylenchidae and subfamily Gymnotylenchinae. This is one of the rarest nematode genera, with poor data on its morphology; and some details of its body structure such as the nature of pharynx and pharyngo-intestinal junction are not well known due to poor illustrations and/or lacking of other reports or redescriptions (). As expected, there is no molecular data for the genus in GenBank database. Recently, two genera are added to the suborder Hexatylina (; ), and in a recent study (), a history of some conducted taxonomic studies of insect-related nematodes is given. In our samplings from several ports of northern forests of Iran, a population belonging to the genus Anguillonema was recovered from rotten wood of a dead trunk of a forest tree.
The objectives of this work were a morphological study of this rare and poorly known genus and a first molecular phylogenetic study using two genomic fragments. Sampling, extracting, and taxonomy: Several soil, wood, bark, and rotten organic material samples were collected from different natural locations and forests of Mazandaran province, northern Iran, during 2015 and 2016. All samples were kept in a cool and dark place. Nematodes were extracted from the collected samples using the tray method () and examined under a Nikon SMZ1000 stereomicroscope. Nematodes were hand-picked and heat-killed by adding boiling 4% formalin solution, transferred to anhydrous glycerin according to, mounted on permanent slides, and examined using a Nikon Eclipse E600 light microscope. Photographs were taken using an Olympus DP72 digital camera attached to an Olympus BX51 microscope powered with differential interference contrast.
Drawings were made using a drawing tube attached to the microscope and were redrawn using CorelDRAW® software version 16. PCR: DNA was extracted from one single female nematode.
The specimen was picked out, studied onto a temporary slide, transferred to a small drop of TE buffer (10 mM Tris-Cl, 0.5 mM EDTA, pH 9.0; 100 QIAGEN Inc., Valencia, CA) on a clean slide and squashed using a clean slide cover glass. The suspension was collected by adding 15 μl of the aforementioned buffer (). The DNA sample was stored at −20°C until using as PCR templates. Primers for amplification of 18S rDNA were forward primer SSU F22 (5′-TCCAAGGAAGGCAGCAGGC-3′) and reverse primer SSU R13 (5′- GGGCATCACAGACCTGTTA-3′) as used. Primers for 28S rDNA D2/D3 amplification were forward primer D2A (5′-ACAAGTACCGTGAGGGAAAGT-3′) and reverse primer D3B (5′-TGCGAAGGAACCAGCTACTA-3′) (). PCR reaction was carried out in a total volume of 30 μl (19.2 μl distilled water, 3 μl 10× PCR buffer, 0.6 μl 10 mM dNTP mixture, 1.2 μl 50 mM MgCl2, 1.2 μl of each primer (10 pmol/μl), 0.6 μl of Taq DNA polymerase (5 unit/μl, CinnaGen, Tehran, Iran), and 3 μl of DNA template.
The thermal cycling program for amplifying two genomic fragments (18S rDNA, 28S rDNA D2/D3) was as follows: denaturation at 95°C for 4 min, followed by 32 cycles of denaturation at 94°C for 30 sec, annealing at 52°C for 40 sec, and extension at 72°C for 80 sec. A final extension was performed at 72°C for 10 min. The PCR products were sequenced in both directions using the same primers with an ABI 3730XL sequencer (Applied Biosystems) at Macrogen (Seoul, South Korea). Newly obtained sequences of the studied species were deposited in GenBank (accession numbers: for partial 18S and for partial 28S rDNA).
Phylogenetic analyses: The newly obtained 18S and 28S rDNA sequences were compared with those of other nematode species available in GenBank using the BLAST homology search program. The selected DNA sequences were aligned using MUSCLE () as implemented in MEGA6 (). The most appropriate model of nucleotide substitution was selected using the Akaike information criterion in MrModeltest 2 ().
The general time reversible model, including a gamma distribution for rates across sites and a proportion of invariant sites (GTR + G + I), was selected. Bayesian inference (BI) was performed using MrBayes v3.1.2 () running the chains for five million generations (nruns = 4). After discarding burn-in samples, the remaining samples were retained for further analyses. The Markov chain Monte Carlo method within a Bayesian framework was used to estimate the posterior probabilities of the phylogenetic trees () using the 50% majority rule. Suitability of the posterior sample size was evaluated using autocorrelation statistics as implemented in TRACER v.1.5 (). A maximum likelihood tree was reconstructed by using RaxmlGUI 1.1 () software using the same nucleotide substitution model as in the BI including 1,000 bootstrap pseudoreplicates. For the 28S rDNA phylogenetic analyses (BI and maximum likelihood), Poikilolaimus oxycerca and Poikilolaimus piniperdae (accession numbers and, respectively), and for the 18S rDNA phylogeny, the species Pseudacrobeles sp., Acrobeloides maximus, and Acrobeles ciliatus (accession numbers,, and, respectively) were used as outgroup taxa.
The resultant files of phylogenetic software were visualized using Dendroscope V.3.2.8 () and redrawn in CorelDRAW software version 16. Female: Vermiform, wide, irregularly ventrally curved after fixation. Cuticle thin, finely annulated with five to six lines in the lateral field. Lip region low, continuous with the body. Amphidial openings not visible. Stylet short, conus as long as the shaft or slightly shorter, small knobs distinct. Dorsal gland orifice close to the stylet knobs.
Corpus muscular, wide, slender, isthmus lacking, pharyngeal glands forming long dorsal overlapping. Phyaryngo-intestinal junction anterior to the nerve ring and excretory pore. Hemizonid indistinct. Intestine simple, rectum and anus functional.
Reproductive system monodelphic-prodelphic, ovary long, usually reaching nerve ring, reflexed once or twice in some individuals, oocytes in multiple rows close to the germinal zone, at two rows distally, oviduct and spermatheca indistint, crustaformeria formed from multiple cells, joining to the uterus, the latter sometimes containing mature egg, vagina with sclerotized walls, vulva a transverse slit with protruding lips and the postvulval uterine sac absent. Tail conical, short, with wide rounded tip dorsally bent in fresh females in water. Diagnosis and relationships: Anguillonema amolensis n.
Is characterized by its long (575 to 820 μm) and wide (30 to 59 μm) females, low lip region continuous with body contour, five to six lines in lateral fields, small stylet with small rounded knobs, monodelphic-prodelphic reproductive system, and conical 15- to 19-μm long tail. It is morphologically close to the type species of the genus, A. Poligraphi, and compared with it, has a continuous head (vs. Offset, according to original drawings), shorter tail (average length of 16.0 ± 1.5 vs. 28 µm, as calculated), smaller c′ (1.2 ± 0.2 vs.
2.1, as calculated from drawing), greater c (45.1 ± 0.7 vs. 23), and broadly rounded tail tip not dorsally bent in mounted specimens (vs. Tail tip dorsally curved, apparently sharp).
Compared with A. Crenati, the new species has shorter body (average length of 700.0 ± 89.3 vs. 1,082 μm), smaller a value (an average of 16.3 ± 4.3 vs. 42), greater V value (an average of 95.7 ± 0.5 vs. 92.6), and shorter tail (c′ = 1.2 ± 0.2 vs.
4, as calculated according to original drawing) not dorsally bent in mounted specimens (vs. Dorsally bent). Molecular phylogenetic analysis: Sequencings of 18S and 28S rDNA D2/D3 fragments of the new species yielded single sequences of 882 and 870 nt (accession numbers and, respectively). The blast search of the GenBank nucleotide database using these sequences revealed they are both unique. A 96% to 98% identity was achieved for BLAST search of partial 18S rDNA sequence for some species belonging to the genera Howardula, Rubzovinema, Deladenus, and Parasitylenchus. The BLAST search using partial 28S rDNA, revealed it has the highest identity (88%) with Howardula phyllotretae (accession number ).
A total number of 49 hexatylenchid species/isolates, and 55 species/isolates of hexatylenchids and anguinids were used for reconstructing of 18S and 28S phylogenetic trees. The multiple alignment of 18S dataset was composed of 1964 total characters with 574 variable characters. The 28S dataset was composed of 431 total characters of which 269 characters were variable. In 18S tree, the new species, representing the only currently sequenced species of the genus has formed a clade with an unidentified isolate (accession number ), both of which forming a clade with two species of Howardula (accession numbers, ) ().
In the 28S phylogenetic tree, the new species has formed a clade with an isolate of Howardula (accession number ) and two isolates of Parasitylenchus (accession numbers and ) (). The monophyletic nature is not seen for most families and subfamilies of Hexatylina in either the 18S or 28S trees, however, genomic sequences are not available for most representatives of the suborder. Discussion The genus Anguillonema belongs to one of the rarest Tylenchomorpha genera and currently includes two species, A. Poligraphi and A. Crenati, both of which are reported in the shape of original descriptions (no other reports or redescriptions are available for these two species).
The “ dubium” status of the genus () and its uncertain taxonomic position was revised by, and the genus was placed inside the family Neotylenchidae, subfamily Gymnotylenchinae. It seems, however, that the type materials of these two known species are not accessible. According to, the suborder Hexatylina contains two superfamilies: Sphaerularioidea and Iotonchioidea.
The first superfamily, contains three families Sphaerulariidae, Allantonematidae, and Neotylenchidae. The latter family, Neotylenchidae, as stated, has “two types of generation, one free-living, fungus- or plant-feeding, another involving a heterosexual female parasitic in the insect haemocoel”. Thus, with regard to the free-living mycetophagous or probably, the plant feeding habit of the recovered new species, the placement of the genus by inside the family Neotylenchidae is confirmed, but the lack of knowledge about other type of generation in the tentative insect host, the assigning of the genus Anguillonema to either of four subfamilies of Neotylenchidae ( Sensu ) or even to a new subfamily was not conducted in this study. However, the assigning of Anguillonema to Gymnotylenchinae by could be logical, as, morphology could support such placement. The suborder Hexatylina ( sensu ) comprises a diverse group of taxa which are separated from each other based on their morphological and/or biological characters. This is an artificial grouping and molecular phylogenetic studies do not infer such classification.
Besides, using the currently available sequences of representatives of several genera of Hexatylina, members of the families and even genera, do not form monophyletic groups in phylogenetic trees using the SSU and large subunit (LSU) rDNA sequences. In our present SSU tree, members of Sphaerularioidea and Iotonchioidea have occupied separate clades within the phylogenetic tree.
For example, the subfamilies of Neotylenchidae are in separate clades, distantly related to each other. The nonmonophyletic nature is also seen for several genera such as Deladenus, Howardula, and Rubzovinema. Serif Webplus X7. In this tree, the new species from the family Neotylenchidae has formed a clade with an unidentified nematode species,, both of which forming a well-supported clade with two species of Howardula (, ) from the family Allantonematidae. Similar to the former phylogenetic analysis by using SSU-ITS1-5.8S-LSU rDNA sequences, the nonmonophyletic nature of families such as Neotylenchidae, Allantonematidae, and Parasitylenchidae is documented. The only currently available sequence of a Gymnotylenchinae member (cf. Gymnotylenchus sp., ) in our SSU tree has placed in a distantly related clade with the new species, and likewise, concerning the uncertainty of its generic identity (the “ cf.” status) and the nonmonophyletic nature of most hexatylenchid taxa, this placement could neither confirm nor reject the placement of the genus Anguillonema under the subfamily Gymnotylenchinae. In our partial LSU tree (), and similar to SSU tree, members of superfamilies Sphaerularioidea and Iotonchioidea have occupied separate clades.
Some genera from different subfamilies (e.g. Psyllotylenchus, Spilotylenchus, Paurodontella and Rubzovinema) formed the clade A. The new species has also formed a clade with a specimen of Howardula () from Allantonematidae and two species of Parasitylenchus (, ) from the family Parasitylenchidae. In conclusion, the fragments studied in this work, 18S rDNA and LSU D2-D3, along with the partial 28S rDNA analysed by, do not infer congruent topologies with the currently available classic taxonomic frameworks for Hexatylina.
Although some genomic or nongenomic fragments remain to be tested for their usefulness in resolving of phylogenetic relations among this group of nematodes, the uncommon nature of these nematodes and paucity of their sequences in databases such as GenBank further complicate phylogenetic studies of these nematodes.