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<mods:title id="4F89E0E99428AF9A5D0D88382CCECA3B">The morphological and molecular identity of Longidoruspiceicola Liskova, Robbins &amp; Brown, 1997 from Romania (Nematoda, Dorylaimida)</mods:title>
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<mods:namePart id="509FB99144373EDEEBC27FD08F4B1993">Groza, Mariana</mods:namePart>
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<mods:namePart id="424CE52810C45A51F4CD605E1FB1232E">Lazarova, Stela</mods:namePart>
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<mods:namePart id="F66D97DDA314606CDBDF3ACF1897E283">Luca, Francesca De</mods:namePart>
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<mods:namePart id="8D803535AA0F007E44403F502351464A">Fanelli, Elena</mods:namePart>
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<mods:namePart id="567D7BDF2EAD67D4D446944548F9CDBD">Milka Elshishka,</mods:namePart>
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<mods:namePart id="A3AF0283655EA7D53DD38E393078EED7">Radoslavov, Georgi</mods:namePart>
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<mods:namePart id="149185977C75D5270BCDC8D12990C598">Hristov, Peter</mods:namePart>
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<mods:namePart id="D4A98630E7915FD1B495A0BC84EB8C47">Coman, Mihaela</mods:namePart>
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<mods:namePart id="4F5AB9D5A808245E71858EA7B91C5CB5">Peneva, Vlada</mods:namePart>
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<paragraph id="B980EEE6489C8215A71F7376009FCA7E" pageId="1" pageNumber="2">
<taxonomicName id="D3B98F31A1BF8858ADB9CFE8DE743EBE" ID-CoL="8GNRW" authority="Liskova, Robbins &amp; Brown, 1997" authorityName="Liskova, Robbins &amp; Brown" authorityYear="1997" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="piceicola">
Longidorus piceicola
<normalizedToken id="DF85754716E0838A7D3C8D86FB16C853" originalValue="Lišková">Liskova</normalizedToken>
, Robbins &amp; Brown, 1997
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<subSubSection id="55CEC47432816B5F81513807173870B2" pageId="1" pageNumber="2" type="materials_examined">
<paragraph id="FE6BD88F1E1E65D66BA34CB9F2C3E707" pageId="1" pageNumber="2">Material examined.</paragraph>
<paragraph id="A9D1947477F94A6501D636536D950136" pageId="1" pageNumber="2">
Eleven females and 21 juveniles, two females and one juvenile from Cernica forest, Ilfov County, Romania on slide numbers NE 35-37 stored at the reference collection of the National Phytosanitary Laboratory, Voluntari, Romania, 9 females and 20 juveniles - at the personal collection of the first author; nine females and 30 juveniles from Bran,
<normalizedToken id="9D17A4B6235D1D06D2A08B097C522C92" originalValue="Braşov">Brasov</normalizedToken>
County, Romania, stored in the nematode collection of IBER, Bulgaria, slide numbers N2-29/2/1-19.
</paragraph>
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<paragraph id="AEA915863E4918E2740AC6AAA8467DAF" pageId="1" pageNumber="2">Description.</paragraph>
<paragraph id="91D99CC62BFA1F9B1491A89C603B57DD" pageId="1" pageNumber="2">Figures 1-7.</paragraph>
<paragraph id="9A335F2D0E785EBFC46B8642AB7CE9CF" pageId="1" pageNumber="2">Measurements See Tables 1-3.</paragraph>
<paragraph id="6335C3B0F788B11CA2CEB37EABC39FBD" pageId="1" pageNumber="2">
Females (Figs 1A, B1-4, G2-4, 5E, 6E, J, O, 7) based on the
<taxonomicName id="500E0B46272EFA1CD676FB826190D2D6" class="Pinopsida" family="Pinaceae" genus="Larix" higherTaxonomySource="CoL" kingdom="Plantae" lsidName="Larix" order="Pinales" pageId="1" pageNumber="2" phylum="Tracheophyta" rank="genus">Larix</taxonomicName>
population, Bran,
<normalizedToken id="71DE11E0F2E68E9903F408E9EC5110F2" originalValue="Braşov">Brasov</normalizedToken>
County.
</paragraph>
<caption id="70F6B2B4EF26840F6C3D44E449B3CA97" pageId="1" pageNumber="2">
<paragraph id="B009B6741D7825DB27C4949ABCF468E2" pageId="1" pageNumber="2">
Figure 1.
<taxonomicName id="7B0F62FC88A43E693EFAE0D647A08B73" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
Female and juveniles: A Neck region - female B1-B4, C Head end with amphidial fovea B1-B3 females, B4 juvenile 4th stage (B2 right and B3 left) C, D, E1, E2, F Anterior ends of first- to fourth-stage juveniles
<normalizedToken id="7FA600D2279CBC6A867C53975B18B76D" originalValue="GK">G-K</normalizedToken>
Pharyngeal bulb of female (G) and first- to fourth-stage juveniles (
<normalizedToken id="30232390DD22ECCB7A8E4D1B8C573FF4" originalValue="HK">H-K</normalizedToken>
).
</paragraph>
</caption>
<paragraph id="58C727F80043C837CFED1A96F245A76C" pageId="1" pageNumber="2">
Habitus spiral shaped, more strongly coiled in posterior part of body. Cuticle 3-4
<normalizedToken id="BC8E3E32C41BEC27ECFBAEA9E187965A" originalValue="μm">μm</normalizedToken>
thick at guide ring region, ca 3
<normalizedToken id="47C8BF9B17A84874EF00EE0885E91B0F" originalValue="μm">μm</normalizedToken>
in mid-body, and 5-6
<normalizedToken id="463FD2CBE780D197E037AF6B2CB3B40F" originalValue="μm">μm</normalizedToken>
on tail posterior to anus. Lip region broadly rounded anteriorly, rounded laterally, almost continuous with rest of body. Amphideal fovea pocket-shaped, varying from not lobed to symmetrically bilobed at base (according to terminology proposed by
<bibRefCitation id="CF5DEC2EBE85E8C43E65482DAFAD93F8" author="Decraemer, W" journalOrPublisher="Nematology" pageId="4" pageNumber="5" pagination="643 - 62" title="Revision of some species of the genus Paralongidorussensu Siddiqi et al. (1993), with a discussion on the relationships within the family Longidoridae (Nematoda: Dorylaimida)." url="https://doi.org/10.1163/156854107782024776" volume="9" year="2007">Decraemer and Coomans 2007</bibRefCitation>
) extending to ca half the distance anterior end-guide ring. Left and right fovea of about equal size (12.7 (11-14)
<normalizedToken id="8C27F3D2A5794A6A97CC3FA614E029D4" originalValue="μm">μm</normalizedToken>
, n = 5), sensillar pouch (fusus) just posterior the guide ring, the distance from the fovea to fusus 24 (23-29
<normalizedToken id="CFF81134200B2ED2F36A120E2CFB27FA" originalValue="μm">μm</normalizedToken>
). Pharyngeal bulb occupying 25 (22-29) % of total pharynx length; dorsal nucleus located at 29.5 (27-32) % (n = 7) of bulb length; ventro-sublateral nuclei at 54 (48-57) % (n = 8) (left) and 54 (52-56.5) % (n = 8) (right); opening of the dorsal gland at 9 (7.5-11) % and opening of the ventro-sublateral glands at 84 (80.5-90.5) % of the distance from anterior end of pharyngeal bulb, respectively. In one female, a small vestigium (5
<normalizedToken id="FEEB3C33326DF4E2BFAE6860568A1FEF" originalValue="μm">μm</normalizedToken>
) observed in wall of slender pharynx. Two nerve rings observed, the first one at 207.2
<normalizedToken id="F73474F1E9B3B95A835079E8B645AB89" originalValue="±">+/-</normalizedToken>
8.8 (193-218)
<normalizedToken id="FD98A511C88D1DE2BBC5995566A9B4D2" originalValue="μm">μm</normalizedToken>
from anterior end, surrounding about mid-odontophore; the second at 329
<normalizedToken id="2A523E0302D64492B4AF241E6691696A" originalValue="±">+/-</normalizedToken>
11.6 (313-344)
<normalizedToken id="AE29C14CE013D4E163CAD2600F439D19" originalValue="μm">μm</normalizedToken>
from anterior end, n = 6, (first at 235.7
<normalizedToken id="52F5CA2BA2694FAEFD9FB1BA31928DBD" originalValue="±">+/-</normalizedToken>
12.7 (215-255) and second at 329.3
<normalizedToken id="2E55B6D13F9B3BD42A1F45FEFFEDED9F" originalValue="±">+/-</normalizedToken>
18.6 (290-343)
<normalizedToken id="4595F84B1C5A3E70039F01807B8B6B38" originalValue="μm">μm</normalizedToken>
from anterior end, n = 7, Cernica forest). Tail bluntly conical, dorsally convex, flat or shallowly concave ventrally. Two pairs of caudal pores. Reproductive system didelphic, two branches of about equal size. Vagina occupies ca 50 % of corresponding body width; pars distalis vaginae and pars proximalis vaginae 13-15
<normalizedToken id="7D1CA15A54C644250656CE3B9E0A1699" originalValue="μm">μm</normalizedToken>
and 15-19
<normalizedToken id="455C42A73569BBD304C663E072FA2E64" originalValue="μm">μm</normalizedToken>
long, respectively. Uteri short, anterior uterus 96.3
<normalizedToken id="B6EA71C256821AD3594C27240473A8BD" originalValue="±">+/-</normalizedToken>
13.5 (80-120)
<normalizedToken id="A614ED8770E32FE7E57118979D4D83CC" originalValue="μm">μm</normalizedToken>
long, posterior 91.0
<normalizedToken id="0B0C62EBFB45055D24BA57926D31E9C9" originalValue="±">+/-</normalizedToken>
10.5 (76-107)
<normalizedToken id="E88BBB6CBAA9BC0297D9508E708057AE" originalValue="μm">μm</normalizedToken>
. Uteri shorter in Cernica population - anterior uterus 80.9
<normalizedToken id="12885D8168806CB6F60BEB144CB15196" originalValue="±">+/-</normalizedToken>
7.0 (70-90)
<normalizedToken id="EB8A6CD64816C8EBA2D4F2AC13670435" originalValue="μm">μm</normalizedToken>
long and posterior 78.3
<normalizedToken id="17ACAF0500FD9C99A8A4397D3FDA054C" originalValue="±">+/-</normalizedToken>
8.3 (70-95)
<normalizedToken id="BACD53281A33BD3F682F36D809B3C0FE" originalValue="μm">μm</normalizedToken>
long. Sphincter between uterus and pars dilatata oviductus well developed. Sperm observed in both uteri of one female.
</paragraph>
<caption id="63ABE230E78443202EB7DA83C04D6401" pageId="1" pageNumber="2">
<paragraph id="79CEE805460E81D93C685523B33B9D56" pageId="1" pageNumber="2">
Figure 2.
<taxonomicName id="9ED42FB61868CEC99FDF65F5DB062CD3" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
Female from Bran locality: A, B Variations in tail shape C Vagina.
</paragraph>
</caption>
<paragraph id="FB973EDB68D4421220402BE3B063070D" pageId="1" pageNumber="2">In the population from Cernica forest two females with reserve odontostyles have been observed (Table 3).</paragraph>
<paragraph id="1226C0D39DBB343A948AE5EB44262A25" pageId="1" pageNumber="2">Male. Not found.</paragraph>
<paragraph id="E94CC5B3CE624F1DF6B710CD1372BDC1" pageId="1" pageNumber="2">
Juveniles (Figs 1
<normalizedToken id="68F15417E494BD606D62AE2C5532698E" originalValue="CF">C-F</normalizedToken>
,
<normalizedToken id="77557FE5E7C82E9AFC521F31CD51A235" originalValue="HK">H-K</normalizedToken>
; 6
<normalizedToken id="438F0266B94CF181EF0943C12CE080E0" originalValue="AD">A-D</normalizedToken>
,
<normalizedToken id="773032129DFA1BA3011DB7AC4D6FACF4" originalValue="FI">F-I</normalizedToken>
,
<normalizedToken id="5F4BA3014B48A16B3D2F26881CA9BF1D" originalValue="KN">K-N</normalizedToken>
, 7).
</paragraph>
<paragraph id="98D662DCEE4AF55FC2262D892766E15F" pageId="1" pageNumber="2">
General morphology similar to adult females. Body habitus similar in all stages, open C- to J-shaped. Tail of all juvenile stages conical, but becoming more rounded and
<normalizedToken id="6E3F37D0B30A7821193ACFE7E8B779C8" originalValue="c">c'</normalizedToken>
decreasing in subsequent stages: tail of first stage juvenile elongate conoid with slightly digitate terminus, in the second stage - elongate conoid, in third - bluntly conoid, variable, with narrow to widely rounded terminus, in fourth - resembling that of female, bluntly conoid (Fig. 5). In several juveniles, the abnormalities in their development did not allow to assign them to a particular stage and the morphometrics are presented separately (Table 3). The lengths of functional and replacement odontostyles used to infer the developmental stages were in contradiction with other measurements such as L, a, b, c etc. which were in correspondence with a different stage, or the functional odontostyle was in the ranges of one stage while the replacement one was not in the ranges of the next stage; in one occasion the length of replacement odontostyle was less than that of the replacement one (Table 3).
</paragraph>
<caption id="F7AC5118EB5F334164D154203E0438F5" pageId="1" pageNumber="2">
<paragraph id="3B80FE5C95A20F815576F4A3CE6F5049" pageId="1" pageNumber="2">
Figure 3.
<taxonomicName id="13B4D672F2481D66498447B1E534BFE8" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
Female from Bran locality: A Neck region, black arrows indicate nerve rings B, C Head end (different magnifications) D, E Amphideal fovea (right and left) F Odontophore G Prerectum H Pharyngeal bulb I, J Variations in tail shape. Scale bars: A, G 80
<normalizedToken id="71501AC43D97593F2E5EEDCC011C869F" originalValue="μm">μm</normalizedToken>
; B, F, H, I, J 40
<normalizedToken id="DB8F4CA63BB4234243E0EE761C756832" originalValue="μm">μm</normalizedToken>
;
<normalizedToken id="775586E22E35DED65C42378DD9DBA3FC" originalValue="CE">C-E</normalizedToken>
20
<normalizedToken id="4E4C4B5C0BF91DA00F607524F52A1559" originalValue="μm">μm</normalizedToken>
.
</paragraph>
</caption>
<caption id="86B9CF8921592C232701B374C4C73039" pageId="1" pageNumber="2">
<paragraph id="E575A202FAE886D12B87444E7188743C" pageId="1" pageNumber="2">
Figure 4.
<taxonomicName id="97E708CF315354570852F6EE418EEA11" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
Female from Bran locality: A Anterior genital branch B Uterus part with sperm C Ovary D Nerve ring E Posterior genital branch F Lateral field and epidermal glans
<normalizedToken id="C73C9DA798D8DF540E562701E26B7427" originalValue="GI">G-I</normalizedToken>
Variations in vagina (different magnifications). Scale bars: A, E 80
<normalizedToken id="09396DE845082A0069A83B5A7E5907F8" originalValue="μm">μm</normalizedToken>
;
<normalizedToken id="C312CBA6F10A8474003A09EC2FF1DDFA" originalValue="BD">B-D</normalizedToken>
,
<normalizedToken id="F00AA175BF1523D5D0059083C3FD7271" originalValue="FH">F-H</normalizedToken>
40
<normalizedToken id="72CBFA8A5B08DD4E219211DA8BABFEE4" originalValue="μm">μm</normalizedToken>
; I 20
<normalizedToken id="9FA5133561E02CECC73577E58721DDE0" originalValue="μm">μm</normalizedToken>
.
</paragraph>
</caption>
<caption id="5A8AC22FA1C2D37FB1847973AF0D92D9" ID-Table-UUID="5A8AC22FA1C2D37FB1847973AF0D92D9" httpUri="http://table.plazi.org/id/5A8AC22FA1C2D37FB1847973AF0D92D9" pageId="1" pageNumber="2">
<paragraph id="412332CA4BD2CC2FF357D435DEC1B0B1" pageId="1" pageNumber="2">
Table 1. Measurements of females and juveniles (J) of
<taxonomicName id="33CA8B0BECD81E97154673F35FE10FC9" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
from Bran,
<normalizedToken id="0F10B537D1B0E298E9141A7FA51BB56B" originalValue="Braşov">Brasov</normalizedToken>
County, Romania (mean
<normalizedToken id="516DA105AB23463516ECCB7C608D5917" originalValue="±">+/-</normalizedToken>
standard deviation, with range). All measurements in micrometers except for body length (mm).
</paragraph>
</caption>
<paragraph id="68CB0DDD75DB6B62F3BC71D46B9BA967" pageId="1" pageNumber="2">
<table id="FBE53D79E5468A5D52D1B14166845ECE" pageId="1" pageNumber="2">
<tr id="47778C828075EFDFB88AE43D6A4B473A" pageId="1" pageNumber="2">
<th id="D9F18D1D94772A6DA806BD1248B900FC" colspan="1" pageId="1" pageNumber="2" rowspan="1">Character</th>
<th id="C457C73B6E5604103BAB6367900AD970" colspan="1" pageId="1" pageNumber="2" rowspan="1">Females</th>
<th id="BC2B28DB9AFB1011DA7E94A9D09FF6A0" colspan="1" pageId="1" pageNumber="2" rowspan="1">J1</th>
<th id="4CE415E3F1C14A6913A6C44D7D23CB72" colspan="1" pageId="1" pageNumber="2" rowspan="1">J2</th>
<th id="5E72987FF865D8BF9E6836819E25D60F" colspan="1" pageId="1" pageNumber="2" rowspan="1">J3</th>
<th id="9CFD9E5E788C42839FEF6038E3BA15FE" colspan="1" pageId="1" pageNumber="2" rowspan="1">J4</th>
</tr>
</table>
</paragraph>
<caption id="C6506DDEC2FC09D95483100B66684C8C" ID-Table-UUID="C6506DDEC2FC09D95483100B66684C8C" httpUri="http://table.plazi.org/id/C6506DDEC2FC09D95483100B66684C8C" pageId="1" pageNumber="2">
<paragraph id="9AF243D3AFC131392FD9ADA49E216AD3" pageId="1" pageNumber="2">
Table 2. Measurements of females and juvenile stages (J) of
<taxonomicName id="8A34341A5EA7F062F01EF40880198E8F" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
from Cernica-Ilfov County, Romania (mean
<normalizedToken id="5D1304E92D78C3E9989C491B454094B1" originalValue="±">+/-</normalizedToken>
standard deviation, with range). All measurements in micrometers except body length (mm).
</paragraph>
</caption>
<paragraph id="C96EE40013AA1B154FC2A35519CD8038" pageId="1" pageNumber="2">
<table id="5ECF43D134B5D59191DEC261358ABA20" pageId="1" pageNumber="2">
<tr id="AE61026C112FC25FFBAC02561928BA59" pageId="1" pageNumber="2">
<th id="0B55E8CB507479656D2C480A406EC00F" colspan="1" pageId="1" pageNumber="2" rowspan="1">Character</th>
<th id="15CAFF92C6BA4EF32617E155EE6C525B" colspan="1" pageId="1" pageNumber="2" rowspan="1">Females</th>
<th id="6444F0F9FD514600549B30EBF6C39CA9" colspan="1" pageId="1" pageNumber="2" rowspan="1">J1</th>
<th id="98A702F7D1BD6DBDBB903CB3E421E8F2" colspan="1" pageId="1" pageNumber="2" rowspan="1">J2</th>
<th id="DAB831FE916656ADA9F3224D1F15F9CB" colspan="1" pageId="1" pageNumber="2" rowspan="1">J3</th>
<th id="2ADCA795ADD53DDE0F2C0001B9EC3B12" colspan="1" pageId="1" pageNumber="2" rowspan="1">J4</th>
</tr>
</table>
</paragraph>
<caption id="146680D064C1206668B60C6B7B7B1757" ID-Table-UUID="146680D064C1206668B60C6B7B7B1757" httpUri="http://table.plazi.org/id/146680D064C1206668B60C6B7B7B1757" pageId="1" pageNumber="2">
<paragraph id="26194DD53271BF4C651233112A94BA3D" pageId="1" pageNumber="2">
Table 3. Measurements of
<taxonomicName id="5A2F1ED4383183B0DAF25CC4BC77DF92" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
females (f) from Cernica, and juveniles (j) from Bran,
<normalizedToken id="CF98430C560A7181367177E3248050A3" originalValue="Braşov">Brasov</normalizedToken>
County, Romania showing different anomalies. All measurements in micrometers except body length (mm).
</paragraph>
</caption>
<paragraph id="BFD5E693D181692B73AFAC8DD6F64521" pageId="1" pageNumber="2">
<table id="FA6E5C3FCE6A184F2EEB14D54778582B" pageId="1" pageNumber="2">
<tr id="28F473D5815855B94FB5049D5850269B" pageId="1" pageNumber="2">
<th id="ACFE41CC17D79630D2A8161D4D30F4C1" colspan="1" pageId="1" pageNumber="2" rowspan="1">Character</th>
<th id="348CC92D23C95BC9F0C886EABD642F47" colspan="1" pageId="1" pageNumber="2" rowspan="1">f</th>
<th id="2BDAF0E6EA7846AC5E6668946BC0FCAB" colspan="1" pageId="1" pageNumber="2" rowspan="1">f</th>
<th id="1AB5C3E552224B68C04627E6C66788D4" colspan="1" pageId="1" pageNumber="2" rowspan="1">j</th>
<th id="4BF7D72C7E704D25F3A1DC5169E28BBD" colspan="1" pageId="1" pageNumber="2" rowspan="1">j</th>
<th id="843F9AC6AC6EECFD89DD7AAFD03705BC" colspan="1" pageId="1" pageNumber="2" rowspan="1">j</th>
<th id="20F716A436FD1F79F1128C753502E270" colspan="1" pageId="1" pageNumber="2" rowspan="1">j</th>
<th id="3897D45EECD8592E02544C41F7967353" colspan="1" pageId="1" pageNumber="2" rowspan="1">j</th>
<th id="9529F6D121116387EEB748CE0422F2F2" colspan="1" pageId="1" pageNumber="2" rowspan="1">j</th>
<th id="A21EC0AB1F192AA6C424EF510CD781C6" colspan="1" pageId="1" pageNumber="2" rowspan="1">j</th>
<th id="A6550AD2EB46157302E69E349C134DB0" colspan="1" pageId="1" pageNumber="2" rowspan="1">j</th>
</tr>
</table>
</paragraph>
</subSubSection>
<subSubSection id="E3776BB778E8B8CFFD0421354893D51F" lastPageId="2" lastPageNumber="3" pageId="1" pageNumber="2" type="sequences and phylogenetic analyses">
<paragraph id="738E1084BED0F6E4F85DB7EF94A276D3" pageId="1" pageNumber="2">Sequences and phylogenetic analyses.</paragraph>
<paragraph id="FDDF893A5370DB9D4C8107C14157BF47" pageId="1" pageNumber="2">
The amplification of the ITS and the D2-D3 expansion domains of the 28S rRNA gene yielded fragments of 1646 and 756 bps, respectively, based on sequencing. The ITS sequences of
<taxonomicName id="9C0B08A5796CC747F5589A1A1ECC24C2" lsidName="L. piceicola" pageId="1" pageNumber="2" rank="species" species="piceicola">L. piceicola</taxonomicName>
from Romania were obtained for the first time in the present study. They showed 98 % similarity (962/984 identities, 9 gaps) when compared with the corresponding sequence of
<taxonomicName id="789A85C9603D171D74FF36F123370008" lsidName="L. intermedius" pageId="1" pageNumber="2" rank="species" species="intermedius">L. intermedius</taxonomicName>
(KT308890) and 86 % with the ITS sequence of
<taxonomicName id="9624C904AE28C485045E567F68B6F9D1" lsidName="L. elongatus" pageId="1" pageNumber="2" rank="species" species="elongatus">L. elongatus</taxonomicName>
Hooper, 1961 (AJ549986, AJ549987). Intraspecific variation for the ITS sequences was low, with only two nucleotides difference and no indels.
</paragraph>
<caption id="100A79F1DD20A593DA6D54B2B76014BA" pageId="1" pageNumber="2">
<paragraph id="A9571BF46ED4FC6B81995A5D4DFCE604" pageId="1" pageNumber="2">
Figure 5.
<taxonomicName id="A3BAC8A40F9A4DEB75D8FEE6EFB0316B" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
Juveniles and female from Bran locality: Variations in tail shape of first (A1-A3), second (B1-B3), third (C1-C4), fourth (D1-D3) juvenile stages and female (E).
</paragraph>
</caption>
<paragraph id="8441B9B765449D0BA7A4823AD964498C" lastPageId="2" lastPageNumber="3" pageId="1" pageNumber="2">
D2-D3 rDNA sequences obtained from both Romanian populations were identical to each other and to the sequence of
<taxonomicName id="596E06C1ABB2B3C280521BDB22A4155E" lsidName="L. piceicola" pageId="1" pageNumber="2" rank="species" species="piceicola">L. piceicola</taxonomicName>
from Slovakia (AY601577,
<bibRefCitation id="C86EA02856F557FBAA6BAFAE796D468C" author="He, Y" journalOrPublisher="Nematology" pageId="5" pageNumber="6" pagination="111 - 124" title="A molecular phylogenetic approach to Longidoridae (Nematoda: Dorylaimida)." url="https://doi.org/10.1163/1568541054192108" volume="7" year="2005">He et al. 2005</bibRefCitation>
). The phylogenetic relationships of
<taxonomicName id="6D3BF0FE0FD509B1D3B07070A02F03B3" lsidName="L. piceicola" pageId="1" pageNumber="2" rank="species" species="piceicola">L. piceicola</taxonomicName>
with several related species is presented in Figure 8.
<taxonomicName id="C777A64CC236F85BC7B2407E83B2E43C" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus intermedius" order="Dorylaimida" pageId="1" pageNumber="2" phylum="Nematoda" rank="species" species="intermedius">Longidorus intermedius</taxonomicName>
revealed sister relationships with
<taxonomicName id="284960C73CE878225854D78E22D3382D" lsidName="L. piceicola" pageId="1" pageNumber="2" rank="species" species="piceicola">L. piceicola</taxonomicName>
and the sequences from both species formed a well-supported clade. In addition, five sequences of
<taxonomicName id="E3FA383EDBB401B4DF1E7A340CFCB45B" lsidName="L. intermedius" pageId="1" pageNumber="2" rank="species" species="intermedius">L. intermedius</taxonomicName>
from Germany (AF480074,
<bibRefCitation id="330AAAE344688DBE0A703324B2D6E085" author="Rubtsova, TV" journalOrPublisher="Russian Journal of Nematology" pageId="6" pageNumber="7" pagination="127 - 136" title="Description of Longidorussturhani sp. n. (Nematoda: Longidoridae) and molecular characterisation of several longidorid species from Western Europe." volume="9" year="2001">Rubtsova et al. 2001</bibRefCitation>
), Russia (KF242311 and KF242312,
<bibRefCitation id="F11507F889451AAB622E0EE6C6111DFD" author="Subbotin, SA" journalOrPublisher="European Journal of Plant Pathology" pageId="6" pageNumber="7" pagination="377 - 390" title="Molecular characterisation and diagnostics of some Longidorus species (Nematoda: Longidoridae) from Russia and other countries using rRNA genes." url="https://doi.org/10.1007/s10658-013-0338-9" volume="138" year="2014">Subbotin et al. 2014</bibRefCitation>
), Spain (KT308868,
<bibRefCitation id="03FA220AB0C65DE5C824E53F168B3DEB" author="Gutierrez Gutierrez, C" journalOrPublisher="Zoological Journal of the Linnean Society" pageId="5" pageNumber="6" pagination="473 - 500" title="Molecular phylogeny of the nematode genus Longidorus (Nematoda: Longidoridae) with description of three new species." url="https://doi.org/10.1111/zoj.12019" volume="167" year="2013">
<normalizedToken id="C5D495AC3DB1F58B829B4B6BF50EF0D8" originalValue="Gutiérrez-Gutiérrez">Gutierrez-Gutierrez</normalizedToken>
et al 2013
</bibRefCitation>
and JX445117,
<bibRefCitation id="DDF6BFDC0C31260E9B29491395EB1B2D" author="Archidona-Yuste, A" journalOrPublisher="PLoS ONE" pageId="4" pageNumber="5" title="Unraveling the biodiversity and molecular phylogeny of needle nematodes of the genus Longidorus (Nematoda: Longidoridae) in olive and a description of six new species" url="https://doi.org/10.1371/journal.pone.0147689" volume="11" year="2016">Archidona-Yuste et al. 2016</bibRefCitation>
), and the
<taxonomicName id="886E576EE0D870168EFACDF84ED571EC" lsidName="L. piceicola" pageId="1" pageNumber="2" rank="species" species="piceicola">L. piceicola</taxonomicName>
<pageBreakToken id="C07BE710702E1B646FC8E1CAFA2B924D" pageId="2" pageNumber="3" start="start">sequence</pageBreakToken>
were realigned separately and pairwise distances estimated. A total of 737 positions was included in the dataset. The between species dissimilarities (p-distances) were 0.3-0.9 % (or 2-6 bp differences). Similarly, the intraspecific p-distances of
<taxonomicName id="9FC8F7417B793FC39161DFB88EF45959" lsidName="L. intermedius" pageId="2" pageNumber="3" rank="species" species="intermedius">L. intermedius</taxonomicName>
from the three European countries were 0.4-0.9 % (i.e. 3-6 bp).
</paragraph>
<caption id="CD94DF4CCC7DBDECC62D9673CA19E345" pageId="2" pageNumber="3">
<paragraph id="CBAFC33B22411D6A93047D2CEAAD3156" pageId="2" pageNumber="3">
Figure 6.
<taxonomicName id="C27BFCF9B94B3503CC76B783FA215CED" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="2" pageNumber="3" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
Juveniles and female from Bran locality:
<normalizedToken id="5419B7C1910D7A9D5ECB13A9C11ECF71" originalValue="AE">A-E</normalizedToken>
Anterior ends of first- to fourth-stage juveniles and female
<normalizedToken id="B16FFA5B4417D4DDF267751A5F7DDAAE" originalValue="FJ">F-J</normalizedToken>
Tails of first to fourth juvenile stages and female
<normalizedToken id="A516DE3E6CE14F34129258FAC78A6C50" originalValue="KM">K-M</normalizedToken>
Genital primordium of first to fourth juvenile stages. O Vagina. Scale bar: 20
<normalizedToken id="3AAA4B41E97A3D36CC78C66B1A54D197" originalValue="μm">μm</normalizedToken>
.
</paragraph>
</caption>
<paragraph id="50DF85F1EF08F4D2932971DE65A6CD0E" pageId="2" pageNumber="3">
The SNPs analysis comparing all D2-D3 sequences of
<taxonomicName id="F9CB9D7C99F8CC636EF4583BDF73EC65" lsidName="L. piceicola" pageId="2" pageNumber="3" rank="species" species="piceicola">L. piceicola</taxonomicName>
and
<taxonomicName id="20717FF725CC651ED13E6C5D35F1E895" lsidName="L. intermedius" pageId="2" pageNumber="3" rank="species" species="intermedius">L. intermedius</taxonomicName>
revealed three parsimony-informative sites (i.e. nucleotide positions with transitions 89T/C, 134T/C and 297A/G) when compared to the reference sequence of
<taxonomicName id="F1F9AD9CC1E16E9D870D3421A7B4E194" lsidName="L. piceicola" pageId="2" pageNumber="3" rank="species" species="piceicola">L. piceicola</taxonomicName>
(AY601577) (Table 4). The most similar sequence to the
<taxonomicName id="4BC1BE5946AE35A71AACE4715D754B65" lsidName="L. piceicola" pageId="2" pageNumber="3" rank="species" species="piceicola">L. piceicola</taxonomicName>
sequence was that of
<taxonomicName id="C731627D9845CCCDCDE5A1BA4137F6C8" lsidName="L. intermedius" pageId="2" pageNumber="3" rank="species" species="intermedius">L. intermedius</taxonomicName>
from Germany, revealing the highest similarity and only two interspecies differentiating nucleotides at positions 89 and 134 compared to the reference sequence (Table 4).
</paragraph>
<caption id="195CE27BE14912802B0C3E8C5B0DAD8B" pageId="2" pageNumber="3">
<paragraph id="027A3C6B751303E4D1FD9EF6D23C0689" pageId="2" pageNumber="3">
Figure 7. Scatter plot of odontostyle (■) and replacement odontostyle (□) against body length of
<taxonomicName id="CBCDBC4C17A16341DBE7721C56BF5084" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="2" pageNumber="3" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
juveniles (J1 to J4) and females from A Cernica forest, Ilfov county and B Bran locality,
<normalizedToken id="FE1AFD842EC08937BB6422623B53013B" originalValue="Braşov">Brasov</normalizedToken>
county.
</paragraph>
</caption>
<caption id="AC58A6A36722922170C02B3867041FFD" pageId="2" pageNumber="3">
<paragraph id="30BE3C2E452561F583432BCE9DCAC1B4" pageId="2" pageNumber="3">Figure 8. Phylogenetic tree using D2-D3 28S rDNA and inferred from a Bayesian analysis with GTR+G model and midpoint rooting. Posterior probabilities ≥ than 0.8 are presented.</paragraph>
</caption>
<caption id="CE6D8D89A277290EFB5CDF45C8D0948A" pageId="2" pageNumber="3">
<paragraph id="5CFAC67C15634142F02BE382042345DF" pageId="2" pageNumber="3">
Table 4. The variable positions in D2-D3 28S rDNA control region sequences of
<taxonomicName id="780BE3517178C42E6ED900538F63670C" class="Enoplea" family="Longidoridae" genus="Longidorus" higherTaxonomySource="CoL" kingdom="Animalia" lsidName="Longidorus piceicola" order="Dorylaimida" pageId="2" pageNumber="3" phylum="Nematoda" rank="species" species="piceicola">Longidorus piceicola</taxonomicName>
and
<taxonomicName id="83FD4599045CC8B58574BE26E3B5CE69" lsidName="L. intermedius." pageId="2" pageNumber="3" rank="species" species="intermedius.">L. intermedius.</taxonomicName>
The
<taxonomicName id="C4D00B440F406806B3633193331D602D" lsidName="L. piceicola" pageId="2" pageNumber="3" rank="species" species="piceicola">L. piceicola</taxonomicName>
sequence from Slovakia (Acc. no AY601577) was used as a reference.
</paragraph>
</caption>
<paragraph id="1BA399FFE6C306F8A59A31D71DC6C9FF" pageId="2" pageNumber="3">
<table id="A73B9CA3187670DA1DE11C9F6FDACAB2" pageId="2" pageNumber="3">
<tr id="C893DD84D7E5F76FBCB4680FAE234D40" pageId="2" pageNumber="3">
<th id="91035278642301C0B5D209B4F0824E90" colspan="12" pageId="2" pageNumber="3" rowspan="1">SNPositions</th>
</tr>
<tr id="92C8A87B1D05F0D1E60F4E2B238CC93E" pageId="2" pageNumber="3">
<th id="4B7EFD87D62BF69F2319E8006CD7B777" colspan="1" pageId="2" pageNumber="3" rowspan="1">89</th>
<th id="E67F3C602F6175D99D05C9FC96A3FEB4" colspan="1" pageId="2" pageNumber="3" rowspan="1">129</th>
<th id="86A90BD42C220C06461879396381F7D2" colspan="1" pageId="2" pageNumber="3" rowspan="1">134</th>
<th id="CA664BE2E597A3C2F94E459548F15FC5" colspan="1" pageId="2" pageNumber="3" rowspan="1">197</th>
<th id="A2F761BB001CE02A98ADC98C5A0A21F2" colspan="1" pageId="2" pageNumber="3" rowspan="1">255</th>
<th id="D806D3B0CD4E72689C5652756B29AD2F" colspan="1" pageId="2" pageNumber="3" rowspan="1">285+1gap</th>
<th id="D07BF60243902A61CB5A1223B79EC464" colspan="1" pageId="2" pageNumber="3" rowspan="1">285+2gap</th>
<th id="7994A9953426D38D17563AEB925FFBCF" colspan="1" pageId="2" pageNumber="3" rowspan="1">297</th>
<th id="65416D8368C52EEAD3D2670CEAC153F2" colspan="1" pageId="2" pageNumber="3" rowspan="1">310</th>
<th id="78C6449E91AC9B1CC262EC79AE33A31F" colspan="1" pageId="2" pageNumber="3" rowspan="1">413</th>
<th id="C49D2FC82D453BAAAC8DFE124FFFBEA9" colspan="1" pageId="2" pageNumber="3" rowspan="1">514</th>
<th id="5EA13E3BD246B065847ABB31000A94BC" colspan="1" pageId="2" pageNumber="3" rowspan="1">584</th>
</tr>
<tr id="577B74AE8554ADEF455A8EA859A0A7D5" pageId="2" pageNumber="3">
<td id="244F0C60032E08660C9F7E1C933C9CE5" colspan="1" pageId="2" pageNumber="3" rowspan="1">AY601577 reference sequence</td>
<td id="15B4CB08285B013524F04AC67858CD0A" colspan="1" pageId="2" pageNumber="3" rowspan="1">T</td>
<td id="B4EC74A7DF5CE908FBE4849AB6440C00" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="206098413E5F66E43B80F0CC44FA1420" colspan="1" pageId="2" pageNumber="3" rowspan="1">T</td>
<td id="C05F3875BD7CF564807CE4C3F4F94919" colspan="1" pageId="2" pageNumber="3" rowspan="1">A</td>
<td id="ED2610FEAE706716664876646B29C239" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="5F257A165EE08303E1D5A2CC13B9CD67" colspan="1" pageId="2" pageNumber="3" rowspan="1">A</td>
<td id="828F2ABE8E1E0A552425CC65D50B5BBF" colspan="1" pageId="2" pageNumber="3" rowspan="1">T</td>
<td id="07275AE71F71B8227D9F31668B9E08CB" colspan="1" pageId="2" pageNumber="3" rowspan="1">G</td>
<td id="4559068031E70C5FFB233BB297FDAD6A" colspan="1" pageId="2" pageNumber="3" rowspan="1">G</td>
<td id="518C526F06A1F2F2B5E7CB2007B1AF54" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
</tr>
<tr id="D6C654615EAE97EA0F13DB21223F7E42" pageId="2" pageNumber="3">
<td id="1187BE89395EF9DC502524CD631AEFBF" colspan="1" pageId="2" pageNumber="3" rowspan="1">
AY601577
<taxonomicName id="83F130401C270B6169199E5829AC6A9B" lsidName="L. piceicola" pageId="2" pageNumber="3" rank="species" species="piceicola">L. piceicola</taxonomicName>
</td>
</tr>
<tr id="214EDB3BFB472F3902E7EBD6915904B1" pageId="2" pageNumber="3">
<td id="541797060BDC01EA13073720CC9C72CF" colspan="1" pageId="2" pageNumber="3" rowspan="1">
KY086070
<taxonomicName id="7531F64BB738A3E21D15498C08DF44B3" lsidName="L. piceicola" pageId="2" pageNumber="3" rank="species" species="piceicola">L. piceicola</taxonomicName>
</td>
</tr>
<tr id="C0B00BFA533559875B54C037B4FCEC4E" pageId="2" pageNumber="3">
<td id="23013130678E53A1BC5D19114FF31C19" colspan="1" pageId="2" pageNumber="3" rowspan="1">
LT669801
<taxonomicName id="CA24EDA444062A27A75C0DA3E2A1C5D2" lsidName="L. piceicola" pageId="2" pageNumber="3" rank="species" species="piceicola">L. piceicola</taxonomicName>
</td>
</tr>
<tr id="A6A00640EBA4A75206A35E100DCADC6A" pageId="2" pageNumber="3">
<td id="CF272D7B5D918C07AE3E0DB2D6B66552" colspan="1" pageId="2" pageNumber="3" rowspan="1">
AF480074
<taxonomicName id="0086633177E3DC7A34F9BDA188DA1C77" lsidName="L. intermedius" pageId="2" pageNumber="3" rank="species" species="intermedius">L. intermedius</taxonomicName>
</td>
<td id="848B484A7B9FF9DFD1B8DF728F56440C" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="91937C5A06BBC45F7A2DC1F39FC8F0AD" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
</tr>
<tr id="E1B141DC4CF5A3BA8ABCB91B90DFFBC9" pageId="2" pageNumber="3">
<td id="159FA2F453ADA99E8D4D120104324027" colspan="1" pageId="2" pageNumber="3" rowspan="1">
JX445117
<taxonomicName id="CF0FE1E962E41B4F9C497E5F293602B7" lsidName="L. intermedius" pageId="2" pageNumber="3" rank="species" species="intermedius">L. intermedius</taxonomicName>
</td>
<td id="2413394B2BC4896A0AA231F1F585E639" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="8152F1ACDBBC80D1CA12D6797CF5906F" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="14410845F10CF01A4CE4C25C615F1555" colspan="1" pageId="2" pageNumber="3" rowspan="1">G</td>
</tr>
<tr id="21FEA195D0BCF386BB499ECE28760BE8" pageId="2" pageNumber="3">
<td id="E2FBEFB31C6060095954A1D83B9CFB00" colspan="1" pageId="2" pageNumber="3" rowspan="1">
KT308868
<taxonomicName id="D61466A190664C03CB21B081855F9AF7" lsidName="L. intermedius" pageId="2" pageNumber="3" rank="species" species="intermedius">L. intermedius</taxonomicName>
</td>
<td id="D1E9254FA1860CD444588595447FBF66" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="2A750BCF2B4617DD85AADE3A05AC3EFB" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="B75DAC3FA0540E8B8556A36832154320" colspan="1" pageId="2" pageNumber="3" rowspan="1">G</td>
</tr>
<tr id="5448F130EE3484BE5EF95D5174F0AA23" pageId="2" pageNumber="3">
<td id="68819A6CB80E90B7E87A05352D51616B" colspan="1" pageId="2" pageNumber="3" rowspan="1">
KF242312
<taxonomicName id="F836AC77B52220F6584AA6AA6FC1F849" lsidName="L. intermedius" pageId="2" pageNumber="3" rank="species" species="intermedius">L. intermedius</taxonomicName>
</td>
<td id="40B444C36BE4E66BE6A0693AD8929C67" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="0FF778573366ADC55C4672590A6ED218" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="B6BBE774A617AFC2A0A8FA4E3DC4CAD1" colspan="1" pageId="2" pageNumber="3" rowspan="1">G</td>
</tr>
<tr id="F551B77BC65874E03174318F783245B6" pageId="2" pageNumber="3">
<td id="810563B4A6491AB4D60776D8445D9946" colspan="1" pageId="2" pageNumber="3" rowspan="1">
KF242311
<taxonomicName id="137AA7E48663B6DEE7CCB79057B1D744" lsidName="L. intermedius" pageId="2" pageNumber="3" rank="species" species="intermedius">L. intermedius</taxonomicName>
</td>
<td id="86C8D48C3C13A4A9F6840C3420AB0ECA" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="2FDE608E878CCF335031269DBAF3C290" colspan="1" pageId="2" pageNumber="3" rowspan="1">C</td>
<td id="4530B1B0771D42B46F85C56C380D5490" colspan="1" pageId="2" pageNumber="3" rowspan="1">G</td>
</tr>
</table>
</paragraph>
</subSubSection>
</treatment>
</document>