Table 1.
Information of the species/populations of Hemicycliophora studied in present paper and those of ingroup and outgroup taxa used in phylogenetic analyses.
| GenBank accession numbers | |||||
|---|---|---|---|---|---|
| Species | Host | Locality | D2-D3 of LSU rDNA | ITS | Reference or identifier |
| H. ahvasiensis n. sp. | Phoenix dactylifera | Khuzestan province, Iran | MT901580, MT901581 | MT901582, MT901583 | Present study |
| H. californica | Salix sp. | Yolo County, CA, USA | KF430518, KF430519 | KF430576 | Subbotin et al. (2014) |
| H. conida | Punica granatum | Gilan province, Iran | – | MT901584 | Present study |
| H. conida | Unknown plant | Belgium | FN433875 | – | I. Tandingan De Ley et al. (unpub.) |
| H. conida | Turf grasses | Football pitch, Madrid, Spain | KF430447 | KF430580 | P. Castillo; Subbotin et al. (2014) |
| H. conida | Unknown plant | Clallam County, WA, USA | KF430448 | KF430579 | Subbotin et al. (2014) |
| H. dulli | Peat | South Africa | MT329669, MT329670 | MT329671, MT329672 | M. Rashidifard (unpub.) |
| H. epicharoides | Ammophila arenaria | Serranova, Brindisi, Italy | KF430512 | – | Subbotin et al. (2014) |
| H. epicharoides | Ammophila arenaria | S. Barrameda, Cádiz, Spain | – | KF430608 | Subbotin et al. (2014) |
| H. epicharoides | Pragmites sp. | Epiros, Greece | – | KF430606 | Subbotin et al. (2014) |
| H. floridensis | Pinus elliotti | Lake City, FL, USA | KF430506 | KF430536 | Subbotin et al. (2014) |
| H. gracilis | Prunus domestica | Hamilton City, Glenn County, CA, USA | KF430480 | KF430562 | Subbotin et al. (2014) |
| H. gracilis | Prunus domestica | Butte City, Glenn County, CA, USA | KF430481 | – | Subbotin et al. (2014) |
| H. gracilis | Unknown plant | Brooklyn Park, MN, USA | KF430482 | – | Subbotin et al. (2014) |
| H. gracilis | Unknown plant | California, USA | – | FN435301 | I. Tandingan De Ley et al. (unpub.) |
| H. gracilis | Unknown plant | Sacramento County, CA, USA | – | MG019827 | Van den Berg et al. (2018) |
| H. halophila | Desmoschoenus spiralis | Taylors Mistake, New Zealand | KF430444, KF430445 | KF430582, KF430583 | Subbotin et al. (2014) |
| H. hellenica | Arundo donax | Filippias, Epirus, Greece | KF430453 | KF430584 | Subbotin et al. (2014) |
| H. iberica | Populus nigra | Arroyo Frío, Jaén, Spain | KF430461 | KF430539, KF430540 | Subbotin et al. (2014) |
| H. iberica | Quercus suber | Hinojos, Huelva, Spain | KF430462 | – | Subbotin et al. (2014) |
| H. iberica | Quercus suber | Santa Elena, Jaén, Spain | KF430463 | KF430541 | Subbotin et al. (2014) |
| H. italiae | Ammophila arenaria | Zapponeta, Foggia, Italy | KF430458 | – | Subbotin et al. (2014) |
| H. labiata | Poa pratensis | South Korea | MK305971, MK305972 | MK305973, MK305974 | Mwamula et al. (2020) |
| H. lutosa | Unknown plant | Gauteng province, South Africa | GQ406240, GQ406241 | GQ406237 | Van den Berg et al. (2010) |
| H. lutosoides | Turf grasses | Madrid, Spain | KF430454 | – | Subbotin et al. (2014) |
| H. lutosoides | Juncus sp. | Cádiz, Spain | – | KF430537, KF430538 | Subbotin et al. (2014) |
| H. obtusa | Pinus pinea | Moguer, Huelva, Spain | KF430521 | KF430578 | Subbotin et al. (2014) |
| H. onubensis1 | Pinus pinea | Moguer, Huelva, Spain | KF430449, KF430450 | KF430587, KF430588 | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. parvana2 | Turf grasses | New Hanover County, NC, USA | KF430501 | – | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. parvana2 | Turf grasses | Carteret County, NC, USA | KF430502 | – | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. parvana2 | Bentgrass | Texas, USA | KC329574 | KC329575 | Ma and Agudelo (2015) |
| H. parvana | Prunus persica | Punta Gorda, FL, USA | MG019825 | – | Van den Berg et al. (2018) |
| H. parvana2 | Andropogon virginicus | Paines Praire, FL, USA | – | KF430524, KF430526 | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. parvana2 | Turf grasses | New Hanover County, NC, USA | – | KF430528 | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. poranga | Poa annua | Monterey County, CA, USA | KF430432, KF430434 | KF430598 | Subbotin et al. (2014) |
| H. poranga | Turf grasses | San Francisco, CA, USA | MG019815 | – | Van den Berg et al. (2018) |
| H. poranga | Unknown plants | Marin County, CA, USA | MG019816 | – | Van den Berg et al. (2018) |
| H. poranga | Salix sp. | Santa Rosa, CA, USA | – | KF430590 | Subbotin et al. (2014) |
| H. poranga | Apium graveolens | Argentina | – | KF430596 | Subbotin et al. (2014) |
| H. poranga | Lepidorrhachis mooreana | San Francisco, CA, USA | – | KF430600 | Subbotin et al. (2014) |
| H. raskii | Grasses | Sacramento County, CA, USA | KF430520 | KF430577 | Subbotin et al. (2014) |
| H. robbinsi3 | Turf grasses | Brunswick, NC, USA | KF430488, KF430492 | – | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. robbinsi3 | Turf grasses | Indian Hills, CA, USA | KF430491 | – | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. robbinsi3 | Turf grasses | San Antonio, TX, USA | – | KF430544 | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. robbinsi3 | Borrichia sp. | St Augustine, FL, USA | – | KF430550 | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. robbinsi3 | Phoenix roebelenii | Fort Lauderdale, FL, USA | – | KF430552 | Subbotin et al. (2014); Van den Berg et al. (2018) |
| H. signata | Grasses | Chemba District, Mozambique | MG019824 | – | Van den Berg et al. (2018) |
| H. similis | Fragaria x ananassa | Cartaya, Huelva, Spain | KF430465 | – | Subbotin et al. (2014) |
| H. subbotini | Cinnamomum camphora | Zhejiang Province, China | MG701275–MG701277 | MG701272, MG701273 | Maria et al. (2018) |
| H. thienemanni | Salix sp. | Moscow, Russia | KF430469–KF430471 | KF430570–KF430572 | Subbotin et al. (2014) |
| H. thienemanni | Populus nigra | Castillo de Locubin, Jaén, Spain | – | KF430568 | Subbotin et al. (2014) |
| H. thornei | Vitis vinifera | La Rambla, Córdoba, Spain | KF430452 | KF430581 | Subbotin et al. (2014) |
| H. typica | Grasses | Gauteng province, South Africa | KF430515 | KF430603 | Subbotin et al. (2014) |
| H. typica | Sugarcane | South Africa | – | GQ406238, GQ406239 | Van den Berg et al. (2010) |
| H. vaccinii | Pinus pinaster | Carnota, Coruña, Spain | – | KF430542 | Subbotin et al. (2014) |
| H. vaccinii | Pinus pinaster | Monteagudo Isl., Pontevedra, Spain | KF430459, KF430460 | – | Subbotin et al. (2014) |
| H. vidua | Camellia sp. | South Carolina, USA | – | JQ708147 | Cordero López et al. (2013) |
| Hemicycliophora sp. | Unknown plant | Iran | KY284835 | – | E. Miraeiz, R. Heydari (unpub.) |
| Hemicycliophora sp. 1 | Grasses | Terovo, Epirus, Greece | AY780974 | KF430602 | Subbotin et al. (2005); Subbotin et al. (2014) |
| Hemicycliophora sp. 2 | Unknown plant | Birdlings Flat, New Zealand | KF430516, KF430517 | KF430609, KF430610 | Subbotin et al. (2014) |
| Hemicycliophora sp. 3 | Zea mays | Tingle Farms, Willcox, AZ, USA | – | KF430573, KF430574 | Subbotin et al. (2014) |
| Hemicycliophora sp. 5 | Turf grasses | Carteret County, NC, USA | – | KF430575 | Subbotin et al. (2014) |
| Hemicycliophora sp. 6 | Nothofagus forest | Kaitoke Waterworks, New Zealand | KF430446 | KF430585, KF430586 | Subbotin et al. (2014) |
| Hemicycliophora sp. 7 | Pinus pinea | Almonte, Huelva, Spain | KF430451 | KF430589 | Subbotin et al. (2014) |
| Hemicycliophora sp. 8 | Unknown plant | Henrieville, UT, USA | KF444173 | – | Subbotin et al. (2014) |
| Hemicycliophora sp. 8 | Turf grasses | Monterey, CA, USA | KF430494 | KF430559 | Subbotin et al. (2014) |
| Hemicycliophora sp. 9 | Trifolium repens | Preveza, Greece | KF430509, KF430511, KF430514 | KF430605 | Subbotin et al. (2014) |
| Hemicycliophora sp. 9 | Agrostis sp. | Jaroslavl region, Russia | – | KF430604 | Subbotin et al. (2014) |
| Hemicycliophora sp. 9 | Unknown plant | Brake, Germany | AY780973 | – | Subbotin et al. (2005); Subbotin et al. (2014) |
| Hemicycliophora sp. 10 | Salix sp. | Yolo County, CA, USA | KF430483, KF430485–KF430486 | KF430566, MG019828 | Subbotin et al. (2014); Van den Berg et al. (2018) |
| Hemicycliophora sp. 11 | Andropogon virginicus | Paines Prairie, FL, USA | KF430493 | KF430557, KF430558 | Subbotin et al. (2014) |
| Hemicycliophora sp. 12 | Grasses | Saint Paul, MN, USA | KF430474 | – | Subbotin et al. (2014) |
| Hemicycliophora sp. 12 | Unknown plant | Brooklyn Park, MN, USA | KF430475 | – | Subbotin et al. (2014) |
| Hemicycliophora sp. 12 | Unknown plant | Sedona, AZ, USA | KF430476 | – | Subbotin et al. (2014) |
| Hemicycliophora sp. 13 | Neoregelia sp. | Los Angeles County, CA, USA | KF430507, KF430508 | – | Subbotin et al. (2014) |
| Hemicycliophora sp. 15 | Unknown plant | Vicinity of Trois–Rivières, Quebec, Canada | MG019819 | – | Van den Berg et al. (2018) |
| Hemicycliophora sp. 16 | Unknown tree | east of Temecula, CA, USA | MG019818 | MG019829 | Van den Berg et al. (2018) |
| Hemicycliophora sp. 17 | Unknown tree | Pismo Beach, San Luis Obispo County, CA, USA | – | MG019830 | Van den Berg et al. (2018) |
| Hemicycliophora sp. 18 | Unknown plant | Vicinity of Quebec City, Quebec, Canada | MG019820 | – | Van den Berg et al. (2018) |
| Gracilacus bilineata | Bambusa sp. | Taiwan | – | EU247525 | Chen et al. (2008) |
| Paratylenchus bukowinensis | Unknown plant | Monopoli, Italy | AY780943 | – | Subbotin et al. (2005) |
| Paratylenchus minutus | Annona squamosa | Taiwan | – | EF126180 | Chen et al. (2009) |
| Paratylenchus nanus | Unknown plant | Niebüll, Germany | AY780946 | – | Subbotin et al. (2005) |
| Trophotylenchulus floridensis | Pinus elliottii | Crystal river, Florida, USA | – | JN112261 | Tanha Maafi et al. (2012) |
Table 2.
Morphometrics of Hemicycliophora ahvasiensis n. sp. from Khuzestan province, Iran.
| Character | Female holotype | Female paratypes | Juvenile |
|---|---|---|---|
| n | 1 | 20 | 1 |
| L | 868.7 | 830.3 ± 48.3 (767–893) | 600 |
| a | 22.2 | 21.5 ± 2.1 (17.9–24.5) | 18.0 |
| b | 5.9 | 5.8 ± 0.3 (5.4–6.5) | 4.8 |
| c | 10.7 | 10.1 ± 1.2 (8.3–11.5) | 9.8 |
| c' | 2.9 | 3.0 ± 0.3 (2.5–3.5) | 2.6 |
| o | 11.5 | 12.2 ± 0.9 (9.4–15.3) | 9.0 |
| DGO | 7.9 | 8.1 ± 0.9 (7.4–10.0) | 5.5 |
| V | 85.5 | 84.1 ± 0.9 (82.6–85.5) | – |
| St | 68.4 | 66.5 ± 2.3 (63.3–71.0) | 60.8 |
| m | 81.5 | 80.6 ± 1.4 (77.4–83.7) | 80.7 |
| Stylet knob height | 4 | 4.1 ± 0.5 (4–5) | 3.8 |
| Stylet knob width | 7 | 6.6 ± 0.7 (6–8) | 6.6 |
| Excretory pore from anterior end | 171 | 168.3 ± 6.1 (159–180) | 168 |
| Diam. at mid-body | 39 | 38.4 ± 3.6 (32–46) | 33 |
| Diam. at anus (ABD) | 27 | 26.8 ± 1.7 (24–29) | 23 |
| Diam. at vulva | 38 | 38.4 ± 2.1 (35–43) | – |
| Vulva-anterior body distance | 744 | 700 ± 43 (653–751) | – |
| Vulva-tail terminus distance | 125 | 129.5 ± 6.0 (113–142) | – |
| Spermatheca-vulva distance | 89 | 87.6 ± 13.2 (74–121) | – |
| Lip diam. | 15 | 15.7 ± 0.9 (14–18) | 14 |
| Lip height | 7 | 6.7 ± 0.7 (6–9) | 6 |
| First body annulus diam. | 16 | 16.9 ± 0.9 (15–19) | 15 |
| Second body annulus diam. | 18 | 18.6 ± 1.1 (16–21) | 16 |
| Pharynx length | 145 | 142.6 ± 4.8 (134–151) | 125 |
| Annulus width | 4 | 4.1 ± 0.3 (3.4–4.7) | 2.8 |
| Tail length | 81 | 83.3 ± 7.9 (74–92) | 61 |
| V-anus distance | 45 | 47.9 ± 9.9 (32–64) | – |
| R | 245 | 221.3 ± 8.6 (212–247) | 216 |
| RSt | 19 | 19.5 ± 1.1 (18–21) | 22 |
| Rph | 41 | 41.1 ± 3.2 (35–48) | 46 |
| Rex | 48 | 47.4 ± 3.6 (42–50) | 58 |
| RV(ant) | 193 | 185.3 ± 8.4 (167–198) | – |
| RV | 52 | 47.8 ± 6.9 (38–59) | – |
| RVan | 15 | 15.0 ± 3.8 (10–22) | – |
| Ran | 37 | 32.9 ± 4.9 (25–47) | – |
| VL/VB | 3.3 | 3.4 ± 0.3 (2.8–3.9) | – |
| Spermatheca length | 29 | 19.8 ± 5.4 (14–29) | – |
| Spermatheca diam. | 15 | 15.5 ± 1.6 (12–22) | – |
| St%L | 7.8 | 7.9 ± 0.4 (7.5–8.4) | 10 |
Figure 1:
Line drawings of Hemicycliophora ahvasiensis n. sp. Female. A: Anterior body region; B: Spermatheca; C: Vulval region; D–F: Variation of posterior body end morphology. (Scale bar = 20 μm).
Figure 2:
Light photomicrographs of Hemicycliophora ahvasiensis n. sp. Female. A: Entire body; B, C: Anterior body region; D: Reproductive system (the arrow indicates the spermatheca); E: Spermatheca; F: Vagina; G: Posterior body region.
Figure 3:
Scanning electron micrographs of Hemicycliophora ahvasiensis n. sp. Female. A: Entire body; B, C: Anterior end showing labial region; D, E: En face view of labial area; F, G: Annuli ornamentation (the arrows indicate the excretory pore); H–L: Posterior body region.
Figure 4:
Scanning electron micrographs of Hemicycliophora ahvasiensis n. sp. Female. A–F: Mid-body annuli ornamentation.
Figure 5:
Light photomicrographs of Hemicycliophora conida Thorne, 1955 from Gilan province, Iran. A–G: Female. A: Anterior body region; B: Pharyngeal region; C: Lateral field at mid-body; D, E: Annuli ornamentation; F, G: Posterior body region; H, I: Male. H: Anterior body region; I: Posterior body region. (Scale bar = 20 μm).
Table 3.
Morphometrics of Hemicycliophora conida Thorne, 1955 from Gilan province, Iran, and comparison with other population from East Azarbaijan province, Iran.
| Reference | Present study | Loof (1984) | |
|---|---|---|---|
| Province | Gilan province | East Azarbaijan province | |
| Character | Female | Male | Female |
| n | 10 | 5 | 11 |
| L | 912.0 ± 21.4 (881–928) | 809.0 ± 13.6 (795–822) | 820–1020 |
| a | 21.2 ± 2.5 (18.6–24.3) | 37.3 ± 5.7 (31.8–43.3) | 26–30 |
| b | 5.3 ± 0.2 (5.2–5.6) | – | 5.0–5.6 |
| c | 11.6 ± 2.0 (10.1–14.5) | 8.2 ± 0.4 (7.8–8.6) | 9.7–13.6 |
| c' | 2.3 ± 0.4 (1.7–2.6) | 5.2 ± 0.5 (4.6–5.6) | – |
| V | 86.7 ± 1.0 (85.5–87.6) | – | 86–89 |
| St | 92.5 ± 2.1 (90–97) | – | 90–103 |
| m | 78.3 ± 4.0 (75.3–84.2) | – | – |
| Stylet knob height | 5.0 ± 0.4 (4.3–5.6) | – | – |
| Stylet knob width | 7.8 ± 0.5 (6.9–8.6) | – | – |
| Excretory pore from anterior end | 178.0 ± 8.9 (169–192) | 140.2 ± 13.4 (127–164) | – |
| Diam. at mid-body | 43.5 ± 5.5 (38–50) | 22 ± 3 (19–25) | – |
| Diam. at anus/cloaca | 35.3 ± 4.3 (30–40) | 19 ± 1 (18–20) | – |
| Diam. at vulva | 46.4 ± 5.8 (40–55) | – | – |
| Vulva-anterior body distance | 791 ± 16 (770–809) | – | – |
| Vulva-tail terminus distance | 124.5 ± 7.8 (115–136) | – | – |
| Spermatheca-vulva distance | 82.2 ± 10.1 (72–96) | – | – |
| Lip diam. | 21.3 ± 2.2 (19–24) | 10.7 ± 1.5 (9–12) | – |
| Lip height | 7.5 ± 0.6 (7–8) | 6.2 ± 0.8 (6–7) | – |
| First body annulus diam. | 23.8 ± 1.9 (20–26) | – | – |
| Second body annulus diam. | 26.1 ± 2.7 (21–30) | – | – |
| Pharynx length | 171.0 ± 4.7 (167–176) | – | – |
| Annulus width | 4.1 ± 0.5 (3.6–5.1) | 1.9 ± 0.1 (1.8–2.0) | – |
| Tail length | 88.0 ± 6.5 (79–94) | 98.7 ± 6.1 (92–104) | – |
| V-anus distance | 42.0 ± 19.7 (28–71) | – | – |
| R | 230.0 ± 9.2 (224–237) | – | 259–286 |
| RSt | 21.0 ± 0.9 (18–23) | – | – |
| Rph | 38.0 ± 0.2 (38–39) | – | – |
| Rex | 41.4 ± 1.4 (39–43) | – | 48–52 |
| RV(ant) | 187.0 ± 3.5 (185–190) | – | 207–226 |
| RV | 46.0 ± 4.9 (37–54) | – | – |
| RVan | 16.0 ± 8.5 (10–22) | – | 11–17 |
| Ran | 27.0 ± 2.8 (25–29) | – | 35–41 |
| VL/VB | 2.7 ± 0.3 (2.2–3.4) | – | 4.0–5.3 |
| Spermatheca length | 22.7 ± 5.9 (12–30) | – | – |
| Spermatheca diam. | 31.8 ± 8.1 (15–39) | – | – |
| Spicules length | – | 54.3 ± 2.1 (52–56) | – |
| Gubernaculum length | – | 20.3 ± 0.6 (20–21) | – |
| Bursa length | – | 41.7 ± 6.4 (37–49) | – |
Figure 6:
Bayesian 50% majority rule consensus tree inferred from analysis of the D2-D3 domains of the LSU rDNA sequences of Hemicycliophora ahvasiensis n. sp. under the GTR + G + I model. (lnL = 6023.6660; freqA = 0.2165; freqC = 0.2342; freqG = 0.3064; freqT = 0.2429; R(a) = 0.4542; R(b) = 1.5000; R(c) = 1.0798; R(d) = 0.4155; R(e) = 4.2300; R(f) = 1; Pinvar = 0.3122; Shape = 0.7157). Bayesian posterior probability values more than 0.50 are given for appropriate clades. New sequences are indicated in bold.
Figure 7:
Bayesian 50% majority rule consensus tree inferred from analysis of the ITS rRNA gene of Hemicycliophora ahvasiensis n. sp. and Iranian population of H. conida under the GTR + G + I model. (lnL = 13525.0293; freqA = 0.2348; freqC = 0.2548; freqG = 0.2520; freqT = 0.2583; R(a) = 1.6876; R(b) = 2.3417; R(c) = 1.8416; R(d) = 0.8470; R(e) = 3.6304; R(f) = 1; Pinvar = 0.1024; Shape = 0.4967). Bayesian posterior probability values more than 0.50 are given for appropriate clades. New sequences are indicated in bold.