Table 1:
Description of Hirschmanniella maritima n. sp. populations isolated from Potamogeton perfoliatus L. plants with GenBank accession numbers of sequences acquired from the large subunit ribosomal DNA (28S rDNA) and the cytochrome c oxidase subunit I (mtCOI).
| Locality and habitat | 28S rDNA | mtCOI | ||
|---|---|---|---|---|
| # individuals from which a sequence was acquired | corresponding GenBank number | # individuals from which a sequence was acquired | corresponding GenBank number | |
| The Gulf of Gdańsk, Jelitkowo Gdańsk, from a 3-m depth, 17.09.2011. Collected with a fyke net set. Type population. | 2* | KC507798 | 1 | PV031916 |
| The Gulf of Gdańsk, Sopot, estuary of the Swelina River, from a 1-m depth, 1.07.2012. The plants were pulled out by hand. | 4 | - | ||
| The Puck Bay, Rewa, the Beka reserve. Plants were washed ashore after an autumn storm. 28.10.2009. | 24** | 11 | PV031913 | |
| 3 | PV031914 | |||
| 1 | PV031915 | |||
| The Puck Bay, Władysławowo, from a 3-m depth, 12.07.2012. Collected with a fyke net set. | 2 | - | ||
| Vistula Lagoon, Kąty Rybackie, from a 1-m depth, 28.06.2011. The plants were pulled out by hand. | - | 1 | PV031914 | |
Figure 1:
Line drawings of Hirschmanniella maritima n. sp. Female: A, B: Anterior end: A: holotype, B: Rewa population. C: Vulval region, holotype. D: posterior genital branch, paratype. E, F: Spermatheca, paratypes: E: empty, F: filled with sperm. G: Posterior part of body, holotype. H - J: Distal part of tail: H: holotype, I: paratype, J: Rewa population. Scale bar = 10 μm.
Figure 2:
Line drawings of Hirschmanniella maritima n. sp. Male, paratype : A: Anterior part of body. B, C: Anterior end. D: Posterior part of body. E: Distal part of tail. F: Male copulatory apparatus. Scale bar = 10 μm.
Figure 3:
Light micrographs Hirschmanniella maritima n. sp. A - J. Female. K - S. Male. A: Anterior part of body, holotype. B: Posterior part of body, holotype. C - E: Anterior end, paratypes. F, G: Vulval region: F: holotype, G: paratype. H - J: Distal part of tail, paratypes. K: Lateral field. L: Posterior part of body. M - O: Anterior part of body. P, Q. Male copulatory apparatus. R, S. Distal part of tail. Scale bar = 10 μm.
Table 2:
Morphometric characteristics of females and males of Hirschmanniella maritima n. sp. Measurements are provided in μm and in the form of mean ± standard deviation, with the range given in parentheses and the coefficient of variation (CV) in italics after the parentheses. All abbreviations are used according to Brzeski (1998).
| Character | Gdańsk | Rewa | Władysławowo | All populations | |||||
|---|---|---|---|---|---|---|---|---|---|
| Holotype female | Paratype females | Paratype males | Females | Males | Females | Males | Females | Males | |
| n | 1 | 14 | 12 | 20 | 22 | 6 | 8 | 41 | 42 |
| L | 3009 | 2355 ± 266.4 (1893–2708) 11.3 | 2122 ± 198.3 (1787–2441) 9.3 | 2688 ± 228.2 (2218–3010) 8.5 | 2619 ± 205.8 (2065–3085) 7.9 | 2531 ± 201.6 (2146–2682) 8.0 | 2248 ± 93.3 (2100–2356) 4.2 | 2559 ± 284.3 (1893–3010) 11.1 | 2406 ± 294.1 (1787–3085) 12.2 |
| a | 62 | 67.0 ± 7.2 (59.2–81.8) 10.8 | 64.5 ± 5.2 (57.3–75.3) 8.1 | 68.6 ± 5.6 (59.5–79.7) 8.2 | 69.7 ± 6.6 (55.3–81.2) 9.5 | 64.5 ± 3.5 (60.4–70.0) 5.4 | 62.6 ± 4.3 (58.2–69.8) 6.9 | 67.3 ± 6.0 (59.2–81.8) 8.9 | 66.9 ± 6.5 (55.3–81.2) 9.8 |
| b | 18.9 | 15.6 ± 1.4 (13.1–17.8) 8.7 | 13.8 ± 1.0 (11.6–15.4) 7.5 | 17.1 ± 1.5 (14.5–19.7) 9.0 | 17.2 ± 1.0 (14.9–18.8) 5.8 | 15.9 ± 1.0 (15.0–17.4) 6.4 | 14.2 ± 0.7 (13.2–15.2) 4.9 | 16.4 ± 1.6 (13.1–19.7) 9.6 | 15.7 ± 1.9 (11.6–18.8) 12.0 |
| b’ | 6.8 | 5.6 ± 0.6 (4.8–6.5) 11.2 | 4.9 ± 0.3 (4.5–5.7) 7.0 | 7.9 ± 0.8 (6.1–9.2) 10.5 | 5.7 ± 0.8 (4.5–7.0) 13.5 | 5.5 ± 0.4 (5.0–5.9) 7.9 | 5.3 ± 0.6 (4.5–6.3) 11.4 | 6.7 ± 1.3 (4.8–9.2) 20.0 | 5.3 ± 0.6 (4.5–7.0) 12.3 |
| c | 14.6 | 15.1 ± 2.0 (12.6–17.8) 13.2 | 15.5 ± 1.2 (13.7–17.9) 7.7 | 14.5 ± 1.2 (12.8–17.4) 8.1 | 17.6 ± 1.5 (14.2–20.6) 8.7 | 14.0 ± 0.5 (13.4–14.8) 3.9 | 15.3 ± 1.5 (13.5–17.4) 9.5 | 14.6 ± 1.5 (12.6–17.8) 9.9 | 16.6 ± 1.8 (13.5–20.6) 10.6 |
| c’ | 6.6 | 5.8 ± 05 (5.1–6.5) 8.2 | 6.0 ± 0.4 (5.4–6.7) 7.0 | 6.7 ± 0.7 (5.2–8.1) 11.0 | 6.1 ± 0.7 (4.7–7.7) 11.2 | 6.6 ± 0.6 (6.2–7.7) 8.8 | 5.9 ± 0.6 (4.7–6.6) 10.6 | 6.4 ± 0.7 (5.1–8.1) 11.7 | 6.0 ± 0.6 (4.7–7.7) 10.0 |
| V | 46.8 | 52.8 ± 1.7 (50.0–55.4) 3.3 | - | 49.8 ± 1.9 (46.8–53.7) 3.9 | - | 52.2 ± 1.7 (49.3–53.5) 3.2 | - | 51.1 ± 2.3 (46.8–55.4) 4.6 | - |
| G1 | 19.2 | 24.0 ± 2.8 (18.8–28.9) 11.8 | - | 22.3 ± 4.8 (14.7–29.8) 21.4 | - | 32.3 ± 3.4 (27.8–36.8) 10.4 | - | 24.9 ± 5.2 (14.7–36.8) 20.8 | - |
| G2 | 16.8 | 23.8 ± 2.6 (18.8–28.8) 10.9 | - | 22.7 ± 5.1 (14.7–28.5) 22.4 | - | 30.7 ± 2.9 (26.1–34.2) 9.4 | - | 24.6 ±4.8 (14.7–34.2) 19.3 | - |
| T | - | - | 44.3 ± 2.8 (38.9–48.0) 6.4 | - | 39.7 ± 4.5 (29.2–51.2) 11.3 | - | 44.9 ± 3.1 (40.4–49.5) 7.0 | - | 42.2 ± 4.4 (29.2–51.2) 10.5 |
| Lip region diameter | 13.2 | 10.5 ± 0.6 (9.6–11.5) 5.8 | 10.3 ± 0.6 (9.7–11.8) 5.5 | 10.5 ± 0.5 (9.5–11.8) 5.0 | 10.3 ± 0.4 (9.5–10.8) 3.5 | 10.9 ± 0.7 (9.6–11.6) 6.3 | 10.2 ± 0.6 (9.2–11.1) 5.7 | 10.6 ± 0.7 (9.5–13.2) 6.6 | 10.3 ± 0.5 (9.2–11.8) 4.5 |
| Lip region height | 5.7 | 4.6 ± 0.4 (3.8–5.2) 9.6 | 4.7 ± 0.6 (3.9–5.9) 12.0 | 4.5 ± 0.4 (3.8–5.2) 9.3 | 4.0 ± 0.4 (3.4–4.9) 9.3 | 4.7 ± 0.2 (4.5–5.1) 5.1 | 4.7 ± 0.3 (4.4–5.2) 5.6 | 4.6 ± 0.4 (3.8–5.7) 9.5 | 4.4 ± 0.5 (3.4–5.9) 12.5 |
| Lip annules number | 5 | 5.3± 0.5(5–6) 8.9 | 5.2±0.4 (5–6) 7.5 | 4.8±0.4 (4–5) 8.7 | 4.7±0.5 (4–5) 9.6 | 5.2±0.4 (5–6) 7.9 | 5.4± 0.5(5–6) 9.6 | 5.1 ± 0.5 (4–6) 10.0 | 5.0 ± 0.5 (4–6) 10.4 |
| Stylet length | 23.2 | 21.3 ± 1.8 (19.3–24.5) 8.6 | 20.9 ± 1.3 (19.4–23.4) 6.2 | 21.5 ± 0.9 (20.0–23.4) 4.3 | 21.3 ± 0.6 (20.4–22.8) 2.8 | 21.6 ± 0.4 (20.9–22.1) 1.9 | 21.5 ± 1.0 (20.2–23.3) 4.5 | 21.5 ± 1.3 (19.3–24.5) 5.8 | 21.2 ± 0.9 (19.4–23.4) 4.3 |
| Conus length | 10.8 | 10.6 ± 0.9 (9.4–12.1) 8.4 | 10.4 ± 0.8 (9.2–12.0) 8.1 | 11.0 ± 1.0 (10.0–12.9) 8.7 | 10.9 ± 0.6 (9.8–11.8) 5.7 | 10.7 ± 0.2 (10.4–11.0) 1.9 | 10.6 ± 0.6 (9.4–11.2) 6.0 | 10.8 ± 0.8 (9.4–12.9) 7.9 | 10.7 ± 0.7 (9.2–12.0) 6,6 |
| Shaft length | 9.5 | 8.3 ± 0.8 (7.0–9.7) 9.6 | 7.9 ± 0.5 (7.3–8.8) 6.6 | 7.8 ± 0.9 (6.7–10.4) 11.2 | 10.4 ± 0.7 (9.0–12.0) 6.4 | 8.1 ± 0.3 (7.6–8.4) 3.7 | 8.3 ± 0.4 (7.7–9.2) 5.2 | 8.0 ± 0.8 (6.7–10.4) 10.3 | 9.3 ± 1.3 (7.3–12.0) 14.5 |
| Stylet knob height | 2.9 | 2.5 ± 0.3 (2.1–3.1) 13.5 | 2.6 ± 0.2 (2.2–3.0) 8.5 | 2.9 ± 0.4 (2.3–3.8) 13.0 | 2.8 ± 0.3 (2.3–3.5) 11.3 | 2.8 ± 0.2 (2.7–3.1) 5.8 | 2.6 ± 0.3 (2.3–3.0) 10.5 | 2.8 ± 0.4 (2.1–3.8) 13.9 | 2.7 ± 0.3 (2.2–3.5) 11.1 |
| Stylet knob width | 5.8 | 5.0 ± 0.5 (4.1–5.9) 10.6 | 4.8 ± 0.3 (4.4–5.3) 5.7 | 4.8 ± 0.6 (3.6–5.8) 12.2 | 4.5 ± 0.3 (4.0–5.2) 7.1 | 5.1 ± 0.3 (4.7–5.5) 6.3 | 4.6 ± 0.2 (4.3–5.0) 5.0 | 4.9 ± 0.5 (3.6–5.9) 11.0 | 4.6 ± 0.3 (4.0–5.3) 6.9 |
| Anterior end to median bulb valve distance | 108.8 | 102.9 ± 6.1 (94.1–115.0) 6.0 | 103.1 ± 5.6 (95–111) 5.4 | 108.5 ± 4.8 (102.1–119.0) 4.4 | 111.2 ± 5.0 (102.1–120.3) 4.5 | 108.8 ± 3.7 (104.4–112.5) 3.4 | 108.5 ± 3.3 (103–112) 3.0 | 106.5 ± 5.7 (94.1–119) 5.4 | 108.3 ± 6.0 (95.0–120.3) 5.5 |
| Anterior end to pharyngo-intestinal junction (PIJ) distance | 159 | 151.1 ± 12.9 (132–172) 8.5 | 153.8 ± 8.4 (136–165) 5.5 | 157.8 ± 8.4 (135–170) 5.3 | 152.4 ± 8.6 (138–173) 5.6 | 159.3 ± 12.5 (143–177) 7.8 | 157.9 ± 4.6 (151–167) 2.9 | 155.8 ± 10.9 (132–177) 7.0 | 153.8 ± 8.0 (136–172.5) 5.2 |
| Pharyngeal overlap length | 286 | 273.6 ± 40.1 (228–377) 14.6 | 277.1 ± 39.1 (226–357) 14.1 | 185.7 ± 31.4 (159–284) 16.9 | 304.9 ± 65.8 (215–423) 21.6 | 302.5 ± 36.3 (259–359) 12.0 | 270.3 ± 53.1 (203–324) 19.7 | 238.2 ± 60.5 (159–377) 25.4 | 283.8 ± 52.5 (203–422.7) 18.5 |
| Anterior end to nerve ring distance | 143.6 | 134 ± 10.4 (120–155) 7.8 | 130.0 ± 8.3 (116–145) 6.4 | 136.5 ± 6.1 (126–143) 4.5 | 141.9 ± 6.2 (133–149) 4.4 | 139.3 ± 8.2 (127–147) 5.9 | 136.3 ± 4.7 (128–141) 3.5 | 136.5 ± 8.5 (120–155) 6.2 | 135.1 ± 8.3 (116–149) 6.2 |
| Anterior end to excretory pore (EP) distance | 180.8 | 167.8 ± 12.8 (144–186) 7.6 | 170.0 ± 9.4 (154–184) 5.5 | 174.8 ± 9.3 (155–190) 5.3 | 176.7 ± 11.7 (159–197) 6.6 | 181.8 ± 11.1 (163–196) 6.1 | 182.1 ± 8.5 (174–200) 4.6 | 173.4 ± 11.8 (144–196) 6.8 | 175.8 ± 11.1 (154–200) 6.3 |
| EP to PIJ distance | 21.8 | 16.7 ± 7.1 (4–25) 42.4 | 16.3 ± 4.4 (7–22) 27.0 | 16.3 ± 6.9 (3.0–30.0) 42.2 | 24.3 ± 10.0 (6.8–46.1) 41.4 | 22.9 ± 3.6 (19.5–28.9) 15.9 | 22.9 ± 5.0 (16.8–30.9) 22.1 | 17.7 ± 6.8 (3–30) 38.2 | 21.7 ± 8.6 (6.8–46.1) 39.5 |
| Max. body diameter | 48.5 | 35.2 ± 2.7 (31.3–40) 7.8 | 33.0 ± 2.4 (29.5–38.2) 7.3 | 39.3 ± 3.2 (34.0–45.2) 8.2 | 37.8 ± 3.3 (31.7–43.8) 8.7 | 39.4 ±4.5 (32.6–44.4) 11.5 | 36.1 ± 2.9 (32.2–40.5) 8.2 | 38.1 ± 4.1 (31.3–48.5) 10.7 | 36.1 ± 3.6 (29.5–43.8) 10.0 |
| Female anterior genital branch length | 578 | 564.3 ± 91.6 (418–729) 16.2 | - | 604.4 ± 105.8 (414–764) 17.5 | - | 814.0 ± 74.1 (709–939) 9.1 | - | 626.0 ± 130.3 (414–939) 20.8 | - |
| Female posterior genital branch length | 505 | 560.9 ± 84.6(424–727) 15.1 | - | 619.9 ± 121.5 (413–761) 19.6 | - | 774.8 ± 82.6 (666–906) 10.7 | - | 619.5 ± 124.8 (413–906) 20.1 | - |
| Anterior spermatheca length | 54.7 | 45.6 ± 12.2 (32.5–74.0) 26.7 | - | 76.0 ± 4.4 (69.5–80.4) 5.8 | - | 54.3 ± 3.2 (51.2–58.8) 5.8 | - | 55.3 ± 15.1 (32.5–80.4) 27.2 | - |
| Anterior spermatheca diameter | 28.8 | 22.7 ± 5.0 (13.2–28.8) 22.1 | - | 34.1 ± 4.7 (26.9–38.3) 13.8 | - | 23.3 ± 3.2 (20–27) 13.6 | - | 25.9 ± 6.5 (13.2–38.3) 25.0 | - |
| Posterior spermatheca length | 47.1 | 43.9 ± 9.3 (32–59) 21.3 | - | 83.1 ± 3.7 (78.6–87.3) 4.5 | - | 55.0 ± 4.3 (47.7–59.2) 7.8 | - | 54.7 ± 16.8 (32.0–87.3) 30.7 | - |
| Posterior spermatheca diameter | 31.5 | 22.1 ±5.3 (11.8–27.8) 24.1 | - | 36.8 ± 3.3 (33.2–40.4) 8.9 | - | 24.3 ± 2.0 (22.1–26.7) 8.2 | - | 26.3 ± 7.2 (11.8–40.4) 27.3 | - |
| Cloaca to most anterior part of testis distance | - | - | 937.9 ± 94.3 (812–1103) 10.1 | - | 1042.7 ± 137.5 (798–1382) 13.2 | - | 1007.8 ± 69.8 (879–1074) 6.9 | - | 1002.2 ± 120.0 (798–1382) 12.0 |
| Anal body diameter | 31.3 | 27.1 ± 2.1 (24.5–30.4) 7.7 | 23.0 ± 2.2 (20.6–25.9) 9.5 | 27.8 ± 2.3 (24.0–31.8) 8.4 | 24.8 ± 2.3 (20.6–28.9) 9.2 | 27.6 ± 3.9 (20.9–31.7) 14.1 | 25.2 ± 1.6 (22.7–28.0) 6.2 | 27.6 ± 2.5 (20.9–31.8) 9.1 | 24.4 ± 2.3 (20.6–28.9) 9.3 |
| Rectum/cloaca length | 24.7 | 25.3± 2.6 (20.6–29.6) 10.3 | 33.5±2.9 (31.4–38.4) 8.8 | 24.5± 4.2 (19.4–35.0) 17.0 | 27.8±3.2 (24.0–35.0) 11.4 | 29.8±1.1 (28.5–31.3) 3.8 | 33.4±2.3 (31.5–36.0) 7.0 | 25.8 ± 3.7 (19.4–35.0) 14.2 | 30.6 ± 4.0 (24–38.4) 13.2 |
| Spicule length | - | - | 38.4 ± 1.1 (36.4–39.4) 2.9 | - | 38.5 ± 2.3 (34.0–43.2) 6.0 | - | 40.8 ± 1.8 (38.4–42.2) 4.4 | - | 38.7 ± 2.2 (34–43.2) 5.6 |
| Gubernaculum length | - | - | 13.1 ± 1.0 (12.0–14.5) 7.3 | - | 12.1 ± 1.0 (10.4–14.6) 8.4 | - | 12.1 ± 0.7 (11.4–12.7) 5.4 | - | 12.3 ± 1.0 (10.4–14.6) 8.3 |
| Tail length | 205.6 | 157 ± 9.0 (150–178) 5.7 | 136.8 ± 9.0 (120–150) 6.6 | 186.6 ± 18.3 (153–215) 9.8 | 149.8 ± 14.3 (115–178) 9.6 | 180.8 ± 13.0 (160–196) 7.2 | 147.5 ± 12.6 (126–159) 8.6 | 176.1 ± 20.3 (150–215) 11.6 | 145.6 ± 13.7 (115–177.5) 9.4 |
Figure 4:
28S rDNA-based Bayesian phylogeny of the genus Hirschmanniella. The new H. maritima n. sp. sequence is indicated in bold. Numbers near nodes indicate posterior probabilities.
Figure 5:
mtCOI-based Bayesian phylogeny of the genus Hirschmanniella and the most closely related genera. The new H. maritima n. sp. sequences are indicated in bold. Numbers near nodes indicate posterior probabilities.
Figure 6:
Micrographs of P. perfoliatus roots infected with H. maritima n. sp. Light (A, B, and E) and transmission electron (C, D, and F) microscopy images. A: Cross section of uninfected root. B: Cross section of infected root with nematode in outer cortex. C: Broken cell walls next to nematode. D: Plant cell protoplast debris next to nematode. F: Nematodes next to degraded inner cortex cells. E: Fungal hyphae in inner cortex of root. Abbreviations: C, cortex; CW, cell wall; E, exodermis; H, hypha; N, nematode; PD, protoplast debris; VC, vascular cylinder. Arrows point to broken cell walls (C); arrowheads point to hyphae (E); asterisks indicate intercellular spaces (A, B, E, and F). Scale bars: = 50 μm (A, B, and E) and = 5 μm (C, D, and F).