Table S1.
Details on taxa used in the molecular analyses.
| GeneBank accession no. | ||||
|---|---|---|---|---|
| Species | Isolate name/geographic origin | ITS rDNA | 28S rDNA | cox1 |
| Steinernema sandneri n. sp. | S17-050, Poland | MW078536 | MW078535 | MW078544 |
| Steinernema affine | B1, England | AF331899 | ||
| Steinernema affine | The Netherlands | AY171298 | ||
| Steinernema akhursti | China | DQ375757 | ||
| Steinernema bicornutum | Serbia | AF331904 | ||
| Steinernema bicornutum | Yugoslavia | AF121048 | ||
| Steinernema cameroonense | OB, Cameroon | JX985267 | ||
| Steinernema carpocapsae | Russia | AY171282 | ||
| Steinernema cholashanense | Tibet, China | EF431959 | EF520284 | |
| Steinernema citrae | 141-C, South Africa | EU740970 | GU004534 | |
| Steinernema costaricense | Costa Rica | EF187017 | ||
| Steinernema feltiae | Bodega Bay, USA | AF331906 | ||
| Steinernema feltiae | SN, USA | AF121050 | ||
| Steinernema feltiae | 3, Portugal | JQ423217 | ||
| Steinernema glaseri | NC, USA | AF331908 | ||
| Steinernema glaseri | NJ, USA | AF122015 | ||
| Steinernema hebeiense | G6, China | DQ105794 | ||
| Steinernema hermaphroditum | VK-2013, India | KC252604 | ||
| Steinernema ichnusae | Sardinia, Italy | EU421129 | EU421130 | |
| Steinernema jollieti | Monsanto, USA | GU569051 | GU569068 | |
| Steinernema jollieti | 73, USA | AY171265 | ||
| Steinernema kraussei | Westphalia, Germany | AY230175 | AF331896 | AY943990 |
| Steinernema kraussei | Altai 35, Russia | AY171270 | ||
| Steinernema kraussei | Nash, UK | AY230176 | ||
| Steinernema kraussei | Italy | AY230174 | ||
| Steinernema kraussei | Iceland | AY171248 | ||
| Steinernema kraussei | 20F, Portugal | JN683825 | ||
| Steinernema kraussei | D, Switzerland | AY171258 | ||
| Steinernema kraussei | Russia | AY171264 | ||
| Steinernema kraussei | HkHm22, Japan | AB243442 | ||
| Steinernema kraussei | Skr-LUB, Lublin, Poland | KY819012 | ||
| Steinernema kraussei | B2, UK | AY230161 | ||
| Steinernema kraussei | 20F, Portugal | JN683829 | ||
| Steinernema kraussei | Quebec, Canada | GU569053 | ||
| Steinernema kraussei | SKR S11-50, Poland | MW647848 | MW647849 | MW647850 |
| Steinernema kushidai | Hamakita, Japan | AB243440 | ||
| Steinernema kushidai | N22, Japan | AY943991 | ||
| Steinernema kushidai | Japan | AF331897 | ||
| Steinernema litorale | AiAt199, Japan | AB243441 | ||
| Species | Isolate name/geographic origin | GeneBank accession no. | ||
| ITS rDNA | 28S rDNA | cox1 | ||
| Steinernema monticolum | Korea, South Korea | AF122017 | EF439651 | |
| Steinernema monticulum | Mt. Chiri, South Korea | AY943994 | ||
| Steinernema nguyeni | F2, South Africa | KP325084 | ||
| Steinernema oregonense | Oregon, USA | AF122019 | ||
| Steinernema oregonense | OS-10, USA | AF331891 | AY943995 | |
| Steinernema sangi | Vietnam | AY355441 | GU569057 | |
| Steinernema scarabaei | New Jersey, USA | AY172023 | ||
| Steinernema scarabaei | Chile | FJ263673 | ||
| Steinernema silvaticum | S16/019, Poland | MG543845 | MG547576 | MG547572 |
| Steinernema silvaticum | B, Germany | AY171255 | ||
| Steinernema silvaticum | B3, UK (type) | AY230162 | ||
| Steinernema texanum | Texas, USA | EF152568 | EF152569 | |
| Steinernema tielingense | LFS65, China | GU994201 | GU994202 | |
| Steinernema weiseri | F, Germany | AY171268 | ||
| Steinernema weiseri | Turkey | GU569059 | GU569075 | |
| Steinernema xinbinense | LFS8, China | JN171593 | ||
| Steinernema xinbinense | LFS40, China | GU994202 | ||
| Steinernema xueshanense | Yunnan, China | FJ666052 | FJ666053 | |
| Caenorhabditis elegans | N2 Bristol, USA | NC001328 | ||
| Caenorhabditis elegans | X03680 | X03680 | ||
Figure 1:
Steinernema sandneri n. sp. A: infective juvenile, anterior region; B: first-generation male, anterior region; C: first-generation female, tail region; D: first-generation female, vulval region; E: infective juvenile, tail region; F: first-generation male, tail region; G: spicule; H: gubernaculum. Scale bars as on images. Lateral views.
Figure 2:
Steinernema sandneri n. sp. Differential interference contrast (A,C,E) and scanning electron (B,D,F) micrographs of infective juveniles. A – amphid openings, An – anus, BP – bacterial pouch, CP – cephalic papillae, EP – excretory pore, ES – esophagus, F – phasmid opening, H – hyaline part, LF – lateral fields, NR – nerve ring. Scale bars as on images.
Figure 3.
Steinernema sandneri n. sp. Scanning electron micrographs of first-generation males. A,B: anterior region with excretory pore (EP), deirids (D), cephalic (CP) and labial papillae (LP). C,D: posterior region with genital papillae (arrows), single preanal papilla (S-PA) and mucron (Mu). Scale bars as on images.
Figure 4:
Steinernema sandneri n. sp. Differential interference contrast micrographs of first- (A,B,E,F,G) and second-generation males (C,D). A,C: anterior region with esophagus (ES), excretory pore (EP) and nerve ring (NR). B,D: posterior region with spicules, gubernaculum, and mucron (Mu). E: spicules. F,G: gubernaculum (lateral and ventral view). Scale bars as on images.
Table 1.
Morphometrics (in μm) of different developmental stages of Steinernema sandneri n. sp. [mean ± SE (range)] [N = 25].
| First generation | Second generation | |||||
|---|---|---|---|---|---|---|
| Males | Females | Males | Females | Infective juveniles | ||
| Character | Holotype | Paratypes | Paratypes | Paratypes | Paratypes | Paratypes |
| Body length [L] | 1,565.3 | 1,461 ± 22.1 (1,205.7–1,635.3) | 4,628 ± 46.4 (4,244.0–5,014.0) | 946 ± 13.8 (817.5–1,093.8) | 2,120 ± 51.5 (1,640.6–2,753.2) | 843.0 ± 13.9 (708.2–964.5) |
| Greatest body width [W] | 143.0 | 155.1 ± 2.7 (123.8–177.7) | 209.6 ± 3.4 (181.3–261.3) | 70.1 ± 1.1 (54.9–79.5) | 126.6 ± 3.3 (88.9–146.6) | 27.4 ± 0.5 (23.0–31.9) |
| Anterior end to excretory pore [EP] | 88.4 | 80.4 ± 1.5 (63.5–92.4) | 84.4 ± 2.1 (61.4–101.6) | 69.8 ± 1.5 (59.0–84.6) | 72.1 ± 1.3 (57.3–88.4) | 55.9 ± 0.8 (44.4–64.2) |
| Anterior end to nerve ring [NR] | 121.9 | 126.0 ± 1.5 (112.0–138.1) | 146.7 ± 1.2 (132.5–157.6) | 97.7 ± 0.8 (86.4–105.8) | 113.7 ± 1.1 (102.5–124.6) | 102.6 ± 1.4 (82.6–117.9) |
| Anterior end to esophagus [ES] | 155.9 | 157.2 ± 1.1 (147.6–169.6) | 184.7 ± 1.0 (173.2–193.9) | 120.5 ± 1.1 (109.0–128.7) | 145.8 ± 1.4 (130.3–158.5) | 138.4 ± 0.5 (122.5–150.5) |
| Testis reflection | 461.6 | 452.1 ± 9.5 (359.3–537.7) | – | 202.6 ± 13.2 (84.9–379.2) | – | – |
| Tail length [T] | 45.1 | 41.2 ± 0.5 (35.4–45.5) | 46.7 ± 1.6 (32.4–60.9) | 42.3 ± 1.0 (31.7–52.1) | 57.5 ± 1.4 (46.5–72.1) | 75.2 ± 1.1 (64.4–86.4) |
| Anal body diameter [ABW] | 50.1 | 54.1 ± 0.6 (49.9–59.2) | 94.0 ± 2.9 (62.1–121.8) | 36.8 ± 0.4 (30.7–40.8) | 54.4 ± 1.5 (43.1–70.7) | 17.3 ± 0.4 (14.6–23.8) |
| Spicule length [SL] | 64.2 | 59.8±0.5 (52.6–65.3) | – | 51.2 ± 0.9 (42.5–60.2) | – | – |
| Gubernaculum length [GL] | 39.2 | 43.6 ± 0.5 (39.1–50.2) | – | 30.4 ± 0.6 (24.2–39.5) | – | – |
| a [L/W] | 10.9 | 9.5 ± 0.1 (8.5–11.0) | 22.2 ± 0.4 (17.4–24.7) | 13.6 ± 0.2 (12.0–16.4) | 16.9 ± 0.4 (14.1–23.2) | 30.9 ± 0.3 (27.2–33.8) |
| b [L/ES] | 10.4 | 9.3 ± 0.1 (8.0–10.2) | 25.1 ± 0.2 (23.5–27.2) | 7.9 ± 0.1 (7.2–9.4) | 14.5±0.3 (12.0–18.1) | 6.1 ± 0.1 (5.5–6.9) |
| c [L/T] | 34.7 | 35.6 ± 0.5 (31.2–41.9) | 102.0 ± 3.8 (75.4–140.3) | 22.7 ± 0.6 (17.2–28.5) | 37.3 ± 1.1 (24.6–50.1) | 11.2 ± 0.1 (10.5–13.2) |
| Hyaline% [(H/T) × 100] | – | – | – | – | – | 33.6 ± 3.9 (22.7–39.9) |
| D% [(EP/ES) × 100] | 56.7 | 51.2 ± 0.9 (42.1–59.3) | 45.7 ± 1.1 (35.5–54.2) | 58.0±1.2 (48.1–71.9) | 49.5 ± 0.9 (36.2–58.1) | 40.4 ± 0.4 (35.8–44.8) |
| E% [(EP/T) × 100] | 196.0 | 195.8 ± 4.0 (160.2–240.9) | 186.0 ± 7.9 (128.1–266.8) | 167.4 ± 1.2 (128.2–222.8) | 127.2 ± 3.9 (101.4–163.9) | 74.4 ± 0.9 (62.6–85.8) |
| SW% [(SL/ABW) × 100] | 128.1 | 110.9 ± 1.5 (97.0–126.9) | – | 139.6 ± 3.2 (105.1–171.4) | – | – |
| GS% [(GL/SL) × 100] | 61.1 | 79.1 ± 1.2 (60.8–82.8) | – | 59.6 ± 1.1 (49.5–68.6) | – | – |
| V% [(Vulva – anterior end/L) × 100] | – | – | 53.7 ± 0.3 (49.0–56.8) | – | 54.3 ± 0.8 (39.3–59.1) | – |
Figure 5:
Steinernema sandneri n. sp. Differential interference contrast (A,C,E) and scanning electron (B,D,F) micrographs of first-generation females. An – anus, EP – excretory pore, ES – esophagus, LP – labial papillae, NR – nerve ring, V – vulva. Scale bars as on images.
Table 2.
Comparative morphometrics of third-stage infective juveniles of S. sandneri n. sp. and related Steinernema spp.
| Morphometric character a | ||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Species | L | W | EP | NR | ES | T | a | b | c | D% | E% | Reference |
| S. kushidai | 589 (424–662) | 26 (22–31) | 46 (42–50) | 76 (70–84) | 111 (106–120) | 50 (44–59) | 22.5 (19–25) | 5.3 (4.9–5.9) | 11.7 (10–13) | 41 (38–44) | 92 (NA) | Mamiya, (1988) |
| S. hebeiense | 658 (610–710) | 26 (23–28) | 48 (43–51) | 78 (73–83) | 107 (100–111) | 66 (63–71) | 26 (24–28) | 6.2 (5.7–6.7) | 10 (9.4–11) | 45 (40–50) | 72 (65–80) | Chen et al. (2006) |
| S. puntauvense | 670 (631–728) | 33 (31–38) | 25 (20–30) | 54 (46–69) | 94 (81–103) | 54 (51–59) | 20 (17–23) | 6.1 (7.1–7.9) | 12 (11–13) | 42 (25–50) | 44 (35–56) | Uribe–Lorío et al. (2007) |
| S. xinbinense | 694 (635–744) | 30 (28–31) | 51 (46–53) | 86 (75–90) | 116 (109–125) | 73 (65–78) | 24 (21–25) | 6.1 (5–7) | 9.7 (8–11) | 44 (40–47) | 71 (65–78) | Ma et al. (2012) |
| S. jollieti | 711 (625–820) | 23 (20–28) | 60 (53–65) | NA | 123 (115–135) | 68 (60–73) | 31 (25–34) | 5.7 (4.9–6.4) | 10.5 (9.0–11.7) | 48 (46–50) | 88 (NA) | Spiridonov et al. (2004b) |
| S. nguyeni | 737 (673–796) | 25 (22–28) | 52 (47–58) | 80 (74–86) | 110 (101–121) | 67 (61–73) | 29 (27–33) | 6.7 (6.2–7.4) | 11 (10–12) | 48 (43–57) | 79 (70–86) | Malan et al. (2016) |
| S. weiseri | 740 (586–828) | 25 (24–29) | 57 (43–65) | 84 (72–92) | 113 (95–119) | 60 (49–68) | 29 (25–33) | 6.6 (5.7–7.2) | 12 (10–14) | 51 (44–55) | 95 (NA) | Mráček et al. (2003) |
| S. sangi | 753 (704–784) | 35 (30–40) | 52 (46–54) | 91 (78–97) | 127 (120–138) | 81 (76–89) | 22 (19–25) | 5.9 (5.6–6.3) | 9.3 (8.7–10.2) | 40 (36–44) | 62 (56–70) | Phan et al. (2001) |
| S. citrae | 754 (623–849) | 26 (23–28) | 56 (49–64) | 98 (83–108) | 125 (118–137) | 71 (63–81) | 30 (25–34) | 6.0 (5.1–7.1) | 15 (13–14) | 44 (39–58) | 110 (85–132) | Stokwe et al. (2011) |
| S. texanum | 756 (732–796) | 30 (29–34) | 59 (52–62) | 92 (84–102) | 115 (111–120) | 73 (60–79) | 25 (22–27) | 6.5 (6.2–7.0) | 10 (9.6–12.5) | 51 (46–53) | 81 (76–88) | Nguyen et al. (2007) |
| S. akhursti | 812 (770–835) | 33 (33–35) | 59 (55–60) | 90 (83–95) | 119 (115–123) | 73 (68–75) | 24 (23–26) | 6.8 (6.6–7.2) | 11 (10–12) | 47 (45–50) | 77 (73–86) | Qiu et al. (2005) |
| S. sandneri n. sp. | 843 (708–965) | 27 (23–32) | 56 (44–64) | 103 (83–118) | 138 (123–151) | 75 (64–86) | 31 (27–34) | 6.1 (5.5–6.9) | 11.2 (11–13.2) | 40 (36–45) | 74 (63–86) | – |
| S. cholashanense | 843 (727–909) | 30 (26–35) | 62 (59–65) | 87 (72–97) | 125 (110–138) | 73 (60–80) | 28 (24–34) | 6.8 (6.1–7.2) | 12 (10–14) | 49 (46–53) | 81 (76–91) | Nguyen et al., (2008) |
| S. feltiae | 849 (766–928) | 29 (22–32) | 63 (58–67) | 113 (108–117) | 136 (130–143) | 86 (81–89) | 30 (27–34) | 6.4 (5.8–6.8) | 10 (9.4–11) | 46 (44–50) | 74 (67–81) | Nguyen et al., (2007) |
| S. silvaticum | 860 (670–975) | 30 (26–35) | 62 (51–73) | 96 (75–109) | 121 (100–141) | 75 (63–86) | 29 (23–33) | 7.3 (6.3–7.7) | 11.4 (9.9–13.1) | 50 (46–56) | – | Sturhan et al. (2005) |
| S. xueshanense | 860 (768–929) | 30 (29–33) | 67 (60–72) | 91 (81–96) | 135 (130–143) | 87 (80–92) | 28 (26–32) | 6.4 (5.8–7.0) | 9.9 (9.0–11) | 50 (46–52) | 78 (70–90) | Mrácˇek et al. (2009) |
| S. ichnusae | 866 (767–969) | 31 (27–35) | 63 (59–68) | 102 (94–108) | 138 (119–148) | 81 (76–89) | 28 (24–32) | 6.3 (5.6–6.9) | 11 (8.8–12) | 46 (42–49) | 77 (68–83) | Tarasco et al. (2008) |
| S. litorale | 909 (834–988) | 31 (28–33) | 61 (54–69) | 96 (89–104) | 125 (114–133) | 83 (72–91) | 29.5 (27–31) | 7.3 (6.7–7.9) | 11 (9.7–11.9) | 49 (44–56) | 73 (68–84) | Yoshida (2004) |
| S. tielingense | 915 (824–979) | 35 (32–38) | 69 (64–73) | 98 (90–105) | 128 (120–135) | 81 (74–85) | 26 (23–28) | 7 (6–8) | 11 (9–13) | 55 (47–61) | 88 (85–94) | Ma et al. (2012) |
| S. kraussei | 951 (797–1,102) | 33 (30–36) | 63 (50–66) | 105 (99–111) | 134 (119–145) | 79 (63–86) | 29 (NA) | 7.1 (NA) | 12.1 (NA) | 47 (NA) | 80 (NA) | Nguyen et al. (2007) |
| S. oregonense | 980 (820–1,110) | 34 (28–38) | 66 (60–72) | NA | 132 (116–148) | 70 (64–78) | 30 (24–37) | 7.6 (6–8) | 14 (12–16) | 50 (40–60) | 100 (90–110) | Liu and Berry (1996) |
2 Notes: Measurements are given in μm and in the form: mean (range). aabbreviations as in Table 1, NA = data not available.
Table 3.
Comparative morphometrics of first-generation males of S. sandneri n. sp. and related Steinernema spp.
| Morphometric character a | ||||||||
|---|---|---|---|---|---|---|---|---|
| Species | SL | GL | W | D% | SW% | GS% | MUC b | n |
| S. sandneri n. sp. | 60 (53–65) | 44 (39–50) | 155 (124–178) | 51 (42–59) | 111 (97–127) | 79 (61-–83) | P | 25 |
| S. akhursti | 90 (85–100) | 64 (58–68) | 131 (115–150) | 56 (52–61) | 180 (140–200) | 71 (65–77) | P | 20 |
| S. cholashanense | 66 (60–71) | 39 (32–45) | 137 (73–204) | 64 (50–85) | 115 (92–144) | 71 (61–85) | P | 20 |
| S. citrae | 65 (57–80) | 44 (32–59) | 103 (87–113) | 58 (47–67) | 198 (156–233) | 68 (48–89) | P | 20 |
| S. costaricense | 92 (81–101) | 46 (41–51) | 128 (89–157) | 53 (51–66) | 160 (150–170) | 49 (45–55) | A | 19 |
| S. feltiae | 70 (65–77) | 41 (34–47) | 75 (60–90) | 60 (51–64) | 113 (99–130) | 59 (52–61) | P | 25 |
| S. hebeiense | 57 (51–63) | 46 (38–50) | 86 (74–98) | 51 (48–59) | 140 (120–170) | 80 (60–90) | A | 20 |
| S. ichnusae | 66 (64–67) | 44 (43–46) | 137 (73–204) | 62 (59–65) | 139 (120–162) | 67 (64–69) | A | 20 |
| S. jollieti | 64 (55–70) | 54 (45–60) | 115 (98–135) | 64 (53–83) | 145 (NA) | 84 (NA) | A | 12 |
| S. kraussei | 49 (42–53) | 33 (29–37) | 128 (110–144) | 53 (NA) | 110 (NA) | 67 (NA) | P | NA |
| S. kushidai | 63 (48–72) | 44 (39–60) | 97 (75–156) | 51 (42–59) | 150 (NA) | 70 (NA) | A | 20 |
| S. litorale | 75 (67–89) | 53 (44–64) | 96 (82–111) | 40 (34–56) | 174 (154–200) | 71 (62–81) | P | 25 |
| S. nguyeni | 66 (58–75) | 43 (30–55) | 82 (58–106) | 48 (38–57) | 215 (185–279) | 66 (46–81) | P | 20 |
| S. oregonense | 71 (65–73) | 56 (52–59) | 138 (105–161) | 73 (64–75) | 151 (NA) | 79 (NA) | A | 20 |
| S. puntauvense | 77 (71–81) | 34 (30–40) | 119 (101–139) | 67 (45–85) | 170 (140–200) | 65 (55–75) | P | 19 |
| S. sangi | 63 (58–80) | 40 (34–46) | 159 (120–225) | 49 (42–63) | 150 (120–160) | 60 (50–70) | P | 20 |
| S. silvaticum | 51 (42–64) | 37 (30–43) | 65 (52–78) | 60 (45–63) | NA | NA | P | 26 |
| S. texanum | 60 (55–66) | 45 (39–53) | 99 (81–116) | 67 (58–73) | 157 (127–203) | 75 (62–84) | A | 20 |
| S. tielingense | 88 (79–98) | 62 (49–70) | 129 (111–159) | 71 (64–78) | 191 (176–212) | 73 (59–82) | A | 20 |
| S. weiseri | 68 (62–72) | 53 (46–57) | 112 (84–138) | 49 (39–60) | 180 (150–240) | 80 (70–85) | A | 20 |
| S. xinbinense | 56 (49–62) | 35 (30–41) | 103 (90–126) | 45 (41–50) | 137 (114–156) | 63 (54–72) | P | 20 |
| S. xueshanense | 76 (66–91) | 49 (41–60) | 144 (97–159) | 80 (73–87) | 152 (93–172) | 64 (58–95) | A | 20 |
3 Notes: Measurements are given in μm and in the form: mean (range). aabbreviations as in Table 1. bMUC = mucron; P = present, A = absent, NA = data not available.
Table S2.
Steinernema sandneri n. sp. – permanent slides description and designation numbers in the collection of Museum and Institute of Zoology, Polish Academy of Sciences, Warsaw, Poland.
| Slide description | Slide ID |
|---|---|
| Slide no. 1 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae) (male), holotype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/1 |
| Slide no. 2 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 55 infective juveniles, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/2 |
| Slide no. 3 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 36 infective juveniles, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/3 |
| Slide no. 4 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 10 (males), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E, in 2017 | MIZ PAN WARSZAWA 2-2021/4 |
| Slide no. 5 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 10 (males), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E, in 2017 | MIZ PAN WARSZAWA 2-2021/5 |
| Slide no. 6 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 10 (males), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E, in 2017 | MIZ PAN WARSZAWA 2-2021/6 |
| Slide no. 7 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 10 (males), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E, in 2017 | MIZ PAN WARSZAWA 2-2021/7 |
| Slide no. 8 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 10 (males), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/8 |
| Slide no. 9 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 5 (females), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/9 |
| Slide no. 10 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 5 (females), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/10 |
| Slide no. 11 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 5 (females), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/11 |
| Slide no. 12 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 5 (females), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E, in 2017 | MIZ PAN WARSZAWA 2-2021/12 |
| Slide no. 13 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 5 (females), first generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/13 |
| Slide no. 14 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 13 (females), second generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/14 |
| Slide no. 15 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 15 (females), second generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/15 |
| Slide no. 16 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 9 (females), second generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/16 |
| Slide no. 17 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 13 (males), second generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/17 |
| Slide no. 18 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 15 (males), second generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/18 |
| Slide no. 19 – Steinernema sandneri n. sp. (Rhabditida: Steinernematidae), 15 (males), second generation, paratype, natural host unknown, isolated from soil samples: 51°46’55”N 22°42’35”E in 2017 | MIZ PAN WARSZAWA 2-2021/19 |
Table S3.
Percentage of similarity (upper triangle) and genetic distance measured by the number of nucleotide substitutions (lower triangle) in the sequences of ITS rDNA regions of S. sandneri n. sp., S. kraussei and S. silvaticum isolates, the closest relatives.
| Species | Acc. no. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | S. sandneri | MW078536 | – | 97.0 | 97.7 | 97.7 | 97.5 | 97.5 | 97.2 | 97.7 | 97.5 | 96.6 | 96.5 | 96.3 | 96.0 | 95.0 | 94.9 | 94.5 |
| 2 | S. kraussei | AY230174 | 21 | – | 98.1 | 98.1 | 97.9 | 97.9 | 98.1 | 97.2 | 95.7 | 98.1 | 98.2 | 97.5 | 97.5 | 95.4 | 95.3 | 94.9 |
| 3 | S. kraussei | AY171270 | 17 | 13 | – | 99.2 | 98.5 | 98.5 | 99.6 | 98.6 | 98.2 | 100 | 99.3 | 99.4 | 98.3 | 96.1 | 96.0 | 95.6 |
| 4 | S. kraussei | AY171248 | 16 | 12 | 5 | – | 99.0 | 99.0 | 99,0 | 99.0 | 97.3 | 99.2 | 98.9 | 98.6 | 97.6 | 95.7 | 95.6 | 95.2 |
| 5 | S. kraussei* | KY819012 | 18 | 14 | 11 | 6 | – | 99.7 | 98.2 | 99.6 | 96.6 | 98.5 | 98.9 | 97.9 | 97.2 | 95.3 | 95.2 | 94.8 |
| 6 | S. kraussei | AY171264 | 18 | 14 | 11 | 6 | 2 | – | 98.2 | 99.9 | 86.6 | 98.5 | 99.2 | 97.9 | 96.9 | 95.3 | 95.2 | 94.8 |
| 7 | S. kraussei | AB243442 | 20 | 13 | 3 | 6 | 13 | 13 | – | 98.2 | 97.8 | 99.6 | 98.9 | 99.0 | 98.3 | 96.3 | 96.1 | 95.7 |
| 8 | S. kraussei | AY230175 | 17 | 14 | 10 | 6 | 3 | 1 | 13 | – | 96.8 | 98.6 | 99.3 | 98.1 | 96.9 | 95.5 | 95.3 | 94.9 |
| 9 | S. kraussei | JN683825 | 17 | 14 | 1 | 6 | 12 | 12 | 4 | 11 | – | 99.2 | 98.5 | 99.5 | 98.3 | 96.1 | 96.0 | 95.6 |
| 10 | S. kraussei | AY171258 | 17 | 13 | 0 | 5 | 11 | 11 | 3 | 10 | 1 | – | 99.3 | 98.6 | 98.3 | 96.1 | 96.0 | 95.6 |
| 11 | S. kraussei | AY230176 | 18 | 12 | 5 | 7 | 8 | 6 | 8 | 5 | 6 | 5 | – | 98.0 | 97.6 | 95.6 | 95.5 | 95.0 |
| 12 | S. kraussei* | MW647848 | 20 | 16 | 3 | 8 | 14 | 14 | 6 | 13 | 4 | 3 | 8 | – | 97.8 | 95.6 | 95.5 | 95.0 |
| 13 | S. kraussei | AY230161 | 28 | 15 | 11 | 15 | 19 | 21 | 11 | 21 | 10 | 11 | 16 | 14 | – | 95.3 | 95.1 | 94.7 |
| 14 | S. silvaticum | MG543845 | 34 | 30 | 26 | 28 | 32 | 32 | 25 | 31 | 24 | 26 | 30 | 29 | 31 | – | 99.9 | 99.4 |
| 15 | S. silvaticum | AY171255 | 35 | 31 | 27 | 29 | 33 | 33 | 26 | 32 | 25 | 27 | 31 | 30 | 32 | 2 | – | 99.6 |
| 16 | S. silvaticum | AY230162 | 37 | 33 | 29 | 31 | 35 | 35 | 28 | 34 | 26 | 29 | 32 | 32 | 34 | 3 | 2 | – |
Table 4.
Percentage of similarity (upper triangle) and genetic distance measured by the number of nucleotide substitutions (lower triangle) in the sequences of ITS rDNA of S. sandneri n. sp. and other closely related Steinernema spp.
| Species | Acc. no. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | S. sandneri n. sp. | MW078536 | – | 97.0 | 94.5 | 92.4 | 90.2 | 94.1 | 94.1 | 94.5 | 89.6 | 89.8 | 89.0 | 89.4 | 90.3 | 88.7 | 85.8 | 76.6 |
| 2 | S. kraussei | AY230174 | 21 | – | 95.2 | 92.1 | 90.4 | 90.5 | 93.8 | 94.9 | 88.6 | 89.2 | 88.6 | 89.1 | 89.4 | 88.0 | 85.2 | 76.0 |
| 3 | S. silvaticum | AY230162 | 37 | 31 | – | 92.0 | 90.4 | 90.2 | 92.6 | 94.6 | 88.8 | 89.4 | 89.2 | 88.9 | 89.6 | 88.0 | 84.8 | 75.8 |
| 4 | S. cholashanense | EF431959 | 51 | 48 | 50 | – | 94.2 | 97.2 | 93.2 | 94.2 | 91.3 | 92.7 | 91.7 | 92.8 | 92.2 | 91.8 | 86.6 | 77.5 |
| 5 | S. oregonense | AF122019 | 59 | 56 | 59 | 33 | – | 93.5 | 92.0 | 92.1 | 89.9 | 91.5 | 89.7 | 91.2 | 90.4 | 90.4 | 86.8 | 76.6 |
| 6 | S. xueshanense | FJ660052 | 67 | 59 | 63 | 17 | 41 | – | 91.3 | 92.1 | 90.3 | 91.2 | 89.0 | 91.2 | 89.8 | 89.9 | 85.6 | 76.9 |
| 7 | S. tielingense | GU994201 | 41 | 42 | 51 | 42 | 48 | 56 | – | 94.8 | 89.9 | 89.5 | 89.7 | 90.5 | 90.1 | 89.2 | 86.2 | 77.2 |
| 8 | S. xinbinense | JN171593 | 35 | 34 | 36 | 35 | 47 | 50 | 34 | – | 90.8 | 90.8 | 90.2 | 91.5 | 91.0 | 90.6 | 86.6 | 77.0 |
| 9 | S. feltiae | AF121050 | 73 | 73 | 74 | 57 | 60 | 61 | 68 | 60 | – | 95.8 | 91.4 | 94.0 | 93.4 | 94.4 | 88.9 | 76.1 |
| 10 | S. ichnusae | EU421129 | 71 | 70 | 71 | 50 | 52 | 55 | 71 | 62 | 28 | – | 91.8 | 95.5 | 93.9 | 85.2 | 88.6 | 76.9 |
| 11 | S. jollieti | AY171265 | 76 | 73 | 70 | 53 | 61 | 62 | 67 | 63 | 54 | 46 | – | 91.1 | 90.0 | 91.1 | 86.1 | 77.3 |
| 12 | S. weiseri | AY171268 | 72 | 69 | 73 | 48 | 52 | 55 | 62 | 55 | 39 | 32 | 51 | – | 94.5 | 96.6 | 89.0 | 77.0 |
| 13 | S. nguyeni | KP325084 | 67 | 69 | 70 | 53 | 60 | 61 | 67 | 61 | 47 | 43 | 56 | 39 | – | 92.9 | 87.9 | 76.4 |
| 14 | S. litorale | AB243441 | 78 | 78 | 80 | 57 | 59 | 61 | 73 | 63 | 38 | 34 | 46 | 24 | 44 | – | 88.7 | 77.2 |
| 15 | S. hebeiense | DQ105794 | 98 | 97 | 102 | 92 | 84 | 90 | 93 | 90 | 76 | 80 | 82 | 77 | 81 | 74 | – | 74.9 |
| 16 | S. monticolum | AF122017 | 140 | 139 | 142 | 133 | 134 | 133 | 132 | 134 | 143 | 135 | 120 | 134 | 132 | 132 | 143 | – |
Table 5.
Percentage of similarity (upper triangle) and genetic distance measured by the number of nucleotide substitutions (lower triangle) in the sequences of D2D3 domain of 28 S rDNA of S. sandneri n. sp. and other closely related Steinernema spp.
| Species | Acc. no. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | S. sandneri n. sp. | MW078535 | – | 98.2 | 98.2 | 98.2 | 96.2 | 97.5 | 97.5 | 96.8 | 96.8 | 96.6 | 96.1 | 97.0 | 95.7 | 92.4 |
| 2 | S. kraussei | AF331896 | 15 | – | 99.8 | 99.8 | 97.2 | 98.2 | 98.9 | 97.6 | 98.2 | 97.9 | 97.4 | 98.1 | 96.2 | 92.6 |
| 3 | S. kraussei | GU569053 | 15 | 2 | – | 99.5 | 97.2 | 98.6 | 99.1 | 97.8 | 98.5 | 98.1 | 97.6 | 98.1 | 96.6 | 92.7 |
| 4 | S. kraussei * | MW647849 | 15 | 2 | 4 | – | 97.2 | 98.2 | 98.9 | 97.7 | 98.2 | 97.8 | 97.3 | 97.0 | 95.7 | 92.9 |
| 5 | S. silvaticum | MG547576 | 32 | 24 | 24 | 24 | – | 97.2 | 96.8 | 96.1 | 96.6 | 96.1 | 95.7 | 96.7 | 94.7 | 91.1 |
| 6 | S. cholashanense | EF520284 | 21 | 14 | 12 | 14 | 24 | – | 98.4 | 97.5 | 98.1 | 97.7 | 97.2 | 97.8 | 96.4 | 92.7 |
| 7 | S. oregonense | AF331891 | 21 | 9 | 7 | 11 | 27 | 14 | – | 97.9 | 98.7 | 98.2 | 98.0 | 98.6 | 96.9 | 93.4 |
| 8 | S. xueshanense | FJ666053 | 27 | 19 | 19 | 19 | 33 | 20 | 18 | – | 98.0 | 97.9 | 97.3 | 98.1 | 96.6 | 92.7 |
| 9 | S. feltiae | AF3311906 | 27 | 15 | 13 | 17 | 29 | 16 | 11 | 17 | – | 99.3 | 98.8 | 99.4 | 97.2 | 93.3 |
| 10 | S. ichnusae | EU421130 | 28 | 18 | 16 | 18 | 32 | 19 | 14 | 17 | 5 | – | 98.6 | 98.9 | 97.2 | 93.3 |
| 11 | S. jollieti | GU569051 | 32 | 22 | 20 | 24 | 36 | 23 | 16 | 22 | 9 | 12 | – | 98.9 | 96.7 | 93.1 |
| 12 | S. weiseri | GU569059 | 26 | 16 | 16 | 18 | 28 | 19 | 12 | 16 | 5 | 8 | 9 | – | 97.1 | 93.7 |
| 13 | S. texanum | EF152569 | 37 | 31 | 29 | 31 | 45 | 30 | 26 | 28 | 24 | 23 | 27 | 25 | – | 93.1 |
| 14 | S. monticolum | EF439651 | 56 | 53 | 53 | 54 | 69 | 56 | 49 | 56 | 51 | 52 | 54 | 49 | 53 | – |
Table 6.
Percentage of similarity (upper triangle) and genetic distance measured by the number of nucleotide substitutions (lower triangle) in the sequences of cox1 gene of S. sandneri n. sp. and other closely related Steinernema spp.
| Species | Acc. no. | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 1 | S. sandneri n. sp. | MW078544 | – | 93.7 | 93.8 | 92.9 | 87.7 | 86.6 | 88.4 | 84.7 | 86.4 | 85.9 | 85.7 |
| 2 | S. kraussei | JN683829 | 36 | – | 94.4 | 94.0 | 87.1 | 86.4 | 87.3 | 84.1 | 85.7 | 84.5 | 85.0 |
| 3 | S. kraussei | AY943990 | 35 | 32 | – | 95.6 | 87.8 | 86.8 | 86.7 | 85.0 | 86.2 | 85.2 | 85.4 |
| 4 | S. kraussei * | MW647850 | 40 | 34 | 25 | – | 86.2 | 86.4 | 86.9 | 83.6 | 85.7 | 84.5 | 85.4 |
| 5 | S. silvaticum | MG547572 | 70 | 73 | 68 | 78 | – | 84.7 | 85.4 | 83.6 | 83.5 | 82.9 | 84.7 |
| 6 | S. oregonense | AY943995 | 76 | 77 | 75 | 77 | 87 | – | 87.8 | 85.5 | 86.6 | 83.4 | 86.8 |
| 7 | S. feltiae | JQ423217 | 66 | 72 | 75 | 74 | 83 | 69 | – | 85.9 | 88.0 | 84.0 | 96.9 |
| 8 | S. jollieti | GU569068 | 87 | 90 | 85 | 93 | 93 | 82 | 80 | – | 85.0 | 84.7 | 84.1 |
| 9 | S. weiseri | GU569075 | 77 | 81 | 78 | 81 | 99 | 76 | 68 | 85 | – | 82.7 | 84.8 |
| 10 | S. kushidai | AY943991 | 80 | 88 | 84 | 88 | 97 | 94 | 91 | 87 | 98 | – | 84.5 |
| 11 | S. monticolum | AY943994 | 81 | 85 | 83 | 83 | 87 | 75 | 74 | 90 | 86 | 88 | – |
Figure 6:
Phylogenetic tree of the phylogenetic relationships of S. sandneri n. sp. with other species of the genus Steinernema based on sequences of the ITS rDNA. Bootstrap values > 50% are indicated at the branching points. The scale bar indicates the number of nucleotide substitutions per site. The evolutionary history was inferred using the Maximum Likelihood method based on the HKY + G model. All positions containing gaps were eliminated. There were a total of 646 positions in the final dataset. Evolutionary analyses were conducted in MEGA6.
Figure 7:
Phylogenetic tree of the phylogenetic relationships of S. sandneri n. sp. with other species of the genus Steinernema based on sequences of the D2D3 regions. Bootstrap values > 50% are indicated at the branching points. The scale bar indicates the number of nucleotide substitutions per site. The evolutionary history was inferred using the Maximum Likelihood method based on the GTR + G model. All positions containing gaps were eliminated. There were a total of 850 positions in the final dataset. Evolutionary analyses were conducted in MEGA6.
Figure 8:
Phylogenetic tree of the phylogenetic relationships of S. sandneri n. sp. with other species of the genus Steinernema based on cox1 gene sequences. Bootstrap values > 50% are indicated at the branching points. The scale bar indicates the number of nucleotide substitutions per site. The evolutionary history was inferred using the Maximum Likelihood method based on the HKY + G + I model. All positions containing gaps were eliminated. There were a total of 567 positions in the final dataset. Evolutionary analyses were conducted in MEGA6.