Figure 1:
Line drawings of first- and second-generation adults and infective juveniles of Steinernema anantnagense n. sp. (A–C) First-generation female: (A) Anterior end; (B) Neck region; (C) Posterior end. (D–F) First-generation male: D) Anterior end; (E) Neck region; (F) Posterior end. (G–I) Second-generation female: (G) Anterior end; (H) Neck region; (I) Posterior end. (J–L) Second-generation male: (J) Anterior end; (K) Neck region; (L) Posterior end. (M–O) Infective juvenile: (M) Anterior end; (N) Neck; (O) Posterior end.
Figure 2:
Light microscope micrographs of first-generation adults and IJ of Steinernema anantnagense n. sp. (A–C) Female: (A) Anterior end; (B) Neck region (arrow pointing to the excretory pore); (C) Posterior end. (D–F) Male: (D) Anterior end; (E) Neck region (arrow pointing to the excretory pore); (F) Posterior end (arrow pointing to the mid-ventral genital papillae). (G–I) Infective juvenile: (G) Anterior end; (H) Neck region (black arrow pointing to the excretory pore, white arrow pointing to the bacteria sac); (I) Posterior end.
Figure 3:
Light microscope micrographs of second-generation adults and IJ of Steinernema anantnagense n. sp. (A–C) Second-generation female: (A) Anterior end; (B) Neck region; (C) Posterior end. (D–F) Second-generation male: D) Anterior end; (E) Neck region; (F) Posterior end (arrow pointing to the mid-ventral genital papillae). (G) Entire infective juvenile (arrow pointing to the bacteria sac).
Figure 4:
Scanning electron microscope micrographs of first-generation adults of Steinernema anantnagense n. sp. (A, B) Lip region of female in frontal and lateral view, respectively (arrows pointing to the amphids); (C) Excretory pore of female; (D) Vulva in ventral view; (E) Female posterior end (arrow pointing to the anus). (F–H) Male posterior end in lateral, ventral and subventral view, respectively (GP: genital papilla, MP: mid-ventral papilla, ph: phasmid). (I, J) Lip region of IJ in frontal and lateral view, respectively (arrows pointing to the amphids); (K) Excretory pore of IJ; (L) Lateral field of IJ; (M, N) Posterior end of IJ in ventral and lateral view, respectively (arrows pointing to the phasmids).
Figure 5:
Scanning electron microscope micrographs of second-generation adults and IJ of Steinernema anantnagense n. sp. (A, B) Lip region of female in frontal and lateral view, respectively (arrows pointing to the amphids); (C) Excretory pore of female; (D) Vulva in ventral view; (E, F) Female posterior end in lateral and ventral view, respectively (arrow pointing to the anus). (G, H) Lip region of male in frontal and lateral view, respectively (arrows pointing to the amphids); (I) Male excretory pore; (J–L) Male posterior end in lateral, ventral and subventral view, respectively (GP: genital papilla, MP: mid-ventral papilla, ph: phasmid).
Table 1.
Morphometrics of the IJs and adult generations of Steinernema anantnagense n. sp. (Steiner_7). All characters are in µm (except n, ratios and percentages) and given as mean ± s.d. (range).
| Characters | First Generation | Second Generation | Infective Juveniles (paratypes) | |||
|---|---|---|---|---|---|---|
| Male (holotype) | Male (paratypes) | Female (paratypes) | Male (paratypes) | Female (paratypes) | ||
| n | 1 | 20 | 20 | 20 | 20 | 20 |
| Body length (L) | 1279 | 1618 ± 246 (1223–1899) | 3765 ± 441 (2327–4872) | 1068 ± 76 (899–1168) | 2081 ± 200 (1761–2437) | 789 ± 35 (749–834) |
| a (L/BD) | 6.6 | 8.3 ± 1.1 (6.4–9.8) | 23 ± 1.2 (17–29) | 19 ± 2.6 (15.1–25.3) | 14.7 ± 1.5 (12.1–17.4) | 22 ± 1.9 (19–24) |
| b (L/NL) | 7.0 | 9.2 ± 1.4 (7.0–11.3) | 17.0 ± 2.4 (13–21) | 7.1 ± 0.6 (5.9–8.2) | 11.0 ± 1.0 (9.6–13.5) | 6.0 ± 0.4 (5.5–6.7) |
| c (L/T) | 35 | 49 ± 9.0 (34–64) | 100 ± 18 (61–122) | 43 ± 5.8 (31–52) | 54 ± 6.6 (43–67) | 13.8 ± 1.8 (12.2–16.4) |
| c’ (T/ABW) | 1.2 | 1.1 ± 0.2 (0.8–1.6) | 0.7 ± 0.1 (0.5–0.8) | 0.9 ± 0.2 (0.6–1.4) | 1.0 ± 0.1 (0.8–1.3) | 1.8 ± 0.2 (1.6–2.1) |
| V (VA/L×100) | – | 53 ± 1.7 (50–58) | – | 53 ± 1.7 (50–57) | – | |
| Max. body diam. (MBD) | 193 | 194 ±11.3 (167–211) | 434 ± 30 (314–409) | 57 ± 7.3 (42–63) | 143 ± 16 (123–173) | 37 ± 3.6 (32–42) |
| Lip region width | 10.2 | 10.1 ± 1.9 (8.2–12.2) | 14.4 ± 1.9 (11.1–18.3) | 8.4 ± 1.6 (6.1–12.1) | 11.4 ± 1.7 (8.9–14.6) | 4.6 ± 2.0 (3.7–5.8) |
| Stoma length | 12.3 | 13.8 ± 1.6 (10.1–16.9) | 16.4 ± 2.4 (12.4–22.1) | 11.7 ± 1.1 (10.1–14.1) | 14.1 ± 2.3 (11.1–18.1) | 10.7 ± 2.0 (8.6–13.8) |
| Procorpus length | 66 | 57 ± 4.1 (51–66) | 65 ± 6.0 (55–76) | 47 ± 3.9 (41–53) | 54 ± 3.8 (49–65) | 43 ± 5.4 (37–52) |
| Metacorpus length | 33 | 34 ± 1.7 (30–36) | 44 ± 4.6 (38–50) | 30 ± 2.4 (25–35) | 37 ± 2.4 (33–43) | 26 ± 2.7 (23–29) |
| Isthmus length | 40 | 42 ± 4.0 (37–50) | 53 ± 6.4 (45–68) | 37 ± 1.5 (35–40) | 49 ± 3.7 (43–55) | 36 ± 2.8 (31–39) |
| Bulb length (BL) | 31 | 30 ± 2.2 (25–35) | 43 ± 4.5 (33–50) | 25 ± 2.7 (21–30) | 35 ± 2.9 (30–39) | 17 ± 1.8 (16–20) |
| Bulb width (EBW) | 26 | 25 ± 1.6 (21–27) | 35 ± 3.5 (27–41) | 21 ± 1.9 (18–25) | 28 ± 2.7 (25–33) | 11 ± 1.7 (9.1–13.9) |
| Pharynx length (PL) | 170 | 162 ± 5.9 (150–172) | 206 ± 14 (182–232) | 139 ± 5.0 (129–146) | 175 ± 6.4 (164–191) | 121 ± 8.0 (109–133) |
| Nerve ring – ant. end (NR) | 109 | 112 ± 6.8 (103–129) | 165 ± 12.1 (143–182) | 91 ± 7.0 (75–104) | 109 ± 11.9 (88–127) | 63 ± 8.4 (54–71) |
| Excretory pore – ant. end (EP) | 114 | 111 ± 10 (88–124) | 112 ± 11.9 (92–140) | 80 ± 9.2 (62–98) | 92 ± 8.1 (82–112) | 55 ± 6.7 (45–62) |
| Width at excretory pore (WEP) | 58 | 61 ± 3.4 (49–65) | 137 ± 11.3 (123–170) | 35 ± 3.6 (30–42) | 60 ± 6.7 (51–74) | 22 ± 2.4 (19–25) |
| Neck length (stoma+pharynx, NL) | 182 | 176 ± 6.4 (165–185) | 223 ± 14 (196–255) | 151 ± 5.0 (142–159) | 189 ± 6.5 (177–204) | 132 ± 8.3 (120–143) |
| Body width at neck base | 79 | 86 ± 5.8 (76–96) | 217 ± 23 (195–264) | 41 ± 3.9 (36–50) | 95 ± 8.5 (82–107) | 29 ± 2.9 (24–32) |
| Testis reflexion | 630 | 631 ± 37 (576–689) | – | 522 ± 32 (481–568) | – | – |
| Vagina length | – | – | 31 ± 3.1 (24–36) | – | 24 ± 3.3 (19–30) | – |
| Body width at vulva | – | – | 386 ± 53 (309–498) | – | 164 ± 25 (124–193) | – |
| Vulva – ant. end (VA) | – | – | 1989 ± 214 (1247–2252) | – | 1105 ± 101 (961–1295) | – |
| Vulva – post. end (VP) | – | – | 1776 ± 242 (1080–2038) | – | 976 ± 110 (801–1161) | – |
| Rectum length | – | – | 35 ± 4.0 (29–44) | – | 21 ± 2.5 (17–24) | 13.8 ± 1.9 (11–16) |
| Anal body diam. (ABD) | 31 | 32 ± 3.9 (25–36) | 86 ± 7.1 (77–110) | 29 ± 4.1 (21–35) | 41 ± 6.8 (31–53) | 20 ± 1.7 (18–22) |
| Tail length (T) | 37 | 34 ± 2.4 (29–39) | 38 ± 4.2 (32–49) | 26 ± 3.9 (21–35) | 49 ± 3.9 (38–53) | 58 ± 6.7 (49–66) |
| Hyaline part of tail (H) | – | – | – | – | – | 16.3 ± 4.1 (11.8–23.7) |
| Spicule length (SL) | 66 | 64 ± 4.6 (56–70) | – | 44 ± 2.6 (40–49) | – | – |
| Gubernaculum length (GL) | 34 | 36 ± 3.8 (31–43) | – | 25 ± 2.3 (21–29) | – | – |
| Stoma length/lip region width | 1.2 | 1.4 ± 0.2 (0.9–1.8) | 1.2 ± 0.2 (0.9–1.8) | 1.4 ± 0.3 (0.9–1.9) | 1.3 ± 0.3 (0.9–1.8) | 2.4 ± 0.5 (1.6–3.0) |
| Nerve ring % (NR/NL×100) | 60 | 64 ± 5.3 (56–78) | 74 ± 7.1 (64–86) | 61 ± 4.6 (52–68) | 58 ± 6.6 (46–67) | 48 ± 7.8 (38–58) |
| Excretory pore % (EP/NL×100) | 62 | 63 ± 6.1 (50–74) | 50 ± 4.6 (42–60) | 53 ± 6.6 (41–68) | 49 ± 4.1 (43–58) | 42 ± 4.4 (35–48) |
| Rectum% (R/ABD×100) | – | – | 0.4 ± 0.1 (0.3–0.6) | 0.5 ± 0.1 (0.3–0.8) | 0.7 ± 0.1 (0.6–0.9) | |
| D% (EP /NL×100) | 62 | 63 ± 6.1 (49–74) | 50 ± 4.9 (43–60) | 53 ± 6.6 (41–68) | 49 ± 4.1 (43–58) | 42 ± 4.4 (35–48) |
| E% (EP/T×100) | 311 | 333 ± 43 (256–403) | 5.7 ± 1.2 (4.5–9.5) | 319 ± 59 (219–460) | 240 ± 36 (201–362) | 96 ± 12.8 (74–113) |
| SW% (SL/ABD×100) | 217 | 208 ± 39 (154–297) | – | 2.3 ± 0.4 (1.8–3.4) | ||
| GS% (GL/SL×100) | 51 | 57 ± 6.7 (46–70) | – | 0.6 ± 0.1 (0.4–0.7) | ||
| H% (H/T ×100) | – | – | – | – | – | 28 ± 5.8 (20–36) |
[i] – = characters absent.
Table 2.
Comparison of morphometrics of the third-stage infective juveniles of Steinernema anantnagense n. sp. with other members of “Feltiae-Kushidai” clade. Measurements are in μm except n, ratio and percentage. Data for new species is in bold.
| Species | Reference | Country | n | L | BD | EP | NR | NL | T | a | b | C | c′ | D% | E% | H% |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| S. akhursti | Qiu et al. (2005) | China | 20 | 770–835 | 33–35 | 55–60 | 83–95 | 115–123 | 68–75 | 23–26 | 6.6–7.2 | 10–12 | 3.3–3.7 | 45–50 | 73–86 | 49–56 |
| S. anantnagense n. sp. | Present Study | India | 20 | 749–834 | 32–42 | 45–62 | 54–71 | 120–143 | 49–66 | 19–24 | 5.5–6.7 | 12–16 | 1.6–2.1 | 35–48 | 74–113 | 20–36 |
| S. africanum | Machado et al. (2022) | Rwanda | 15 | 690–802 | 25–32 | 54–68 | 87–132 | 123–167 | 52–72 | 23–30 | 4.3–6.3 | 10–15 | 2.9–4.2 | 34–46 | 79–129 | 28–39 |
| S. cholashanense | Nguyen et al. (2008) | China | 20 | 727–909 | 26–35 | 59–65 | 72–97 | 110–138 | 60–80 | 24–34 | 6.1–7.2 | 10–14 | 3.5–5.0 | 46–53 | 76–91 | 33–47 |
| S. citrae | Stokwe et al. (2011) | South Africa | 20 | 623–849 | 23–28 | 49–64 | 83–108 | 118–137 | 63–81 | 25–34 | 5.1–7.1 | 13–14 | 13–17 | 39–58 | 85–132 | 37–50 |
| S. feltiae | Nguyen et al. (2007) | Russia | 25 | 766–928 | 22–32 | 58–67 | 108–117 | 130–143 | 81–89 | 27–34 | 5.8–6.8 | 9.4–11 | 4.5–5.1 | 44–50 | 67–81 | 37–51 |
| S. hebeiense | Chen et al. (2006) | China | 20 | 610–710 | 23–28 | 43–51 | 73–83 | 100–111 | 63–71 | 24–28 | 5.7–6.7 | 9.4–11 | NA | 40–50 | 65–80 | 32–50 |
| S. ichnusae | Tarasco et al. (2008) | Italy | 20 | 767–969 | 27–35 | 59–68 | 94–108 | 119–148 | 76–89 | 24–32 | 5.6–6.9 | 9–12 | 4.2–5.1 | 42–49 | 68–83 | 44–50 |
| S. jollieti | Spiridonov et al. (2004) | USA | 12 | 625–820 | 20–28 | 53–65 | NA | 115–135 | 60–73 | 25–34 | 4.9–6.4 | 9–12 | NA | 46–50 | NA | 46–60 |
| S. kraussei | Nguyen et al. (2007) | Germany | 25 | 797–1102 | 30–36 | 50–66 | 99–111 | 119–145 | 63–86 | NA | NA | NA | NA | NA | NA | 35–40 |
| S. kushidai | Mamiya (1998) | Japan | 20 | 424–662 | 22–31 | 42–50 | 70–84 | 106–120 | 44–59 | 19–25 | 4.9–5.9 | 10–13 | NA | 38–44 | NA | NA |
| S. litorale | Yoshida (2004) | Japan | 25 | 834–988 | 28–33 | 54–69 | 89–104 | 114–133 | 72–91 | 27–31 | 6.7–7.9 | 10–11.9 | 3.8–5.4 | 44–56 | 68–84 | NA |
| S. nguyeni | Malan et al. (2016) | South Africa | 20 | 673–796 | 22–28 | 47–58 | 74–86 | 101–121 | 61–73 | 27–33 | 6.2–7.4 | 10–12 | 2.8–4.8 | 43–57 | 70–86 | 20–31 |
| S. oregonese | Liu and Berry (1996) | USA | 20 | 820–1110 | 28–38 | 60–72 | NA | 116–148 | 64–78 | 24–37 | 6–8 | 12–16 | NA | 40–60 | 90–110 | 30–33 |
| S. populi | Tian et al. (2022) | China | 25 | 973–1172 | 33–41 | 70–86 | 98–113 | 134–159 | 55–72 | 28–33 | 6.8–7.5 | 15–20 | 2.4–3.3 | 47–61 | 105–140 | 26–44 |
| S. puntauvense | Uribe-Lorío et al. (2007) | Costa Rica | 19 | 631–728 | 31–38 | 20–30 | 46–69 | 81–103 | 51–59 | 17–23 | 7.1–7.9 | 11–13 | NA | 25–50 | 35–56 | 52–55 |
| S. sandneri | Lis et al. (2021) | Poland | 25 | 708–965 | 23–32 | 44–64 | 83–118 | 123–151 | 64–86 | 27–34 | 5.5–6.9 | 11–13 | NA | 36–45 | 63–86 | 23–40 |
| S. sangi | Phan et al. (2001) | Vietnam | 20 | 704–784 | 30–40 | 46–54 | 78–97 | 120–138 | 76–89 | 19–25 | 5.6–6.3 | 9–10 | NA | 36–44 | 56–70 | 44–52 |
| S. silvaticum | Sturhan et al. (2005) | Germany | 26 | 670–975 | 26–35 | 51–73 | 75–109 | 100–141 | 63–86 | 23–33 | 6.3–7.7 | 10–13 | 3.1–4.9 | 46–56 | – | 37–53 |
| S. tielingense | Ma et al. (2012) | China | 20 | 824–979 | 32–38 | 64–73 | 90–105 | 120–135 | 74–85 | 27–31 | 6.7–7.9 | 10–12 | 3.5–4.6 | 44–56 | 68–84 | 53–64 |
| S. texanum | Nguyen et al. (2007) | USA | 20 | 732–796 | 29–34 | 52–62 | 84–102 | 111–120 | 60–79 | 22–27 | 6.2–7.0 | 10–13 | 3.3–4.6 | 46–53 | 76–88 | 53–69 |
| S. xinbinense | Ma et al. (2012) | China | 20 | 635–744 | 28–31 | 46–53 | 75–90 | 109–125 | 65–78 | 21–25 | 5–7 | 8–11 | 3–5 | 40–47 | 65–78 | 30–42 |
| S. xueshanense | Mrácek et al. (2009) | China | 20 | 768–929 | 29–33 | 60–72 | 81–96 | 130–143 | 80–92 | 26–32 | 5.8–7.0 | 9–11 | 3.8–5.1 | 46–52 | 70–90 | 46–55 |
| S. weiseri | Mrácek et al. (2003) | Czech Republic | 20 | 586–828 | 24–29 | 43–65 | 72–92 | 95–119 | 49–68 | 25–33 | 5.7–7.2 | 10–14 | 3.2–4.1 | 44–55 | NA | 18–24 |
[i] NA = Not available; P = Present; A = Absent.
Table 3.
Comparison of morphometrics of the first-generation males of Steinernema anantnagense n. sp. with other members of “Feltiae-Kushidai” clade. Measurements are in μm except n, ratio and percentage. Data for new species is in bold.
| Species | n | L | BD | EP | NR | NL | T | SL | GL | a | B | c | c′ | D% | SW% | GS% | Mucron |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| S. akhursti | 20 | 1350–1925 | 115–150 | 93–113 | 120–163 | 168–205 | 30–40 | 85–100 | 58–68 | NA | NA | NA | NA | 52–61 | 140–200 | 65–77 | P |
| S. anantnagense n. sp. | 20 | 1223–1899 | 167–211 | 88–124 | 103–129 | 165–185 | 29–39 | 56–70 | 31–43 | 6–10 | 7–11 | 34–64 | 0.8–1.6 | 49–74 | 154–297 | 46–70 | P |
| S. africanum | 15 | 977–1400 | 65–131 | 69–109 | 79–104 | 132–147 | 34–46 | 65–76 | 32–49 | 9–12 | 7–12 | 25–34 | 0.9–1.1 | 52–74 | 144–197 | 49–68 | P |
| S. cholashanense | 20 | 1070–1778 | 73–204 | 75–135 | 91–126 | 135–173 | 29–43 | 60–71 | 32–45 | 9–24 | 8–11 | 36–51 | 0.6–0.9 | 50–85 | 92–144 | 61–85 | P |
| S. citrae | 20 | 1028–1402 | 87–113 | 64–92 | 92–119 | 123–155 | 17–31 | 57–80 | 32–59 | NA | NA | NA | NA | 47–67 | 156–233 | 48–89 | P |
| S. feltiae | 25 | 1414–1817 | 60–90 | 110–126 | NA | 164–180 | 37–43 | 65–77 | 34–47 | NA | NA | NA | NA | 51–64 | 99–130 | 52–61 | P |
| S. hebeiense | 20 | 1036–1450 | 74–98 | 58–73 | 78–93 | 118–132 | 24–35 | 51–63 | 38–50 | 12–17 | 8–11 | 30–49 | 0.6–0.9 | 48–59 | 120–170 | 60–90 | A |
| S. ichnusae | 20 | 1151–1494 | 73–204 | 94–108 | NA | 135–173 | 33–48 | 64–67 | 43–46 | 20–29 | 7–9 | 29–39 | 0.8–0.9 | 59–65 | 120–162 | 64–69 | A |
| S. jollieti | 12 | 1296–1952 | 98–135 | 83–110 | NA | 110–168 | 24–38 | 55–70 | 45–60 | 12–19 | 8–14 | 53–86 | NA | 53–83 | NA | NA | A |
| S. kraussei | 20 | 1200–1600 | 110–144 | 73–99 | 95–122 | 137–178 | 36–44 | 42–53 | 29–37 | 11 | 9 NA | NA | NA | NA | NA | NA | P |
| S. kushidai | 20 | 1200–1900 | 75–156 | 71–105 | 120–137 | 156–189 | 30–40 | 48–72 | 39–60 | NA | NA | NA | NA | 42–59 | NA | NA | A |
| S. litorale | 25 | 1230–1514 | 82–111 | 77–107 | 94–128 | 133–163 | 26–41 | 67–89 | 44–64 | 12–16 | 8–10 | 33–56 | 0.6–0.9 | 34–56 | 154–200 | 62–81 | P |
| S. nguyeni | 20 | 818–1171 | 58–106 | 47–71 | 70–103 | 112–144 | 18–25 | 58–75 | 30–55 | 11–15 | 7–10 | 38–53 | 0.6–0.8 | 38–57 | 185–279 | 46–81 | P |
| S. oregonense | 20 | 1560–1820 | 105–161 | 95–139 | 101–133 | 139–182 | 24–32 | 65–73 | 52–59 | NA | NA | NA | 0.6 NA | 64–75 | NA | NA | A |
| S. populi | 25 | 1258–1514 | 66–95 | 95–121 | 107–143 | 131–177 | 39–68 | 57–77 | 38–60 | 15–20 | 8–10 | 20–33 | 0.8–1.5 | 59–78 | 107–160 | 58–82 | P/A |
| S. puntauvense | 19 | 1010–1931 | 101–139 | 68–114 | 104–128 | 130–159 | 28–40 | 71–81 | 30–40 | NA | NA | NA | NA | 45–85 | 140–200 | 55–75 | P |
| S. sandneri | 25 | 1206–1635 | 124–178 | 64–92 | 112–138 | 148–170 | 35–46 | 53–65 | 39–50 | 9–11 | 8–10 | 31–42 | NA | 42–59 | 97–127 | 61–83 | P |
| S. sangi | 20 | 1440–2325 | 120–225 | 67–99 | 109–166 | 150–221 | 27–42 | 58–80 | 34–46 | NA | NA | NA | NA | 42–63 | 120–160 | 50–70 | P |
| S. silvaticum | 26 | 975–1270 | 52–78 | 71–92 | 90–126 | 116–168 | 20–47 | 42–64 | 30–43 | 14–20 | 8–9 | 24–55 | 0.8–1.4 | 45–63 | NA | NA | P |
| S. tielingense | 20 | 1430–2064 | 111–159 | 94–133 | 96–132 | 145–173 | 22–33 | 79–98 | 49–70 | 11–18 | 9–13 | 57–85 | 0.3–0.6 | 64–78 | 176–212 | 59–82 | A |
| S. texanum | 20 | 1197–1406 | 81–116 | 79–100 | 94–114 | 123–147 | 19–30 | 55–66 | 39–53 | NA | NA | NA | NA | 58–73 | 127–203 | 62–84 | A |
| S. xinbinense | 20 | 1133–1440 | 90–126 | 57–75 | 91–120 | 138–159 | 30–41 | 49–62 | 30–41 | 11–13 | 7–9 | 31–39 | 0.7–1.0 | 41–50 | 114–156 | 54–72 | P |
| S. xueshanense | 20 | 1313–2040 | 97–159 | 113–137 | NA | 151–175 | 29–48 | 66–91 | 41–60 | NA | NA | NA | NA | 73–87 | 93–172 | 58–95 | A |
| S. weiseri | 20 | 990–1395 | 84–138 | 57–84 | 94–115 | 134–154 | 19–32 | 62–72 | 46–57 | 9–12 | 7–10 | 36–64 | 0.6–0.9 | 39–60 | 150–240 | 70–85 | A |
[i] NA = Not available; P = Present; A = Absent.
Table 4.
Comparison of morphometrics of the first-generation females of Steinernema anantnagense n. sp. with other members of “Feltiae-Kushidai” clade. Measurements are in μm except n, ratio and percentage. Data for new species is in bold.
| Species | L | BD | EP | NR | NL | T | A | B | c | c′ | V | ABD | D% | Mucron |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| S. akhursti | 5625–9000 | 200–270 | 113–138 | 150–175 | 213–258 | 38–63 | 30* | 32* | 149* | 0.6* | 48–53 | 68–100 | NA | P |
| S. anantnagense n. sp. | 2327–4872 | 314–409 | 92–140 | 143–182 | 196–255 | 32–49 | 17–29 | 13–21 | 61–122 | 0.5–0.8 | 50–58 | 77–110 | 43–60 | P |
| S. africanum | 2469–4215 | 154–194 | 67–111 | 79–130 | 170–201 | 35–55 | 13–27 | 13–24 | 51–104 | 0.7–1.0 | 50–57 | 37–70 | 32–62 | P |
| S. cholashanense | 3232–6363 | 156–332 | 111–148 | 176–223 | 181–231 | 46–70 | 13–23 | 18–32 | 62–119 | 0.6–1.0 | 50–57 | 54–105 | 29–65 | P |
| S. citrae | 2038–4019 | 137–212 | 54–90 | 130–179 | 189–220 | 33–60 | NA | NA | NA | NA | 50–59 | 43–79 | 27–46 | P |
| S. feltiae | 3095–3774 | 170–254 | 68–97 ** | 70–97 ** | 197–304 | 39–70 | 14–20 | 12–17 | 49–88 | 0.7–1.2 * | 44–57 | 47–62 | 40–54 * | P |
| S. hebeiense | 3972–4254 | 142–245 | 48–95 | 88–123 | 133–158 | 133–158 | 17–25 | 21–29 | 67–129 | 0.5–0.9 | 50–57 | 45–65 | 36–66 | A |
| S. ichnusae | 4547–6186 | 242–323 | 106–156 | NA | 215–262 | 51–79 | 17–24 | 21–26 | 68–113 | 0.6–1.0 | 51–57 | 70–94 | 47–63 | P |
| S. jollieti | 3746–6030 | 219–298 | 96–136 | NA | 184–310 | 31–55 | 15–24 | 19–31 | 72–185 | NA | 44–56 | NA | 52 | P |
| S. kraussei | 2500–5400 | 153–288 | 66–99 | 127–146 | 178–205 | 33–59 | 17 | 22 | 88 | NA | 54 | 39–50 | 45 | P |
| S. kushidai | 2100–4700 | 54–59 | 78–105 | 111–144 | 204–255 | 30–45 | NA | NA | NA | NA | 54–59 | 54–84 | 37–46 | A |
| S. litorale | 3930–5048 | 175–215 | 65–105 | 130–165 | 185–213 | 25–60 | 21–26 | 20–26 | 78–157 | 0.5–0.9 | 0.5–0.9 | 55–75 | 33–57 | P |
| S. nguyeni | 2290–5361 | 130–216 | 49–98 | 84–139 | 137–194 | 20–67 | 15–30 | 15–30 | 53–165 | 0.6–1.1 | 52–63 | 130–216 | 30–56 | A |
| S. oregonense | 4400–6200 | 217–268 | 217–268 | 129–162 | 186–220 | 28–46 | NA | NA | NA | NA | 46–56 | 42–79 | 43–57 | A |
| S. populi | 4038–13762 | 217–531 | 90–178 | 150–213 | 213–278 | 41–88 | 18–36 | 19–50 | 75–182 | 0.5–0.9 | 45–60 | 60–157 | 36–65 | A |
| S. puntauvense | 3687–8335 | 181–221 | 51–85 | 123–146 | 141–206 | 41–66 | NA | NA | NA | NA | 51–55 | 57–102 | 25–45 | P |
| S. sandneri | 4244–5014 | 181–261 | 61–102 | 132–158 | 173–194 | 32–61 | 17–25 | 24–27 | 75–140 | NA | 49–57 | 62–122 | 36–54 | P |
| S. sangi | 4830–7200 | 270–360 | 80–121 | 140–170 | 216–240 | 36–62 | NA | NA | NA | NA | 43–530 | 84–140 | 35–51 | P |
| S. silvaticum | 1520–3290 | 50–175 | 50–175 | 50–175 | 121–188 | 33–79 | 15–41 | 10–18 | 34–80 | 1.0–1.8 | 44–57 | 26–53 | 33–79 | A |
| S. texanum | 2720–3623 | 130–202 | 78–107 | 111–135 | 160–189 | 30–52 | NA | NA | NA | NA | 50–55 | 50–71 | NA | A |
| S. tielingense | 4028–8538 | 200–307 | 82–103 | 111–144 | 186–263 | 40–69 | 17–32 | 21–45 | 72–158 | 0.5–0.9 | 49–54 | 56–92 | 32–49 | A |
| S. xinbinense | 3025–5121 | 159–200 | 70–87 | 106–141 | 167–192 | 30–53 | 19–25 | 17–26 | 79–123 | 0.5–0.8 | 46–57 | 50–67 | 38–45 | P |
| S. xueshanense | 4181–8181 | 182–343 | 117–148 | NA | 196–274 | 43–66 | NA | NA | NA | NA | 52–62 | 38–72 | NA | P |
| S. weiseri | 3780–5940 | 202–263 | 75–86 | 108–154 | 162–226 | 38–59 | 17–29 | 22–31 | 87–156 | 0.5–0.8 | 50–58 | 51–80 | NA | P |
[i] NA = Not available; P = Present; A = Absent.
Table 5.
Pairwise distances in base pairs of the ITS rRNA regions among closely related Steinernema species and Steinernema anantnagense n. sp. Data for new species is in bold.
| Species (ITS rRNA) | S. anantnagense n. sp. OQ407490 | S. akhursti DQ375757 | S. kushidaiAB243440 | S. cholashanense EF431959 | S. oregonense AY230180 | S. sangi AY355441 | S. texanum EF152568 | S. xueshanense FJ666052 | S. populi MZ367621 | S. jollieti AY171265 | S. xinbinense JN171593 | S. weiseri KJ696685 | S. tielingense GU994201 | S. africanum ON041031 | S. kraussei AY230175 | S. citrae EU754718 | S. silvaticum AY230162 | S. litorale AB243441 | S. ichnusae EU421129 | S. nguyeni KP325084 | S. feltiae AY230169 | S. hebeiense DQ105794 | S. monticola AB698756 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| S. anantnagense n. sp. OQ407490 | 19 | 53 | 72 | 72 | 72 | 72 | 73 | 74 | 74 | 80 | 83 | 85 | 86 | 86 | 87 | 87 | 90 | 91 | 92 | 101 | 116 | 117 | |
| S. akhursti DQ375757 | 97 | 51 | 75 | 74 | 73 | 77 | 76 | 65 | 75 | 82 | 86 | 89 | 89 | 90 | 87 | 91 | 93 | 95 | 94 | 102 | 119 | 115 | |
| S. kushidai AB243440 | 92 | 93 | 95 | 89 | 96 | 103 | 90 | 94 | 89 | 97 | 104 | 101 | 103 | 108 | 110 | 104 | 107 | 110 | 110 | 122 | 128 | 130 | |
| S. cholashanense EF431959 | 89 | 88 | 85 | 61 | 74 | 58 | 58 | 106 | 69 | 68 | 72 | 71 | 72 | 83 | 70 | 82 | 73 | 74 | 84 | 75 | 112 | 116 | |
| S. oregonense AY230180 | 89 | 88 | 86 | 91 | 47 | 51 | 33 | 99 | 19 | 31 | 43 | 46 | 39 | 52 | 42 | 45 | 56 | 49 | 52 | 51 | 94 | 115 | |
| S. sangi AY355441 | 89 | 88 | 85 | 88 | 93 | 57 | 58 | 98 | 57 | 54 | 43 | 45 | 60 | 53 | 60 | 61 | 48 | 48 | 54 | 47 | 88 | 103 | |
| S. texanum EF152568 | 89 | 88 | 83 | 91 | 92 | 91 | 53 | 95 | 61 | 60 | 66 | 67 | 59 | 75 | 55 | 69 | 74 | 69 | 77 | 68 | 110 | 118 | |
| S. xueshanense FJ666052 | 89 | 88 | 86 | 91 | 95 | 91 | 92 | 99 | 41 | 43 | 46 | 54 | 47 | 60 | 44 | 54 | 57 | 53 | 60 | 54 | 86 | 114 | |
| S. populi MZ367621 | 89 | 90 | 86 | 83 | 84 | 84 | 85 | 84 | 105 | 103 | 110 | 111 | 112 | 109 | 113 | 115 | 117 | 116 | 112 | 124 | 145 | 123 | |
| S. jollieti AY171265 | 88 | 88 | 86 | 89 | 97 | 91 | 91 | 94 | 83 | 48 | 52 | 54 | 55 | 64 | 55 | 58 | 63 | 56 | 62 | 58 | 95 | 115 | |
| S. xinbinense JN171593 | 88 | 87 | 85 | 90 | 96 | 92 | 91 | 94 | 84 | 93 | 45 | 50 | 32 | 58 | 40 | 38 | 58 | 57 | 57 | 50 | 92 | 116 | |
| S. weiseri KJ696685 | 87 | 86 | 83 | 89 | 94 | 94 | 90 | 93 | 82 | 92 | 93 | 17 | 59 | 32 | 60 | 65 | 26 | 27 | 34 | 27 | 80 | 114 | |
| S. tielingense GU994201 | 87 | 86 | 84 | 89 | 93 | 93 | 90 | 92 | 82 | 92 | 93 | 98 | 63 | 46 | 65 | 66 | 30 | 28 | 47 | 32 | 88 | 117 | |
| S. africanum ON041031 | 87 | 86 | 84 | 89 | 94 | 91 | 91 | 93 | 82 | 92 | 95 | 91 | 91 | 64 | 45 | 35 | 72 | 68 | 64 | 62 | 98 | 124 | |
| S. kraussei AY230175 | 86 | 86 | 82 | 87 | 92 | 92 | 88 | 91 | 82 | 90 | 91 | 95 | 93 | 90 | 65 | 69 | 44 | 46 | 19 | 40 | 89 | 120 | |
| S. citrae EU754718 | 86 | 86 | 82 | 89 | 94 | 91 | 92 | 94 | 82 | 92 | 94 | 91 | 90 | 93 | 90 | 55 | 73 | 70 | 64 | 60 | 98 | 118 | |
| S. silvaticum AY230162 | 86 | 86 | 83 | 87 | 93 | 91 | 89 | 92 | 81 | 91 | 95 | 90 | 90 | 95 | 90 | 92 | 76 | 70 | 68 | 68 | 101 | 126 | |
| S. litorale AB243441 | 86 | 85 | 82 | 89 | 92 | 93 | 88 | 91 | 80 | 90 | 91 | 96 | 96 | 89 | 93 | 89 | 88 | 38 | 45 | 34 | 79 | 118 | |
| S. ichnusae EU421129 | 86 | 85 | 82 | 88 | 93 | 93 | 89 | 92 | 81 | 92 | 92 | 96 | 96 | 90 | 93 | 89 | 89 | 94 | 45 | 25 | 87 | 122 | |
| S. nguyeni KP325084 | 85 | 85 | 82 | 87 | 92 | 92 | 88 | 91 | 81 | 90 | 91 | 95 | 93 | 90 | 97 | 90 | 90 | 93 | 93 | 41 | 86 | 117 | |
| S. feltiae AY230169 | 84 | 84 | 80 | 88 | 92 | 93 | 89 | 92 | 79 | 91 | 92 | 96 | 95 | 91 | 94 | 91 | 90 | 95 | 96 | 94 | 79 | 119 | |
| S. hebeiense DQ105794 | 81 | 80 | 78 | 82 | 85 | 86 | 82 | 86 | 75 | 85 | 86 | 88 | 86 | 84 | 86 | 85 | 84 | 88 | 86 | 87 | 88 | 135 | |
| S. monticola AB698756 | 78 | 79 | 76 | 78 | 79 | 81 | 78 | 79 | 77 | 79 | 79 | 79 | 78 | 77 | 78 | 78 | 77 | 78 | 77 | 78 | 78 | 74 |
[i] Below diagonal: percentage similarity; above diagonal: total character difference.
Table 6.
Pairwise distances in base pairs of the D2D3 fragment of 28S rRNA regions among closely related Steinernema species and Steinernema anantnagense n. sp. Data for new species is in bold.
| Species (D2D3 rRNA) | S. anantnagense n. sp. OQ407498 | S. akhursti KF289902 | S. weiseri FJ165549 | S. oregonense GU569055 | S. puntauvense EF187018 | S. feltiae AF331906 | S. ichnusae EU421130 | S. africanum OM423154 | S. kushidai AF331897 | S. tielingense GU994202 | S. populi MZ367685 | S. xueshanense FJ666053 | S. kraussei KC631424 | S. jollieti GU569051 | S. cholashanense EF520284 | S. texanum EF152569 | S. xinbinense GU994204 | S. citrae MF540676 | S. silvaticum KC631426 | S. sangi MF620997 | S. nguyeni KR815816 | S. monticola GU395647 |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| S. anantnagense n. sp. OQ407498 | 5 | 12 | 13 | 13 | 13 | 15 | 16 | 17 | 18 | 18 | 18 | 19 | 19 | 23 | 24 | 25 | 25 | 27 | 30 | 31 | 35 | |
| S. akhursti KF289902 | 99 | 16 | 16 | 17 | 17 | 17 | 18 | 18 | 22 | 20 | 22 | 22 | 21 | 26 | 27 | 29 | 27 | 28 | 27 | 33 | 38 | |
| S. weiseri FJ165549 | 98 | 98 | 12 | 5 | 5 | 7 | 6 | 22 | 13 | 18 | 15 | 16 | 9 | 18 | 21 | 19 | 18 | 24 | 32 | 22 | 37 | |
| S. oregonense GU569055 | 98 | 98 | 98 | 11 | 11 | 13 | 14 | 24 | 10 | 27 | 17 | 8 | 17 | 13 | 22 | 14 | 25 | 19 | 33 | 29 | 38 | |
| S. puntauvense EF187018 | 98 | 98 | 99 | 99 | 0 | 4 | 5 | 23 | 10 | 23 | 16 | 13 | 10 | 15 | 20 | 16 | 17 | 21 | 30 | 21 | 40 | |
| S. feltiae AF331906 | 98 | 98 | 99 | 99 | 100 | 4 | 5 | 23 | 10 | 23 | 16 | 13 | 10 | 15 | 20 | 16 | 17 | 21 | 30 | 21 | 40 | |
| S. ichnusae EU421130 | 98 | 98 | 99 | 98 | 99 | 99 | 5 | 25 | 14 | 25 | 17 | 15 | 10 | 19 | 20 | 20 | 17 | 25 | 32 | 21 | 40 | |
| S. africanum OM423154 | 98 | 98 | 99 | 98 | 99 | 99 | 99 | 21 | 13 | 22 | 19 | 16 | 8 | 18 | 22 | 19 | 12 | 26 | 30 | 15 | 37 | |
| S. kushidai AF331897 | 98 | 98 | 97 | 97 | 97 | 97 | 97 | 97 | 26 | 30 | 26 | 26 | 29 | 28 | 36 | 29 | 30 | 34 | 32 | 33 | 42 | |
| S. tielingense GU994202 | 98 | 97 | 98 | 99 | 99 | 99 | 98 | 98 | 97 | 27 | 20 | 10 | 18 | 12 | 25 | 12 | 22 | 20 | 36 | 26 | 44 | |
| S. populi MZ367685 | 98 | 97 | 98 | 96 | 97 | 97 | 97 | 97 | 96 | 96 | 27 | 30 | 27 | 30 | 29 | 31 | 32 | 38 | 40 | 37 | 44 | |
| S. xueshanense FJ666053 | 98 | 97 | 98 | 98 | 98 | 98 | 98 | 98 | 97 | 97 | 96 | 20 | 22 | 20 | 25 | 22 | 30 | 28 | 41 | 33 | 45 | |
| S. kraussei KC631424 | 98 | 97 | 98 | 99 | 98 | 98 | 98 | 98 | 97 | 99 | 96 | 97 | 21 | 13 | 27 | 11 | 27 | 16 | 37 | 31 | 45 | |
| S. jollieti GU569051 | 98 | 97 | 99 | 98 | 99 | 99 | 99 | 99 | 96 | 98 | 96 | 97 | 97 | 23 | 24 | 22 | 20 | 31 | 35 | 24 | 42 | |
| S. cholashanense EF520284 | 97 | 97 | 98 | 98 | 98 | 98 | 98 | 98 | 96 | 98 | 96 | 97 | 98 | 97 | 27 | 10 | 27 | 19 | 38 | 30 | 45 | |
| S. texanum EF152569 | 97 | 96 | 97 | 97 | 97 | 97 | 97 | 97 | 95 | 97 | 96 | 97 | 96 | 97 | 96 | 32 | 32 | 37 | 39 | 35 | 45 | |
| S. xinbinense GU994204 | 97 | 96 | 98 | 98 | 98 | 98 | 97 | 98 | 96 | 98 | 96 | 97 | 99 | 97 | 99 | 96 | 30 | 17 | 39 | 34 | 47 | |
| S. citrae MF540676 | 97 | 96 | 98 | 97 | 98 | 98 | 98 | 98 | 96 | 97 | 96 | 96 | 96 | 97 | 96 | 96 | 96 | 37 | 42 | 11 | 49 | |
| S. silvaticum KC631426 | 96 | 96 | 97 | 98 | 97 | 97 | 97 | 97 | 95 | 97 | 95 | 96 | 98 | 96 | 98 | 95 | 98 | 95 | 41 | 41 | 50 | |
| S. sangi MF620997 | 96 | 96 | 96 | 96 | 96 | 96 | 96 | 96 | 96 | 95 | 95 | 94 | 95 | 95 | 95 | 95 | 95 | 94 | 95 | 46 | 46 | |
| S. nguyeni KR815816 | 96 | 96 | 97 | 96 | 97 | 97 | 97 | 98 | 96 | 97 | 95 | 96 | 96 | 97 | 96 | 95 | 95 | 99 | 95 | 94 | 51 | |
| S. monticola GU395647 | 95 | 95 | 95 | 95 | 95 | 95 | 95 | 95 | 94 | 94 | 94 | 94 | 94 | 94 | 94 | 94 | 94 | 93 | 93 | 94 | 93 |
[i] Below diagonal: percentage similarity; above diagonal: total character difference.
Figure 6:
Maximum-likelihood phylogenetic tree between the newly described Steinernema anantnagense n. sp. and other closely related species of Steinernema species based on nucleotide sequences of: (A) the Internal Transcribed Spacer (ITS1-5.8S-ITS2) rRNA, flanked by primers 18S and 26S, and (B) the D2-D3 expansion segments of the large subunit (28S) of rRNA flanked by primers D2F and 536. Numbers at nodes represent bootstrap values based on 100 replications. Bars represent average nucleotide substitutions per sequence position. NCBI accession numbers of the nucleotide sequences used for the analyses are shown next to the species names. The scale bar shows the number of substitutions per site.
Figure 7:
Maximum-likelihood phylogenetic tree between the newly described Steinernema anantnagense n. sp. and other closely related species of Steinernema species based on the nucleotide sequences of: (A) the COI region of the mitochondrial gene, flanked by primers LCO-1490 and HCO-2198, and (B) the mitochondrial 12S rRNA gene, flanked by primers 505F and 506R. Numbers at nodes represent bootstrap values based on 100 replications. Bars represent average nucleotide substitutions per sequence position. NCBI accession numbers of the nucleotide sequences used for the analyses are shown next to the species names (accession numbers in bold font are the sequences newly generated in this study). The scale bar shows the number of substitutions per site.
Figure 8:
Phylogenetic reconstruction based on core genome sequences of Xenorhabdus bacterial strains. 1719910 nucleotide positions (1501 core genes) were used in the analysis. Numbers at the nodes represent SH-like branch supports. Bar represents 0.05 nucleotide substitutions per sequence position. Accession numbers of the genome sequences used for the reconstruction are shown in Table S3.