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Multigene Sequence-Based and Phenotypic Characterization Reveals the Occurrence of a Novel Entomopathogenic Nematode Species, Steinernema anantnagense n. sp.

Journal of Nematology
Volume 55 (2023): Issue 1 (February 2023)
By:
Open Access
|Jul 2023

Figures & Tables

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.
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.
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).
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).
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).
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).

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.
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.
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.
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.

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_

SpeciesReferenceCountrynLBDEPNRNLTabCc′D%E%H%
S. akhurstiQiu et al. (2005)China20770–83533–3555–6083–95115–12368–7523–266.6–7.210–123.3–3.745–5073–8649–56
S. anantnagense n. sp.Present StudyIndia20749–83432–4245–6254–71120–14349–6619–245.5–6.712–161.6–2.135–4874–11320–36
S. africanumMachado et al. (2022)Rwanda15690–80225–3254–6887–132123–16752–7223–304.3–6.310–152.9–4.234–4679–12928–39
S. cholashanenseNguyen et al. (2008)China20727–90926–3559–6572–97110–13860–8024–346.1–7.210–143.5–5.046–5376–9133–47
S. citraeStokwe et al. (2011)South Africa20623–84923–2849–6483–108118–13763–8125–345.1–7.113–1413–1739–5885–13237–50
S. feltiaeNguyen et al. (2007)Russia25766–92822–3258–67108–117130–14381–8927–345.8–6.89.4–114.5–5.144–5067–8137–51
S. hebeienseChen et al. (2006)China20610–71023–2843–5173–83100–11163–7124–285.7–6.79.4–11NA40–5065–8032–50
S. ichnusaeTarasco et al. (2008)Italy20767–96927–3559–6894–108119–14876–8924–325.6–6.99–124.2–5.142–4968–8344–50
S. jollietiSpiridonov et al. (2004)USA12625–82020–2853–65NA115–13560–7325–344.9–6.49–12NA46–50NA46–60
S. krausseiNguyen et al. (2007)Germany25797–110230–3650–6699–111119–14563–86NANANANANANA35–40
S. kushidaiMamiya (1998)Japan20424–66222–3142–5070–84106–12044–5919–254.9–5.910–13NA38–44NANA
S. litoraleYoshida (2004)Japan25834–98828–3354–6989–104114–13372–9127–316.7–7.910–11.93.8–5.444–5668–84NA
S. nguyeniMalan et al. (2016)South Africa20673–79622–2847–5874–86101–12161–7327–336.2–7.410–122.8–4.843–5770–8620–31
S. oregoneseLiu and Berry (1996)USA20820–111028–3860–72NA116–14864–7824–376–812–16NA40–6090–11030–33
S. populiTian et al. (2022)China25973–117233–4170–8698–113134–15955–7228–336.8–7.515–202.4–3.347–61105–14026–44
S. puntauvenseUribe-Lorío et al. (2007)Costa Rica19631–72831–3820–3046–6981–10351–5917–237.1–7.911–13NA25–5035–5652–55
S. sandneriLis et al. (2021)Poland25708–96523–3244–6483–118123–15164–8627–345.5–6.911–13NA36–4563–8623–40
S. sangiPhan et al. (2001)Vietnam20704–78430–4046–5478–97120–13876–8919–255.6–6.39–10NA36–4456–7044–52
S. silvaticumSturhan et al. (2005)Germany26670–97526–3551–7375–109100–14163–8623–336.3–7.710–133.1–4.946–56–37–53
S. tielingenseMa et al. (2012)China20824–97932–3864–7390–105120–13574–8527–316.7–7.910–123.5–4.644–5668–8453–64
S. texanumNguyen et al. (2007)USA20732–79629–3452–6284–102111–12060–7922–276.2–7.010–133.3–4.646–5376–8853–69
S. xinbinenseMa et al. (2012)China20635–74428–3146–5375–90109–12565–7821–255–78–113–540–4765–7830–42
S. xueshanenseMrácek et al. (2009)China20768–92929–3360–7281–96130–14380–9226–325.8–7.09–113.8–5.146–5270–9046–55
S. weiseriMrácek et al. (2003)Czech Republic20586–82824–2943–6572–9295–11949–6825–335.7–7.210–143.2–4.144–55NA18–24

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. OQ407498S. akhursti KF289902S. weiseri FJ165549S. oregonense GU569055S. puntauvense EF187018S. feltiae AF331906S. ichnusae EU421130S. africanum OM423154S. kushidai AF331897S. tielingense GU994202S. populi MZ367685S. xueshanense FJ666053S. kraussei KC631424S. jollieti GU569051S. cholashanense EF520284S. texanum EF152569S. xinbinense GU994204S. citrae MF540676S. silvaticum KC631426S. sangi MF620997S. nguyeni KR815816S. monticola GU395647
S. anantnagense n. sp. OQ407498 51213131315161718181819192324252527303135
S. akhursti KF28990299 1616171717181822202222212627292728273338
S. weiseri FJ1655499898 125576221318151691821191824322237
S. oregonense GU569055989898 11111314241027178171322142519332938
S. puntauvense EF18701898989999 0452310231613101520161721302140
S. feltiae AF33190698989999100 452310231613101520161721302140
S. ichnusae EU421130989899989999 52514251715101920201725322140
S. africanum OM42315498989998999999 211322191681822191226301537
S. kushidai AF3318979898979797979797 26302626292836293034323342
S. tielingense GU994202989798999999989897 272010181225122220362644
S. populi MZ36768598979896979797979696 2730273029313238403744
S. xueshanense FJ6660539897989898989898979796 20222025223028413345
S. kraussei KC631424989798999898989897999697 211327112716373145
S. jollieti GU56905198979998999999999698969797 2324222031352442
S. cholashanense EF5202849797989898989898969896979897 27102719383045
S. texanum EF152569979697979797979795979697969796 323237393545
S. xinbinense GU99420497969898989897989698969799979996 3017393447
S. citrae MF5406769796989798989898969796969697969696 37421149
S. silvaticum KC631426969697989797979795979596989698959895 414150
S. sangi MF62099796969696969696969695959495959595959495 4646
S. nguyeni KR8158169696979697979798969795969697969595999594 51
S. monticola GU395647959595959595959594949494949494949493939493

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_

SpeciesLBDEPNRNLTABcc′VABDD%Mucron
S. akhursti5625–9000200–270113–138150–175213–25838–6330*32*149*0.6*48–5368–100NAP
S. anantnagense n. sp.2327–4872314–40992–140143–182196–25532–4917–2913–2161–1220.5–0.850–5877–11043–60P
S. africanum2469–4215154–19467–11179–130170–20135–5513–2713–2451–1040.7–1.050–5737–7032–62P
S. cholashanense3232–6363156–332111–148176–223181–23146–7013–2318–3262–1190.6–1.050–5754–10529–65P
S. citrae2038–4019137–21254–90130–179189–22033–60NANANANA50–5943–7927–46P
S. feltiae3095–3774170–25468–97 **70–97 **197–30439–7014–2012–1749–880.7–1.2 *44–5747–6240–54 *P
S. hebeiense3972–4254142–24548–9588–123133–158133–15817–2521–2967–1290.5–0.950–5745–6536–66A
S. ichnusae4547–6186242–323106–156NA215–26251–7917–2421–2668–1130.6–1.051–5770–9447–63P
S. jollieti3746–6030219–29896–136NA184–31031–5515–2419–3172–185NA44–56NA52P
S. kraussei2500–5400153–28866–99127–146178–20533–59172288NA5439–5045P
S. kushidai2100–470054–5978–105111–144204–25530–45NANANANA54–5954–8437–46A
S. litorale3930–5048175–21565–105130–165185–21325–6021–2620–2678–1570.5–0.90.5–0.955–7533–57P
S. nguyeni2290–5361130–21649–9884–139137–19420–6715–3015–3053–1650.6–1.152–63130–21630–56A
S. oregonense4400–6200217–268217–268129–162186–22028–46NANANANA46–5642–7943–57A
S. populi4038–13762217–53190–178150–213213–27841–8818–3619–5075–1820.5–0.945–6060–15736–65A
S. puntauvense3687–8335181–22151–85123–146141–20641–66NANANANA51–5557–10225–45P
S. sandneri4244–5014181–26161–102132–158173–19432–6117–2524–2775–140NA49–5762–12236–54P
S. sangi4830–7200270–36080–121140–170216–24036–62NANANANA43–53084–14035–51P
S. silvaticum1520–329050–17550–17550–175121–18833–7915–4110–1834–801.0–1.844–5726–5333–79A
S. texanum2720–3623130–20278–107111–135160–18930–52NANANANA50–5550–71NAA
S. tielingense4028–8538200–30782–103111–144186–26340–6917–3221–4572–1580.5–0.949–5456–9232–49A
S. xinbinense3025–5121159–20070–87106–141167–19230–5319–2517–2679–1230.5–0.846–5750–6738–45P
S. xueshanense4181–8181182–343117–148NA196–27443–66NANANANA52–6238–72NAP
S. weiseri3780–5940202–26375–86108–154162–22638–5917–2922–3187–1560.5–0.850–5851–80NAP

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 GenerationSecond GenerationInfective Juveniles (paratypes)
Male (holotype)Male (paratypes)Female (paratypes)Male (paratypes)Female (paratypes)
n12020202020
Body length (L)12791618 ± 246 (1223–1899)3765 ± 441 (2327–4872)1068 ± 76 (899–1168)2081 ± 200 (1761–2437)789 ± 35 (749–834)
a (L/BD)6.68.3 ± 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)7.09.2 ± 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)3549 ± 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)1.21.1 ± 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)193194 ±11.3 (167–211)434 ± 30 (314–409)57 ± 7.3 (42–63)143 ± 16 (123–173)37 ± 3.6 (32–42)
Lip region width10.210.1 ± 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 length12.313.8 ± 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 length6657 ± 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 length3334 ± 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 length4042 ± 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)3130 ± 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)2625 ± 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)170162 ± 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)109112 ± 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)114111 ± 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)5861 ± 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)182176 ± 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 base7986 ± 5.8 (76–96)217 ± 23 (195–264)41 ± 3.9 (36–50)95 ± 8.5 (82–107)29 ± 2.9 (24–32)
Testis reflexion630631 ± 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)3132 ± 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)3734 ± 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)6664 ± 4.6 (56–70)–44 ± 2.6 (40–49)––
Gubernaculum length (GL)3436 ± 3.8 (31–43)–25 ± 2.3 (21–29)––
Stoma length/lip region width1.21.4 ± 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)6064 ± 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)6263 ± 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)6263 ± 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)311333 ± 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)217208 ± 39 (154–297)–2.3 ± 0.4 (1.8–3.4)
GS% (GL/SL×100)5157 ± 6.7 (46–70)–0.6 ± 0.1 (0.4–0.7)
H% (H/T ×100)–––––28 ± 5.8 (20–36)

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_

SpeciesnLBDEPNRNLTSLGLaBcc′D%SW%GS%Mucron
S. akhursti201350–1925115–15093–113120–163168–20530–4085–10058–68NANANANA52–61140–20065–77P
S. anantnagense n. sp.201223–1899167–21188–124103–129165–18529–3956–7031–436–107–1134–640.8–1.649–74154–29746–70P
S. africanum15977–140065–13169–10979–104132–14734–4665–7632–499–127–1225–340.9–1.152–74144–19749–68P
S. cholashanense201070–177873–20475–13591–126135–17329–4360–7132–459–248–1136–510.6–0.950–8592–14461–85P
S. citrae201028–140287–11364–9292–119123–15517–3157–8032–59NANANANA47–67156–23348–89P
S. feltiae251414–181760–90110–126NA164–18037–4365–7734–47NANANANA51–6499–13052–61P
S. hebeiense201036–145074–9858–7378–93118–13224–3551–6338–5012–178–1130–490.6–0.948–59120–17060–90A
S. ichnusae201151–149473–20494–108NA135–17333–4864–6743–4620–297–929–390.8–0.959–65120–16264–69A
S. jollieti121296–195298–13583–110NA110–16824–3855–7045–6012–198–1453–86NA53–83NANAA
S. kraussei201200–1600110–14473–9995–122137–17836–4442–5329–37119 NANANANANANAP
S. kushidai201200–190075–15671–105120–137156–18930–4048–7239–60NANANANA42–59NANAA
S. litorale251230–151482–11177–10794–128133–16326–4167–8944–6412–168–1033–560.6–0.934–56154–20062–81P
S. nguyeni20818–117158–10647–7170–103112–14418–2558–7530–5511–157–1038–530.6–0.838–57185–27946–81P
S. oregonense201560–1820105–16195–139101–133139–18224–3265–7352–59NANANA0.6 NA64–75NANAA
S. populi251258–151466–9595–121107–143131–17739–6857–7738–6015–208–1020–330.8–1.559–78107–16058–82P/A
S. puntauvense191010–1931101–13968–114104–128130–15928–4071–8130–40NANANANA45–85140–20055–75P
S. sandneri251206–1635124–17864–92112–138148–17035–4653–6539–509–118–1031–42NA42–5997–12761–83P
S. sangi201440–2325120–22567–99109–166150–22127–4258–8034–46NANANANA42–63120–16050–70P
S. silvaticum26975–127052–7871–9290–126116–16820–4742–6430–4314–208–924–550.8–1.445–63NANAP
S. tielingense201430–2064111–15994–13396–132145–17322–3379–9849–7011–189–1357–850.3–0.664–78176–21259–82A
S. texanum201197–140681–11679–10094–114123–14719–3055–6639–53NANANANA58–73127–20362–84A
S. xinbinense201133–144090–12657–7591–120138–15930–4149–6230–4111–137–931–390.7–1.041–50114–15654–72P
S. xueshanense201313–204097–159113–137NA151–17529–4866–9141–60NANANANA73–8793–17258–95A
S. weiseri20990–139584–13857–8494–115134–15419–3262–7246–579–127–1036–640.6–0.939–60150–24070–85A

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. OQ407490S. akhursti DQ375757S. kushidaiAB243440S. cholashanense EF431959S. oregonense AY230180S. sangi AY355441S. texanum EF152568S. xueshanense FJ666052S. populi MZ367621S. jollieti AY171265S. xinbinense JN171593S. weiseri KJ696685S. tielingense GU994201S. africanum ON041031S. kraussei AY230175S. citrae EU754718S. silvaticum AY230162S. litorale AB243441S. ichnusae EU421129S. nguyeni KP325084S. feltiae AY230169S. hebeiense DQ105794S. monticola AB698756
S. anantnagense n. sp. OQ407490 19537272727273747480838586868787909192101116117
S. akhursti DQ37575797 517574737776657582868989908791939594102119115
S. kushidai AB2434409293 95899610390948997104101103108110104107110110122128130
S. cholashanense EF431959898885 61745858106696872717283708273748475112116
S. oregonense AY23018089888691 4751339919314346395242455649525194115
S. sangi AY3554418988858893 57589857544345605360614848544788103
S. texanum EF152568898883919291 5395616066675975556974697768110118
S. xueshanense FJ66605289888691959192 9941434654476044545753605486114
S. populi MZ3676218990868384848584 105103110111112109113115117116112124145123
S. jollieti AY171265888886899791919483 485254556455586356625895115
S. xinbinense JN17159388878590969291948493 4550325840385857575092116
S. weiseri KJ6966858786838994949093829293 17593260652627342780114
S. tielingense GU994201878684899393909282929398 634665663028473288117
S. africanum ON04103187868489949191938292959191 6445357268646298124
S. kraussei AY2301758686828792928891829091959390 65694446194089120
S. citrae EU754718868682899491929482929491909390 557370646098118
S. silvaticum AY23016286868387939189928191959090959092 76706868101126
S. litorale AB2434418685828992938891809091969689938988 38453479118
S. ichnusae EU421129868582889393899281929296969093898994 452587122
S. nguyeni KP32508485858287929288918190919593909790909393 4186117
S. feltiae AY2301698484808892938992799192969591949190959694 79119
S. hebeiense DQ105794818078828586828675858688868486858488868788 135
S. monticola AB69875678797678798178797779797978777878777877787874
DOI: https://doi.org/10.2478/jofnem-2023-0029 | Journal eISSN: 2640-396X | Journal ISSN: 0022-300X
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Language: English
Submitted on: Apr 6, 2023
Published on: Jul 13, 2023
Published by: Society of Nematologists, Inc.
In partnership with: Paradigm Publishing Services
Publication frequency: 1 issue per year
Keywords:
Species description,
nematodes,
molecular characterization,
phylogenetic systematics,
taxonomy,
biological control,
sustainable agriculture
Related subjects:
Life sciences,
Life sciences, other

© 2023 Aashaq Hussain Bhat, Ricardo A. R. Machado, Joaquín Abolafia, Tarique Hassan Askary, Vladimír Půža, Alba Nazaret Ruiz-Cuenca, Aasha Rana, Samy Sayed, Laila A. Al-Shuraym, published by Society of Nematologists, Inc.
This work is licensed under the Creative Commons Attribution 4.0 License.

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