Molecular Phylogeny and Species Delimiting for the Genus Hoplolaimus (Nematoda: Tylenchida) with Description of Hoplolaimus floridensis sp. n. and Notes on Biogeography of the Genus in the United States
Abstract
1. Introduction
2. Results
2.1. Hoplolaimus floridensis sp. n. (Figure 1, Figure 2, Figure A1, Figure A2I and Figure A4D,I,J; Table A1)
2.2. Hoplolaimus galeatus (Figure A2A–G,I, Figure A3A–G,K and Figure A4A–C,G–I; Table A2)
2.3. Hoplolaimus galeatus from Nichols (Figure A2K and Figure A3K; Table A3)
2.4. Hoplolaimus concaudajuvencus (Figure A2J, Figure A3J, Figure A4E,K and Figure A5, Table A4)
2.5. Peltamigratus christiei (Figure A2L, Figure A3L and Figure A4F,L)
2.6. Molecular Characterization, Phylogenetic Relationships, and Molecular Species Delimiting Hoplolaimus Species
2.6.1. D2–D3 of the 28S rRNA Gene
2.6.2. ITS of the rRNA Gene
2.6.3. COI Gene
2.6.4. Combined rRNA and COI Gene Alignment
2.7. Distribution Map of Hoplolaimus Species in the United States
3. Discussion
3.1. Identification of Hoplolaimus galeatus and H. stephanus syn. n.
3.2. Identification of Hoplolaimus floridensis sp. n. and H. tylenchiformis
3.3. Identification of Hoplolaimus concaudajuvencus
3.4. Identification of Hoplolaimus seinhorsti
3.5. Unidentified Hoplolaimus Species
3.6. Subgenera of Hoplolaimus
3.7. Biogeography of Hoplolaimus Species in the United States
3.8. Hoplolaimus Species from Florida
4. Materials and Methods
4.1. Nematode Samples and Extraction
4.2. Morphological Study and Species Identification
4.3. DNA Extraction, PCR, and Sequencing
4.4. Phylogenetic and Sequence Analysis
4.5. Distribution Map of Hoplolaimus Species in the United States
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Appendix A
Population | Citra, Florida (CD3650; 11152021-04429) | Gainesville, Florida (CD3630; 10222021-3125) | |||
---|---|---|---|---|---|
Character | Holotype Female | Paratype Females | Paratype Males | Females | Males |
n | 1 | 14 | 17 | 15 | 10 |
L (length) | 1575.0 | 1596.9 ± 127.6 (1385.0–1827.0) | 1347.8 ± 119.2 (1150.0–1545.0) | 1543.5 ± 163.4 (1305.0–1893.0) | 1251.0 ± 120.6 (1050.0–1400.0) |
a | 35.0 | 31.9 ± 3.2 (26.6–36.0) | 29.9 ± 3.0 (23.0–34.0) | 32.5 ± 3.8 (26.5–39.4) | 27.3 ± 1.3(25.0–28.7) |
b | 9.1 | 9.1 ± 1.2 (6.7–11.2) | 7.6 ± 0.6 (6.6–8.8) | 8.6 ± 1.6 (7.0–12.6) | 6.9 ± 1.0 (4.8–8.2) |
c | 68.5 | 67.6 ± 9.0 (55.0–86.8) | 43.3 ± 5.3 (34.8–51.8) | 67.2 ± 12.4 (48.2–94.7) | 43.3 ± 5.4 (35.0–50.0) |
V% | 55.5 | 54.6 ± 1.7 (51.3–57.0) | – | 54.9 ± 1.9 (52.2–59.0) | - |
Stylet length | 48.0 | 46.6 ± 1.7 (43.0–50.0) | 44.0 ± 1.2 (42.0–46.0) | 47.7 ± 1.3 (46.0–50.0) | 44.4 ± 1.5(43.0–47.0) |
Anterior body-pharyngeal gland lobe distance | 174.0 | 176.9 ± 13.9 (150.0–207.0) | 177.9 ± 14.7 (155.0–200.0) | 181.0 ± 15.4 (145.0–205.0) | 183.0 ± 16.5 (165.0–220.0) |
Excretory pore-anterior body distance | 150.0 | 160.0 ± 13.3 (140.0–180.0) | - | 167.5 ± 11.6 (135.0–185.0) | - |
Mid-body diameter (MBW) | 45.0 | 50.3 ± 2.9 (45.0–56.0) | 45.2 ± 3.1 (40.0–50.0) | 47.6 ± 3.4 (43.0–55.0) | 45.8 ± 4.0 (40.0–50.0) |
Tail length | 23.0 | 23.9 ± 2.8 (16.0–28.0) | 31.3 ± 1.7 (28.0–35.0) | 23.4 ± 3.0 (20.0–28.0) | 28.3 ± 1.9 (25.0–30.0) |
Tail annuli | 10.0 | 9.0 ± 1.5 (7.0–12.0) | – | 10.5 ± 1.7 (8.0–14.0) | - |
Spicule length | - | - | 49.0 ± 5.1 (41.0–60.0) | – | 50.3 ± 2.3(45.0–52.0) |
Gubernaculum length | - | - | 25.1 ± 2.7 (21.0–32.0) | – | 18.6 ± 4.5 (15.0–25.0) |
Population | Topotype Population, Virginia, Arlington (CD3582) (This Study) | Type Population Sher [29] | ||
---|---|---|---|---|
Character | Females | Males | Females | Males |
n | 10 | 3 | 20 | 10 |
L | 1165.7 ± 92.4 (1038.0–1300.0) | 1086.7 ± 2.9 (1085.0–1090.0) | 1240.0–1940.0 | 1050.0–1560.0 |
a | 28.1 ± 2.4 (25.5–32.1) | 26.9 ± 0.4 (26.4–27.2) | 25.0–34.0 | 25.0–32.0 |
b | 7.0 ± 0.6 (6.0–8.1) | 7.0 ± 0.9 (6.3–8.0) | 7.6–10.8 | 8.3–10.3 |
c | 55.3 ± 5.1 (46.6–62.0) | 39.9 ± 0.5 (39.4–40.3) | 42.0–82.0 | 28.0–40.0 |
V% | 56.1 ± 1.1 (51.1–57.7) | - | 52.0–60.0 | 52.0–60.0 |
Stylet length | 43.3 ± 1.4 (41.0–45.0) | 40.3 ± 0.6 (40.0–41.0) | 43.0–52.0 | 43.0–52.0 |
Anterior body-pharyngeal gland lobe distance | 166.0 ± 9.4 (150.0–175.0) | 156.7 ± 18.9 (135.0–170.0) | 100.0–160.0 | 100.0–170.0 |
Excretory pore-anterior body distance | - | - | - | - |
Mid-body diameter (MBW) | 41.7 ± 4.5 (38.0 -50.0) | 40.3 ± 0.6 (40.0–41.0) | - | - |
Tail length | 21.2 ± 2.0 (18.0–25.0) | 27.2 ± 0.3 (27.0–27.5) | - | - |
Tail annuli | 10.3 ± 1.2 (9.0–12.0) | - | - | - |
Spicule length | - | 44.0 ± 1.7 (42.0–45.0) | - | 40.0–52.0 |
Gubernaculum length | - | 22.0 ± 1.4 (21.0–23.0) | - | 20.0–28.0 |
Population | Topotype Population, Nichols, South Carolina (CD4603) (This Study) | Type Population Sher [29] | ||
---|---|---|---|---|
Character | Females | Males | Females | Males |
n | 9 | 5 | 20 | 10 |
L | 1139.4 ± 84.2 (1040.0–1302.0) | 966.6 ± 86.8 (850.0–1055.0) | 1001.0–1450.0 | 870.0–1008.0 |
a | 25.0 ± 2.5 (22.6–29.8) | 22.9 ± 1.3 (21.5–25.0) | 22.0–32.0 | 26.0–33.0 |
b | 6.3 ± 0.7 (5.5–7.9) | 6.3 ± 0.9 (5.2–7.5) | 7.3–9.8 | 7.3–8.9 |
c | 64.9 ± 12.4 (46.2–81.3) | 35.2 ± 4.1 (31.5–42.0) | 50.0–80.0 | 30.0–38.0 |
V% | 56.9 ± 2.6 (52.7–61.2) | - | 52.0–61.0 | - |
Stylet length | 44.1 ± 2.2 (40.0–46.0) | 42.2 ± 1.3 (40.0–43.0) | 43.0–50.0 | 39.0–44.0 |
Anterior body-pharyngeal gland lobe distance | 182.2 ± 12.5 (165.0–200.0) | 155.0 ± 9.4 (140.0–165.0) | - | - |
Excretory pore-anterior body distance | 142.1 ± 4.8 (135.0–150.0) | 123.0 ± 6.0 (120.0–132.0) | - | - |
Mid-body diameter (MBW) | 45.8 ± 3.1 (41.0–51.0) | 42.2 ± 4.1 (38.0–41.0) | - | - |
Tail length | 18.1 ± 3.6 (15.0–25.0) | 27.6 ± 2.4 (25.0–31.0) | - | - |
Head annuli | 5 | 5 | 4 | 4 |
Spicule length | - | 40.8 ± 1.3 (40.0–43.0) | - | 30.0–38.0 |
Gubernaculum length | - | 22.4 ± 0.2 (22.0–25.0) | - | 13.0–20.0 |
Population | Topotype Population from Pensacola (CD3711; 02152022-01216) (Stenotaphrum secundatum) (This Study) | Type Population (Cynodon dactylon) Golden & Minton [30] | ||
---|---|---|---|---|
Character | Females | Males | Females | Males |
n | 10 | 7 | 20 | 20 |
L | 1614.7 ± 117.1 (1427.0–1815.0) | 1376.0 ± 114.9 (1262.0–1575.0) | 1606.0 (1120.0–2041.0) | 1423.0 (1118.0–1608.0) |
a | 31.9 ± 3.8 (27.3–38.6) | 27.7 ± 2.9 (24.1–33.8) | 28.6 (23.9–31.9) | 25.9 (20.0–30.7) |
b | 8.3 ± 1.1 (6.7–10.1) | 7.9 ± 0.5 (7.3–8.8) | 8.6 (6.3–11.0) | 7.2 (6.2–8.8) |
c | 69.5 ± 8.4 (53.5–84.3) | 48.8 ± 10.3 (36.3–63.0) | 65.1 (49.5–85.9) | 37.9 (33.5–45.6) |
V% | 56.1 ± 2.9 (52.0–61.3) | - | 54.4 (50.0–57.0) | - |
Head annuli | 6.0 ± 0.8 (5.0–7.0) | - | - | - |
Stylet length | 53.3 ± 2.1 (50.0–56.0) | 48.0 ± 4.2 (43.0–52.0) | 54.9 (50.4–56.6) | 50.0 (46.5–53.8) |
Anterior body-pharyngeal gland lobe distance | 196.0 ± 19.3 (175.0–225.0) | 174.3 ± 14.8 (155.0–200.0) | - | - |
Excretory pore-anterior body distance | 163.4 ± 17.0 (135.0–185.0) | 139.3 ± 17.4 (125.0–165.0) | - | - |
Mid-body diameter (MBW) | 51.2 ± 6.5 (43.0–65.0) | 50.0 ± 4.8 (43.0–56.0) | - | - |
Tail length | 23.5 ± 3.0 (21.0–30.0) | 29.1 ± 6.0 (22.0–40.0) | - | - |
Spicule length | - | 49.9 ± 4.4 (45.0–56.0) | - | 50.0 (45.0–56.0) |
Gubernaculum length | - | 22.0 ± 3.6 (18.0–25.0) | - | 26.0 (23.0–28.0) |
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Species | Location | Associated Plants | Sample Code | GenBank Accession Number | Source or Reference | ||
---|---|---|---|---|---|---|---|
ITS rRNA Gene | D2–D3 of 28S rRNA Gene | COI Gene | |||||
H. concaudajuvencus | USA, Florida, Escambia County, Pensacola | Stenotaphrum secundatum, Lolium sp. | CD3711; 02152022-01216 | PV863258, PV863260, PV863261, PV863263 | PV863155, PV863156, PV863158, PV863159 | PV857622-PV857625 | L. Violett, R.N. Inserra |
H. concaudajuvencus | USA, Florida, Escambia County, Pensacola | S. secundatum | CD3738; 03292022-02584 | PV863259, PV863262 | PV863157, PV863160 | PV857620, PV857621 | L. Violett, R.N. Inserra |
H. galeatus | USA, Virginia, Arlington cemetery | Grasses | CD3582 | PV863244, PV863245 | PV863179, PV863181, PV863182 | PV857650, PV857651 | M. Kantor |
H. galeatus | USA, Virginia, Arlington cemetery | Grasses | CD3583 | PV863251 | PV863178 | PV857652 | M. Kantor |
H. galeatus (=H. stephahus syn. n.) | USA, South Carolina, Marion County, Nicols | Unknown | CD4603; sample 5 | PV863240-PV863243 | PV863161-PV863164 | PV857615-PV857618 | M. Kantor |
H. galeatus | USA, Michigan, Leelanau County | Hop | CD3921 | - | PV863184 | - | M. Quintanilla-Tornel |
H. galeatus | USA, Maryland, Prince George’s County, Upper Marlboro | Grasses | CD3580 | PV863246-PV863248 | PV863167, PV863168, PV863172 | PV857640, PV857645, PV857646 | M. Kantor |
H. galeatus | USA, Maryland, Prince George’s County, Upper Marlboro | Grasses | CD3361 | - | PV863170 | - | M. Kantor |
H. galeatus | USA, Iowa | Corn | CD1056 | PV863256 | - | PV857653 | G. Tylka |
H. galeatus | USA, South Carolina | Unknown | CD3641 | - | PV863169, PV863171, PV863177, PV863180, PV863183 | PV857644 | Z. Handoo |
H. galeatus | USA, North Dakota | Pea | CD1897 | PV863255 | PV863185 | PV857654 | G.P. Yan |
H. galeatus | USA, Minnesota, Wright County, Monticello | Grasses | CD1098 | - | - | PV857655 | D. Mollov |
H. galeatus | USA, Pennsylvania, Centre County, Rock Springs | Corn | CD4574 | 3PV863252-PV863254 | PV863173, PV863175, PV863176 | PV857641-PV857643 | M. Kantor |
H. galeatus | USA, Delaware, Sussex County, Ellendale | Corn | CD4575 | PV863249, PV863250 | PV863165, PV863166, PV863174 | PV857647-PV857649 | M. Kantor |
H. seinhorsti | Israel | Unknown | 721 | - | DQ328752 | PV857656-PV857658 | N. Vovlas; Subbotin et al. [15] |
H. seinhorsti | India, Uttar Pradesh | Grasses | CD2086 | PV863266 | PV863204 | PV857659 | S.A. Subbotin |
H. floridensis sp. n. | USA, Florida, Alachua County, Gainesville | S. secundatum | CD3630; 10222021-3125 | PV863231-PV863233, PV863235 | PV863189, PV863195, PV863196, PV863198, PV863200 | PV857626-PV857630 | R.N. Inserra |
H. floridensis sp. n. | USA, Florida, Marion County, Citra | Cynodon dactylon | CD3650; 11152021-04429 | PV863228-PV863230, PV863234 | PV863190, PV863199, PV863202, PV863203 | PV857631-PV857633 | W.W. Crow, R.N. Inserra |
H. floridensis sp. n. | USA, Florida, Collier County, Chokoloskee | Grasses | CD3700; 12202021-03392 | PV863239 | PV863192, PV863194, PV863197 | PV857639 | J.S. Stanley, R.N. Inserra |
H. floridensis sp. n. | USA, Florida, Collier County, Everglades City | Grasses | CD3701; 12202021-03390 | PV863236-PV863238 | PV863186, PV863187 | PV857636-PV857638 | J.S. Stanley, R.N. Inserra |
H. floridensis sp. n. | USA, Florida, Hendry County, Clewiston | Zoysia sp. + S. secundatum | CD3712; 02172022-01316 | PV863224-PV863226 | PV863188, PV863193 | - | J.S. Stanley, R.N. Inserra |
H. floridensis sp. n. | USA, Florida, Collier County, Immokalee | S. secundatum | CD3739; 05202022-04589 | PV863227 | PV863191, PV863201 | PV857634, PV857635 | J. S. Stanley, R.N. Inserra |
Hoplolaimus sp. A | USA, California, Riverside County, Temecula, Spring Canyon | Unknown | CD1822 | PV863257 | PV863205 | PV857619 | S.A. Subbotin |
Peltamigratus christiei | USA, Florida, Okeechobee County, Okeechobee | Palm | CD1495; 06232022-05749 | PV863264, PV863265 | PV863206 | PV857660, PV857661 | R. Bloom; R.N. Inserra |
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Subbotin, S.A.; Kantor, M.; Consoli, E.; Lyndby, N.H.; Michaud, A.; Handoo, Z.; Inserra, R.N. Molecular Phylogeny and Species Delimiting for the Genus Hoplolaimus (Nematoda: Tylenchida) with Description of Hoplolaimus floridensis sp. n. and Notes on Biogeography of the Genus in the United States. Int. J. Mol. Sci. 2025, 26, 8501. https://doi.org/10.3390/ijms26178501
Subbotin SA, Kantor M, Consoli E, Lyndby NH, Michaud A, Handoo Z, Inserra RN. Molecular Phylogeny and Species Delimiting for the Genus Hoplolaimus (Nematoda: Tylenchida) with Description of Hoplolaimus floridensis sp. n. and Notes on Biogeography of the Genus in the United States. International Journal of Molecular Sciences. 2025; 26(17):8501. https://doi.org/10.3390/ijms26178501
Chicago/Turabian StyleSubbotin, Sergei A., Mihail Kantor, Erika Consoli, Niclas H. Lyndby, Amy Michaud, Zafar Handoo, and Renato N. Inserra. 2025. "Molecular Phylogeny and Species Delimiting for the Genus Hoplolaimus (Nematoda: Tylenchida) with Description of Hoplolaimus floridensis sp. n. and Notes on Biogeography of the Genus in the United States" International Journal of Molecular Sciences 26, no. 17: 8501. https://doi.org/10.3390/ijms26178501
APA StyleSubbotin, S. A., Kantor, M., Consoli, E., Lyndby, N. H., Michaud, A., Handoo, Z., & Inserra, R. N. (2025). Molecular Phylogeny and Species Delimiting for the Genus Hoplolaimus (Nematoda: Tylenchida) with Description of Hoplolaimus floridensis sp. n. and Notes on Biogeography of the Genus in the United States. International Journal of Molecular Sciences, 26(17), 8501. https://doi.org/10.3390/ijms26178501