Table 1.
Morphometric data of H. multicinctus and reference populations.
| Species | H. multicinctus | H. multicinctus | H. multicinctus | H. multicinctus | H. multicinctus | H. multicinctus |
|---|---|---|---|---|---|---|
| Locality/publication | Plantain, Cordoba–Quindío–HM1 n = 19 | Plantain, Rozo–Palmira–HM2 n = 20 | Banana, Brazil HM3–* n = 24 | Banana, Brazil 233–HM4 n = 21 |
Goodey (1940), Golden (1956) n = 2 |
Uzma et al. (2015) n = not defined |
| L | 0.6 ± 0.1 (0.4–0.8) | 0.6 ± 0.04 (0.5-0.7) | 0.5 ± 0.05 (0.5-0.6) | 0.5 ± 0.1 (0.4-0.6) | 0.54 (0.46-0.68) | 0.47-0.53 |
| a | 26.1 ± 2.8 (22.5-31.1) | 24.4 ± 1.5 (22.1-27.4) | 24.7 ± 1.9 (22.4-30.3) | 24.5 ± 2.0 (21.2-27.9) | 24.47 (23.8-28.5) | 24.0-30.0 |
| c | 45.6 ± 5.1 (36.1-55.2) | 44.6 ± 5.6 (36.5-59.8) | 49.2 ± 7.0 (37.7-64.1) | 50.7 ± 7.4 (39.6-69.6) | 53.8 (48.0-63.0) | 35.0-46.0 |
| c′ | 0.9 ± 0.1 (0.7-1.1) | 0.9 ± 0.1 (0.7-1.0) | 0.8 ± 0.1 (0.6-1.1) | 0.8 ± 0.1 (0.4-1.0) | 0.8-1.0 | 0.8-1.0 |
| V% | 68.4 ± 1.6 (65.9-71.7) | 67.2 ± 1.7 (64.1-70.0) | 68.9 ± 2.3 (65.0-73.1) | 69.1 ± 1.1 (66.2-71.1) | 69.0 (64.6-71.8) | 65.0-69.0 |
| DGO | 8.6 ± 0.6 (8.0-10.0) | 9.5 ± 0.5 (9.0-10.0) | 9.1 ± 1.0 (7.0-11.0) | 9.3 ± 1.1 (7.0-11.0) | – | – |
| Stylet length | 23.4 ± 1.3 (21.0-26.0) | 23.7 ± 0.7 (23.0-25.0) | 23.5 ± 0.9 (22.0-25.0) | 23.1 ± 0.8 (22.0-24.0) | 20.0-24.0 | 22.0-24.0 |
| Number of head annuli | 4.0 ± 0.6 (3.0-5.0) | 4.0 ± 0.5 (4.0-5.0) | 4.0 ± 0.5 (4.0-5.0) | 4.0 ± 0.0 (4.0-4.0) | 3.0-5.0 | 3.0-4.0 |
| Tail length | 13.0 ± 1.9 (10.0-16.0) | 12.9 ± 1.2 (10.0-15.0) | 11.3 ± 1.7 (8.0-15.0) | 10.5 ± 1.9 (7.0-15.0) | – | – |
| Number of tail annuli | 9.0 ± 1.2 (7.0-12.0) | 9.3 ± 1.3 (7.0-12.0) | 8.0 ± 1.9 (5.0-12.0) | 7.7 ± 1.5 (5.0-10.0) | 6.0-13.0 | 6.0-12.0 |
| Position of phasmid | 3.0 ± 1.2 (2.0-6.0) | 3.8 ± 1.0 (3.0-6.0) | 5.0 ± 2.1 (2.0-7.0) | 5.0 ± 1.2 (3.0-7.0) | 1.0-6.0 | 2.0-6.0 |
Table 4.
Morphometric data of H. californicus and reference populations.
| Species | H. californicus | H. californicus | H. californicus | H. californicus | H. californicus |
|---|---|---|---|---|---|
| Locality/publication | Plantain, Delfina–Buenaventura–HC1 n = 12 | Banana, Brazil HC2 n = 19 |
Van Den Berg and Heyns (1975) n = 25 |
Uzma et al. (2015) n = not defined |
Sher (1966) n = 16 |
| L | 0.6 ± 0.05 (0.5-0.6) | 0.6 ± 0.1 (0.5-0.7) | 0.7 (0.6-0.8) | 0.58-0.78 | 0.6 (0.5-0.7) |
| a | 23.5 ± 1.0 (22.0-25.2) | 24.6 ± 1.4 (21.5-26.6) | 28.8 (23.4-35.0) | 27-32 | 24.1 (21.1-26.2) |
| c | 35.5 ± 4.7 (27.2-45.8) | 34.9 ± 3.0 (30.3-40.2) | 39.0 (30.8-53.0) | 32.0-50.0 | 38.7 (31.2-48.1) |
| c′ | 1.0 ± 0.1 (0.8-1.1) | 1.1 ± 0.1 (1.0-1.3) | 1.1 (0.7-1.5) | 0.8-1.3 | 1.2 (0.9-2.1) |
| V% | 62.1 ± 1.7 (59.3-64.8) | 65.6 ± 3.0 (61.3-75.5) | 63.0 (60.0-64) | 59-64 | 64 (60-67) |
| DGO | 11.4 ± 1.2 (10.0-14.0) | 11.4 ± 2.4 (8.0-14.0) | – | – | 10 (8-14) |
| Stylet length | 22.4 ± 0.9 (20.0-23.0) | 23.5 ± 1.0 (21.0-25.0) | 24.1 (22.8-27.2) | 24-27 | 27 (26-29) |
| Number of head annuli | 4.7 ± 0.7 (4.0-6.0) | 4.2 ± 0.5 (4.0-6.0) | 3.0-5.0 | 4 | 5-7 |
| Tail length | 15.9 ± 1.0 (13.0-19.0) | 16.5 ± 1.5 (14.0-20.0) | 17.9 (15.1-25.0) | – | 16 (13-21) |
| Number of tail annuli | 7.8 ± 1.1 (6.0-9.0) | 7.8 ± 0.9 (6.0-9.0) | 10.0-17.0 | 8 | 9 (6-12) |
| Position of phasmid | 6.8 ± 1.3 (5.0-8.0) | 6.0 ± 2.7 (2.0-8.0) | 3.0-14.0 | 2 | 1-11 |
Figure 1:
Helicotylenchus multicinctus. A: Habitus post-mortem of a female, B and C: Anterior region of female body, D and E: Posterior region of female body, F: Medium region of female body, G: Posterior region of male body. vu = Vulva, dgo = Dorsal esophageal Gland Orifice, an = Anus, lf = Lateral field, sptc = Spermatheca, spc = Spicule.
Table 2.
Morphometric data of H. dihystera and reference populations.
| Species | H. dihystera | H. dihystera | H. dihystera | H. dihystera | H. dihystera | |
|---|---|---|---|---|---|---|
| Locality/publication | Banana, Córdoba–Quindío–HD1 | Plantain, Córdoba–Quindío–HD2 | Sher (1961) | Uzma et al. (2015) | Wount and Yates | |
| n = 10 | n = 15 | n = 12 | n = not defined | n = 10 | n = 10 | |
| L | 0.62 ± 0.08 (0.53-0.77) | 0.61 ± 0.06 (0.48-0.69) | 0.67 (0.61-0.86) | 0.59-0.79 | 0.67 ± 0.05 (0.57-0.73) | 0.71 ± 0.04 (0.61-0.75) |
| a | 25.5 ± 1.94 (22.93-29.69) | 24.0 ± 1.83 (20.50-28.13) | 29.5 (26.0-34.0) | 27-35 | 29.3 ± 2.21 (26.0-33.0) | 29.5 ± 1.35 (27.0-32.0) |
| c | 43.8 ± 8.36 (25.12-55.45) | 45.3 ± 4.49 (39.0-56.3) | 48.0 (40.0-65.0) | 35- 49 | 46.0 ± 4.69 (37.0-53.0) | 43.0 ± 4.67 (35.0-50.0) |
| c′ | 1.0 ± 0.17 (0.80-1.31) | 0.9 ± 0.07 (0.71-0.94) | 1.0-1.3 | 0.8-1.2 | 1.11 ± 0.11 (1.0-1.3) | 1.09 ± 0.11 (0.9-1.3) |
| V% | 64.6 ± 1.19 (62.24-66.35) | 64.5 ± 1.24 (61.94-66.12) | 63.0 (60.0-66.0) | 60-65 | 65 ± 1.32 (63.0-67.0) | 63.0 ± 0.84 (61.0-64.0) |
| DGO | 14.7 ± 1.38 (13.0-16.0) | 14.9 ± 1.57 (13.0-17.0) | – | – | 11.5 ± 1.63 (8.5-13.5) | 10.0 ± 1.25 (7.0-11.0) |
| Stylet length | 24.4 ± 0.97 (23.0-26.0) | 23.8 ± 0.77 (23.0-25.0) | 26.0 (24.5-27.5) | 24-29 | 27.9 ± 1.08 (25.5-29.0) | 26.1 ± 0.74 (25.0-27.0) |
| Number of head annuli | 4.0 ± 0.0 (4.0-4.0) | 4.0 ± 0.26 (4.0-5.0) | 3.0-4.0 | 4-5 | – | – |
| Tail length | 14.5 ± 3.06 (11.0-21.0) | 13.5 ± 1.51 (12.0-16.0) | – | – | 14.5 ± 1.48 (12.0-17.0) | 16.7 ± 2.50 (12.0-21.0) |
| Number of tail annuli | 8.0 ± 1.88 (5.0-12.0) | 8.0 ± 0.68 (7.0-10.0) | 8-12 | 6-12 | – | – |
| Position of phasmid | 8.0 ± 0.99 (7.0-10.0) | 9.0 ± 1.91 (6.0-11.0) | 6-12 | 5-11 | – | – |
Figure 2:
Helicotylenchus dihystera, H. erythrinae, and H. californicus. A: Habitus post mortis of a female in H. dihystera, B: Anterior region of female body in H. dihystera, C: Posterior region of female body in H. dihystera, D: Habitus post mortis of a female in H. erythrinae, E: Anterior region of female body in H. erythrinae, F: Posterior region of female body in H. erythrinae, G: Spermatheca of female in H. erythrinae, H: Habitus post mortis of female in H. californicus, I and J: Anterior region of female body in H. californicus, K: Posterior region of female body in H. californicus with short ventral projection and sharply pointed tail, L: Posterior region of female body in H. californicus with short ventral projection and blunt tail, M: Posterior region of male body in H. californicus.
Table 3.
Morphometric data of H. erythrinae and reference populations.
| Species | H. erythrinae | H. erythrinae | H. erythrinae | H. erythrinae | H. erythrinae | |
|---|---|---|---|---|---|---|
| Locality/publication | Banana, Calarcá–Quindío (La Julia)–HE1 | Plantain, Calarcá –Quindío (Resta)–HE2 | Van Den Berg and Heyns (1975) | Uzma et al. (2015) | Wount and Yates | |
| n = 15 | n = 25 | n = 7 | n = not defined | n = 10 | n = 10 | |
| L | 0.53 ± 0.1 (0.4-0.7) | 0.6 ± 0.1 (0.46-0.68) | 0.5 (04-0.6) | 0.48-0.61 | 0.87 ± 0.06 | 0.69 ± 0.05 |
| a | 25.03 ± 2.0 (22.7-29.7) | 25.90 ± 2.2 (22.86-30.91) | 25.1 (21.1-26.7) | 23.0-26.0 | 29.7 ± 1.64 (27.0-32.0) | 28.2 ± 1.87 (24.0-30.0) |
| c | 25.93 ± 4.1 (19.2-31.5) | 28.09 ± 3.0 (21.82-32.94) | 28.9 (23.3-30.3) | 27.0-34.0 | 43.0 ± 5.99 (31.0 -51.0) | 48.0 ± 10.55 (35.0-74.0) |
| c′ | 1.46 ± 0.2 (1.1-1.7) | 1.41 ± 0.15 (1.13-1.73) | 1.4 (1.3-1.4) | 1.0-1.6 | 1.13 ± 0.106 (1.0-1.3) | 1.03 ± 0.206 (0.60-1.30) |
| V% | 63.8 ± 1.8 (60.1-67.6) | 63.7 ± 2.16 (60.82-68.54) | 64.0 (61.0-67.0) | 60.0-65.0 | 62.0 ± 2.85 (58.0-68.0) | 63.0 ± 2.22 (59.0-65.0) |
| DGO | 10.36 ± 2.6 (5.0-8.0) | 12 ± 1.97 (8.0-15.0) | – | – | 9.6 ± 2.07 (75.0-13.0) | 9.1 ± 0.94 (7.0-10.0) |
| Stylet length | 22.7 ± 1.0 (21.0-25.0) | 22.8 ± 0.83 (21.0-24.0) | 24.6 (21.7-26.8) | 23.0-26.0 | 28.3 ± 1.23 (26.0-30.0) | 27.9 ± 0.97 (25.5-29.0) |
| Number of head annuli | 4.0 ± 0.0 (4.0-4.0) | 4.0 ± 0.32 (3.0-5.0) | 4.0 | 4.0-5.0 | – | – |
| Tail length | 20.53 ± 2.2 (17.0-25.0) | 20.65 ± 2.39 (17.0-26.0) | 17.9 | – | 20.6 ± 2.8 (16.0-24.0) | 14.7 ± 2.71 (9.0-18.0) |
| Number of tail annuli | 10.0 ± 2.8 (7.0-17) | 11.0 ± 1.01 (10.0-13.0) | 10.0-14.0 | 6.0-12.0 | 13.4 ± 0.97 (12.0-15.0) | 10.2 ± 2.25 (7.0-15.0) |
| Position of phasmid | 6.0 ± 0.9 (4.0-7.0) | 3.0 ± 1.01 (1.0-5.0) | 9 | 4/2 | – | – |
Table 5.
Eigenvalues and percent total variance accounted for each principal component.
| Principal component | Eigenvalues | Cumulative total | % of Total variance | Cum. % of total variance |
|---|---|---|---|---|
| 1 | 4.92 | 4.92 | 37.88 | 37.88 |
| 2 | 3.75 | 8.68 | 28.88 | 66.76 |
| 3 | 2.16 | 10.84 | 16.63 | 83.39 |
| 4 | 1.04 | 11.88 | 7.99 | 91.39 |
| 5 | 0.35 | 12.23 | 2.70 | 94.09 |
| 6 | 0.29 | 12.52 | 2.22 | 96.31 |
| 7 | 0.22 | 12.74 | 1.68 | 97.99 |
| 8 | 0.16 | 12.89 | 1.23 | 99.22 |
| 9 | 0.06 | 12.95 | 0.43 | 99.65 |
| 10 | 0.04 | 12.99 | 0.29 | 99.94 |
| 11 | 0.01 | 13 | 0.06 | 100 |
| 12 | 1.5128E–7 | 13 | 1.16369E–6 | 100 |
| 13 | –4.65844E–10 | 13 | –3.58342E–9 | 100 |
Table 6.
Correlations between the first four component principals and the morphometric parameters of females in Helicotylenchus spp.
| Vector | Diagnostic character | 1 | 2 | 3 | 4 |
|---|---|---|---|---|---|
| 1 | L | –0.25 | 0.21 | 0.02 | 0.66 |
| 2 | a | 0.09 | –0.01 | –0.57 | 0.35 |
| 3 | c | –0.34 | –0.33 | –0.01 | 0.02 |
| 4 | c′ | 0.36 | 0.28 | –0.12 | –0.11 |
| 5 | V % | –0.06 | –0.48 | –0.13 | 0.01 |
| 6 | DGO | –0.22 | 0.35 | –0.25 | –0.27 |
| 7 | Stylet length | –0.31 | –0.05 | –0.39 | –0.03 |
| 8 | Number of head annuli | 0.01 | 0.05 | 0.61 | 0.07 |
| 9 | Number of tail annuli | 0.37 | 0.07 | –0.09 | 0.33 |
| 10 | Position of phasmid | –0.29 | 0.31 | –0.09 | –0.40 |
| 11 | Maximum body diameter | –0.39 | 0.22 | 0.02 | 0.15 |
| 12 | Tail length | 0.32 | 0.35 | –0.10 | –0.05 |
| 13 | Anal body diameter | –0.24 | 0.37 | 0.17 | 0.22 |
Figure 3:
Populations of H. multicinctus (HM), H. dihystera (HD), H. erythrinae (HE), and H. californicus (HC) from Colombia and Brazil can be assigned to its corresponding species based on morphometric data. The two first axes of a principal components analysis (PCA) are shown. HM1 to HM2 from Colombia and HM3 to HM4 from Brazil; HD1 to HD3 from Colombia; HE1 to HE3 from Colombia; and HC1 from Colombia and HC2 from Brazil.
Figure 4:
Dendrogram obtained by a conglomerates analysis to classify the Colombian and Brazilian populations of H. multicinctus (HM), H. dihystera (HD), H. erythrinae (HE), and H. californicus (HC). HM1 to HM2 from Colombia and HM3 to HM4 from Brazil; HD1 to HD3 from Colombia; HE1 to HE3 from Colombia; and HC1 from Colombia and HC2 from Brazil.
Table 7.
Information of sequences D2 to D3 of ribosomal DNA downloaded from GenBank and obtained in the present study for Helicotylenchus.
| Species name | Location | Plant-host | GenBank accession number | Reference or source |
|---|---|---|---|---|
| H. dihystera | Colombia, Córdoba, Quindío | Musa AAA (Banana) | MT321729; MT321730 | Present study |
| H. dihystera | USA, Florida, Ft Lauderdale | Schefflera arboricola | HM014245 | Subbotin et al. (2011) |
| H. dihystera | USA, Florida, Goulds | Bromeliads | HM014246 | Subbotin et al. (2011) |
| H. dihystera | USA, Florida, Ft Pierce | Ficus | HM014247 | Subbotin et al. (2011) |
| H. dihystera | USA, Hawaii, Kawai | Grasses | HM014248 | Subbotin et al. (2011) |
| H. dihystera | USA, Georgia | – | HM014249 | Subbotin et al. (2011) |
| H. dihystera | USA, Hawaii, Kawai | Grasses | HM014250 | Subbotin et al. (2011) |
| H. dihystera | USA, Georgia | – | HM014251 | Subbotin et al. (2011) |
| H. dihystera | USA, California, Kern county | Vitis vinifera | KM506834 | Subbotin et al. (2015) |
| H. dihystera | USA, Texas (plant shipment) | – | KM506835 | Subbotin et al. (2015) |
| H. dihystera | USA, California, Merced | Grasses | KM506836 | Subbotin et al. (2015) |
| H. dihystera | China, Fujian Province | Musa sp. | KF486503 | Xiao et al. (2014) |
| H. leiocephalus | USA, Nebraska, Lincoln, Nine mile Prairie | Amorpha canescens | HM014268; HM014269 | Subbotin et al. (2011) |
| H. pseudorobustus | USA, Kansas, Konza Prairie, Manhattan | Koeleria pyramidata | HM014266; HM014267 | Subbotin et al. (2011) |
| H. pseudorobustus | New Zealand, MAF farm, Kaitoke | Lolium perenne, Trifolium repens | HM014278 | Subbotin et al. (2011) |
| H. pseudorobustus | New Zealand, Rotorua | Lolium perenne | HM014279; HM014280 | Subbotin et al. (2011) |
| H. pseudorobustus | China, Beijing | – | DQ328747 | Subbotin et al. (2011) |
| H. pseudorobustus | USA, California, Fresno | – | DQ328748 | Subbotin et al. (2011) |
| H. pseudorobustus | Italy, Ancona | – | DQ328750 | Subbotin et al. (2011) |
| H. pseudorobustus | Germany, Münster, BBA glasshouse | – | DQ328751 | Subbotin et al. (2011) |
| Helicotylenchus spIV-2 | USA, Florida, Gainesville, Winter garden | Calathea sp. | KM506844 | Subbotin et al. (2015) |
| H. multicinctus | Colombia, Calarcá, Quindío | Musa AAA (Banana) | MT321731 | Present study |
| H. multicinctus | Colombia, Rozo, Palmira, Valle del Cauca | Musa AAB (Plantain) | MT321732; MT321733; MT321734 | Present study |
| H. multicinctus | South Africa, Lambani | Musa sp. | HM014290; HM014291 | Subbotin et al. (2011) |
| H. multicinctus | USA, Florida, Ft. Pierce | Ficus benjamina | HM014292 | Subbotin et al. (2011) |
| H. multicinctus | Sudan | Musa sp. | DQ328745; DQ328746 | Subbotin et al. (2011) |
| H. multicinctus | China, Fujian Province | Musa sp. | KF443214 | Xiao et al. (2014) |
| H. californicus | Colombia, La Delfina, Buenaventura, Valle del Cauca | Musa AAB (Plantain) | MT321735; MT321736; MT321737; MT321738 | Present study |
| H. erythrinae | Colombia, La Celia, Risaralda | Musa AAB (Plantain) | MT321739 | Present study |
| H. labiodiscinus | USA, Kansas, Manhattan | Poa pratensis | HM014293 | Subbotin et al. (2011) |
| H. labiodiscinus | USA, Kansas, Manhattan | Andropogon gerardii | HM014294 | Subbotin et al. (2011) |
| H. labiodiscinus | USA, Kansas, Manhattan | Schizachyrium scoparium | HM014295 | Subbotin et al. (2011) |
| H. martini | South Africa, Lambani | Grasses | HM014304; HM014305 | Subbotin et al. (2011) |
| H. brevis | South Africa, North West Province | Solanum mauritianum | HM014299; HM014300 | Subbotin et al. (2011) |
| H. vulgaris | Italy, Ancona | – | DQ328759; DQ328760; DQ328761 | Subbotin et al. (2011) |
| H. vulgaris | USA, Kansas, University of Arkansas | – | FJ485650 | Subbotin et al. (2011) |
Figure 5:
Maximum likelihood phylogeny of Helicotylenchus. The tree was estimated using the D2 to D3 expansion segment of 28S RNAr and 250 bootstraps under GTR + Γ model. The outgroup (Rotylenchus magnus) is shown in gray font; the sequences that were obtained in this study appear in bold typeface. Values at the nodes represent the bootstrap support. Were provided bootstrap support for nodes with values > 80. The scale represents the number of substitutions per site.
Figure 6:
Bayesian phylogeny of Helicotylenchus based on D2 to D3 expansion segment of 28S RNAr. The phylogeny is a consensus tree from a posterior distribution of 1,600 trees that were inferred under GTR + Γ model in MrBayes. The outgroup (Rotylenchus magnus) is shown in gray font; the sequences that were obtained in this study appear in bold typeface. Values at the nodes represent the posterior probability. Were provided posterior probabilities for nodes with values > 0.5. The scale represents the number of substitutions per site.