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Morphological and molecular characterization of Heterodera dunensis n. sp. (Nematoda: Heteroderidae) from Gran Canaria, Canary Islands

Phougeishangbam Rolish Singh
Phougeishangbam Rolish Singh
, 
Gerrit Karssen
Gerrit Karssen
, 
Marjolein Couvreur
Marjolein Couvreur
 e 
Wim Bert
Wim Bert
   | 24 nov 2020
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Pubblicato online: 24 nov 2020
Pagine: 1 - 14
DOI: https://doi.org/10.21307/jofnem-2020-098
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© 2020 Phougeishangbam Rolish Singh et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1:

Light microscopy and scanning electron microscopy images of second-stage juveniles of Heterodera dunensis n. sp. A to D: En face view, E: Anterior part up to pharyngeal gland end, F to H: Labial region showing stylet and labial annuli, I: Total body, J to L: Anterior part showing median bulb, hemizonid, and secretory-excretory pore, M to Q: Tail region showing hyaline portion and anus.
Light microscopy and scanning electron microscopy images of second-stage juveniles of Heterodera dunensis n. sp. A to D: En face view, E: Anterior part up to pharyngeal gland end, F to H: Labial region showing stylet and labial annuli, I: Total body, J to L: Anterior part showing median bulb, hemizonid, and secretory-excretory pore, M to Q: Tail region showing hyaline portion and anus.

Figure 2:

Light microscopy and scanning electron microscopy images of cysts and vulva cones of Heterodera dunensis n. sp. A, E: In toto, lemon-shape cysts showing protruded vulva cones and bent necks, B to D, F to H: Vulva cones showing cone fenestration and vulva slits, I to L: Vulva cones showing anus, bullae, and underbridge.
Light microscopy and scanning electron microscopy images of cysts and vulva cones of Heterodera dunensis n. sp. A, E: In toto, lemon-shape cysts showing protruded vulva cones and bent necks, B to D, F to H: Vulva cones showing cone fenestration and vulva slits, I to L: Vulva cones showing anus, bullae, and underbridge.

Figure 3:

Line illustrations of cysts and second-stage juveniles (J2) of Heterodera dunensis n. sp. A: Whole cysts, B, C: Vulva cones showing cone fenestration, vulva slit, bullae, underbridge, and anus, D, F: Anterior part of J2 showing lip region, pharynx, hemizonid, and secretory excretory pore, E: Stylet of J2, G to I: Tail region of J2 showing lateral field differentiation, anus, phasmid, and tail hyaline portion.
Line illustrations of cysts and second-stage juveniles (J2) of Heterodera dunensis n. sp. A: Whole cysts, B, C: Vulva cones showing cone fenestration, vulva slit, bullae, underbridge, and anus, D, F: Anterior part of J2 showing lip region, pharynx, hemizonid, and secretory excretory pore, E: Stylet of J2, G to I: Tail region of J2 showing lateral field differentiation, anus, phasmid, and tail hyaline portion.

Figure 4:

Phylogenetic relationships of Heterodera dunensis n. sp. with 33 known Heterodera species. Bayesian 50% majority-rule consensus tree as inferred from the analysis of D2-D3 of 28S rDNA sequences under GTR + G model. Posterior probabilities of more than 0.5 are given for appropriate clades.
Phylogenetic relationships of Heterodera dunensis n. sp. with 33 known Heterodera species. Bayesian 50% majority-rule consensus tree as inferred from the analysis of D2-D3 of 28S rDNA sequences under GTR + G model. Posterior probabilities of more than 0.5 are given for appropriate clades.

Figure 5:

Phylogenetic relationships of Heterodera dunensis n. sp. with 54 known Heterodera species. Bayesian 50% majority-rule consensus tree as inferred from the analysis of partial ITS of rDNA sequences under GTR + I + G model. Posterior probabilities of more than 0.5 are given for appropriate clades.
Phylogenetic relationships of Heterodera dunensis n. sp. with 54 known Heterodera species. Bayesian 50% majority-rule consensus tree as inferred from the analysis of partial ITS of rDNA sequences under GTR + I + G model. Posterior probabilities of more than 0.5 are given for appropriate clades.

Figure 6:

Phylogenetic relationships of Heterodera dunensis n. sp. with 29 known Heterodera species. Bayesian 50% majority-rule consensus tree as inferred from the analysis of COI of mtDNA sequences under GTR + I + G model. Posterior probabilities of more than 0.5 are given for appropriate clades.
Phylogenetic relationships of Heterodera dunensis n. sp. with 29 known Heterodera species. Bayesian 50% majority-rule consensus tree as inferred from the analysis of COI of mtDNA sequences under GTR + I + G model. Posterior probabilities of more than 0.5 are given for appropriate clades.

Comparison of important characters of seventeen Heterodera species of the Schachtii group.

Character Cyst length (L) Cyst width (W) L/W ratio Fenestral length Fenestral width Vulval slit length J2 body length J2 stylet length J2 tail length J2 tail hyaline length Hyaline% of the tail
H. agrostis 429-800 320-541 1.2-2.6 34-54 16-24 39-48 384-472 25-26 46-61 27-40 59-66
H. betae 830-878 455-518 1.6-1.9 44-55 38-43 48-57 547-607 29-31 70-74 38-42 54-57
H. cajani 448-670 209-422 1.4-2.1 27-65 25-40 43-55 420-519 22-27 42-52 23-31 55-60
H. ciceri 570-930 350-550 1.6-2.4 32-52 20-37 43-60 440-585 27-30 53-72 31-42 58-58
H. daverti 650-749 380-491 1.4-1.5 42-54 31-40 47-52 457-476 25-26 54-57 30-33 56-58
H. dunensis n. sp. 316-610 241-478 1.2-1.9 37-51 34-53 35-63 426-520 27-31 35-45 16-23 46-51
H. galeopsidis 576-797 408-556 1.4-1.5 41-50 31-38 39-50 485-553 26-28 61-75 35-40 57-53
H. glycines 474-709 327-535 1.3-1.7 34-58 16-41 38-50 386-471 21-23 39-51 22-30 56-59
H. lespedezae 678-719 371-522 1.4-1.8 43-59 35-41 45-47 457-481 24-25 54-56 26-30 48-54
H. medicaginis 568-728 364-570 1.4-1.5 39-55 30-40 39-55 417-512 24-26 41-60 22-33 54-55
H. mediterranea 430-690 240-570 1.2-1.6 38-45 37-42 42-48 360-430 25-27 38-45 19-26 50-58
H. rosii 537-1173 403-634 1.0-1.7 48-65 40-45 45-59 430-662 27-34 58-77 37-45 64-58
H. schachtii 768-815 512-529 1.5-1.6 35-38 25-31 41-44 436-489 25-26 45-49 24-27 53-55
H. sonchophila 732-1032 381-616 1.6 37-58 29-50 42-56 437-492 24-27 47-56 26-30 55-54
H. spiraeae 467-861 283-566 1.1-1.7 33-60 20-45 39-45 371-446 22-23 39-49 21-27 54-55
H. swarupi 520-700 320-475 1.6-2.1 45 35 41 400-440 21-23 39-54 20-29 51-54
H. trifolii 608-841 341-536 1.3-1.8 43-53 33-44 40-53 492-613 25-28 60-72 32-37 53-51

Morphometric data of Heterodera dunensis n. sp. from fixed specimens mounted in glycerin, except for cysts that were not fixed.

Cysts
 n 21
 Length without neck (L) 482  ±  75 (316-610)
 Maximum cyst width (W) 360 ± 72.1 (241-478)
 Neck length 110 ± 31.9 (55-180)
 L/W 1.4 ± 0.2 (1.2-1.9)
Vulval area
 n 13
 Fenestral length 42.1 ± 4.3 (37-51)
 Mean semifenestral width 41.8 ± 5.2 (34-53)
 Vulva bridge width 8.1 ± 1.6 (5.2-11)
 Vulva slit length 52.1 ± 7.3 (35-63)
 Underbridge length 63.4 ± 9.8 (44-83)
 Underbridge width 18.2 ± 6.2 (8.8-29)
 Vulva-anus distance 58.4 ± 5.7 (50-68)
Juveniles
 n 21
 Body length (L) 471 ± 25.3 (426-520)
 a = L/MBD 20.1 ± 1.4 (19-23)
 b = L/Anterior end to pharynx-intestine junction 3.6 ± 0.4 (3.1-4.8)
 b′ = L/Anterior end to the end of pharyngeal gland 2.7 ± 0.2 (2.4-3.1)
 c = L/Tail length 11.6 ± 0.6 (11-12)
 c′ = Tail length/ABD 33.6 ± 2.6 (27-39)
 Stylet length 29.1 ± 1.1 (27-31)
 Stylet knob width 6.4 ± 0.4 (5.9-7)
 Lip region height 4.8 ± 0.3 (4.3-5.3)
 Lip region width 9.1 ± 0.4 (8.6-10)
 Dorsal gland opening from stylet base 3.7 ± 0.4 (3.2-4.4)
 Anterior end to median valve 74.2 ± 2.4 (70-79)
 Anterior end to secretory-excretory pore 109 ± 3.9 (101-117)
 Anterior end to pharynx-intestine junction 131 ± 11.1 (103-155)
 Anterior end to pharyngeal gland end 177 ± 9.2 (156-193)
 Maximum body diameter (MBD) 23.5 ± 1.4 (21-26)
 Anal body diameter (ABD) 14.1 ± 0.8 (13-16)
 Tail length 40.7 ± 2.4 (35-45)
 Hyaline tail length 20.6 ± 1.9 (16-23)
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