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DNA barcoding evidence for the North American presence of alfalfa cyst nematode, Heterodera medicaginis Cover

DNA barcoding evidence for the North American presence of alfalfa cyst nematode, Heterodera medicaginis

Journal of Nematology
Volume 51 (2019): Issue 1 (January 2019)
By: Thomas Powers,  Andrea Skantar,  Tim Harris,  Rebecca Higgins,  Peter Mullin,  Saad Hafez,  Zafar Handoo,  Tim Todd and  Kirsten Powers  
Open Access
|Apr 2019

Figures & Tables

Figure 1

Maximum likelihood tree inferred from the COI gene. Each new specimen is represented by an identification number, species name, and location. GenBank accessions are labeled by accession number, name, and the label ‘GB’ (GenBank). Bootstrap support values (%) are labeled in red, the species nodes of H. schachtii, H. medicaginis, and H. glycines are circled in blue.
Maximum likelihood tree inferred from the COI gene. Each new specimen is represented by an identification number, species name, and location. GenBank accessions are labeled by accession number, name, and the label ‘GB’ (GenBank). Bootstrap support values (%) are labeled in red, the species nodes of H. schachtii, H. medicaginis, and H. glycines are circled in blue.

Figure 2

Maximum likelihood tree inferred from the ITS1 gene. Bootstrap support values above 50% are labeled in red. GenBank accession specimen from Russia is boxed in yellow.
Maximum likelihood tree inferred from the ITS1 gene. Bootstrap support values above 50% are labeled in red. GenBank accession specimen from Russia is boxed in yellow.

Figure 3

Phylogenetic relationships of Heterodera species as inferred from analysis of a partial Hsp90 gene using Bayesian inference. Posterior probabilities more than 50% are given next to nodes. Newly obtained sequences are shown in bold.
Phylogenetic relationships of Heterodera species as inferred from analysis of a partial Hsp90 gene using Bayesian inference. Posterior probabilities more than 50% are given next to nodes. Newly obtained sequences are shown in bold.

Figure 4

Heterodera medicaginis juvenile and male specimens. (A–G), juvenile specimens; (H–J), male specimen. (A) NID 7095, entire body; (B) NID 7243, entire body; (C) NID 7095, anterior; (D) NID 7243, anterior; (E) NID 7095, tail; (F) NID 7243, tail; (G) NID 7243, lateral lines; (H) PNID 169028, anterior; (I) PNID 169028, tail; (J) PNID 169028, entire body.
Heterodera medicaginis juvenile and male specimens. (A–G), juvenile specimens; (H–J), male specimen. (A) NID 7095, entire body; (B) NID 7243, entire body; (C) NID 7095, anterior; (D) NID 7243, anterior; (E) NID 7095, tail; (F) NID 7243, tail; (G) NID 7243, lateral lines; (H) PNID 169028, anterior; (I) PNID 169028, tail; (J) PNID 169028, entire body.

Figure 5

(A–D) light micrograph of vulval cones of H. medicaginis showing fenestra (A), bullae (B,D) and underbridge (B,D).
(A–D) light micrograph of vulval cones of H. medicaginis showing fenestra (A), bullae (B,D) and underbridge (B,D).

Figure 6

SEM micrographs of Heterodera medicaginis cysts from Kansas. (A) entire cyst 1, partial view of cyst 2; (B) entire cyst 3; (C) anterior region and excretory pore of cyst 1; (D) anterior region of cyst 3; (E) vulva and anus of cyst 2; (F) vulva of cyst 2 (left) and cyst 1 (right).
SEM micrographs of Heterodera medicaginis cysts from Kansas. (A) entire cyst 1, partial view of cyst 2; (B) entire cyst 3; (C) anterior region and excretory pore of cyst 1; (D) anterior region of cyst 3; (E) vulva and anus of cyst 2; (F) vulva of cyst 2 (left) and cyst 1 (right).

Morphological data on juveniles, all measurements in µm_

Specimen No.LengthStyletTailHyaline
144025–25
242125–21
3425254525
4420255025
5421244322.5
6420234623
7447254528
8475245225.5
9462254522.5
1039525.54525
1149125–25.5
12485255021
13413254825
14410264020
15415254523
NID 7095462.5255027
NID 7096442.5245130
NID 7243477.52552.530
NID 7244455265025
NID 7245450264922.5
NID 7246425254827.5
NID 7247452.52547.527.5
NID 8876472275228
NID 8877470265032
Mean445.025.147.825.3
SD27.20.83.23.0
Maximum4912752.532
Minimum395234020
Gerber and Maas (1982) n = 100 Heterodera medicaginis 462 (417–512)25 (24–26)51.8 (41–60)28.5 (22–33)
Mulvey and Golden (1983) H. glycines 439 ± 6.7 (375–490)23 ± 0.01 (22–24)50.4 ± 1.0 (42–59)26.6 ± 0.7 (20–33)
Mulvey and Golden (1983) H. schachtii 452 ± 49.725.6 ± 0.1148.5 ± 0.7327.1 ± 0.61

Collection data for specimens used in this study_ Specimens 1 to 15 were examined as fixed specimens_

Specimen IDSpeciesLocalityHostMarkerGenBank Accession No.
1 H. medicaginis Kearny County, KansasAlfalfa N/A
2 H. medicaginis Kearny County, KansasAlfalfa N/A
3 H. medicaginis Kearny County, KansasAlfalfa N/A
4 H. medicaginis Kearny County, KansasAlfalfa N/A
5 H. medicaginis Kearny County, KansasAlfalfa N/A
6 H. medicaginis Kearny County, KansasAlfalfa N/A
7 H. medicaginis Kearny County, KansasAlfalfa N/A
8 H. medicaginis Kearny County, KansasAlfalfa N/A
9 H. medicaginis Kearny County, KansasAlfalfa N/A
10 H. medicaginis Kearny County, KansasAlfalfa N/A
11 H. medicaginis Kearny County, KansasAlfalfa N/A
12 H. medicaginis Kearny County, KansasAlfalfa N/A
13 H. medicaginis Kearny County, KansasAlfalfa N/A
14 H. medicaginis Kearny County, KansasAlfalfa N/A
15 H. medicaginis Kearny County, KansasAlfalfa N/A
P169028 H. medicaginis Kearny County, KansasAlfalfa18SAY912048
N838 H. schachtii Goshen County, WyomingSoybeanCOIMK093062
N839 H. schachtii Goshen County, WyomingSoybeanCOIMK093063
N864 H. schachtii Goshen County, WyomingSoybeanCOIMK093064
N4143 Meloidodera sp.Big Thicket National Preserve, TexasBeechCOIMK093163
N4178 Meloidodera sp.Big Thicket National Preserve, TexasWater oakCOIMK093159
N7083 H. avenae Rio Grande County, ColoradoBarleyCOIMK093164
N7095 H. medicaginis Kearny County, KansasAlfalfaCOIMK093160
N7096 H. medicaginis Kearny County, KansasAlfalfaCOIMK093162
N7243 H. medicaginis Kearny County, KansasAlfalfaCOI ITS1MK093168 MK093180
N7244 H. medicaginis Kearny County, KansasAlfalfaCOI ITS1MK093169 MK093181
N7245 H. medicaginis Kearny County, KansasAlfalfaCOIMK093170
N7246 H. medicaginis Kearny County, KansasAlfalfaCOI ITS1MK093171 MK093182
N7247 H. medicaginis Kearny County, KansasAlfalfaCOIMK093172
N7248 H. trifolii Rio Arriba County, New MexicoVineyardCOIMK093173
N7250 H. trifolii Rio Arriba County, New MexicoVineyardCOI ITS1MK093174 MK093183
N7251 H. avenae Akershus, NorwayOatsCOIMK093175
N7253 H. avenae Akershus, NorwayOatsCOIMK093176
N8306 H. schachtii Platte County, WyomingBeanCOIMK093165
N8876 H. medicaginis Kearny County, KansasAlfalfaCOIMK093166
N8877 H. medicaginis Kearny County, KansasAlfalfaCOIMK093161
N8881 Heterodera sp.Dąbrówka, PolandPeat meadowCOIMK093167
P119063 H. medicaginis Carbon County, MontanaAlfalfaCOIMK093053
P119064 H. schachtii Big Horn County, MontanaAlfalfaCOIMK093054
P125039 H. schachtii Platte County, WyomingBeanCOIMK093055
P125063 H. schachtii Laramie County, WyomingSugar beetCOIMK093056
P145076 H. schachtii Platte County, WyomingSugar beetCOIMK093057
P150049 H. avenae MontanaBarleyCOIMK093058
P150069 Vittatidera zeaphila TennesseeCornCOIMK093060
P164043 H. avenae MontanaBarleyCOIMK093059
P200034 Cactodera sp.Sheridan County, NebraskaPotatoes-wheatCOIMK093061
P231089 H. glycines UnknownSoybeanCOIMK093049
P213017 H. glycines UnknownSoybeanCOIMK093050
P231088 H. glycines UnknownSoybeanCOIMK093051
P231091 H. schachtii UnknownSoybeanCOIMK093052
P243062 H. glycines Richardson County, NebraskaSoybeanCOIMK093086
P243063 H. glycines Richardson County, NebraskaSoybeanCOIMK093087
P243064 H. glycines Richardson County, NebraskaSoybeanCOIMK093088
P243069 H. glycines Seward County, NebraskaSoybeanCOIMK093077
P243080 H. glycines Lancaster County, NebraskaSoybeanCOIMK093078
P243083 H. glycines Lancaster County, NebraskaSoybeanCOIMK093079
P243085 H. glycines Cuming County, NebraskaSoybeanCOIMK093080
P243086 H. glycines Cuming County, NebraskaSoybeanCOIMK093081
P243094 H. glycines NebraskaSoybeanCOIMK093084
P243096 H. glycines NebraskaSoybeanCOIMK093085
P244003 H. glycines Cedar County, NebraskaSoybeanCOIMK093095
P244004 H. glycines Cedar County, NebraskaSoybeanCOIMK093096
P244020 H. glycines Holt County, NebraskaSoybeanCOIMK093089
P244021 H. glycines Holt County, NebraskaSoybeanCOIMK093090
P244041 H. glycines Buffalo County, NebraskaSoybeanCOIMK093093
P244042 H. glycines Buffalo County, NebraskaSoybeanCOIMK093094
P244060 H. glycines Burt County, NebraskaSoybeanCOIMK093091
P244062 H. glycines Burt County, NebraskaSoybeanCOIMK093092
P244065 H. glycines Stanton County, NebraskaSoybeanCOIMK093082
P244066 H. glycines Stanton County, NebraskaSoybeanCOIMK093083
P244072 H. glycines Cass County, NebraskaSoybeanCOIMK093097
P244074 H. glycines Cass County, NebraskaSoybeanCOIMK093098
P244076 H. schachtii Big Horn County, MontanaPinto beanCOIMK093065
P244077 H. schachtii Big Horn County, MontanaPinto beanCOIMK093066
P244078 H. schachtii Big Horn County, MontanaBlack beanCOIMK093067
P244079 H. schachtii Big Horn County, MontanaBlack beanCOIMK093068
P244081 H. schachtii Park County, WyomingPinto beanCOIMK093069
P244082 H. schachtii Park County, WyomingPinto beanCOIMK093070
P244084 H. schachtii Platte County, WyomingPinto beanCOIMK093071
P244085 H. schachtii Platte County, WyomingPinto beanCOIMK093072
P244086 H. schachtii Platte County, WyomingPinto beanCOIMK093073
P244087 H. schachtii Platte County, WyomingPinto beanCOIMK093074
P244088 H. schachtii Platte County, WyomingPinto beanCOIMK093075
P244089 H. schachtii Platte County, WyomingPinto beanCOIMK093076
P245019 H. glycines AlabamaSoybeanCOIMK093127
P245023 H. glycines AlabamaSoybeanCOIMK093128
P245025 H. glycines AlabamaSoybeanCOIMK093129
P245027 H. glycines AlabamaSoybeanCOIMK093130
P245029 H. glycines AlabamaSoybeanCOIMK093131
P245033 H. glycines AlabamaSoybeanCOIMK093119
P245035 H. glycines AlabamaSoybeanCOIMK093132
P245038 H. glycines AlabamaSoybeanCOIMK093133
P245041 H. glycines AlabamaSoybeanCOIMK093120
P245043 H. glycines AlabamaSoybeanCOIMK093134
P245044 H. glycines Cedar County, NebraskaSoybeanCOIMK093109
P245046 H. glycines Cedar County, NebraskaSoybeanCOIMK093110
P245089 H. glycines Oconee County, GeorgiaSoybeanCOIMK093111
P245090 H. glycines Oconee County, GeorgiaSoybeanCOIMK093112
P245094 H. glycines Burke County, GeorgiaSoybeanCOIMK093135
P245095 H. glycines Burke County, GeorgiaSoybeanCOIMK093136
P246006 H. glycines Boone County, MissouriSoybeanCOIMK093121
P246008 H. glycines Boone County, MissouriSoybeanCOIMK093137
P246010 H. glycines Livingston County, MissouriSoybeanCOIMK093138
P246014 H. glycines Knox County, MissouriSoybeanCOIMK093113
P246016 H. glycines Knox County, MissouriSoybeanCOIMK093114
P246018 H. glycines Atchison County, MissouriSoybeanCOIMK093139
P246021 H. glycines Atchison County, MissouriSoybeanCOIMK093122
P246033 H. glycines Fulton County, OhioSoybeanCOIMK093123
P246035 H. glycines Fulton County, OhioSoybeanCOIMK093140
P246042 H. glycines Sandusky County, OhioSoybeanCOIMK093124
P246044 H. glycines Sandusky County, OhioSoybeanCOIMK093115
P246047 H. glycines Sandusky County, OhioSoybeanCOIMK093116
P247094 H. glycines Lee County, ArkansasSoybeanCOIMK093141
P247096 H. glycines Lee County, ArkansasSoybeanCOIMK093142
P247099 H. glycines Lee County, ArkansasSoybeanCOIMK093143
P248001 H. glycines Lee County, ArkansasSoybeanCOIMK093144
P248004 H. glycines Washington County, ArkansasSoybeanCOIMK093145
P248006 H. glycines Washington County, ArkansasSoybeanCOIMK093146
P248009 H. glycines McLeod County, MinnesotaSoybeanCOIMK093147
P248010 H. glycines McLeod County, MinnesotaSoybeanCOIMK093125
P248013 H. glycines Wilken County, MinnesotaSoybeanCOIMK093117
P248016 H. glycines Wilken County, MinnesotaSoybeanCOIMK093118
P248017 H. glycines Nicollet County, MinnesotaSoybeanCOIMK093148
P248019 H. glycines Nicollet County, MinnesotaSoybeanCOIMK093149
P248023 H. glycines Dodge County, MinnesotaSoybeanCOIMK093099
P248024 H. glycines Dodge County, MinnesotaSoybeanCOIMK093100
P248026 H. glycines Red Lake County, MinnesotaSoybeanCOIMK093101
P248027 H. glycines Red Lake County, MinnesotaSoybeanCOIMK093102
P248029 H. glycines Redwood County, MinnesotaSoybeanCOIMK093150
P248031 H. glycines Redwood County, MinnesotaSoybeanCOIMK093151
P248035 H. glycines Dakota County, MinnesotaSoybeanCOIMK093103
P248036 H. glycines Dakota County, MinnesotaSoybeanCOIMK093104
P248040 H. glycines Waseca County, MinnesotaSoybeanCOIMK093105
P248041 H. glycines Waseca County, MinnesotaSoybeanCOIMK093106
P248058 H. cf. urticae Faulkner County, ArkansasChickweedCOIMK093155
P248059 H. cf. urticae Faulkner County, ArkansasChickweedCOIMK093156
P248060 H. trifolii Washington County, ArkansasCloverCOIMK093157
P248061 H. trifolii Washington County, ArkansasCloverCOIMK093158
P248066 H. glycines Gentry County, MissouriSoybeanCOIMK093107
P248068 H. glycines Gentry County, MissouriSoybeanCOIMK093108
P248071 H. glycines Mercer County, MissouriSoybeanCOIMK093152
P248073 H. glycines Mercer County, MissouriSoybeanCOIMK093153
P248086 H. glycines Sussex County, DelawareSoybeanCOIMK093154
P248087 H. glycines Sussex County, DelawareSoybeanCOIMK093126
105A13 H. medicaginis Kearny County, KansasAlfalfaCOIMK093177
105A14 H. medicaginis Kearny County, KansasAlfalfaCOIMK093178
105A15 H. medicaginis Kearny County, KansasAlfalfaCOIMK093179
Hsp90_3530 H. medicaginis Kearny County, KansasAlfalfaHsp90MH798843
Hsp90_3531 H. medicaginis Kearny County, KansasAlfalfaHsp90MH798844
Hsp90_3532 H. medicaginis Kearny County, KansasAlfalfaHsp90MH798845
Hsp90_3533 H. medicaginis Kearny County, KansasAlfalfaHsp90MH798846
Hsp90_2996 H. trifolii Alexandria, EgyptSugar beetHsp90MK095224
Hsp90_2997 H. trifolii Alexandria, EgyptSugar beetHsp90MK095225
Hsp90_2994 H. trifolii Alexandria, EgyptSugar beetHsp90MK095222
Hsp90_2995 H. trifolii Alexandria, EgyptSugar beetHsp90MK095223
Hsp90_2998 H. trifolii Alexandria, EgyptSugar beetHsp90MK095226
HetITS105 H. medicaginis Kearny County, KansasAlfalfaITSMK093184
HetITS106 H. medicaginis Kearny County, KansasAlfalfaITSMK093185
HetITS107 H. medicaginis Kearny County, KansasAlfalfaITSMK093186
HetITS108 H. medicaginis Kearny County, KansasAlfalfaITSMK093187
HetITS109 H. medicaginis Kearny County, KansasAlfalfaITSMK093188
HetITS110 H. medicaginis Kearny County, KansasAlfalfaITSMK093189
HetITS111 H. medicaginis Kearny County, KansasAlfalfaITSMK093190
HetITS112 H. medicaginis Kearny County, KansasAlfalfaITSMK093191
HetITS113 H. medicaginis Kearny County, KansasAlfalfaITSMK093192

Host trial for alfalfa cyst nematodes_

Crop
AlfalfaCornSoybeanTomato
Month 1
Root weight (g)0.040.290.210.16
Heterodera
Females and cysts0000
Cysts/g0000
Heterodera infective juveniles
Root incubation70000
J2/g1,750000
Meloidogyne J2 infective juveniles
Root incubation2104103,100
J2/g5,250144819,375
Heterodera/Meloidogyne males
Root incubation1300200
Males/g3,2500950
Month 2
Root weight (g)0.690.950.540.23
Heterodera
Females and cysts52000
Cysts/g75000
Heterodera infective juveniles
Root incubation292000
J2/g423000
Meloidogyne J2 infective juveniles
Root incubation0030812,040
J2/g0057052,348
Heterodera/Meloidogyne males
Root incubation136056520
Males/g19701042,261
DOI: https://doi.org/10.21307/jofnem-2019-016 | Journal eISSN: 2640-396X | Journal ISSN: 0022-300X
Journal RSS Feed
Language: English
Page range: 1 - 17
Submitted on: Sep 4, 2018
Published on: Apr 24, 2019
Published by: Society of Nematologists, Inc.
In partnership with: Paradigm Publishing Services
Publication frequency: 1 issue per year
Keywords:
Alfalfa cyst nematode,
COI DNA barcode,
Detection,
Diagnosis,
Taxonomy
Related subjects:
Life sciences,
Life sciences, other

© 2019 Thomas Powers, Andrea Skantar, Tim Harris, Rebecca Higgins, Peter Mullin, Saad Hafez, Zafar Handoo, Tim Todd, Kirsten Powers, published by Society of Nematologists, Inc.
This work is licensed under the Creative Commons Attribution 4.0 License.

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