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Molecular identification of Bursaphelenchus cocophilus associated to oil palm (Elaeis guineensis) crops in Tibu (North Santander, Colombia)

Greicy Andrea Sarria
Greicy Andrea Sarria
, 
Donald Riascos-Ortiz
Donald Riascos-Ortiz
, 
Hector Camilo Medina
Hector Camilo Medina
, 
Yuri Mestizo
Yuri Mestizo
, 
Gerardo Lizarazo
Gerardo Lizarazo
 und 
Francia Varón De Agudelo
Francia Varón De Agudelo
   | 30. Nov. 2020
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Online veröffentlicht: 30. Nov. 2020
Seitenbereich: 1 - 12
DOI: https://doi.org/10.21307/jofnem-2020-117
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© 2020 Greicy Andrea Sarria et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1:

Foliar symptoms caused by Bursaphelenchus cocophilus, causal agent of red ring disease on oil palm. A: General aspect of diseased oil palm and some chlorosis on upper leaves. B: Thin leaflets and leaf chlorosis of the upper third. C: Shortening of young leaves. D-F: Progressive collapse of chlorotic leaves and leaf rotting.
Foliar symptoms caused by Bursaphelenchus cocophilus, causal agent of red ring disease on oil palm. A: General aspect of diseased oil palm and some chlorosis on upper leaves. B: Thin leaflets and leaf chlorosis of the upper third. C: Shortening of young leaves. D-F: Progressive collapse of chlorotic leaves and leaf rotting.

Figure 2:

Symptoms caused by Bursaphelenchus cocophilus, the red ring nematode, in oil palm production in Colombia. A: Final piece of stem tissue from symptomatic and advance zone of the lesion. B: Small necrotic dots that form a discontinuous ring. C, D: Continuous necrotic area forming a ring of reddish brown color. E-G: Symptoms at the base of petioles of leaves.
Symptoms caused by Bursaphelenchus cocophilus, the red ring nematode, in oil palm production in Colombia. A: Final piece of stem tissue from symptomatic and advance zone of the lesion. B: Small necrotic dots that form a discontinuous ring. C, D: Continuous necrotic area forming a ring of reddish brown color. E-G: Symptoms at the base of petioles of leaves.

Figure 3:

Microphotographs Bursaphelenchus cocophilus. A, B: Previous segment observed estomatostilet and male tail detail. Scale bar = 20 µm. C: Long slim male of Bursaphelenchus cocophilus. Scale bar = 100 µm.
Microphotographs Bursaphelenchus cocophilus. A, B: Previous segment observed estomatostilet and male tail detail. Scale bar = 20 µm. C: Long slim male of Bursaphelenchus cocophilus. Scale bar = 100 µm.

Figure 4:

Phylogenetic tree obtained by the statistical method of maximum likelihood based on the general time reversible (GTR) model of the consensus sequences of the D2-D3 partial segment of some species of the genus Bursaphelechus. The sequences of this work are indicated in bold. The numbers on the nodes indicate bootstrap values >70%. The species Aphelenchoides besseyi (AY508109.1) is included as outgroup.
Phylogenetic tree obtained by the statistical method of maximum likelihood based on the general time reversible (GTR) model of the consensus sequences of the D2-D3 partial segment of some species of the genus Bursaphelechus. The sequences of this work are indicated in bold. The numbers on the nodes indicate bootstrap values >70%. The species Aphelenchoides besseyi (AY508109.1) is included as outgroup.

Population of Bursaphelenchus cocophilus according to sampling and extraction method.

Extraction method (Decantation) Sampling method
With facial paper Without facial paper Palms knocked down Palms standing (Drill)
Maximum Population 1,295 720 X
Maximum population 2 5 X
Absolute density 150 150 X
Absolute density 0.86 4 X
Absolute frequency 58.8 71.4 X
Absolute frequency 42.9 85.7 X

Average number of nematodes (Bursaphelenchus cocophilus) in five grams of fresh tissue from different plant organs according to extraction and sampling method.

Extraction method (Decantation) Sampling method
Tissue class With facial paper Without facial paper Palms knocked down Palms standing (Drill)
Peduncle 10 8 X
Petiolar Base 16 501 X
Spear leaf base 5 0 X
Inflorescences 110 – X
Stem 469 51 X
Stem 1 4 X

Morphometric data of males of a B. cocophilus population characterized in the present study and others of reference populations

Locality/Publication Tibu-North Santander, Colombia oil palm (Present study) n=12 Brazil Coconut Red Ring Disease After (Lordello and Zamith, 1954) n=5 Trinidad coconut Red Ring Disease (Basil, 1960) n=10 Venezuela oil palm Red Ring Disease (Gerber et al., 1989) n=10 Venezuela oil palm Little leaf (Gerber et al., 1989) n=10
Body length 964.67±70.07 (807.09-1062.67) 820-1420 1020 (840-1160) 1017±77 (841-1111) 866±71 (789-965)
a 83.89±10.68 (69.53-99.78) 92-143 120 (100-179) 129.7±13.3 (113.8-150.0) 99.3±11.8 (78.9-116.1)
c 29.33±1.72 (27.02-32.25) 22-47 28 (24-35) 26.1±2.2 (22.5-29.2) 22.4±2.8 (18.8-26.5)
c′ 3.70±0.31 (3.19-4.31) – – 5.6±0.5 (4.8-6.4) 5.4±0.8 (4.7-6.8)
T 96.58±0.20 (96.30-96.90) – – – –
Max. Body diam. 11.62±1.34 (9.97-14.22) – – 8±1.0 (7.0-9.0) 9.0±1.0 (7.0-10.0)
Stylet 10.90±0.88 (9.07-12.12) 10.7-13.8 12-13 11±1.0 (11-12) 12.0±1.0 (11.0-12.0)
Lip region width 4.61±0.52 (3.81-5.22) – 5.5 – –
Lip region height 3.13±0.43 (2.43-365) – 3.0 – –
Median bulb length 12.16±0.79 (10.75-13.04) – – – –
Median bulb diam. 5.99±0.84 (4.66-7.64) – – – –
Tail length 32.83±2.35 (28.94-37.90) – – 39.0±4.0 (32.0-46.0) 39.0±4.0 (33.0-46.0)
Cloacal or anal body diam. 8.90±0.71 (7.75-10.16) – – 7.0±1.0 (5.0-8.0) 7.0±1.0 (6.0-9.0)
Spicule length 15.21±1.41 (13.29-17.32) – – 12.0±1.0 (11.0-13.0) 11±1.0 (10.0-13.0)

Information of partial sequences D2-D3 of ribosomal DNA downloaded from GenBank and obtained in the present study for Bursaphelenchus.

Isolate Species name Location Plant-host Insect host GenBank accession number Reference or source
1 B. cocophilus Tibu, Norte de Santander, Colombia Elaeis guineensis R. palmarum MN612640 Present study
2 B. cocophilus Tibu, Norte de Santander, Colombia Elaeis guineensis R. palmarum MN612641 Present study
3 B. cocophilus Tibu, Norte de Santander, Colombia Elaeis guineensis R. palmarum MN612642 Present study
4 B. cocophilus Tibu, Norte de Santander, Colombia Elaeis guineensis R. palmarum MN612643 Present study
136 B. abruptus USA None Anthophora abrupta AY508073 Ye et al. (2007)
137 B. abietinus Austria Pityokteines vorontzowi Abies alba AY508074 Ye et al. (2007)
170 B. anatolius Turkey Halictus sp. None AY508093 Ye et al. (2007)
S12 B. cocophilus Brazil Cocos nucifera R. palmarum KT156772 Silva et al. (2016)
NT25 B. cocophilus Colombia Cocos nucifera R. palmarum KT156775 Silva et al. (2016)
NT26 B. cocophilus Colombia Cocos nucifera R. palmarum KT156776 Silva et al. (2016)
153 B. fungivorus Germany Greenhouse soil Unknown AY508082 Ye et al. (2007)
154 B. hellenicus Greece Tomicus piniperda Pinus brutia AY508083 Ye et al. (2007)
168 B. mucronatus Germany Picea abies Monochamus galloprovincialis AY508091 Ye et al. (2007)
173 B. poligraphi Germany Picea abies Polygraphus poligraphus AY508096 Ye et al. (2007)
171 B. platzeri USA None Carpohilus humeralis AY508094 Ye et al. (2007)
174 B. seani USA None Anthophora bomboides AY508097 Ye et al. (2007)
176 B. seani USA None Anthophora bomboides AY508099 Ye et al. (2007)
180 B. sexdentati Italy Pinus pinaster Unknow AY508103 Ye et al. (2007)
1057 J B. taphrorychi Poland Fagus sylvatica Taphrorychus bicolor MF422699 Tomalak et al. (2017)
98 Aphelenchoides besseyi USA Fragaria ananassa – AY508109 Ye et al. (2007)
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