Survey of Pulse Crop Field for Plant-Parasitic Nematodes in the Canadian Prairies

Gouvea-Pereira, F.; Tenuta, M.; Risula, D.; Harding, M. W.

doi:10.2478/jofnem-2025-0040

Figures & Tables

Pulse growing areas in the Canadian Prairie provinces (gray shading) and locations of commercial fields sampled in 2014 and 2015 (n = 94).

Mean population densities for plant samples (nematodes per gram) and soil (nematodes per 100 g dry soil mass) positive for taxa of plant-parasitic nematodes from commercial lentil fields in the Canadian Prairies sampled in 2014 and 2015_

Taxa	Above-Ground Crop
	Pods		Stems/Leaves		Soil Under Crop

	Mean	Max^a	Mean	Max	Mean	Max
Anguina	-^b	-	<1	n/a^c	-	-
Aphelenchoides	<1	1	<1	n/a	43	169
Aphelenchidae	<1	1	<1	1	18	41
Ditylenchus	<1	n/a	-	-	16	40
Paratylenchus	-	-	-	-	355	901
Pratylenchus	-	-	-	-	7	n/a
Telotylenchinae	-	-	-	-	23	84

Sequencing of the ITS, 18S and the D2–D3 region of the 28S rDNA gene of Paratylenchus individuals recovered from soil samples_

					Sequencing
Province	Crop	Sample Type	Field ID	No of individual nematodes analyzed	Identity	Accession	Query Cover %	Size (bp)	Identity %	Primer
Alberta	Pea	Pea soil	33	1	P. projectus	MW413605	97	567	99.8	ITS (ITSF-ITSR)
Saskatchewan	Pea	Pea soil	62	2	P. projectus	MW413656	99–100	646–708	99.69–100	28S (D2A-D3B)
Saskatchewan	Lentil	Lentil soil	120	1	P. projectus	MW413605	99	579	99.65	ITS (ITSF-ITSR)
Saskatchewan	Lentil	Lentil soil	130	1	P. projectus	KJ636434	100	852	99.77	18S (18SF-18SR)
Saskatchewan	Lentil	Lentil soil	130	1	P. projectus	MW413605	97	584	97.36	ITS (ITSF-ITSR)

Primers used for PCR reactions and sequencing in this study_

Primer Name	Specificity	Product length (bp)^*	Target gene or fragment	Primer Sequence 5′-3′	Reference
AB28-TW81	Universal	700–1100	ITS 1&2	ATA TGC TTA AGT TCA GCG GGT GGT TCC GTA GGT GAA CCT GC	Howlett et al. (1992) and Joyce et al. (1994)
D2A-D3B	Universal	800–1000	(D2–D3) 28S	ACA AGT ACC GTG AGG GAA AGT T TCG GAA GGA ACC AGC TAC TA	De Ley et al. (2005)
ITSF-ITSR	Universal	500–700	ITS 1	TTG ATT ACG TCC CTG CCC TTT ACG AGC CGA GTG ATC CAC CG	Vrain et al. (1992) and Cherry (1997)
18SF-18SR	Universal	900	18S	TTG GAT AAC TG TGG TTT AAC TAG ATT TCA CCT CTC ACG CAA CA	Qiao et al. (2016)
U831-Dipsaci_hsp90R	D. dipsaci	190	hsp90	AAY AAR ACM AAG CCN TYT GGA C GWG TTA WAT AAC TTG GTC RGC	Madani et al. (2015)
U831-Weischeri_hsp90R	D. weischeri	200	hsp 90	AAY AAR ACM AAG CCN TYT GGA C AGC ACT AAA ATT AAG YGT AAA GG	Madani et al. (2015)
PNEG-D3B	P. neglectus	290	(D3) 26S rDNA	ATG AAA GTG AAC ATG TCC TC TCG GAA GGA ACC AGC TAC TA	Al-Banna et al. (2004)
PPEN-D3B	P. penetrans	278	(D3) 26S rDNA	TAA AGA ATC CGC AAG GAT AC TCG GAA GGA ACC AGC TAC TA	Al-Banna et al. (2004)
PSCR-D3B	P. scribneri	286	(D3) 26S rDNA	AAA GTG AAC GTT TCC ATT TC TCG GAA GGA ACC AGC TAC TA	Al-Banna et al. (2004)
PTHO-D3B	P. thornei	288	(D3) 26S rDNA	GAA AGT GAA GGT ATC CCT CG TCG GAA GGA ACC AGC TAC TA	Al-Banna et al. (2004)
1770–1772	A. besseyi	325	SSU rDNA	GCG GGA TTC GTG GTT C*T CGA CAT GCC GAA ACA TGA G	Rybarczyk-Mydłowska et al. (2012)
1496–1499	A. ritzemabosi	347	SSU rDNA	CGC TGG TGG GTT TCG A CCC GCT AAG AAA TGA TCA C*C	Rybarczyk-Mydłowska et al. (2012)
AFragF1-AFragR1	A. fragariae	169	ITS1	GCA AGT GCT ATG CGA TCT TCT GCC ACA TCG GGT CAT TAT TT	McCuiston et al. (2007)

Species identity by species-specific PCR, RFLP and sequencing of the ITS, 18S and D2-D3 regions of the 28S rDNA gene of Ditylenchus individuals recovered from soil and plant sample types_

							Sequencing
Province	Crop	Sample Type	Field ID	No of individual nematodes analyzed	Species-specific PCR Identity	RFLP Identity	Identity	Accession	Query Cover %	Size (bp)	Identity %	Gene (Primer)
Manitoba	Pea	Pea pods	7	7	D. dipsaci	D. dipsaci	D. dipsaci	MG384731	100	368	98.00	ITS (TW81-AB28)
Manitoba	Pea	Thistle flowers	8	1	D. weischeri	a	D. weischeri	MG386859	100	676	99.41	ITS (AB28-TW81)
Manitoba	Pea	Thistle flowers	16	8	D. weischeri	D. weischeri	D. weischeri	MG386878	100	457	99.34	ITS (TW81-AB28)
Manitoba	Pea	Thistle flowers	20	1			D. weischeri	MG386851	100	471	98.73	ITS (TW81-AB28)
Manitoba	Pea	Thistle stems and leaves	20	1			D. weischeri	MG386873	100	555	100	ITS (TW81-AB28)
Manitoba	Pea	Pea pods	23	1	D. weischeri
Manitoba	Pea	Thistle flowers	24	8	D. weischeri	D. weischeri	D. weischeri	MG386859	98–100	676–695	99.70–99.86	ITS (TW81-AB28)
Saskatchewan	Pea	Thistle flowers	25	3	D. weischeri
Saskatchewan	Pea	Thistle stems and leaves	25	1	D. weischeri
Manitoba	Pea	Thistle flowers	28	16	D. weischeri	D. weischeri
Alberta	Pea	Pea pods	31	1	D. weischeri	D. weischeri
Alberta	Pea	Pea stems and leaves	31	1		D. weischeri
Saskatchewan	Pea	Thistle flowers	39	1	D. weischeri
Saskatchewan	Pea	Thistle stems and leaves	39	3	D. weischeri
Alberta	Pea	Thistle stems and leaves	40	1	D. weischeri
Alberta	Pea	Thistle soil	41	1	D. weischeri
Alberta	Pea	Thistle stems and leaves	44	1			D. weischeri	MG386859	99	700	99.57	ITS (TW81-AB28)
Alberta	Pea	Pea soil	57	1	D. weischeri
Alberta	Pea	Thistle soil	57	3	D. weischeri
Alberta	Pea	Thistle flowers	59	5	D. weischeri		D. weischeri	MG386825	100	680	97.79	ITS (AB28-TW81)
Saskatchewan	Pea	Thistle soil	70	5	D. weischeri		D. weischeri	MG386878	98	504	100	ITS (ITSF-ITSR)
Alberta	Chickpea	Thistle stems and leaves	76	8	D. weischeri		D. weischeri	MG386825 MG386869	99–100	642–679	98.8–99.00	ITS (AB28-TW81)
Saskatchewan	Pea	Thistle stems and leaves	148	2	D. weischeri
Alberta	Pea	Thistle soil	165	3	D. weischeri
Saskatchewan	Pea	Thistle stems and leaves	186	5	D. weischeri		D. weischeri	MG551907	99	694	99.85	28S (D2A-D3B)
Saskatchewan	Pea	Thistle stems and leaves	190	1	D. weischeri

Species characterization by species-specific PCR and sequencing of the ITS, 18S and D2-3 regions of the 28S rDNA gene of Pratylenchus individuals recovered from soil samples_

						Sequencing
Province	Crop	Sample Type	Field ID	No of individual nematodes analyzed	Species-specific PCR Identity	Identity	Accession	Query Cover %	Size (bp)	Identity %	Primer
Alberta	Pea	Pea soil	98	1	^a	P. neglectus	KY424172	100	547	99.82	18S (18SF-18SR)
Alberta	Pea	Pea soil	131	1		P. neglectus	JQ303332	100	865	99.65	18S (18SF-18SR)
Alberta	Pea	Thistle soil	50	7	P. neglectus	P. neglectus	MT261132	100	758	99.08	28S (D2A-D3B)
Alberta	Chickpea	Chickpea soil	69	1	P. neglectus
Alberta	Chickpea	Thistle soil	90	3	P. neglectus
Saskatchewan	Pea	Pea soil	185	185-1	unknown	P. neglectus	JQ303332	99	823	99.76	18S (18SF-18SR)
Saskatchewan	Pea	Pea soil	185	185-4	P. neglectus
Saskatchewan	Pea	Pea soil	185	185-5	P. neglectus
Saskatchewan	Pea	Pea soil	185	185-6	P. neglectus
Saskatchewan	Pea	Thistle soil	184	184-2	unknown	P. neglectus	KM200579	100	667	99.40	28S (D2A-D3B)
Saskatchewan	Pea	Thistle soil	184	184-5	P. neglectus
Saskatchewan	Pea	Thistle soil	192	192-1	P. neglectus

Mean population densities for plant samples (nematodes per g) and soil (nematodes per 100 g dry soil mass) positive for taxa of plant-parasitic nematodes from commercial chickpea fields in the Canadian Prairies sampled in 2014 and 2015_

Taxa	Above-Ground Crop		Above-Ground Thistle

	Stems/Leaves		Flowers		Stems/Leaves		Soil Under Crop		Soil Under Thistle

	Mean	Max^a	Mean	Max	Mean	Max	Mean	Max	Mean	Max
Aphelenchoides	-^b	-	<1	n/a	<1	n/a	36	108	35	n/a
Aphelenchidae	<1	n/a^c	-	-	-	-	18	25	5	n/a
Ditylenchus	-	-	-	-	125	n/a	8	16	-	-
Helicotylenchus	-	-	-	-	-	-	27	n/a	-	-
Hoplolaimus	-	-	-	-	-	-	4	n/a	-	-
Paratylenchus	-	-	-	-	-	-	35	72	-	-
Pratylenchus	-	-	-	-	-	-	16	27	90	127
Telotylenchinae	-	-	-	-	-	-	11	23	-	-

Mean population densities for plant samples (nematodes per gram) and soil (nematodes per 100 g dry soil mass) positive for taxa of plant-parasitic nematodes from commercial pea fields in the Canadian Prairies sampled in 2014 and 2015_

Taxa	Above-Ground Crop				Above-Ground Thistle
	Pods		Stems/Leaves		Flowers		Stems/Leaves		Soil Under Crop		Soil Under Thistle
	Mean	Max^a	Mean	Max	Mean	Max	Mean	Max	Mean	Max	Mean	Max
Anguina	<1	n/a^b	1	n/a	–^c	–	–	–	–	–	–	–
Aphelenchoides	<1	1	5	25	<1	<1–1	<1	10	32	159	28	97
Aphelenchidae	<1	n/a	2	12	–	–	1	3	22	79	31	91
Ditylenchus	1	2	4	14	55	<1–300	27	37	28	91	82	332
Helicotylenchus	–	–	–	–	–	–	–	–	115	506	110	328
Hoplolaimus	–	–	–	–	–	–	–	–	–	–	2	n/a
Longidorus	–	–	–	–	–	–	–	–	–	–	7	n/a
Merliniinae	–	–	–	–	–	–	–	–	10	n/a	–	–
Paratrichodorus	–	–	8	n/a	–	–	–	–	–	–	–	–
Paratylenchus	–	–	–	–	–	–	–	–	131	1024	66	420
Pratylenchus	–	–	–	–	–	–	–	–	106	630	87	176
Subanguina	<1	n/a	6	11	–	–	1	1	8	n/a	–	–
Telotylenchinae	–	–	1	n/a	–	–	–	–	74	659	106	980
Xiphinema	–	–	–	–	–	–	–	–	13	18	10	15

Number of fields surveyed and plant and soil samples collected from commercial farms in the Canadian Prairie provinces_

			Sample Types
			Crop			Canada Thistle			Total

Province		Fields	Pods	Stems/Leaves	Soil	Flowers	Stems/Leaves	Soil
Saskatchewan	Pea	31	31	31	31	25	31	31	180
Saskatchewan	Lentil	13	13	13	13	0	0	0	39
Saskatchewan	Chickpea	3	2	3	3	0	0	0	8
Alberta	Pea	33	25	32	33	17	27	28	162
Alberta	Chickpea	7	7	7	7	5	6	3	35
Manitoba	Pea	7	7	7	7	7	7	7	42

Total		94	85	93	94	54	71	69	466

Proportion of fields and samples (above ground plant tissue and soil for crops and thistles) positive for taxa of plant-parasitic nematodes, and chi-square tests for comparison of crop to thistle samples, in commercial fields in the Canadian Prairies_

Taxa	Fields	Sample
Taxa	Fields	Above-Ground Crop	Above-Ground Thistle	χ²	p-value	Soil Under Crop	Soil Under Thistle	χ²	p-value

		*proportion*				*proportion*
Anguina	0.03	0.02	–^a	–	–	–	–	–	–
Aphelenchoides	0.76	0.24	0.14	4.68	0.03^*	0.46	0.48	0.07	0.79
Aphelenchidae	0.54	0.12	0.05	4.43	0.03^*	0.35	0.20	4.25	0.03^*
Ditylenchus	0.52	0.07	0.22	15.71	<.0001^*	0.19	0.22	0.17	0.68
Helicotylenchus	0.21	–	–	–	–	0.16	0.19	0.23	0.63
Hoplolaimus	0.02	–	–	–	–	–	0.01	–	–
Longidorus	0.01	–	–	–	–	–	0.01	–	–
Merliniinae	0.01	–	–	–	–	0.01	–	–	–
Paratrichodorus	0.01	0.01	–	–	–	–	–	–	–
Paratylenchus	0.49	–	–	–	–	0.38	0.36	0.07	0.79
Pratylenchus	0.20	–	–	–	–	0.16	0.12	0.63	0.43
Subanguina	0.05	0.02	0.03	–	–	–	–	–	–
Telotylenchinae	0.64	0.01	–	–	–	0.47	0.50	0.43	0.62
Xiphinema	0.06	–	–	–	–	0.03	0.04	–	–

Number Examined	94	178	125			94	69

Survey of Pulse Crop Field for Plant-Parasitic Nematodes in the Canadian Prairies

Figures & Tables

Figure 1:

Mean population densities for plant samples (nematodes per gram) and soil (nematodes per 100 g dry soil mass) positive for taxa of plant-parasitic nematodes from commercial lentil fields in the Canadian Prairies sampled in 2014 and 2015_

Sequencing of the ITS, 18S and the D2–D3 region of the 28S rDNA gene of Paratylenchus individuals recovered from soil samples_

Primers used for PCR reactions and sequencing in this study_

Species identity by species-specific PCR, RFLP and sequencing of the ITS, 18S and D2-D3 regions of the 28S rDNA gene of Ditylenchus individuals recovered from soil and plant sample types_

Species characterization by species-specific PCR and sequencing of the ITS, 18S and D2-3 regions of the 28S rDNA gene of Pratylenchus individuals recovered from soil samples_

Mean population densities for plant samples (nematodes per g) and soil (nematodes per 100 g dry soil mass) positive for taxa of plant-parasitic nematodes from commercial chickpea fields in the Canadian Prairies sampled in 2014 and 2015_

Mean population densities for plant samples (nematodes per gram) and soil (nematodes per 100 g dry soil mass) positive for taxa of plant-parasitic nematodes from commercial pea fields in the Canadian Prairies sampled in 2014 and 2015_

Number of fields surveyed and plant and soil samples collected from commercial farms in the Canadian Prairie provinces_

Proportion of fields and samples (above ground plant tissue and soil for crops and thistles) positive for taxa of plant-parasitic nematodes, and chi-square tests for comparison of crop to thistle samples, in commercial fields in the Canadian Prairies_

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