Detection of the root-lesion nematode, Pratylenchus penetrans (Cobb), in a nematode community using real-time PCR

Pratylenchus scribneri is a plant-parasitic root-lesion nematode causing economic damage to various crops worldwide. Identifying root-lesion nematodes to species using traditional morphological methods is an arduous task requiring extensive training on nematode taxonomy and years of experience. Thus, molecular methods for P. scribneri detection and identification were developed. Conventional and real-time polymerase chain reaction (PCR) assays with new species-specific primers were used in this study, which exclusively amplified DNA of P. scribneri but not DNA from other Pratylenchus spp. or non-Pratylenchus spp. tested. Compared with conventional PCR that was able to detect an equivalent to 1/4 of the DNA of a single nematode, real-time PCR was more sensitive and could amplify an equivalent to 1/128 of the DNA of one nematode. Both conventional and real-time PCR assays successfully identified P. scribneri and distinguished it from P. penetrans and P. neglectus isolated from field samples collected from various locations in North Dakota and Minnesota. The Blast-search based on the sequence information confirmed the reliability of the PCR assays for species identification. This is the first report of P. scribneri identification using a real-time PCR assay. The developed PCR assays are suitable for use in diagnostic laboratories and detection of field infestations with this nematode species.

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Development of Real-Time and Conventional PCR Assays for Identifying a Newly Named Species of Root-Lesion Nematode (Pratylenchus dakotaensis) on Soybean

International Journal of Molecular Sciences ◽

10.3390/ijms22115872 ◽

2021 ◽

Vol 22 (11) ◽

pp. 5872

Author(s):

Intiaz Amin Chowdhury ◽

Guiping Yan

Keyword(s):

North Dakota ◽

Real Time ◽

Real Time Pcr ◽

Detection Sensitivity ◽

Root Lesion Nematode ◽

Conventional Pcr ◽

Cycle Number ◽

Lesion Nematode ◽

Pcr Assays ◽

Species Specific

A rapid and accurate PCR-based method was developed in this study for detecting and identifying a new species of root-lesion nematode (Pratylenchus dakotaensis) recently discovered in a soybean field in North Dakota, USA. Species-specific primers, targeting the internal transcribed spacer region of ribosomal DNA, were designed to be used in both conventional and quantitative real-time PCR assays for identification of P.dakotaensis. The specificity of the primers was evaluated in silico analysis and laboratory PCR experiments. Results showed that only P.dakotaensis DNA was exclusively amplified in conventional and real-time PCR assays but none of the DNA from other control species were amplified. Detection sensitivity analysis revealed that the conventional PCR was able to detect an equivalent to 1/8 of the DNA of a single nematode whereas real-time PCR detected an equivalent to 1/32 of the DNA of a single nematode. According to the generated standard curve the amplification efficiency of the primers in real-time PCR was 94% with a R2 value of 0.95 between quantification cycle number and log number of P.dakotaensis. To validate the assays to distinguish P.dakotaensis from other Pratylenchus spp. commonly detected in North Dakota soybean fields, 20 soil samples collected from seven counties were tested. The PCR assays amplified the DNA of P.dakotaensis and discriminated it from other Pratylenchus spp. present in North Dakota soybean fields. This is the first report of a species-specific and rapid PCR detection method suitable for use in diagnostic and research laboratories for the detection of P.dakotaensis.

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EFFECTS OF SOIL MANAGEMENT ON NUMBERS OF THE ROOT-LESION NEMATODE PRATYLENCHUS PENETRANS IN SOILS OF ONTARIO PEACH ORCHARDS

Canadian Journal of Plant Science ◽

10.4141/cjps73-033 ◽

1973 ◽

Vol 53 (1) ◽

pp. 181-185

Author(s):

C. F. MARKS ◽

W. J. SAIDAK ◽

P. W. JOHNSON

Keyword(s):

Cover Crops ◽

Sandy Loam ◽

Management Practice ◽

Soil Management ◽

Pratylenchus Penetrans ◽

Root Lesion Nematode ◽

Entire Area ◽

Red Fescue ◽

Rate Of Increase ◽

Lesion Nematode

The use of herbicides and cover crops in peach orchards influenced the numbers of the root-lesion nematode, Pratylenchus penetrans, in Fox sandy loam soils. Plots treated over the entire area with the herbicide combination of paraquat (1,1′-dimethyl-4,4′-bipyridinium ion) and linuron (3-(3,4-dichlorophenyl)-1-methoxy-1-methylurea) had the smallest number of P. penetrans in the soil. The soil management practice used by many Ontario growers, clean cultivation until 1 July followed by a weed cover, resulted in the largest numbers of nematodes in the soil. Creeping red fescue (Festuca rubra L.) as a cover crop retarded the rate of increase of P. penetrans numbers in the soil but Sudan grass (Sorghum vulgare cult sudanense Hitchc.) did not. Weed control practices that permitted a temporary re-establishment of weed covers, did not retard the increase of P. penetrans numbers. Use of paraquat plus linuron to limit weed growth in the tree rows coupled with a permanent cover of creeping red fescue between the rows appears to be an effective way of retarding increases of P. penetrans numbers in peach orchards. Soil management systems that incorporate these features may be of practical value to Ontario peach growers.

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THE PATHOGENICITY OF PRATYLENCHUS PENETRANS TO CELERY

Canadian Journal of Plant Science ◽

10.4141/cjps63-011 ◽

1963 ◽

Vol 43 (1) ◽

pp. 70-74 ◽

Cited By ~ 13

Author(s):

J. L. Townshend

Keyword(s):

Plant Nutrients ◽

Silica Sand ◽

Pratylenchus Penetrans ◽

Root Lesion Nematode ◽

Primary Parasite ◽

Lesion Nematode

Celery seedlings, grown aseptically in silica sand with plant nutrients, were inoculated with surface-sterilized specimens of the root lesion nematode Pratylenchus penetrans (Cobb, 1917) Filip. & Stek., 1941. The reactions of invaded roots were studied microscopically. The epidermis, cortex, and endodermis of young celery roots showed different degrees of discoloration after invasion of P. penetrans, with the endodermis most severely affected. Pratylenchus penetrans was a primary parasite and pathogen of celery.

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Tagetes patula as an effective catch crop for long-term control of Pratylenchus penetrans

Nematology ◽

10.1163/1568541044038632 ◽

2004 ◽

Vol 6 (6) ◽

pp. 877-881 ◽

Cited By ~ 18

Author(s):

Albartus Evenhuis ◽

Gerard Korthals ◽

Leendert Molendijk

Keyword(s):

Population Density ◽

Economic Loss ◽

Soil Fumigation ◽

Economic Damage ◽

Tagetes Patula ◽

Pratylenchus Penetrans ◽

Root Lesion Nematode ◽

Rate Of Increase ◽

Chemical Soil ◽

Lesion Nematode

AbstractTagetes patula is well known to be able to diminish Pratylenchus penetrans (Cobb) populations. In a field experiment, the increase of the P. penetrans soil population density after growing T. patula was significantly less than after chemical soil fumigation with metam sodium. The effect of T. patula on P. penetrans population densities lasted longer than the effect of chemical soil fumigation. Strawberries were grown for 3 consecutive years after T. patula without damage by the root lesion nematode. Assuming a linear rate of increase of the P. penetrans population density, we predict that strawberries could be grown for about 7 successive years without economic damage due to the root lesion nematode. Strawberry yield was greater from the crop grown 3 years after T. patula than that after soil fumigation 4 years previously. About 2 t strawberry yield increase is necessary to allow the growth of Tagetes every 5th year, without economic loss. The possibility of growing both strawberries and Tagetes in the same year should be investigated.

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Root lesion nematode, Pratylenchus penetrans, in ‘Fire King’ lilies: attack and control with aldicarb

Netherlands Journal of Plant Pathology ◽

10.1007/bf01987031 ◽

1978 ◽

Vol 84 (6) ◽

pp. 217-225 ◽

Cited By ~ 1

Author(s):

P. W. Th. Maas ◽

P. Mantel ◽

J. Boontjes

Keyword(s):

Pratylenchus Penetrans ◽

Root Lesion Nematode ◽

Lesion Nematode ◽

In Fire ◽

And Control

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GREENHOUSE EVALUATION OF PRUNUS ROOTSTOCKS FOR RESISTANCE OR TOLERANCE TO THE ROOT LESION NEMATODE (PRATYLENCHUS PENETRANS)

Acta Horticulturae ◽

10.17660/actahortic.1998.465.91 ◽

1998 ◽

pp. 723-736 ◽

Cited By ~ 2

Author(s):

W. McFadden-Smith ◽

N.W. Miles ◽

J.W. Potter

Keyword(s):

Pratylenchus Penetrans ◽

Root Lesion Nematode ◽

Greenhouse Evaluation ◽

Lesion Nematode

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Interaction of Verticillium albo-atrum and the Root Lesion Nematode, Pratylenchus penetrans, in Tomato Roots at Controlled Inoculum Densities

Phytopathology ◽

10.1094/phyto-62-362 ◽

1972 ◽

Vol 62 (3) ◽

pp. 362 ◽

Cited By ~ 20

Author(s):

J. J. Conroy

Keyword(s):

Pratylenchus Penetrans ◽

Root Lesion Nematode ◽

Tomato Roots ◽

Lesion Nematode ◽

Verticillium Albo Atrum

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Nematicidal Activities of 4-Hydroxyphenylacetic Acid and Oidiolactone D Produced by the Fungus Oidiodendron sp.

Zeitschrift für Naturforschung C ◽

10.1515/znc-2011-1-205 ◽

2011 ◽

Vol 66 (1-2) ◽

pp. 31-34 ◽

Cited By ~ 5

Author(s):

Kouhei Ohtani ◽

Shozo Fujioka ◽

Atsumi Shimada ◽

Yasuo Kimura

Keyword(s):

Bursaphelenchus Xylophilus ◽

Pine Wood Nematode ◽

Pratylenchus Penetrans ◽

Root Lesion Nematode ◽

Pine Wood ◽

Spectroscopic Analyses ◽

Hydroxyphenylacetic Acid ◽

Lesion Nematode

Two nematicides, 4-hydroxyphenylacetic acid (4-HPA) (1) and oidiolactone D (2), were isolated from cultures of the fungus Oidiodendron sp., and their structures were identified by spectroscopic analyses. Compound 2 showed nematicidal activities against the root-lesion nematode, Pratylenchus penetrans, and the pine wood nematode, Bursaphelenchus xylophilus. Compound 1 was also active against these two nematodes but to a lesser extent.

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