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Development of a species-specific PCR to detect the cereal cyst nematode Heterodera latipons

(2013) NEMATOLOGY. 15(6). p.709-717
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Abstract
Several Heterodera species can reduce the yield of wheat and barley, among which H. avenae, H. filipjevi and H. latipons are economically the most important. Their identification, based on morphological characteristics, is not straightforward but can be made easier using molecular techniques. In this study, we developed species-specific primers for the detection of H. latipons. The actin gene of eight Heterodera species was partially sequenced and, after purifying and sequencing the PCR products, all sequences were aligned to find unique sites. The alignment showed moderate to very high similarities between the species. However, a small fragment of the actin gene was suitable for the construction of a potentially useful species-specific primer for H. latipons. The optimised PCR was subsequently tested with several populations of 14 Heterodera species and a single population of Punctodera punctata. Heterodera latipons was represented by 16 populations originating from six different countries. The primer set (Hlat-act), designed using AlleleID 7.73, was shown to be very specific. To test its sensitivity further, the PCR was conducted on DNA extracted from five second-stage juveniles (J2) of H. latipons mixed with five or 100 J2 belonging to H. avenae. The PCR was able to detect up to 1:10 dilution of the DNA obtained from five J2. The results showed that a specific and sensitive H. latipons species-specific PCR was constructed.
Keywords
species-specific primer, RIBOSOMAL DNA, sequences, molecular detection, diagnostics, actin gene, MOLECULAR CHARACTERIZATION, AVENAE GROUP, MORPHOLOGICAL CHARACTERIZATION, IDENTIFICATION, COMPLEX, RFLP, RDNA, MORPHOMETRICS, POLYMORPHISM

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MLA
Toumi, Fateh, Lieven Waeyenberge, Nicole Viaene, et al. “Development of a Species-specific PCR to Detect the Cereal Cyst Nematode Heterodera Latipons.” NEMATOLOGY 15.6 (2013): 709–717. Print.
APA
Toumi, F., Waeyenberge, L., Viaene, N., Dababat, A., Nicol, J., Ogbonnaya, F., & Moens, M. (2013). Development of a species-specific PCR to detect the cereal cyst nematode Heterodera latipons. NEMATOLOGY, 15(6), 709–717.
Chicago author-date
Toumi, Fateh, Lieven Waeyenberge, Nicole Viaene, Amer Dababat, Julie Nicol, Francis Ogbonnaya, and Maurice Moens. 2013. “Development of a Species-specific PCR to Detect the Cereal Cyst Nematode Heterodera Latipons.” Nematology 15 (6): 709–717.
Chicago author-date (all authors)
Toumi, Fateh, Lieven Waeyenberge, Nicole Viaene, Amer Dababat, Julie Nicol, Francis Ogbonnaya, and Maurice Moens. 2013. “Development of a Species-specific PCR to Detect the Cereal Cyst Nematode Heterodera Latipons.” Nematology 15 (6): 709–717.
Vancouver
1.
Toumi F, Waeyenberge L, Viaene N, Dababat A, Nicol J, Ogbonnaya F, et al. Development of a species-specific PCR to detect the cereal cyst nematode Heterodera latipons. NEMATOLOGY. 2013;15(6):709–17.
IEEE
[1]
F. Toumi et al., “Development of a species-specific PCR to detect the cereal cyst nematode Heterodera latipons,” NEMATOLOGY, vol. 15, no. 6, pp. 709–717, 2013.
@article{4413384,
  abstract     = {Several Heterodera species can reduce the yield of wheat and barley, among which H. avenae, H. filipjevi and H. latipons are economically the most important. Their identification, based on morphological characteristics, is not straightforward but can be made easier using molecular techniques. In this study, we developed species-specific primers for the detection of H. latipons. The actin gene of eight Heterodera species was partially sequenced and, after purifying and sequencing the PCR products, all sequences were aligned to find unique sites. The alignment showed moderate to very high similarities between the species. However, a small fragment of the actin gene was suitable for the construction of a potentially useful species-specific primer for H. latipons. The optimised PCR was subsequently tested with several populations of 14 Heterodera species and a single population of Punctodera punctata. Heterodera latipons was represented by 16 populations originating from six different countries. The primer set (Hlat-act), designed using AlleleID 7.73, was shown to be very specific. To test its sensitivity further, the PCR was conducted on DNA extracted from five second-stage juveniles (J2) of H. latipons mixed with five or 100 J2 belonging to H. avenae. The PCR was able to detect up to 1:10 dilution of the DNA obtained from five J2. The results showed that a specific and sensitive H. latipons species-specific PCR was constructed.},
  author       = {Toumi, Fateh and Waeyenberge, Lieven and Viaene, Nicole and Dababat, Amer and Nicol, Julie and Ogbonnaya, Francis and Moens, Maurice},
  issn         = {1388-5545},
  journal      = {NEMATOLOGY},
  keywords     = {species-specific primer,RIBOSOMAL DNA,sequences,molecular detection,diagnostics,actin gene,MOLECULAR CHARACTERIZATION,AVENAE GROUP,MORPHOLOGICAL CHARACTERIZATION,IDENTIFICATION,COMPLEX,RFLP,RDNA,MORPHOMETRICS,POLYMORPHISM},
  language     = {eng},
  number       = {6},
  pages        = {709--717},
  title        = {Development of a species-specific PCR to detect the cereal cyst nematode Heterodera latipons},
  url          = {http://dx.doi.org/10.1163/15685411-00002713},
  volume       = {15},
  year         = {2013},
}

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