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.
Molecular polymorphism and morphometrics of species of the Heterodera avenae group in Syria and Turkey.
Journal of Nematology37146-154.
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Variation in spacer ribosomal DNA in some cyst-forming species of plant parasitic nematodes.
Fundamental and Applied Nematology16177-184.
Ribosomal DNA sequence analysis of Heterodera filipjevi and H. latipons isolates from Russia and comparisons with other nematode isolates.
Russian Journal of Nematology7121-125.
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Phylum-wide analysis of SSU rDNA reveals deep phylogenetic relationships among nematodes and accelerated evolution toward crown clades.
Molecular Biology and Evolution231792-1800.
Molecular characterization of cyst nematode species (Heterodera spp.) from the Mediterranean Basin using RFLPs and sequences of ITS-rDNA.
Journal of Phytopathology152229-234.
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Synonymy of Afenestrata with Heterodera supported by phylogenetics with molecular and morphological characterisation of H. koreana comb. n. and H. orientalis comb. n. (Tylenchida: Heteroderidae).
Global knowledge and its application for the integrated control and management of nematodes on wheat. In:
Integrated management and biocontrol of vegetable and grain crops nematodesVol. 2.
Dordrecht, The NetherlandsSpringer pp.
Genetic and phenotypic diversity in the graminaceous cyst nematode complex, inferred from PCR-RFLP of ribosomal DNA and morphometric analysis.
European Journal of Plant Pathology109227-241.
The ClustalX Windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools.
Nucleic Acids Research244876-4882.
ToumiF.HassanG.WaeyenbergeL.ViaeneN.DababatA.NicolM.J.OgbonnayaC.F.Al-AssasK.Abou Al-FadilT.MoensM. (2012). Distribution of cereal cyst nematodes (Heterodera spp.) in wheat and barley fields in northeastern regions of Syria. Proceedings of the third workshop of the international cereal cyst nematode initiative 22-23 October 2012 Adana Turkey CIMMYT p. 310.
TytgatT.VanholmeB.De MeutterJ.ClaeysM.CouvreurM.VanhoutteI.GheysenG.CriekingeW.BorgonieG.CoomansA. (2004).
A new class of ubiquitin extension proteins secreted by the dorsal pharyngeal gland in plant parasitic cyst nematodes.
The American Phytopathological Society17846-852.
WaeyenbergeL.ViaeneN.SubbotinS.A.MoensM. (2009). Molecular identification of Heterodera spp. an overview of fifteen years of research. Proceedings of the first workshop of the International Cereal Cyst Nematode Initiative 21-23 October 2009 Antalya Turkey pp. 109-114.
Separation of three species of Ditylenchus and some host races of D. dipsaci by restriction fragment length polymorphism.
Journal of Nematology25555-563.