Fateh Toumi, Lieven Waeyenberge, Nicole Viaene, Amer Dababat, Julie M. Nicol, Francis Ogbonnaya and Maurice Moens
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 ofPunctodera 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. latiponsspecies-speciﬁc PCR was constructed.
Trackback from your site.