Genetic differentiation of Helicoverpa armigera (Hübner) and H. assulta (Guenée) (Lepidoptera: Noctuidae) based on AFLP markers

Abstract

Here we use amplified fragment length polymorphism (AFLP) to assess genetic differentiation of Helicoverpa armigera and H. assulta. The results indicated that both species‐specific fingerprints and cluster analysis showed the ability of AFLP technique to discriminate the two sibling species; among a total 1963 AFLP markers amplified from nine primer combinations: 777 (39.6%) were H. armigera‐specific, 602 (30.7%) were H. assulta‐specific, and 584 (29.7%) were common bands. The mean number of H. armigera‐specific bands was significantly more than that of H. assulta‐specific bands for nine primer combinations (P < 0.05); the intraspecific distance of H. armigera and H. assulta was 0.123 0 and 0.110 7 respectively, and the interspecific distance was 0.178 3. In addition, the percentage of polymorphic loci and estimated average heterozygosity were used to estimate genetic diversity of the two species. This study therefore demonstrates that AFLP analysis is a sensitive and reliable technique to study genetic differentiation and genetic relationships between species and provides sufficient molecular markers for future linkage map construction, location and eventual cloning of genes involved in traits differentiation.

A Multiplex Real-Time PCR Assay to Diagnose and Separate Helicoverpa armigera and H. zea (Lepidoptera: Noctuidae) in the New World

“…Other rapid molecular methods that do not require sequencing have proven successful in separating H . armigera from other species of Helicoverpa present in the Old World [ 41 – 42 ]. Behere et al [ 18 ] developed a PCR-RFLP assay using two regions of mtDNA, COI, and cytochrome b (Cyt b ), to distinguish between H .…”

Molecular markers to discriminate among four pest species ofHelicoverpa(Lepidoptera: Noctuidae)

“…H. armigera can be differentiated from H. punctigera based on a DNA-PCR of the ITS region (Pearce, 2003), allozyme electrophoresis (Daly & Gregg, 1985) and immunoassay (Trowell et al , 2000). Distinguishing between H. assulta and H. armigera is possible based on PCR-RFLP (Kranthi et al , 2005) and AFLP markers (Ming & Wang, 2006). Monoclonal antibodies raised against H. zea can distinguish, inter alia , H. armigera from H. punctigera (Greenstone et al , 1991).…”

A simple and reliable method for discriminating between Helicoverpa armigera and Helicoverpa assulta (Lepidoptera: Noctuidae)

“…The H. armigera ‐specific DNA marker we developed allows a simple, fast and reliable discrimination between the two sister species of Helicoverpa . Compared with other diagnostic methods previously reported (Orui et al , 2000; Kranthi et al , 2005; Ming & Wang, 2006; Behere et al , 2008), the method reported here has the following advantages. First, it is very simple, consisting of only two straightforward steps (PCR amplification and electrophoresis separation), with no need for further treatment such as restriction enzyme digestion.…”

“…Although they have different host preferences, populations of these two species often occur in sympatry in China (Ming et al , 2007) owing to the common practice of intercropping agricultural systems (Wang et al , 2004; Wu et al , 2008). Moreover, the two insects are morphologically very similar, almost indistinguishable at the egg, larval and pupal stages, and only well‐trained entomologists can identify them by some morphological characteristics in the adult stage (Ming & Wang, 2006). These discriminable characters consist of slight differences of vein color in the forewing, different numbers of the diverticula on the basal pouch of everted vesicae, and some anatomical features of the adult genitalia (Matthews, 1999).…”

“…Certainly, an independent diagnostic method, preferably based on nuclear DNA, is desired. Ming and Wang (2006) developed an amplified fragment length polymorphism (AFLP) method for differentiating H. armigera and H. assulta , which was derived largely from the nuclear genome. However, their method was established mainly for evaluating the genetic differentiation between H. armigera and H. assulta , and for producing species‐specific markers for genetic mapping and cloning of linked genes (Ming & Wang, 2006); hence it is not specifically intended for rapid species discrimination.…”