Immatures of Helicoverpa zae and Heliothis virescens are difficult to separate morphologically, especially when they occur sympatrically. PCR assays using primers targeted to the mitochondria genes encoding 16S rRNA were developed to precisely identify eggs and larvae of these two species. After partial sequencing of mitochondria DNA from both species using conserved primers, specific primers for H. zea and H. virescens were designed and tested. The primers amplified species-specific fragments from HV and HZ with sizes 200bp and 150 bp, respectively, without cross-reaction between the two species. The two-species identification can be conducted in one PCR reaction. These results will be very helpful for development of cotton IPM programs in regions where both species occur together in early season.
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