Random amplified polymorphic DNA markers for discriminating \u3ci\u3eCochliomyia hominivorax\u3c/i\u3e from \u3ci\u3eC. macellaria\u3c/i\u3e (Diptera: Calliphoridae)

The screwworm, Cochliomyia hominivorax (Coquerel), is one of the most important pests of livestock in the Western Hemisphere. During early immature stages it is morphologically very similar (first instars are virtually indistinguishable) to the secondary screwworm, C. macellaria (Fabricius). Here, the utility of the random amplified polymorphic DNA–polymerase chain reaction (RAPD–PCR) was explored as a technique for developing molecular genetic markers for these two species. Of the 120 arbitrary primers screened, 21 primers produced markers that were further investigated. Seven of the 21 primers produced clear and reproducible markers that were tested with DNA of five individuals from four populations of each species; five of these primers showed 12 RAPD markers that differentiated the species in all populations. Analyses of data from these seven primers also suggested that intraspecific polymorphisms exist that could be useful in distinguishing populations of screwworms. Some population genetic tools, such as genetic distance, cluster analysis and bootstrapping, were used to statistically explore these polymorphisms. The resulting statistics showed 100% support for the ability of RAPD–PCR to discriminate between the two species. Bootstrapping with data from one of the genetic distance calculations produced a tree with all individual screwworms in the correct populations, indicating that RAPD–PCR has promise for displaying intraspecific genetic variation that could be used in establishing the general geographic origin of screwworm samples

Hydrolase and Oxido-Reductase Activities in \u3ci\u3eDiuraphis noxia\u3c/i\u3e and \u3ci\u3eRhopalosiphum padi\u3c/i\u3e (Hemiptera: Aphididae)

Four hydrolases and five oxido-reductases were examined using native stacking polyacrylamide gel electrophoresis. Homogenate of Russian wheat aphid, Diuraphis noxia (Mordvilko), bird cherry-oat aphid, Rhopalosiphum padi (L.), Arapahoe (aphid-susceptible) and Halt (aphid-resistant) wheat, Triticum aestivum L., and powdery mildew-infected Erysiphe graminis DC. ex Merat f. sp. tritici Em. Marchal, Arapahoe wheat leaves were assayed for enzyme activities. Pectinesterase, polygalacturonase (or pectinase), cellulase, and amylase activities were examined in the hydrolase group. Catalase, peroxidase, catechol oxidase, superoxide dismutase, and ascorbate oxidase activities were examined in the group of oxido-reductases. The two aphid species had the same hydrolases but different oxido-reductases. Although pectinesterase and cellulase enzymes were present in D. noxia and R. padi, the banding patterns were different. Polygalacturonase and d-amylase were not detected from either aphid species. In the oxido-reductase group, catalase was detected from D. noxia, wheras peroxidase was detected from R. padi. Superoxide dismutase and ascorbate oxidase activities also were detected from both aphids. Enzyme assays using aphid head tissue that included salivary glands but excluded aphid foregut supported the enzyme assays using whole aphids. Peroxidase activity was detected from the salivary tissue of R. padi, but not D. noxia, and catalase activity was detected from D. noxia salivary tissue, but not R. padi. We suggest that the salivary enzyme difference between the 2 aphid species (i.e., catalase and peroxidase) is important in the type of damage symptom formation on susceptible wheat plants

Comparison on DNA patterns of different ecotypes of European corn borer (Ostrinia nubilalis Hbn.) in Hungary

For a molecular genetic study on Hungarian populations of European corn borer L5 stage larvae were collected from 14 places of three different regions of the country (uni- and bivoltine ecotypes). Additionally, the study included larvae from Egypt, too (multivoltine ecotype). Molecular examinations of European corn borer larvae using the study of mitochondrial cytochrome b (cyt b) revealed that by single strand conformation polimorphism (SSCP) the populations found in Hungary represented the same haplotype. Even the Egyptian sample showed no genetic divergence. Some minor deviatons were found in the case of a sample from Székkutas, but that did not prove the genetic divergence of the bivoltine ecotype either, since the other samples of South-East Hungary did not display this kind of genetic variation. On the basis of our investigations it can be said that the univoltine and bivoltine generations, have uniform genetic complements