Development of a bioassay method to test activity of cry insecticidal proteins against Diatraea spp. (Lepidoptera: Crambidae) sugarcane stem borers.

The genus Diatraea (Lepidoptera: Crambidae) includes stem borers representing the most critical sugarcane pests in the Americas. Colombia's most widely distributed and damaging Diatraea species include Diatraea saccharalis, D. indigenella, D. busckella, and D. tabernella. The reduced efficacy of biological tools commonly used in controlling several species highlights the importance of evaluating alternative management strategies, such as transgenic plants expressing insecticidal proteins from the bacterium Bacillus thuringiensis (Bt). The selection of optimal Bt insecticidal proteins for Diatraea control depends on bioassays with purified Bt proteins. Because there is no described artificial diet for borer species other than D. saccharalis and availability of most purified Bt toxins is restricted, this study aimed at developing a bioassay method using fresh corn tissue and providing proof of concept by testing susceptibility to the Cry1Ac insecticidal protein from Bt. Toxicity was evaluated with a single Cry1Ac dose applied directly to corn discs. Stem borer mortality after seven days was higher than 90% for all four tested Diatraea species, while control mortality was below 8%. In addition, we observed that Cry1Ac caused more than 90% weight inhibition in all survivors and delayed development. These results validate the use of this method to determine mortality and growth inhibition due to the consumption of the Cry1Ac protein in each of the Diatraea species. Furthermore, this method could be used to assess other entomopathogenic substances to control these insect pests.

Resistance to Diatraea (Lepidoptera: Crambidae) stem borers in sugarcane is independent of species.

Evaluation of host-plant resistance on sugarcane to the sugarcane stem borers of Diatraea spp. is normally conducted in Colombia under field conditions, where environmental variations make the study of the insect-plant relationships difficult. Additionally, several species (i.e., D. saccharalis, D. indigenella, D. tabernella, and D. busckella), which are predominant in Colombia, can overlap in their distribution, raising the question of whether different varieties have the same responses to different pest species. The present study conducted evaluations of host-plant resistance under screen house conditions using two contrasting varieties (CC 93-3895, resistant, and CC 93-3826, susceptible) that were infested with the above-mentioned borer species. Observations of pest injury were conducted on internodes, leaves, and spindles. Survival and size (body mass) of the individuals recovered were analyzed and a Damage Survival Ratio (DSR) was proposed. The resistant CC 93-3895 exhibited less stalk injury, less emergence holes on internodes, and lower DSR; additionally, recovery of pest individuals was lower in comparison with CC 93-3826, independent of the borer species. Insect-plant interactions are discussed, as no previous information was available for three of the species tested (i.e., D. tabernella, D. indigenella, and D. busckella). This screen house protocol is proposed to characterize host-plant resistance among several cultivars from the Colombian sugarcane germplasm bank, using CC 93-3826 and CC 93-3895 as contrasting controls and D. saccharalis as the species model.

Biology of the Most Widely Distributed Sugarcane Stem Borers, Diatraea spp. (Lepidoptera: Crambidae), in Colombia.

Most studies on insect biology and ecology of sugarcane borers have focused on Diatraea saccharalis (Fabricius), the most widely distributed species in the Americas. Little information is available on the biology of other borer species present in Colombia, such as D. indigenella Dyar & Heinrich, D. busckella Dyar & heinrich, and D. tabernella Dyar, that present greater expansion and damage in sugarcane-growing regions. The biology of all four species was accordingly studied under laboratory conditions. Diatraea saccharalis presented the shortest development time (39.4 days) and D. busckella the longest (58.2 days). Immature survival was higher for D. saccharalis (83%) and D. tabernella (77%), with the latter also presenting the highest pupal weight (256.6 mg). Observations on reproduction indicate that D. tabernella develops a larger number of egg masses per female (67.3) as compared with D. saccharalis (28.7). All three species spent more time in the pupal stage and resulted in greater pupal size than D. saccharalis; in particular, D. indigenella showed longer female longevity than D. saccharalis. High immature survival rate and greater reproductive success in D. tabernella could potentially generate a larger population in the field, whereas D. busckella takes longer to complete its development, thus increasing the chances of causing greater injury to sugarcane plants. Discussion on biology, ecology, and pest management of these little-known species is done using as model the better-known D. saccharalis.