The whitefly, Bemisia tabaci, an insect of the order Hemiptera which attacks more than 600 species of plants, is one of the most important agricultural pests around the world. The insecticidal Cry proteins from Bacillus thuringiensis (Bt) are useful biological pesticides, and some are toxic to Hemipteran insects. In this study, Colombian native isolates of Bt were functionally characterized at molecular and biological levels. The strains contained between one and five different crystal shapes: round, triangular, amorphous, bipyramidal and squared. The strains presented between three to seven bands of proteins in their electrophoretic pattern that were organized into six groups according to their possible biological activity on insect pests. Cry1Aa, cry1Ab, cry1Ac, cry1B and cry1C genes were identified for PCR in the different Bt isolates. Bioassays were performed on tomato leaves whose surface was spread with 3 μg · ml−1 crude extract of Bt toxins. Second instar larvae of whitefly, which were placed on top of leaves and exposed to the toxins for 7 days, exhibited mortalities from 18 to 69%. The lethal concentration 50 (LC50) of ZBUJTL39, Bt kurstaki HD1 and ZCUJTL9 strains were 1.83, 1.85 and 2.16 μg · ml−1, respectively (p < 0.05). These results show that the native Bt strain ZBUJTL39, which contained the genes cry1Aa, cry1Ab, cryCa and cryBa could eventually be used for the development of an integrated management program together with other tools for the control of B. tabaci.
In this short communication describing experiments carried out on the larvae of two insects, Unaspis euonymi Comstock (feeding on Euonymus japonicus Thunb.) and Dynaspidiotus britannicus Newstead (feeding on Laurus nobilis L.), we evaluate for the first time the efficiency of using DNA insecticides in the control of sap-sucking insects, including armored scale insects. Over a period of 10 days, high insect mortality was detected in both U. euonymi and D. britannicus, accompanied by a significant decrease in the concentration of target RNAs. At the same time, no visible changes were observed when the leaves of the host plants were subjected to treatment with DNA insecticides for one month. The results show the high efficiency of DNA insecticides used against hemipteran insect pests. It is noteworthy that the high efficiency of DNA insecticides and their low cost in comparison with RNA preparations provides a safe and extremely promising potential vehicle for the control of sap-sucking insects.
5.8S ribosomal RNA plays an important role in protein synthesis and eukaryotic ribosome translocation. Contact DNA insecticides based on antisense fragments of 5.8S ribosomal RNA gene of gypsy moth Lymantria dispar L. showed prospective insecticidal activity on its larvae. The most pronounced insecticidal effect was found for antisense fragments 10 and 11 nucleotides long (oligoRIBO-10 and oligoRIBO-11), whereas 12 nucleotides long fragment (oligoRIBO-12) caused the lowest level of insect mortality. This data corresponds to results obtained earlier using rabbit reticulocyte and wheat germ extracts, where maximum inhibition of protein synthesis was observed when a relevant oligomer 10-11 nucleotides long was used, whilst longer chain lengths resulted in reduced inhibition. Using oligoRIBO-11 fragment we have shown penetration of antisense oligonucleotides to insect cells through insects’ exoskeletons. MALDI technique registered the penetration of the oligoRIBO-11 fragment into insect cells after 30 min and a significant response of insect cells to the applied oligonucleotide after 60 min, which indicates not only that the oligonucleotide enters the insect cells, but also the synthesis of new substances in response to the applied DNA fragment. Contact DNA insecticides developed from the L. dispar 5.8S ribosomal RNA gene provide a novel biotechnology for plant protection using unmodified antisense oligonucleotides.
Obviously, the moment has come in agriculture and forestry when we must decide to gradually abandon (where possible) non-selectively acting chemical insecticides, taking into consideration the overall decrease in the total biomass of insects, especially pollinators, and the increased number of diseases and human deaths directly or indirectly associated with chemical insecticides. Yet with the world facing the rapid growth of human populations, the annual reduction of cultivated areas, and substantial losses from insect pests, most experts believe that no serious alternative to chemical insecticides exists. However, there is definitely room to create more well-tailored chemical insecticides. And there is hope, in the form of effective DNA insecticides able to provide an adequate level of safety for non-target organisms. In this short communication describing experiments carried out on the larvae of Ceroplastes japonicus Green (feeding on Ilex aquifolium Linnaeus), we show for the first time the enormous potential for the use of DNA insecticides in the control of soft scale insects and how they could replace non-selective organophosphate insecticides.