Computational and biological characterization of fusion proteins of two insecticidal proteins for control of insect pests

Abstract

Sucking pests pose a serious agricultural challenge, as available transgenic technologies such as Bacillus thuringiensis crystal toxins (Bt) are not effective against them. One approach is to produce fusion protein toxins for the control of these pests. Two protein toxins, Hvt (ω-atracotoxin from Hadronyche versuta) and onion leaf lectin, were translationally fused to evaluate the negative effects of fusion proteins on Phenacoccus solenopsis (mealybug), a phloem-feeding insect pest. Hvt was cloned both N-terminally (HL) and then C-terminally (LH) in the fusion protein constructs, which were expressed transiently in Nicotiana tabacum using a Potato Virus X (PVX) vector. The HL fusion protein was found to be more effective against P. solenopsis, with an 83% mortality rate, as compared to the LH protein, which caused 65% mortality. Hvt and lectin alone caused 42% and 45%, respectively, under the same conditions. Computational studies of both fusion proteins showed that the HL protein is more stable than the LH protein.Together, these results demonstrate that translational fusion of two insecticidal proteins improved the insecticidal activity relative to each protein individually and could be expressed in transgenic plants for effective control of sucking pests.

Effects of a Reserve Protein on Spodoptera frugiperda Development: A Biochemical and Molecular Approach to the Entomotoxic Mechanism

“…Exploring proteins that are multifunctional, including talisin, may be an advantageous path in the search for molecules that can be used in more efficient strategies for pest control. The pyramiding of multiple defense genes in a plant is a promising strategy to increase its protection in order to avoid or delay the development of insect pest resistance [14,56]. The use of entomotoxic genes [57,58] is only one of the existing options for pest control.…”

“…There is an obvious need to better understand how plant bioactive compounds interact and affect insects, as this knowledge would help to uncover the causes of insect pests' development and their resistance mechanisms. A very promising alternative seems to be the expression of proteins with different modes of action [14]. In this context, this work aimed to explore the multifunctional properties of talisin, focusing on the investigation of talisin intake effects on S. fugiperda development and on their main digestive enzymes' activity, as well as on their expression pattern.…”

NHA1 is a cation/proton antiporter essential for the water-conserving functions of the rectal complex inTribolium castaneum

“…Finally, we performed dynamic stability estimations of NHA1 in the presence of either Na + or K + ions, by submitting the protein for 10 ns molecular dynamics (MD) simulation using AMBER18 (63) on the Computerome 2.0 high-performance computing cluster. System preparation including minimization and equilibrium processe were performed as previously described (64). The CPPTRAJ module incorporated in AMBER suite was used for trajectory analysis according to their root mean square deviation (RMSD), radius of gyration (RoG), and hydrogen bond calculations.…”

“…Finally, we performed dynamic stability estimations of NHA1 in the presence of either Na + or K + ions, by submitting the protein for 10 ns molecular dynamics (MD) simulation using AMBER18 (63) on the Computerome 2.0 high-performance computing cluster. System preparation including minimization and equilibrium processe were performed as previously described (64).…”

Biology, ecology, and management of cotton mealybug Phenacoccus solenopsisTinsley (Hemiptera: Pseudococcidae)

“…207,208 Feeding on N. tabacum infected with potato virus X as a vector, RNAi mortality of P. solenopsis was 28.6% and 31.9% in Bursicon (PsBur)-and V-ATPase (PsV-ATPase)-silenced plants. 209 Fusion protein toxins from Hadronyche versuta (Hvt) (ω-atracotoxin) and onion leaf lectin induced 100% mortality after 120 h. 210,211 Allium cepa and Leptochloa fusca lectin genes induced >81.25% mortality under laboratory and glasshouse conditions. 212 Knockdown of genes induces sublethal effects in P. solenopsis.…”