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The development of novel anti-infectives against Kinetoplastids pathogens targeting proteins is a big problem occasioned by the antigenic variation in these parasites. This is also a global concern due to the zoonosis of these parasites, as they infect both humans and animals. Therefore, we need not only to create novel antibiotics, but also to speed up the development pipeline for these antibiotics. This may be achieved by using novel drug targets for Kinetoplastids drug discovery. In this study, we focused our attention on motifs of rRNA molecules that have been created using homology modeling. The RNA is the most ambiguous biopolymer in the kinetoplatid, which carries many different functions. For instance, tRNAs, rRNAs, and mRNAs are essential for gene expression both in the pro-and eukaryotes. However, all these types of RNAs have sequences with unique 3D structures that are specific for kinetoplastids only and can be used to shut down essential biochemical processes in kinetoplastids only. All these features make RNA very potent targets for antibacterial drug development. Here, we combine in silico methods combined with both computational biology and structure prediction tools to address our hypothesis. In this study, we outline a systematic approach for identifying kinetoplastid rRNA-ligand interactions and, more specifically, techniques that can be used to identify small molecules that target particular RNA. The high-resolution optimized model structures of these kineoplastids were generated using RNA 123, where all the stereochemical conflicts were solved and energies minimized to attain the best biological qualities. The high-resolution optimized model’s structures of these kinetoplastids were generated using RNA 123 where all the stereochemical conflicts were solved and energies minimized to attain the best biological qualities. These models were further analyzed to give their docking assessment reliability. Docking strategies, virtual screening, and fishing approaches successfully recognized novel and myriad macromolecular targets for the myxobacterial natural products with high binding affinities to exploit the unmet therapeutic needs. We demonstrate a sensible exploitation of virtual screening strategies to 18S rRNA using natural products interfaced with classical maximization of their efficacy in phamacognosy strategies that are well established. Integration of these virtual screening strategies in natural products chemistry and biochemistry research will spur the development of potential interventions to these tropical neglected diseases.

The banana aphid,Pentalonia nigronervosa, is the sole insect vector of banana bunchy top virus (BBTV), the causal agent of banana bunchy top disease. The aphid acquires and transmits BBTV while feeding on infected banana plants. RNA interference (RNAi) enables the generation of pest and disease-resistant crops, however, its effectiveness relies on the identification of pivotal gene sequences to target and silence. Acetylcholinesterase (AChE) is an essential enzyme responsible for the hydrolytic metabolism of the neurotransmitter acetylcholine in animals. In this study, the AChE gene of the banana aphid was targeted for silencing by RNAi through transgenic expression of AChE dsRNA in banana and plantain plants. The efficacy of dsRNA was first assessed using an artificial feeding assay. In vitro aphid feeding on a diet containing 7.5% sucrose, and sulfate complexes of trace metals supported aphid growth and reproduction. When AChE dsRNA was included in the diet, a dose of 500 ng/μL was lethal to the aphids. Transgenic banana cv. Cavendish Williams and plantain cvs. Gonja Manjaya and Orishele expressing AChE dsRNA were regenerated and assessed for transgene integration and copy number. When aphids were maintained on elite transgenic events, there was a 67.8%, 46.7%, and 75.6% reduction in aphid populations growing on Cavendish Williams, Gonja Manjaya, and Orishele cultivars, respectively, compared to those raised on nontransgenic control plants. These results suggest that RNAi targeting an essential aphid gene could be a useful means of reducing both aphid infestation and potentially the spread of the disease they transmit.

Banana bunchy top virus (BBTV) is one of the world’s invasive species. Banana aphid (Pentalonia nigronervosa) is found in all banana producing areas and it is the insect pest known to transmit BBTV causing banana bunchy top disease (BBTD) in bananas and plantains (Musa spp.) and can cause a significant yield loss of up to 100% in severe cases. Controlling the spread of BBTD has been very challenging since there is no known endogenous gene in the Musa germplasm that could confer resistance to BBTV. Excessive dependence on insecticides for disease control is detrimental to the environment and off-target-organisms. The objective of this study was to use RNA interference (RNAi) targeting the acetylcholinesterase (AChE) gene in banana aphid to develop resistance against aphids in farmer preferred plantain cultivars. This could help sustain smallholder farmers in areas where BBTD is an epidemic. To achieve this, plantain cultivars were initiated using plant tissue culture techniques and rapidly multiplied using Temporary Immersion Bioreactor. This was followed by generation of embryogenic cell suspension (ECS), Agrobacterium-mediated transformation of banana and plantain ECS using a RNAi plasmid construct and molecular characterization of putative transgenic lines. Agro-infected ECS of banana and plantain cultivars were regenerated on selective medium and produced several transgenic lines. Molecular characterization confirmed the presence of transgene in about 80% transgenic lines. Preliminary glasshouse screening of transgenic lines showed reduction in population of banana aphids in comparison to control non-transgenic plants. This is the first report on using RNAi targeting AChE gene for developing transgenic plantain that are resistant to banana aphids.