Abstract
Gram-negative binding proteins (GNBPs) are important in the innate immune system of insects in recognition of fungi pathogen, such as Beauveria bassiana. However, this information in aphids is not clear, which might be exploited to develop a novel aphid control strategy based on integrating RNAi and B. bassiana. Here, we firstly identified two GNBPs, ApGNBP1 and ApGNBP2, using the model aphid Acyrthosiphon pisum, and observed that two ApGNBPs were highly expressed in hemolymph and fat body as well as upon the injection of β-1,3-glucan and heat-killed B. bassiana. Intriguingly, RNAi-based silencing of ApGNBP1 but not ApGNBP2 decreased the activity of immune-related phenoloxidase. This led to the increased virulence of B. bassiana in A. pisum upon silencing of ApGNBP1, and the synergetic effects were also observed in other two aphids: Myzus persicae and Aphis citricidus. Importantly, no negative effects were detected in aphid predator Propylaea japonica under the co-application of the RNAi in targeting ApGNBP1 and B. bassiana. Taking together, we conclude that GNBP1 could be an optimal target in aphid control by combining RNAi, entomopathogenic fungi and ladybeetle predator.
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Funding
This research was supported by National Natural Science Foundation of China–Major International (Regional) Joint Research Project (32020103010), Fundamental Research Funds for the Central Universities (XDJK2020C067), the Chongqing Postdoctoral Science Foundation (cstc2020jcyj-bshX0090) and 111 Project (B18044). Olivier Christiaens is a recipient of a Postdoctoral Fellowship from the Research Foundation–Flanders (FWO Vlaanderen).
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Supplementary file2 (JPG 158 kb Figure S1. The expression level of ApGNBP at different developmental stages of A. pisum. The relative expression of ApGNBP1 (A) and ApGNBP2 (B) at different developmental stages. Substages were discriminated within each developmental stage. Three substages were determined for the nymph of different instars: ES, early stage; MS, middle stage; LS, late stage. Four substages were determined for the adult: NE, newly emerged; EP: early production; MP, middle production; LP, late production. The mean (± SEM) expression level was based on four or five biological replicates. Lowercase letters above each bar indicate significant differences among different treatments using ANOVA followed by Tukey's honestly significant difference test.)
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Supplementary file3 (JPG 128 kb Figure S2. Reduced Expression of GNBP1 in aphids after ingestion of dsRNA. Relative expression of MpGNBP1 (A) and AcGNBP1 (B) at 48 h after M. persicae and A. citridicus ingested dsRNA. The mean (± SEM) expression level was based on four biological replicates. The statistical differences between the compared groups are denoted by an exact p-values and asterisks.)
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Supplementary file4 (JPG 200 kb Figure S3. Effects of survival of dsRNA-fed A. pisum, infected by P. aeruginosa or M. luteus. (A) Experimental design for dsRNA-fed aphids infected by bacterium. Percent survival of dsRNA-fed A. pisum within 7 d since infection by (B) P. aeruginosa and (C) M. luteus. Sixty biological replicates were analyzed for each treatment to record survival. The comparison of survival curves was analyzed using the log-rank (Mantel–Cox) test. n.s., no significant difference. *p < 0.05.)
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Supplementary file5 (JPG 133 kb Figure S4. Survival of P. japonica 3rd instar larvae (A) and adults (B) inoculated with B. bassiana after feeding on dsRNA-fed aphids. Thirty biological replicates were analyzed for each treatment to record survival. The survival curves were compared and analyzed using the log-rank (Mantel–Cox) test. n.s., no significant difference.)
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Supplementary file6 (JPG 979 kb Figure S5. Sequence alignment between dsApGNBP1 and PjGNBP1 (A) or PjGNBP2 (B) using Jalview.)
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Ye, C., Wang, ZW., Sheng, YL. et al. GNBP1 as a potential RNAi target to enhance the virulence of Beauveria bassiana for aphid control. J Pest Sci 95, 87–100 (2022). https://doi.org/10.1007/s10340-021-01388-x
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DOI: https://doi.org/10.1007/s10340-021-01388-x