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Alighting aphids probe a new host plant by intracellular test punctures for suitability. These induce immediate calcium signals that emanate from the punctured sites and might be the first step in plant recognition of aphid feeding and the subsequent elicitation of plant defence responses. Calcium is also involved in the transmission of non-persistent plant viruses that are acquired by aphids during test punctures. Therefore, we wanted to determine whether viral infection alters calcium signalling. For this, calcium signals triggered by aphids were imaged on transgenic Arabidopsis plants expressing the cytosolic FRET-based calcium reporter YC3.6-NES and infected with the non-persistent viruses cauliflower mosaic (CaMV) and turnip mosaic (TuMV), or the persistent virus, turnip yellows (TuYV). Aphids were placed on infected leaves and calcium elevations were recorded by time-lapse fluorescence microscopy. Calcium signal velocities were significantly slower in plants infected with CaMV or TuMV and signal areas were smaller in CaMV-infected plants. Transmission tests using CaMV-infected Arabidopsis mutants impaired in pathogen perception or in the generation of calcium signals revealed no differences in transmission efficiency. A transcriptomic meta-analysis indicated significant changes in expression of receptor-like kinases in the BAK1 pathway as well as of calcium channels in CaMV- and TuMV-infected plants. Taken together, infection with CaMV and TuMV, but not with TuYV, impacts aphid-induced calcium signalling. This suggests that viruses can modify plant responses to aphids from the very first vector/host contact.

(1) Background: Genistein is a naturally occurring flavonoid with a rich spectrum of biological activities, including plant-herbivore interactions. The aim of the study was to evaluate the effect of exogenous application of genistein on aphid behavior during probing in plant tissues. (2) Methods: Vicia faba, Brassica rapa ssp. pekinensis, and Avena sativa were treated transepidermally with a 0.1% ethanolic solution of genistein, and the probing behavior of generalist aphid species Aphis fabae, Myzus persicae, and Rhopalosiphum padi was monitored on their respective treated and untreated host plants using electropenetrography (=electrical penetration graph technique, EPG); (3) Results: Genistein did not deter aphid probing activities in non-phloem tissues. In A. fabae and R. padi, a trend towards reduction and in M. persicae a trend towards increase in phloem sap ingestion occurred on genistein-treated plants, but these trends were not statistically significant. (4) Conclusions: Genistein is not a deterrent chemical against generalist aphid species studied; therefore, it is not recommended for practical application.

The enantiomerically enriched γ- and δ-decalactones (4a and 4b) were prepared from corresponding racemic primary-secondary 1,4- and 1,5-diols (1a and 1b), as products of enzymatic oxidation catalyzed by different alcohol dehydrogenases. The results of biotransformations indicated that the oxidation processes catalyzed by alcohol dehydrogenase (HLADH), both isolated from horse liver and recombinant in Escherichia coli, were characterized by the highest degree of conversion with moderate enantioselectivity of the reaction. Useful, environmentally friendly extraction procedure of decalactones (4a and 4b) based on hydrodistillation using a Deryng apparatus was developed. Both racemic lactones (4a and 4b), as well as their enantiomerically enriched isomers, were tested for feeding deterrent activity against Myzus persicae. The effect of these compounds on probing, feeding and settling behavior of M. persicae was studied in vivo. The deterrent activity of decalactones (4a and 4b) against aphids depended on the size of the lactone ring and the enantiomeric purity of the compounds. δ-Decalactone (4b) appeared inactive against M. persicae while γ-decalactone (4a) restrained aphid probing at ingestional phase. Only (-)-(S)-γ-decalactone (4a) had strong and durable (i.e. lasting for at least 24 hours) limiting effect, expressed at phloem level.

Substances that alter insect behavior have attracted a lot of attention as potential crop protection agents. Naringenin (5,7,4′-trihydroxyflavanone) is a naturally occurring bioactive flavanone. We evaluated the influence of naringenin on aphid activities during individual phases of probing and feeding and the effect of structural modifications of naringenin on its activity towards aphids. We monitored the probing behavior of Myzus persicae (Sulz.) (Hemiptera: Aphididae) using the Electrical Penetration Graph (EPG) technique. The chemical modifications were the substitution of hydrogen atoms with methyl, ethyl or pentyl groups and the replacement of the carbonyl group in naringenin and its derivatives with an oxime moiety. Depending on the substituents, the activity of naringenin-derived compounds varied in potency and mode of action. Naringenin was an attractant of moderate activity, which enhanced sap ingestion. The naringenin derivative with two methyl groups—7,4′-di-O-methylnaringenin—was a deterrent, which hindered aphid probing in non-phloem tissues. Naringenin oxime derivatives with methyl substituents—7,4′-di-O-methylnaringenin oxime, 7-O-methylnaringenin oxime, and 5,7,4′-tri-O-methylnaringenin oxime—and the derivative with a pentyl substituent—7-O-pentylnaringenin oxime—were strong attractants which stimulated aphid probing in non-phloem tissues and the ingestion of phloem sap.

Apigenin and luteolin are products of the phenylpropanoid pathway, where apigenin serves as a substrate for the synthesis of luteolin. Apigenin and luteolin are highly bioactive flavones; therefore, in search of prospective biopesticides, the receptiveness of the polyphagous green peach aphid Myzus persicae (Sulzer) (Hemiptera: Aphididae) to apigenin and luteolin was studied. The flavones were applied as 0.1% ethanolic solutions to the host plant leaf surface, and aphid probing and feeding activities were monitored using the Electrical Penetration Graph (EPG) technique. The structural difference between apigenin and luteolin, which was the number of hydroxyl groups in the molecule, had an impact on the activity of these flavones. On apigenin-treated plants, the duration of the first probe was three times as short as on the control and five times as short as on the luteolin-treated plants; the duration of the time to the first ingestion phase within the successful probe was shorter than on the control and luteolin-treated plants; the mean duration of xylem sap ingestion bouts and the proportion of xylem phase in all probing activities were the highest; and the duration of salivation before phloem sap ingestion was the longest. Aphids’ response to luteolin-treated plants was less distinct as compared to apigenin-treated plants.

Bemisia tabaci MEAM1 (Middle East-Asia Minor 1) feeding alters antioxidative enzyme activity in some plant species. Infestation of B. tabaci nymphs decreases Myzus persicae performance on systemic, but not local leaves of tobacco plants. However, it is unclear if B. tabaci nymphs induced antioxidant activities contributing to the aphid resistance. We investigated the relationship between antioxidants induced by nymphs of B. tabaci feeding on tobacco and aphid resistance. The activities of catalase (CAT), peroxidase (POD), superoxide dismutase (SOD) and the concentration of hydrogen peroxide (H2O2) were assayed in tobacco leaves at different feeding times following infestation of B. tabaci nymphs. The infestation altered the activities of CAT and POD, but had no significant effect on SOD activity. The highest CAT activity was observed at 15 d after infestation. This was 98.2% greater than control systemic leaves, but 32.6% lower than the control in local leaves. Higher POD activity was recorded in local vs. systemic leaves after 15 d of infestation. POD activity was 71.0% and 112.9% higher in local and systemic leaves, respectively, than in the controls. The changes of CAT, but not POD or SOD activity were correlated to levels of aphid resistance. H2O2 levels were higher in local than in systemic leaves in contrast to CAT activity. Tobacco curly shoot virus mediated virus-induced gene silencing was employed to determine if CAT activation was involved in the aphid resistance induced by B. tabaci nymphs. B. tabaci induced CAT activity decreased when the Cat1 expression was silenced. The performance assay indicated that Cat1 silencing made B. tabaci infested plants a more suitable host for aphids than infested control plants. The aphid survival rate was reduced by 40.4% in infested control plants, but reduced by only 26.1% in Cat1-silenced plants compared to uninfested controls. Also, qPCR results showed that silencing of Cat1 led to the suppression of the B. tabaci mediated PR-2a expression. Aphid resistance in plants infested with B. tabaci nymphs is associated with enhanced antioxidant activities in which CAT may play a dominant role. This resistance probably acted via interactions with SA-mediated defense responses.

Corn leaf aphid, Rhopaloshium maidis Fitch (Hemiptera: Aphididae), is a prevalent maize pest, causing significant yield losses by affecting all crop stages. To mitigate such losses, current management relies on the application of chemical insecticides, posing environmental degradation risks and promoting insecticide-resistant aphid populations. These challenges necessitate the development of environment friendly and effective alternatives. One such promising alternative is silicon (Si) supplementation, which alleviates a variety of abiotic and biotic stresses in plants including insect herbivory, though it’s specific effects on corn leaf aphids are poorly understood. Against this back drop, this study investigated the effect of Si supplementation in mitigating corn leaf aphid infestations in maize. Plants were treated with three Si doses (T1: 500, T2: 750 and T3: 1000 mg L⁻ 1 ) and infested by corn leaf aphids. Then, performance of aphids was determined by measuring their feeding preferences, fecundity and mortality rates. Si accumulation in maize plants was also determined, with the highest concentration observed in T3 plants, followed by T2 and T1 plants. Correspondingly, aphid number were lowest in T3 (0.65 in free-choice and 0.30 in paired-choice tests) and T2 (0.78 in free-choice and 0.40 in paired-choice tests). Aphid fecundity was significantly reduced in T3 (6.85 nymphs, representing a 78.94% reduction compared to the control) and aphid mortality was highest in T3 (43.3%), and T2 (30.0%). Si supplementation also enhanced antioxidant enzyme activities (superoxide dismutase, catalase and peroxidase) contributing to increased plant resistance against aphid infestation. The results indicate that Si application effectively reduces corn leaf aphid infestation and represents a sustainable strategy for maize pest management.

The regulatory role of small RNAs (sRNAs) in various biological processes is an active area of investigation; however, there has been limited information available on the role of sRNAs in plant-insect interactions. This study was designed to identify sRNAs in cotton-melon aphid (Aphis gossypii) during the Vat-mediated resistance interaction with melon (Cucumis melo). The role of miRNAs was investigated in response to aphid herbivory, during both resistant and susceptible interactions. sRNA libraries made from A. gossypii tissues feeding on Vat⁺ and Vat⁻ plants revealed an unexpected abundance of 27 nt long sRNA sequences in the aphids feeding on Vat⁺ plants. Eighty-one conserved microRNAs (miRNAs), twelve aphid-specific miRNAs, and nine novel candidate miRNAs were also identified. Plant miRNAs found in the aphid libraries were most likely ingested during phloem feeding. The presence of novel miRNAs was verified by qPCR experiments in both resistant Vat⁺ and susceptible Vat⁻ interactions. The comparative analyses revealed that novel miRNAs were differentially regulated during the resistant and susceptible interactions. Gene targets predicted for the miRNAs identified in this study by in silico analyses revealed their involvement in morphogenesis and anatomical structure determination, signal transduction pathways, cell differentiation and catabolic processes. In this study, conserved and novel miRNAs were reported in A. gossypii. Deep sequencing data showed differences in the abundance of miRNAs and piRNA-like sequences in A. gossypii. Quantitative RT-PCR revealed that A. gossypii miRNAs were differentially regulated during resistant and susceptible interactions. Aphids can also ingest plant miRNAs during phloem feeding that are stable in the insect.

Bacterial symbionts are broadly distributed among insects, influencing their bioecology to different degrees. Aphids carry a number of secondary symbionts that can influence aphid physiology and fitness attributes. Spiroplasma is seldom reported as an aphid symbiont, but a high level of infection has been observed in one population of the tropical aphid Aphis citricidus . We used sister isolines of Spiroplasma -infected (Ac-BS) and Spiroplasma -free (Ac-B) aphids reared on sweet orange (optimum host) and orange jasmine (suboptimum host) to demonstrate the effects of Spiroplasma infection in the aphid proteome profile. A higher number of proteins were differently abundant in aphids feeding on orange jasmine, indicating an impact of host plant quality. In both host plants, the majority of proteins affected by Spiroplasma infection were heat shock proteins, proteins linked to cell function and structure, and energy metabolism. Spiroplasma also induced changes in proteins involved in antimicrobial activity, carbohydrate processing and metabolism, amino acid synthesis and metabolism in aphids feeding on orange jasmine. We discuss on how the aphid host proteome is differentially affected by Spiroplasma infection when the host is exploiting host plants with different nutritional values.