Proteome changes in pepper (Capsicum annuum L.) leaves induced by the green peach aphid (Myzus persicae Sulzer).

BACKGROUND: Aphid attack induces defense responses in plants activating several signaling cascades that led to the production of toxic, repellent or antinutritive compounds and the consequent reorganization of the plant primary metabolism. Pepper (Capsicum annuum L.) leaf proteomic response against Myzus persicae (Sulzer) has been investigated and analyzed by LC-MS/MS coupled with bioinformatics tools. RESULTS: Infestation with an initially low density (20 aphids/plant) of aphids restricted to a single leaf taking advantage of clip cages resulted in 6 differentially expressed proteins relative to control leaves (3 proteins at 2 days post-infestation and 3 proteins at 4 days post-infestation). Conversely, when plants were infested with a high density of infestation (200 aphids/plant) 140 proteins resulted differentially expressed relative to control leaves (97 proteins at 2 days post-infestation, 112 proteins at 4 days post-infestation and 105 proteins at 7 days post-infestation). The majority of proteins altered by aphid attack were involved in photosynthesis and photorespiration, oxidative stress, translation, protein folding and degradation and amino acid metabolism. Other proteins identified were involved in lipid, carbohydrate and hormone metabolism, transcription, transport, energy production and cell organization. However proteins directly involved in defense were scarce and were mostly downregulated in response to aphids. CONCLUSIONS: The unexpectedly very low number of regulated proteins found in the experiment with a low aphid density suggests an active mitigation of plant defensive response by aphids or alternatively an aphid strategy to remain undetected by the plant. Under a high density of aphids, pepper leaf proteome however changed significantly revealing nearly all routes of plant primary metabolism being altered. Photosynthesis was so far the process with the highest number of proteins being regulated by the presence of aphids. In general, at short times of infestation (2 days) most of the altered proteins were upregulated. However, at longer times of infestation (7 days) the protein downregulation prevailed. Proteins involved in plant defense and in hormone signaling were scarce and mostly downregulated.

Phytohormone responses in pepper (Capsicum annuum L.) leaves under a high density of aphid infestation.

The time course response of selected phytohormones has been evaluated in sweet pepper plants (Capsicum annuum L.) submitted to a high density (200 aphids/plant) of aphid (Myzus persicae Sulzer) infestation. Abscisic acid (ABA), salicylic acid (SA), indole-3-acetic acid (IAA), and jasmonates (JAs), including jasmonic acid (JA), jasmonoyl-l-isoleucine (JA-Ile), and cis-OPDA have been simultaneously identified and quantitated by UHPLC-MS/MS in pepper leaf tissue harvested at 3, 8 hours post-infestation (hpi), 1, 2, 4 and 7 days post-infestation (dpi). Infested plants showed a reduction in stem length at 7 dpi and in the number of leaves and leaf width from 4 dpi onwards. JA and JA-Ile significantly increased very early (from 3 hpi) while SA only accumulated at 7 dpi. Despite the high density of infestation, the aphid-induced accumulation of JAs was much lower than the burst typically induced by chewing herbivores. On the other side, ABA peaked in aphid-infested plants at 2 and 4 dpi, while IAA content did not change significantly at any time point. Growth inhibition may be partially explained by the high levels of JAs found in aphid-infested plants. The possibility that the obtained results support the hypothesis of the aphid manipulation of plant metabolism is discussed.

Local and systemic hormonal responses in pepper (Capsicum annuum L.) leaves under green peach aphid (Myzus persicae Sulzer) infestation.

This study examined the temporal changes in the leaf content of defence-involved phytohormones in pepper (Capsicum annuum L.) plants responding to the green peach aphid (Myzus persicae Sulzer) infestation, at both local and systemic level. Aphid infestation did not alter the content of cis-12-oxo-phytodienoic acid, the jasmonic acid (JA) precursor, even though endogenous levels of JA and its bioactive isoleucine-conjugated form (JA-Ile) significantly increased from 8 to 96 h in local infested leaves. Systemic effects in jasmonates were only showed at 48 h for JA, and 8 and 48 h in the case of JA-Ile. SA accumulated only in local infested leaves after 96 h of infestation, when the level of JA-Ile decreased in these leaves. This suggests a possible antagonistic interaction between JA and SA pathways, although other pathways may be also involved. Endogenous level of indole-3-acetic acid was higher in systemic relative to local infested leaves at 3 and 24 h, although no significant changes in its content were found compared to control leaves. Abscisic acid content was lower in local infested relative to control leaves at 24 h, but was higher at 48 h when it also increased systemically. The possible roles of the studied phytohormones in plant defence responses against aphids are discussed.

Changes in the free amino acid composition of Capsicum annuum (pepper) leaves in response to Myzus persicae (green peach aphid) infestation. A comparison with water stress.

Amino acids play a central role in aphid-plant interactions. They are essential components of plant primary metabolism, function as precursors for the synthesis of defense-related specialized metabolites, and are major growth-limiting nutrients for aphids. To quantify changes in the free amino acid content of pepper (Capsicum annuum L.) leaves in response to green peach aphid (Myzus persicae Sulzer) feeding, plants were infested with a low (20 aphids/plant) or a high (200 aphids/plant) aphid density in time-course experiments ranging from 3 hours to 7 days. A parallel experiment was conducted with pepper plants that had been subjected to water stress. Factor Analysis of Mixed Data revealed a significant interaction of time x density in the free amino acid response of aphid-infested leaves. At low aphid density, M. persicae did not trigger a strong response in pepper leaves. Conversely, at high density, a large increase in total free amino acid content was observed and specific amino acids peaked at different times post-infestation. Comparing aphid-infested with water-stressed plants, most of the observed differences were quantitative. In particular, proline and hydroxyproline accumulated dramatically in response to water stress, but not in response to aphid infestation. Some additional differences and commonalities between the two stress treatments are discussed.

The Down syndrome-related protein kinase DYRK1A phosphorylates p27(Kip1) and Cyclin D1 and induces cell cycle exit and neuronal differentiation.

A fundamental question in neurobiology is how the balance between proliferation and differentiation of neuronal precursors is maintained to ensure that the proper number of brain neurons is generated. Substantial evidence implicates DYRK1A (dual specificity tyrosine-phosphorylation-regulated kinase 1A) as a candidate gene responsible for altered neuronal development and brain abnormalities in Down syndrome. Recent findings support the hypothesis that DYRK1A is involved in cell cycle control. Nonetheless, how DYRK1A contributes to neuronal cell cycle regulation and thereby affects neurogenesis remains poorly understood. In the present study we have investigated the mechanisms by which DYRK1A affects cell cycle regulation and neuronal differentiation in a human cell model, mouse neurons, and mouse brain. Dependent on its kinase activity and correlated with the dosage of overexpression, DYRK1A blocked proliferation of SH-SY5Y neuroblastoma cells within 24 h and arrested the cells in G1 phase. Sustained overexpression of DYRK1A induced G0 cell cycle exit and neuronal differentiation. Furthermore, we provide evidence that DYRK1A modulated protein stability of cell cycle-regulatory proteins. DYRK1A reduced cellular Cyclin D1 levels by phosphorylation on Thr286, which is known to induce proteasomal degradation. In addition, DYRK1A phosphorylated p27(Kip1) on Ser10, resulting in protein stabilization. Inhibition of DYRK1A kinase activity reduced p27(Kip1) Ser10 phosphorylation in cultured hippocampal neurons and in embryonic mouse brain. In aggregate, these results suggest a novel mechanism by which overexpression of DYRK1A may promote premature neuronal differentiation and contribute to altered brain development in Down syndrome.