Leonor Ramirez

Phosphatidic acid production in chitosan-elicited tomato cells, via both phospholipase D and phospholipase C/diacylglycerol kinase, requires nitric oxide

Nitric oxide (NO) and the lipid second messenger phosphatidic acid (PA) are involved in plant defense responses during plant-pathogen interactions. NO has been shown to be involved in the induction of PA production in response to the pathogen associated molecular pattern (PAMP) xylanase in tomato cells. It was shown that NO is critical for PA production induced via phospholipase C (PLC) in concerted action with diacylglycerol kinase (DGK) but not for the xylanase-induced PA via phospholipase D (PLD). In order to study whether this is a general phenomenon during PAMP perception or if it is particular for xylanase, we studied the effect of the PAMP chitosan in tomato cell suspensions. We observed a rapid NO production in tomato cells treated with chitosan. Chitosan induced the formation of PA by activating both PLD and PLC/DGK. The activation of either phospholipase-mediated signaling pathway was inhibited in cells treated with the NO scavenger cPTIO. This indicates that NO is required for PA generation via both the PLD and PLC/DGK pathway during plant defense response in chitosan elicited cells. Responses downstream PA were studied. PLC inhibitors neomycin and U73122 inhibited chitosan-induced ROS production. Differences between xylanase and chitosan-induced phospholipid signaling pathways are discussed.

Nitric oxide, nitrosyl iron complexes, ferritin and frataxin: A well equipped team to preserve plant iron homeostasis

Iron is a key element in plant nutrition. Iron deficiency as well as iron overload results in serious metabolic disorders that affect photosynthesis, respiration and general plant fitness with direct consequences on crop production. More than 25% of the cultivable land possesses low iron availability due to high pH (calcareous soils). Plant biologists are challenged by this concern and aimed to find new avenues to ameliorate plant responses and keep iron homeostasis under control even at wide range of iron availability in various soils. For this purpose, detailed knowledge of iron uptake, transport, storage and interactions with cellular compounds will help to construct a more complete picture of its role as essential nutrient. In this review, we summarize and describe the recent findings involving four central players involved in keeping cellular iron homeostasis in plants: nitric oxide, ferritin, frataxin and nitrosyl iron complexes. We attempt to highlight the interactions among these actors in different scenarios occurring under iron deficiency or iron overload, and discuss their counteracting and/or coordinating actions leading to the control of iron homeostasis.

Glutathione and ascorbic acid protect Arabidopsis plants against detrimental effects of iron deficiency

Iron is an essential micronutrient required for a wide variety of cellular functions in plant growth and development. Chlorosis is the first visible symptom in iron-deficient plants. Glutathione (GSH) and ascorbic acid (ASC) are multifunctional metabolites playing important roles in redox balancing. In this work, it was shown that GSH and ASC treatment prevented chlorosis and the accumulation of reactive oxygen species induced by iron deficiency in Arabidopsis leaves. In iron deficiency, GSH and ASC increased the activity of the heme protein ascorbate peroxidase at a similar level to that found in iron-sufficient seedlings. GSH was also able to preserve the levels of the iron-sulfur protein ferredoxin 2. GSH content decreased 25% in iron-deficient Arabidopsis seedlings, whereas the ASC levels were not affected. Taken together, these results showed that GSH and ASC supplementation protects Arabidopsis seedlings from iron deficiency, preserving cell redox homeostasis and improving internal iron availability.

Nitric oxide and frataxin: two players contributing to maintain cellular iron homeostasis.

BACKGROUND Nitric oxide (NO) is a signalling and physiologically active molecule in animals, plants and bacteria. The specificity of the molecular mechanism(s) involved in transducing the NO signal within and between cells and tissues is still poorly understood. NO has been shown to be an emerging and potent signal molecule in plant growth, development and stress physiology. The NO donor S-nitrosoglutathion (GSNO) was shown to be a biologically active compound in plants and a candidate for NO storage and/or mobilization between plant tissues and cells. NO has been implicated as a central component in maintaining iron bioavailavility in plants. SCOPE AND CONCLUSIONS Iron is an essential nutrient for almost all organisms. This review presents an overview of the functions of NO in iron metabolism in animals and discusses how NO production constitutes a key response in plant iron sensing and availability. In plants, NO drives downstream responses to both iron deficiency and iron overload. NO-mediated improvement of iron nutrition in plants growing under iron-deficient conditions represents a powerful tool to cope with soils displaying low iron availability. An interconversion between different redox forms based on the iron and NO status of the plant cells might be the core of a metabolic process driving plant iron homeostasis. Frataxin, a recently identified protein in plants, plays an important role in mitochondria biogenesis and in maintaining mitochondrial iron homeostasis. Evidence regarding the interaction between frataxin, NO and iron from analysis of frataxin knock-down Arabidopsis thaliana mutants is reviewed and discussed.

Abscisic acid enhances the immune response in Apis mellifera and contributes to the colony fitness

The primary food of adult honey bees (Apis mellifera) is honey prepared by bees from nectar, provided by plants in order to stimulate the bee’s pollination service. Nectar consists of carbohydrates, amino acids and water, as well as other minor compounds whose proportion varies among plant species and whose biological implications in the honey bee physiology require intense research. Several environmental stressors are causing the decline of bee colonies, and thereby, we tried to connect the nutritional quality of bee’s diet with the strength of the bee’s immune system. The phytohormone abscisic acid (ABA) is present in nectar, honey and adult honey bees. It has been demonstrated that ABA stimulates innate immune defences in animal cells. However, the influence of ABA on A. mellifera’s health and fitness is unknown. Here, we show that honey bees fed with an ABA supplement in field experiments resulted in (i) the appearance of ABA in larvae and adult bees, (ii) enhanced haemocyte response to non-self recognition, (iii) improved wound healing and granulocyte and plasmatocyte activation and (iv) maximum adult bee population after the winter and increased pesticide tolerance. The results indicate that the naturally occurring compound ABA has a positive influence in honey bee immunity. ABA emerges as a potent booster of immune defence in A. mellifera and may be useful in addressing the colony losses threatening apiculture and pollination service worldwide.

The tomato phosphatidylinositol-phospholipase C2 (SlPLC2) is required for defense gene induction by the fungal elicitor xylanase.

The tomato [Solanum lycopersicum (Sl)] phosphatidylinositol-phospholipase C (PI-PLC) gene family is composed of six members, named SlPLC1 to SlPLC6, differentially regulated upon pathogen attack. We have previously shown that the fungal elicitor xylanase rapidly induces nitric oxide (NO), which is required for PI-PLCs activity and downstream defense responses in tomato cell suspensions. Here, we show that all six SlPLC genes are expressed in tomato cell suspensions. Treatment of the cells with xylanase induces an early increase in SlPLC5 transcript levels, followed by a raise of the amount of SlPLC2 transcripts. The production of NO is required to augment SlPLC5 transcript levels in xylanase-treated tomato cells. Xylanase also induces SlPLC2 and SlPLC5 transcript levels in planta. We knocked-down the expression of SlPLC2 and SlPLC5 by virus-induced gene silencing. We found that SlPLC2 is required for xylanase-induced expression of the defense-related genes PR1 and HSR203J.

Decoding plant responses to iron deficiency: Is nitric oxide a central player?

Plants respond to iron deprivation by inducing a series of physiological and morphological responses to counteract the nutrient deficiency. These responses include: (i) the acidification of the extracellular medium, (ii) the reduction of ferric ion and (iii) the increased transport of ferrous ion inside of root cells. This iron transport system is present in strategy I plants and is strictly regulated; at low iron concentration the responses are induced whereas upon iron supply they are repressed. The mechanisms related with this process has been extensively studied, however, the specific cellular effectors involved in sensing iron deficiency, the cascade of components participating in signal transduction, and the way iron is metabolized and delivered, are yet poorly understood. Recently, it has been proposed nitric oxide (NO) as a signaling molecule required for plant responses to iron deficiency. NO is produced rapidly in the root epidermis of tomato plants that are growing under iron deficient conditions. Furthermore, it was demonstrated that NO is required for the expression and activity of iron uptake components in roots during iron deprivation. Here we propose and discuss a working hypothesis to understand the way NO is acting in plants responses to iron deficiency. We specifically highlight the cross talk between NO and plant hormones, and the interaction between NO, iron and glutathione for the formation of dinitrosyl iron complexes (DNICs). Finally, a potential role of DNICs in iron mobilization is proposed. Addendum to: Graziano M, Lamattina L. Nitric oxide accumulation is required for molecular and physiological responses to iron deficiency in tomato roots. Plant J 2007; 52:949-60.

Cellular immunity in Apis mellifera: studying hemocytes brings light about bees skills to confront threats

Honey bee colonies are threatened by different stress factors around the world. Considerable efforts are being devoted to understanding honey bee defences to confront different kinds of stress factors. Despite the importance of honey bee hemocytes in resisting disease, detailed information about their role in response to challenge is still scarce. This manuscript combines the results obtained in studies which focused on the cellular defences of Apis mellifera, aiming to show how the understanding of the cellular components of the immune system is central to developing new strategies to enhance bees’ fitness.

Abscisic acid enhances cold tolerance in honeybee larvae

The natural composition of nutrients present in food is a key factor determining the immune function and stress responses in the honeybee (Apis mellifera). We previously demonstrated that a supplement of abscisic acid (ABA), a natural component of nectar, pollen, and honey, increases honeybee colony survival overwinter. Here we further explored the role of ABA in in vitro-reared larvae exposed to low temperatures. Four-day-old larvae (L4) exposed to 25°C for 3 days showed lower survival rates and delayed development compared to individuals growing at a standard temperature (34°C). Cold-stressed larvae maintained higher levels of ABA for longer than do larvae reared at 34°C, suggesting a biological significance for ABA. Larvae fed with an ABA-supplemented diet completely prevent the low survival rate due to cold stress and accelerate adult emergence. ABA modulates the expression of genes involved in metabolic adjustments and stress responses: Hexamerin 70b, Insulin Receptor Substrate, Vitellogenin, and Heat Shock Proteins 70. AmLANCL2, the honeybee ABA receptor, is also regulated by cold stress and ABA. These results support a role for ABA increasing the tolerance of honeybee larvae to low temperatures through priming effects.

Dietary Supplementation of Honey Bee Larvae with Arginine and Abscisic Acid Enhances Nitric Oxide and Granulocyte Immune Responses after Trauma

Many biotic and abiotic stressors impact bees’ health, acting as immunosupressors and contribute to colony losses. Thus, the importance of studying the immune response of honey bees is central to develop new strategies aiming to enhance bees’ fitness to confront the threats affecting them. If a pathogen breaches the physical and chemical barriers, honey bees can protect themselves from infection with cellular and humoral immune responses which represent a second line of defense. Through a series of correlative studies we have previously reported that abscisic acid (ABA) and nitric oxide (NO) share roles in the same immune defenses of Apis mellifera (A. mellifera). Here we show results supporting that the supplementation of bee larvae’s diet reared in vitro with l-Arginine (precursor of NO) or ABA enhanced the immune activation of the granulocytes in response to wounding and lipopolysaccharide (LPS) injection.