Browsing by Author "Kumari, Aprajita"

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Adaptive reprogramming during early seed germination requires temporarily enhanced fermentation-A critical role for alternative oxidase regulation that concerns also microbiota effectiveness
(Frontiers Media S.A., 2021) Bharadwaj, Revuru; Noceda, Carlos; Mohanapriya, Gunasekharan; Kumar, Sarma Rajeev; Thiers, Karine Leitão Lima; Costa, José Hélio; Macedo, Elisete Santos; Kumari, Aprajita; Gupta, Kapuganti Jagadis; Srivastava, Shivani; Adholeya, Alok; Oliveira, Manuela; Velada, Isabel; Sircar, Debabrata; Sathishkumar, Ramalingam; Arnholdt-Schmitt, Birgit
Plants respond to environmental cues via adaptive cell reprogramming that can affect whole plant and ecosystem functionality. Microbiota constitutes part of the inner and outer environment of the plant. This Umwelt underlies steady dynamics, due to complex local and global biotic and abiotic changes. Hence, adaptive plant holobiont responses are crucial for continuous metabolic adjustment at the systems level. Plants require oxygen-dependent respiration for energy-dependent adaptive morphology, such as germination, root and shoot growth, and formation of adventitious, clonal, and reproductive organs, fruits, and seeds. Fermentative paths can help in acclimation and, to our view, the role of alternative oxidase (AOX) in coordinating complex metabolic and physiological adjustments is underestimated. Cellular levels of sucrose are an important sensor of environmental stress. We explored the role of exogenous sucrose and its interplay with AOX during early seed germination. We found that sucrose-dependent initiation of fermentation during the first 12 h after imbibition (HAI) was beneficial to germination. However, parallel upregulated AOX expression was essential to control negative effects by prolonged sucrose treatment. Early downregulated AOX activity until 12 HAI improved germination efficiency in the absence of sucrose but suppressed early germination in its presence. The results also suggest that seeds inoculated with arbuscular mycorrhizal fungi (AMF) can buffer sucrose stress during germination to restore normal respiration more efficiently. Following this approach, we propose a simple method to identify organic seeds and low-cost on-farm perspectives for early identifying disease tolerance, predicting plant holobiont behavior, and improving germination. Furthermore, the research strengthens the view that AOX can serve as a powerful functional marker source for seed hologenomes.
Alternative oxidase (AOX) senses stress levels to coordinate auxin-induced reprogramming from seed germination to somatic embryogenesis- A role relevant for seed vigor prediction and plant robustness
(Frontiers Media S.A., 2019) Mohanapriya, Gunasekaran; Bharadwaj, Revuru; Noceda, Carlos; Costa, José Hélio; Kumar, Sarma Rajeev; Sathishkumar, Ramalingam; Thiers, Karine Leitão Lima; Macedo, Elisete Santos; Silva, Sofia; Annicchiarico, Paolo; Groot, Steven P.C.; Kodde, Jan; Kumari, Aprajita; Gupta, Kapuganti Jagadis; Arnholdt-Schmitt, Birgit
Somatic embryogenesis (SE) is the most striking and prominent example of plant plasticity upon severe stress. Inducing immature carrot seeds perform SE as substitute to germination by auxin treatment can be seen as switch between stress levels associated to morphophysiological plasticity. This experimental system is highly powerful to explore stress response factors that mediate the metabolic switch between cell and tissue identities. Developmental plasticity per se is an emerging trait for in vitro systems and crop improvement. It is supposed to underlie multi-stress tolerance. High plasticity can protect plants throughout life cycles against variable abiotic and biotic conditions. We provide proof of concepts for the existing hypothesis that alternative oxidase (AOX) can be relevant for developmental plasticity and be associated to yield stability. Our perspective on AOX as relevant coordinator of cell reprogramming is supported by real-time polymerase chain reaction (PCR) analyses and gross metabolism data from calorespirometry complemented by SHAM-inhibitor studies on primed, elevated partial pressure of oxygen (EPPO)–stressed, and endophyte-treated seeds. In silico studies on public experimental data from diverse species strengthen generality of our insights. Finally, we highlight ready-to-use concepts for plant selection and optimizing in vivo and in vitro propagation that do not require further details on molecular physiology and metabolism. This is demonstrated by applying our research & technology concepts to pea genotypes with differential yield performance in multilocation fields and chickpea types known for differential robustness in the field. By using these concepts and tools appropriately, also other marker candidates than AOX and complex genomics data can be efficiently validated for prebreeding and seed vigor prediction.
Alternative oxidase is an important player in the regulation of nitric oxide levels under normoxic and hypoxic conditions in plants
(Oxford University Press, 2019) Kumari, Aprajita; Pathak, Pradeep Kumar; Bulle, Mallesham; Igamberdiev, Abir U; Gupta, Kapuganti Jagadis
Plant mitochondria possess two different pathways for electron transport from ubiquinol: the cytochrome pathway and the alternative oxidase (AOX) pathway. The AOX pathway plays an important role in stress tolerance and is induced by various metabolites and signals. Previously, several lines of evidence suggested that the AOX pathway prevents overproduction of superoxide and other reactive oxygen species. More recent evidence suggests that AOX also plays a role in regulation of nitric oxide (NO) production and signalling. The AOX pathway is induced under low phosphate, hypoxia, pathogen infections and elicitor treatments. The induction of AOX under aerobic conditions in response to various stresses can reduce electron transfer through complexes III and IV and thus prevents the leakage of electrons to nitrite and the subsequent accumulation of NO. Excess of NO under various stresses can inhibit complex IV, thus the AOX pathway minimizes nitrite-dependent NO synthesis that would arise from enhanced electron leakage in the cytochrome pathway. By preventing NO generation, AOX can reduce peroxynitrite formation and tyrosine nitration. In contrast to its function under normoxia, AOX has a specific role under hypoxia, where AOX can facilitate nitrite-dependent NO production. This reaction drives the phytoglobin-NO cycle to increase energy efficiency under hypoxia.
Alternative oxidase plays a role in minimizing ROS and RNS produced under salinity stress in Arabidopsis thaliana
(John Wiley & Sons, 2022) Manbir; Singh, Pooja; Kumari, Aprajita; Gupta, Kapuganti Jagadis
Under stress conditions, the overproduction of different reactive oxygen species (ROS) and reactive nitrogen species (RNS) causes imbalance in the redox homeostasis of the cell leading to nitro-oxidative stress in plants. Alternative oxidase (AOX) is a conserving terminal oxidase of the mitochondrial electron transport chain, which can minimize the ROS. Still, the role of AOX in the regulation of RNS during nitro-oxidative stress imposed by salinity stress is not known. Here, we investigated the role of AOX in minimizing ROS and RNS induced by 150 mM NaCl in Arabidopsis using transgenic plants overexpressing (AOX OE) and antisense lines (AOX AS) of AOX. Imposing NaCl treatment leads to a 4-fold enhanced expression of AOX accompanied by enhanced AOX capacity in WT Col-0. Further AOX-OE seedlings displayed enhanced growth compared with the AOX-AS line under stress. Examination of NO levels by DAF-FM fluorescence and chemiluminescence revealed that AOX overexpression leads to reduced levels of NO. The total NR activity was elevated under NaCl, but no significant change was observed in wild-type (WT), AOX OE, and AS lines. The total ROS, superoxide, H2O2 levels, and lipid peroxidation were higher in the AOX-AS line than in WT and AOX-OE lines. The peroxynitrite levels were also higher in the AOX-AS line than in WT and AOX-OE lines; further, the expression of antioxidant genes was elevated in AOX-AS. Taken together, our results suggest that AOX plays an important role in the mitigation of ROS and RNS levels and enhances plant growth, thus providing tolerance against nitro-oxidative stress exerted by NaCl.
Chemiluminescence detection of nitric oxide from roots, leaves, and root mitochondria
(Springer, 2016) Wany, Aakanksha; Gupta, Alok Kumar; Kumari, Aprajita; Gupta, Shika; Mishra, Sonal; Jaintu, Ritika; Pathak, Pradeep K.; Gupta, Kapuganti Jagadis
NO is a free radical with short half-life and high reactivity; due to its physiochemical properties it is very difficult to detect the concentrations precisely. Chemiluminescence is one of the robust methods to quantify NO. Detection of NO by this method is based on reaction of nitric oxide with ozone which leads to emission of light and amount of light is proportional to NO. By this method NO can be measured in the range of pico moles to nano moles range. Using direct chemiluminescence method, NO emitted into the gas stream can be detected whereas using indirect chemiluminescence oxidized forms of NO can also be detected. We detected NO using purified nitrate reductase, mitochondria, cell suspensions, and roots; detail measurement method is described here.
Current approaches to measure nitric oxide in plants
(Oxford University Press, 2019) Vishwakarma, Abhaypratap; Wany, Aakanksha; Pandey, Sonika; Bulle, Mallesham; Kumari, Aprajita; Kishorekumar, Reddy; Igamberdiev, Abir U.; Mur, Luis A.J.; Gupta, Kapuganti Jagadis
Nitric oxide (NO), is now established as an important signalling molecule in plants where they influence growth, development and responses to stress. Despite of extensive research, the most appropriate methods to measure and localise these signalling radicals are debated and still needs investigation. Many confounding factors such as presence of other reactive intermediates, scavenging enzymes and compartmentation, influence how accurately each can be measured. Further, these signalling radicals have short half-lives ranging from seconds to minutes based on the cellular redox condition. Hence, it is necessary to use the sensitive and specific methods in order to understand the contribution of each signalling molecule to various biological processes. In this review, we will provide current state knowledge on NO measurement in plant samples, via various methods. We will also discuss advantages, limitations and wider applications of each method.
Detection of nitric oxide from chickpea using DAF fluorescence and chemiluminescence methods
(John Wiley & Sons, 2022) Kumari, Aprajita; Bhatoee, Manbir; Singh, Pooja; Kaladhar, Vemula Chandra; Yadav, Nidhi; Paul, Debarati; Loake, Gary J.; Gupta, Kapuganti Jagadis
The free radical nitric oxide (NO) has emerged as an important signal molecule in plants, due to its involvement in various plant growth, development, and stress responses. For elucidating the role of NO, it is very important to precisely determine, localize, and quantify NO levels. Due to a relatively short half-life and its rapid, complex reactivity with other radicals, together with its capacity to diffuse from the source of production, the quantification of NO in whole plants, tissues, organelles, and extracts is notoriously difficult. Hence, it is essential to employ sensitive procedures for precise detection of NO. Currently available methods can fulfill many requirements to precisely determine NO, but each method has several advantages and pitfalls. In this article, we describe a detailed procedure for the measurement of NO by diaminofluorescein (DAF) in cell-permeable forms (DAF-FM-DA). In this method, the tissues are immersed in DAF-FM DA, leading to their diffusion from the plasma membrane to the inside of the cell, where intracellular esterases cleave the ester bonds, leading to DAF-FM release. The resulting DAF-FM reacts with intracellularly generated NO and forms highly fluorescent triazolofluorescein (DAF-FMT), which can be localized and monitored by fluorescence or confocal microscopy, and can also be detected via fluorimetry and flow cytometry. DAF dyes are very popular as they are non-invasive, relatively easy to handle, and commercially available. Another precise and very sensitive method is chemiluminescence detection of NO, where NO reacts with ozone (O3), leading to emission of a quantum of light from which NO can be calculated. Using chickpea seedlings, we describe in detail the measurement of NO using DAF-FM-DA and chemiluminescence methods. © 2022 Wiley Periodicals LLC. Basic Protocol 1: Measurement of nitric oxide from chickpea seedlings using DAF-FM DA fluorescence with fluorescence and confocal microscopy Basic Protocol 2: Chemiluminescence detection of nitric oxide from chickpea seedlings
A discrete role for alternative oxidase under hypoxia to increase nitric oxide and drive energy production
(Elsevier B.V., 2018) Vishwakarma, Abhaypratap; Kumari, Aprajita; Mur, Luis A.J.; Gupta, Kapuganti Jagadis
Alternative oxidase (AOX) is an integral part of the mitochondrial electron transport and can prevent reactive oxygen species (ROS) and nitric oxide (NO) production under non-stressed, normoxic conditions. Here we assessed the roles of AOX by imposing stress under normoxia in comparison to hypoxic conditions using AOX over expressing (AOX OE) and anti-sense (AOX AS) transgenic Arabidopsis seedlings and roots. Under normoxic conditions stress was induced with the defence elicitor flagellin (flg22). AOX OE reduced NO production whilst this was increased in AOX AS. Moreover AOX AS also exhibited an increase in superoxide and therefore peroxynitrite, tyrosine nitration suggesting that scavenging of NO by AOX can prevent toxic peroxynitrite formation under normoxia. In contrast, during hypoxia interestingly we found that AOX is a generator of NO. Thus, the NO produced during hypoxia, was enhanced in AOX OE and suppressed in AOX AS. Additionally, treatment of WT or AOX OE with the AOX inhibitor SHAM inhibited hypoxic NO production. The enhanced levels of NO correlated with expression of non-symbiotic haemoglobin, increased NR activity and ATP production. The ATP generation was suppressed in nia1,2 mutant and non symbiotic haemoglobin antisense line treated with SHAM. Taken together these results suggest that hypoxic NO generation mediated by AOX has a discrete role by feeding into the haemoglobin-NO cycle to drive energy efficiency under conditions of low oxygen tension.
The emerging roles of nitric oxide and its associated scavengers-phytoglobins-in plant symbiotic interactions
(Oxford University Press, 2024) Pathak, Pradeep Kumar; Yadav, Nidhi; Kaladhar, Vemula Chandra; Jaiswal, Rekha; Kumari, Aprajita; Igamberdiev, Abir U.; Loake, Gary J.; Gupta, Kapuganti Jagadis
A key feature in the establishment of symbiosis between plants and microbes is the maintenance of the balance between the production of the small redox-related molecule, nitric oxide (NO), and its cognate scavenging pathways. During the establishment of symbiosis, a transition from a normoxic to a microoxic environment often takes place, triggering the production of NO from nitrite via a reductive production pathway. Plant hemoglobins [phytoglobins (Phytogbs)] are a central tenant of NO scavenging, with NO homeostasis maintained via the Phytogb–NO cycle. While the first plant hemoglobin (leghemoglobin), associated with the symbiotic relationship between leguminous plants and bacterial Rhizobium species, was discovered in 1939, most other plant hemoglobins, identified only in the 1990s, were considered as non-symbiotic. From recent studies, it is becoming evident that the role of Phytogbs1 in the establishment and maintenance of plant–bacterial and plant–fungal symbiosis is also essential in roots. Consequently, the division of plant hemoglobins into symbiotic and non-symbiotic groups becomes less justified. While the main function of Phytogbs1 is related to the regulation of NO levels, participation of these proteins in the establishment of symbiotic relationships between plants and microorganisms represents another important dimension among the other processes in which these key redox-regulatory proteins play a central role.
The functional role of nitric oxide in plant mitochondrial metabolism
(Elsevier B.V., 2016) Gupta, Alok Kumar; Kumari, Aprajita; Mishra, Sonal; Wany, Aakanksha; Gupta, Kapuganti Jagadis
In recent years, mitochondrial nitric oxide (NO) production has attracted increasing attention. Mitochondria generate NO using nitrite as a substrate. Cytochrome c oxidase and other components of the electron transport chain also contribute to NO generation. Accumulating evidence indicates that mitochondria are scavengers of NO. Furthermore, several genes encoding mitochondrial proteins, as well as mitochondrial proteins, are regulated by NO. In this chapter, we provided an overview of the mechanisms of NO generation and scavenging in mitochondria and of the NO-dependent regulation of proteins and genes encoding mitochondrial proteins. In addition, the functional roles of NO in mitochondrial metabolism, such as inhibition of aconitase, production of ATP and induction of alternative oxidase are presented.
Greenhouse and field experiments revealed that clove oil can effectively reduce bacterial blight and increase yield in pomegranate
(John Wiley & Sons, 2021) Kumar, Pavan; Lokesh, Veeresh; Doddaraju, Pushpa; Kumari, Aprajita; Singh, Pooja; Meti, Bharati S.; Sharma, Jyotsana; Gupta, Kapuganti Jagadis; Manjunatha, Girigowda
Bacterial blight in pomegranate is a devastating disease caused by bacterial pathogen Xanthomonas axonopodis pv. punicae (XAP), recording huge damage to pomegranate crop worldwide. Antibiotics and copper-based chemicals are being used for the management of this blight, while in this present work, we investigated the effect of eugenol and clove oil either singly or in combination with copper oxychloride (COC) on the induction of plant defense responses and concomitant prevention of bacterial blight. Our results provided evidence that clove oil (0.2%–1%) and eugenol (0.1% and 0.2%) successfully inhibit the growth of XAP in paper disk diffusion assay. Strikingly under the greenhouse condition, clove oil (0.2%) as foliar application 24 h before XAP inoculation recorded the lowest disease severity of 7.34%, whereas eugenol (0.2%) recorded maximum disease severity of 14.56%. However, the combination of clove oil (0.2%) and copper oxychloride (0.3%) recorded the least disease severity of 2.38%. A similar trend was observed in field conditions. Prophylactic application of clove oil leads to enhanced nitrate reductase activity and nitric oxide production which was further enhanced in clove oil pre-treated plants challenged with XAP. Strikingly, the total ROS and H2O2 levels were reduced in response to clove oil application. Clove oil also induced the systemic response by inducing expression levels of defense genes. The reduction of disease severity by clove oil and COC combination also reflected on total yield recording via large-scale field experiments where maximum yield of 14.04 tonnes/acre was observed, whereas streptocycline application recorded 11.12 tonnes/acre. Application of COC and clove oil resulted in a high remunerative value of ₹ 1:5.6, compared to streptocycline (1:4.85) and control (1:1.85). The present study revealed that clove oil as a plant derivative and eugenol as a synthetic option can be effectively used for the successful management of bacterial blight in pomegranate.
Interaction of nitric oxide with the components of plant mitochondrial electron transport chain
(Oxford University Press, 2018) Gupta, Kapuganti Jagadis; Kumari, Aprajita; Florez-Sarasa, Igor; Fernie, Alisdair R; Igamberdiev, Abir U
Mitochondria are not only major sites for energy production but also participate in several alternative functions, among these generation of nitric oxide (NO) and its different impacts on this organelle is increasing attention. The inner mitochondrial membrane contains the chain of protein complexes, and electron transfer via oxidation of various organic acids and reducing equivalents leads to generation of proton gradient that results in energy production. Recent evidence suggests that these complexes are sources and targets for NO. Complex I and rotenone-insensitive NAD(P)H dehydrogenases regulate hypoxic NO production, while complex I also participates in the formation of supercomplex with complex III under hypoxia. Complex II is a target for NO which by inhibiting Fe-S centres regulates ROS generation, and complex III is one of the major sites for NO production and the produced NO participates in the phytoglobin-NO cycle that leads to the maintenance of redox level and limited energy production under hypoxia. Expression of the alternative oxidase (AOX) is induced by NO under various stress conditions, and evidence exists that AOX can regulate mitochondrial NO production. Complex IV is another major site for NO production which can also be linked to ATP generation via the phytoglobin-NO cycle. Inhibition of complex IV by NO can prevent oxygen depletion at the frontier of anoxia. The NO production and action on various complexes play a major role in NO signalling and energy metabolism.
Isolation and measurement of respiration and structural studies of purified mitochondria from heterotrophic plant tissues
(John Wiley & Sons, 2021) Pandey, Sonika; Kumari, Aprajita; Singh, Pooja; Gupta, Kapuganti Jagadis
Mitochondria are the power houses of eukaryotic cells. These organelles contain various oxidoreductase complexes. Electron transfer from different reducing equivalents channeled via these complexes drives proton translocation across the inner mitochondrial membrane, leading to ATP generation. Plant mitochondria contain alternative NAD(P)H dehydrogenases, alternative oxidase, and uncoupling protein, and TCA cycle enzymes are located in their matrix. Apart from ATP production, mitochondria are also involved in synthesis of vitamins and cofactors and participate in fatty acid, nucleotide, photorespiratory, and antioxidant metabolism. Recent emerging evidence suggests that mitochondria play a role in redox signaling and generation of reactive oxygen and nitrogen species. For mitochondrial studies, it is essential to isolate physiologically active mitochondria with good structural integrity. In this article, we explain a detailed procedure for isolation of mitochondria from various heterotrophic tissues, such as germinating chickpea seeds, potato tubers, and cauliflower florets. This procedure requires discontinuous Percoll gradient centrifugation and can give a good yield of mitochondria, in the range of 4 to 8 mg per 50 g tissue with active respiratory capacity. After MitoTracker staining, isolated mitochondria can be visualized by using a confocal microscope. The structure of mitochondria can be monitored by scanning electron microscopy.
Isolation of physiologically active and intact mitochondria from chickpea
(Springer, 2017) Pandey, Sonika; Kumari, Aprajita; Gupta, Kapuganti Jagadis
Chickpea is an important leguminous crop that belongs to Fabaceae family, highly valued for its nutritious seeds. Seeds contain reserve food for the developing embryos. Mitochondria are crucial organelle for generation of chemical energy in the form of ATP which is required for achieving metabolically active state; therefore, investigating mitochondrial function and respiration rate is crucial for exploring various metabolic and physio-biochemical changes that occur during seed germination. Here we describe a method for isolation of mitochondria from germinating seeds of two chickpea varieties, i.e., Desi and Kabuli. Structure of Mitotracker-stained isolated mitochondria was observed by confocal microscopy and respiration rate was measured using an oxygen microsensor.
Measurement of oxygen status in Arabidopsis leaves undergoing the hypersensitive response during Pseudomonas infection
(Springer, 2017) Kumari, Aprajita; Preston, Gail M.; Gupta, Kapuganti Jagadis
When plants are infected with pathogens they defend themselves via various processes. Once such process is the development of the hypersensitive response (HR), a kind of programmed cell death (PCD), in which localized cell death takes place in order to prevent pathogen spread to other part of tissue. The Arabidopsis and Pseudomonas syringae system is one of the best known examples to study the HR. Here we used the VisiSens™ oxygen-imaging system to investigate oxygen distributions in Arabidopsis leaves infected with an avirulent strain of P. syringae and undergoing the hypersensitive response. Using this method we observed a change in oxygen status at 6 h post-infection and a drop in oxygen levels at 24 h after infection.
Measurement of reactive oxygen species and nitric oxide from tomato plants in response to abiotic and biotic stresses
(Springer Nature Publishing AG, 2024) Samant, Sanjib Bal; Manbir; Rekha; Swain, Jagannath; Singh, Pooja; Kumari, Aprajita; Gupta, Kapuganti Jagadis
Nitric oxide (NO) is a free radical molecule that has been known to influence several cellular processes such as plant growth, development, and stress responses. NO together with reactive oxygen species (ROS) play a role in signaling process. Due to extremely low half-life of these radicals in cellular environment, it is often difficult to precisely monitor them. Each method has some advantages and disadvantages; hence, it is important to measure using multiple methods. To interpret the role of each signaling molecule in numerous biological processes, sensitive and focused methods must be used. In addition to this complexity, these Reactive Oxygen Species (ROS) and NO react with each other leads to nitro-oxidative stress in plants. Using tomato as a model system here, we demonstrate stepwise protocols for measurement of NO by chemiluminescence, DAF fluorescence, nitrosative stress by western blot, and ROS measurement by NBT and DAB under stress conditions such as osmotic stress and Botrytis infection. While describing methods, we also emphasized on benefits, drawbacks, and broader applications of these methods.
Measurement of respiration and internal oxygen in germinating Cicer arietinum L. seeds using optic microsensor
(Springer, 2017) Pandey, Sonika; Kumari, Aprajita; Bharadwaj, Chellapilla; Gupta, Kapuganti Jagadis
Internal oxygen concentrations vary in different tissues depending on tissue size, developmental stage, and their location. Respiratory rate of tissue also determines internal oxygen levels. For studying various signaling pathways it is essential to establish a correlation between respiration and internal oxygen. Seed germination is associated with increase in respiration which can dictate the internal oxygen and subsequent production of reactive oxygen species. Using optic oxygen microsensor we made an attempt to measure respiratory rate and internal oxygen. We found that microsensor is able to sense internal oxygen and it is also possible to measure oxygen levels in a close vial that contains seeds. Step-by-step protocol is described here along with illustration.
Metabolism and signalling in pea (Pisum sativum) leaves exposed to drought and subsequent recovery
(John Wiley & Sons, 2026) Pandey, Jayendra; Mantena, Chakradhar; Kumari, Aprajita; Singh, Pooja; Foyer, Christine H.; Gupta, Kapuganti Jagadis; Subramanyam, Rajagopal
Uncovering the metabolic and molecular mechanisms involved in plant responses to drought and subsequent recovery, is essential to identify drought tolerance mechanisms that can be used to improve crop plants. Here we combine plant physiology and biochemistry, with gene expression, quantitative proteomics and metabolite profiling to identify the genetic and metabolic networks that operate in plants experiencing and recovering from drought. Network analysis of transcripts, proteins and metabolites revealed that certain biological processes such as the tricarboxylic acid cycle and lipid metabolism had a strong impact on the overall control of leaf responses to drought and recovery. The stimulation of carbohydrate oxidation pathways is demonstrated to be a key node in the generation of energy and precursors required to support diverse survival pathways of defence.
Metabolomic response to high light from pgrl1 and pgr5 mutants of Chlamydomonas reinhardtii
(Springer Nature Publishing AG, 2023) Chouhan, Nisha; Marriboina, Sureshbabu; Kumari, Aprajita; Singh, Pooja; Yadav, Ranay Mohan; Gupta, Kapuganti Jagadis; Subramanyam, Rajagopal
Chlamydomonas (C.) reinhardtii metabolomic changes in cyclic electron flow-dependent mutants are still unknown. Here, we used mass spectrometric analysis to monitor the changes in metabolite levels in wild-type, cyclic electron-deficient mutants pgrl1 and pgr5 grown under high-light stress. A total of 55 metabolites were detected using GC-MS analysis. High-light stress-induced selective anaplerotic amino acids in pgr5. In addition, pgr5 showed enhancement in carbohydrate, polyamine, and polyol metabolism by 2.5-fold under high light. In response to high light, pgr5 triggers an increase in several metabolites involved in regulating osmotic pressure. Among these metabolites are glycerol pathway compounds such as glycerol-3-phosphate and glyceryl-glycoside, which increase significantly by 1.55 and 3.07 times, respectively. In addition, pgr5 also enhanced proline and putrescine levels by 2.6- and 1.36-fold under high light. On the other hand, pgrl1-induced metabolites, such as alanine and serine, are crucial for photorespiration when subjected to high-light stress. We also observed a significant increase in levels of polyols and glycerol by 1.37- and 2.97-fold in pgrl1 under high-light stress. Both correlation network studies and KEGG pathway enrichment analysis revealed that metabolites related to several biological pathways, such as amino acid, carbohydrate, TCA cycle, and fatty acid metabolism, were positively correlated in pgrl1 and pgr5 under high-light stress conditions. The relative mRNA expression levels of genes related to the TCA cycle, including PDC3, ACH1, OGD2, OGD3, IDH3, and MDH4, were significantly upregulated in pgrl1 and pgr5 under HL. In pgr5, the MDH1 level was significantly increased, while ACS1, ACS3, IDH2, and IDH3 levels were reduced considerably in pgrl1 under high-light stress. The current study demonstrates both pgr5 and prgl1 showed a differential defense response to high-light stress at the primary metabolites and mRNA expression level, which can be added to the existing knowledge to explore molecular regulatory responses of prg5 and pgrl1 to high-light stress.
Moving nitrogen to the centre of plant defence against pathogens
(Oxford University Press, 2017) Mur, Luis A.J.; Simpson, Catherine; Kumari, Aprajita; Gupta, Alok Kumar; Gupta, Kapuganti Jagadis
Background Plants require nitrogen (N) for growth, development and defence against abiotic and biotic stresses. The extensive use of artificial N fertilizers has played an important role in the Green Revolution. N assimilation can involve a reductase series (NO–3→ NO–2 → NH+4) followed by transamination to form amino acids. Given its widespread use, the agricultural impact of N nutrition on disease development has been extensively examined. Scope: When a pathogen first comes into contact with a host, it is usually nutrient starved such that rapid assimilation of host nutrients is essential for successful pathogenesis. Equally, the host may reallocate its nutrients to defence responses or away from the site of attempted infection. Exogenous application of N fertilizer can, therefore, shift the balance in favour of the host or pathogen. In line with this, increasing N has been reported either to increase or to decrease plant resistance to pathogens, which reflects differences in the infection strategies of discrete pathogens. Beyond considering only N content, the use of NO-3 or NH+4 fertilizers affects the outcome of plant–pathogen interactions. NO-3 feeding augments hypersensitive response- (HR) mediated resistance, while ammonium nutrition can compromise defence. Metabolically, NO–3 enhances production of polyamines such as spermine and spermidine, which are established defence signals, with NH+4 nutrition leading to increased γ-aminobutyric acid (GABA) levels which may be a nutrient source for the pathogen. Within the defensive N economy, the roles of nitric oxide must also be considered. This is mostly generated from NO–2 by nitrate reductase and is elicited by both pathogen-associated microbial patterns and gene-for-gene-mediated defences. Nitric oxide (NO) production and associated defences are therefore NO-3 dependent and are compromised by NH+4. Conclusion:This review demonstrates how N content and form plays an essential role in defensive primary and secondary metabolism and NO-mediated events.