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In plants, glyoxalase enzymes are activated under stress conditions to mitigate the toxic effects of hyperaccumulated methylglyoxal (MG), a highly reactive carbonyl compound. Until recently, a glutathione-dependent bi-enzymatic pathway involving glyoxalase I (GLYI) and glyoxalase II (GLYII) was considered the primary MG-detoxification system. Recently, a new glutathione-independent glyoxalase III (GLYIII) mediated direct route was also reported in plants. However, the physiological significance of this new pathway remains to be elucidated across plant species. This study identified the full complement of 22 glyoxalases in tomato. Based on their strong induction under multiple abiotic stresses, SlGLYI4, SlGLYII2 and SlGLYIII2 were selected candidates for further functional characterisation. Stress-inducible overexpression of both glutathione-dependent (SlGLYI4 + SlGLYII2) and independent (SlGLYIII2) pathways led to enhanced tolerance in both sets of transgenic plants under abiotic stresses. However, SlGLYIII2 overexpression (OE) plants outperformed the SlGLYI4 + SlGLYII2 OE counterparts for their stress tolerance under abiotic stresses. Further, knockdown of SlGLYIII2 resulted in plants with exacerbated stress responses than those silenced for both SlGLYI4 and SlGLYII2. The superior performance of SlGLYIII2 OE tomato plants for better growth and yield under salt and osmotic treatments could be attributed to better GSH/GSSG ratio, lower reactive oxygen species levels, and enhanced antioxidant potential, indicating a prominent role of GLYIII MG-detoxification pathway in abiotic stress mitigation in this species.

Methylglyoxal (MG), a toxic compound produced as a by-product of several cellular processes, such as respiration and photosynthesis, is well known for its deleterious effects, mainly through glycation of proteins during plant stress responses. However, very little is known about its impact on fruit ripening. Here, we found that MG levels are maintained at high levels in green tomato (Solanum lycopersicum L.) fruits and decline during fruit ripening despite a respiratory burst during this transition. We demonstrate that this decline is mainly mediated through a glutathione-dependent MG detoxification pathway and primarily catalyzed by a Glyoxalase I enzyme encoded by the SlGLYI4 gene. SlGLYI4 is a direct target of the MADS-box transcription factor RIPENING INHIBITOR (RIN), and its expression is induced during fruit ripening. Silencing of SlGLYI4 leads to drastic MG overaccumulation at ripening stages of transgenic fruits and interferes with the ripening process. MG most likely glycates and inhibits key enzymes such as methionine synthase and S-adenosyl methionine synthase in the ethylene biosynthesis pathway, thereby indirectly affecting fruit pigmentation and cell wall metabolism. MG overaccumulation in fruits of several nonripening or ripening-inhibited tomato mutants suggests that the tightly regulated MG detoxification process is crucial for normal ripening progression. Our results underpin a SlGLYI4-mediated regulatory mechanism by which MG detoxification controls fruit ripening in tomato.

Methylglyoxal (MG), a toxic compound produced as a byproduct in several cellular processes such as respiration and photosynthesis, is well investigated for its deleterious effects, mainly through glycation of proteins during plant stress responses. However, very little is known about its impact on fruit ripening. In the present study, we report that MG levels are maintained at high level in green tomato fruits, which declines during fruit ripening inspite of a respiratory burst during this transition. We demonstrate that this decline is mainly mediated by glutathione-dependent MG detoxification pathway and primarily catalyzed by glyoxalase enzyme encoded bySlGLY14gene.SlGLYI4is a direct target of MADS-RIN and is induced during fruit ripening. Silencing of this gene leads to drastic MG overaccumulation at ripening-stages in the transgenic fruits and interferes with the ripening process. Further investigations show that MG plausibly glycates and inhibits key enzymes such as methionine synthase (MS) and S-adenosyl methionine synthase (SAMS) of ethylene biosynthesis pathway, thereby indirectly affecting fruit pigmentation and cell was metabolism. MG overaccumulation in several non-ripening or inhibited- ripening tomato mutant fruits suggests the tightly regulated MG detoxification process is crucial for normal ripening program. Overall, we underpin aSlGLYI4-mediated novel regulatory mechanism of MG detoxification controlling fruit ripening in tomato.

Zinc plays a very critical role and function in all organisms. Its deficiency can cause a serious issue. In

Environmental factors play a significant role in controlling growth, development and defense responses of plants. Changes in the abiotic environment not only significantly alter the physiological and molecular pathways in plants, but also result in attracting the insect pests that carry a payload of viruses. Invasion of plants by viruses triggers the RNA silencing based defense mechanism in plants. In counter defense the viruses have gained the ability to suppress the host RNA silencing activities. A new paradigm has emerged, with the recognition that plant viruses also have the intrinsic capacity to modulate host plant response to environmental cues, in an attempt to favour their own survival. Thus, plant-virus interactions provide an excellent system to understand the signals in crosstalk between biotic (virus) and abiotic stresses. In this review, we have summarized the basal plant defense responses to pathogen invasion while emphasizing on the role of RNA silencing as a front line of defense response to virus infection. The emerging knowledge indicates overlap between RNA silencing with the innate immune responses during antiviral defense. The suppressors of RNA silencing serve as Avr proteins, which can be recognized by the host R proteins. The defense signals also function in concert with the phytohormones to influence plant responses to abiotic stresses. The current evidence on the role of virus induced host tolerance to abiotic stresses is also discussed.

Glyoxalase (GLY) system, comprising of GLYI and GLYII enzymes, has emerged as one of the primary methylglyoxal (MG) detoxification pathways with an indispensable role during abiotic and biotic stresses. MG homeostasis is indeed very closely guarded by the cell as its higher levels are cytotoxic for the organism. The dynamic responsiveness of MG-metabolizing GLY pathway to both endogenous cues such as, phytohormones, nutrient status, etc., as well as external environmental fluctuations (abiotic and biotic stresses) indicates that a tight regulation occurs in the cell to maintain physiological levels of MG in the system. Interestingly, GLY pathway is also manipulated by its substrates and reaction products. Hence, an investigation of signalling and regulatory aspects of GLY pathway would be worthwhile. Herein, we have attempted to converge all known factors acting as signals or directly regulating GLYI/II enzymes in plants. Further, we also discuss how crosstalk between these different signal molecules might facilitate the regulation of glyoxalase pathway. We believe that MG detoxification is controlled by intricate mechanisms involving a plethora of signal molecules.

Endosomal trafficking plays an integral role in various eukaryotic cellular activities and is vital for higher-order functions in multicellular organisms. RAB GTPases are important proteins that influence various aspects of membrane traffic, which consequently influence many cellular functions and responses. Compared to yeast and mammals, plants have evolved a unique set of plant-specific RABs that play a significant role in their development. RABs form the largest family of small guanosine triphosphate (GTP)-binding proteins, and are divided into eight sub-families named RAB1, RAB2, RAB5, RAB6, RAB7, RAB8, RAB11 and RAB18. Recent studies on different species suggest that RAB proteins play crucial roles in intracellular trafficking and cytokinesis, in autophagy, plant microbe interactions and in biotic and abiotic stress responses. This review recaptures and summarizes the roles of RABs in plant cell functions and in enhancing plant survival under stress conditions.

Heat stress, owing to the recent climate change events, has emerged as one of the major environmental factors posing a grave threat to plant survival and crop productivity. Oxidative damage due to increased generation of reactive oxygen and dicarbonyl species in response to heat, is one of the serious consequences of high temperature stress. Methylglyoxal (MG), is one such cytotoxic dicarbonyl metabolite whose levels are known to increase in response to high temperature and various other stress conditions in plants. In fact, it is considered as a general consequence of stress in plants. The synergistic co-activation of reactive oxygen species scavenging and the MG detoxification pathways thus, plays a key role in regulation of stress responses for plant acclimation. Glyoxalase enzymes are the major MG detoxifying proteins in the living organisms. In plants, glyoxalases are encoded as multigenic families, known to be involved in plant growth and development, although, chiefly associated with multiple stress inducible behaviour and tolerance. While their role in salinity, drought and heavy metal stresses has been well-studied, their significance under high temperature stress has received limited coverage. In this review, we emphasize on the dynamic role of glyoxalases under high temperature stress for conferring thermotolerance. Through analysis of heat responsive transcriptomes and proteomes of various plants, we provide evidence for a hitherto, unexplored role of glyoxalases as a mechanism of mounting thermotolerance responses. We also, show that ‘priming’ induced stress/cross stress tolerance in plants involves the activation of glyoxalase pathway.

Methylglyoxal (MG) is a metabolically generated highly cytotoxic compound that accumulates in all living organisms, from Escherichia coli to humans, under stress conditions. To detoxify MG, nature has evolved reduced glutathione (GSH)-dependent glyoxalase and NADPH-dependent aldo-keto reductase systems. But both GSH and NADPH have been reported to be limiting in plants under stress conditions, and thus detoxification might not be performed efficiently. Recently, glyoxalase III (GLY III)-like enzyme activity has been reported from various species, which can detoxify MG without any cofactor. In the present study, we have tested whether an E. coli gene, hchA, encoding a functional GLY III, could provide abiotic stress tolerance to living systems. Overexpression of this gene showed improved tolerance in E. coli and Saccharomyces cerevisiae cells against salinity, dicarbonyl, and oxidative stresses. Ectopic expression of the E. coli GLY III gene (EcGLY-III) in transgenic tobacco plants confers tolerance against salinity at both seedling and reproductive stages as indicated by their height, weight, membrane stability index, and total yield potential. Transgenic plants showed significantly increased glyoxalase and antioxidant enzyme activity that resisted the accumulation of excess MG and reactive oxygen species (ROS) during stress. Moreover, transgenic plants showed more anti-glycation activity to inhibit the formation of advanced glycation end product (AGE) that might prevent transgenic plants from stress-induced senescence. Taken together, all these observations indicate that overexpression of EcGLYIII confers salinity stress tolerance in plants and should be explored further for the generation of stress-tolerant plants.

Methylglyoxal (MG), a reactive carbonyl compound, is generated during metabolism in living systems. However, under stress, its levels increase rapidly leading to cellular toxicity. Although the generation of MG is spontaneous in a cell, its detoxification is essentially catalyzed by the glyoxalase enzymes. In plants, modulation of MG content via glyoxalases influences diverse physiological functions ranging from regulating growth and development to conferring stress tolerance. Interestingly, there has been a preferred expansion in the number of isoforms of these enzymes in plants, giving them high plasticity in their actions for accomplishing diverse roles. Future studies need to focus on unraveling the interplay of these multiple isoforms of glyoxalases possibly contributing towards the unique adaptability of plants to diverse environments.

Zinc plays a very critical role and function in all organisms. Its deficiency can cause a serious issue. In Oryza sativa, the ZRT/IRT transporter-like proteins play a role in the zinc metal uptake and transport. Few OsZIPs genes have been validated and characterized for their biological functions and most of OsZIPs are not well physiologically, biochemically and phenotypically characterized. In the current study, they analyzed for their function through subcellular localization, phylogenetic analysis, homology modeling, expression analysis, protein-protein interaction (PPI) network prediction, and prediction of their binding sites. Hierarchical clustering of OsZIP genes based on different anatomical parts and developmental stages also orthologs prediction was identified. The presence of SNPs, SSRs, ESTs, FSTs, MPSS, and SAGE tags were analyzed for useful development of markers. SNPs were identified in all OsZIPs genes and each gene was further classified based on their number and position in the 3'UTR and 5'UTR regions of the gene-specific sequences. Binding clusters and their location on the protein sequences were predicted. We found Changing in residues number and position which were due to partial overlapping and sequence alignment, but they share the same mechanism of binding and transporting Zinc. A wide range of CRISPR Cas9 gRNAs was designed based on single nucleotide polymorphism (SNP) for each OsZIP transporter gene for well-function identification and characterization with genome-wide association studies. Hence this study would provide useful information, understanding, and predicting molecular insights for the future studies that will help for improvement of nutritional quality of rice varieties. Communicated by Ramaswamy H. Sarma

Serotonin (Ser) and melatonin (Mel) serve as master regulators of plant growth and development by influencing diverse cellular processes. The enzymes namely, tryptophan decarboxylase (TDC) and tryptamine 5-hydroxylase (T5H) catalyse the formation of Ser from tryptophan. Subsequently, serotonin N-acetyl transferase (SNAT) and acetyl-serotonin methyltransferase (ASMT) form Mel from Ser. Plant genomes harbour multiple genes for each of these four enzymes, all of which have not been identified. Therefore, to delineate information regarding these four gene families, we carried out a genome-wide analysis of the genes involved in Ser and Mel biosynthesis in Arabidopsis, tomato, rice and sorghum. Phylogenetic analysis unravelled distinct evolutionary relationships among these genes from different plants. Interestingly, no gene family except ASMTs showed monocot- or dicot-specific clustering of respective proteins. Further, we observed tissue-specific, developmental and stress/hormone-mediated variations in the expression of the four gene families. The light/dark cycle also affected their expression in agreement with our quantitative reverse transcriptase-PCR (qRT-PCR) analysis. Importantly, we found that miRNAs (miR6249a and miR-1846e) regulated the expression of Ser and Mel biosynthesis under light and stress by influencing the expression of OsTDC5 and OsASMT18, respectively. Thus, this study may provide opportunities for functional characterization of suitable target genes of the Ser and Mel pathway to decipher their exact roles in plant physiology.

Plant growth promotion by microbes is a cumulative phenomenon involving multiple traits, many of which are not explored yet. Hence, to unravel microbial mechanisms underlying growth promotion, we have analyzed the genomes of two potential growth-promoting microbes, viz. Pseudomonas sp. CK-NBRI-02 (P2) and Bacillus marisflavi CK-NBRI-03 (P3) for the presence plant-beneficial traits. Besides known traits, we found that microbes differ in their ability to metabolize methylglyoxal (MG), a ubiquitous cytotoxin regarded as general consequence of stress in plants. P2 exhibited greater tolerance to MG and possessed better ability to sustain plant growth under dicarbonyl stress. Whereas under salinity, only P3 showed a dose-dependent induction in MG detoxification activity in accordance with concomitant increase in MG levels, contributing to enhanced salt tolerance. Further, salt-stressed transcriptomes of both the strains showed differences with respect to MG, ion and osmolyte homeostasis, with P3 being more responsive to stress. Importantly, application of either strain altered MG levels and subsequently MG detoxification machinery in Arabidopsis, probably to strengthen plant defense response and growth. We therefore, suggest a crucial role of microbial MG resistance in plant growth promotion and that it should be considered as a beneficial trait while screening microbes for stress mitigation in plants. This article is protected by copyright. All rights reserved.

Marker-free transgenic lines of rice are developed with enhanced trehalose accumulation that is associated with improved grain yield under salinity, sodicity, and drought stresses., Edaphic factors such as salinity, sodicity, and drought adversely affect crop productivity, either alone or in combination. Despite soil sodicity being reported as an increasing problem worldwide, limited efforts have been made to address this issue. In the present study, we aimed to generate rice with tolerance to sodicity in conjunction with tolerance to salinity and drought. Using a fusion gene from E. coli coding for trehalose-6-phosphate synthase/phosphatase (TPSP) under the control of an ABA-inducible promoter, we generated marker-free, high-yielding transgenic rice (in the IR64 background) that can tolerate high pH (~9.9), high EC (~10.0 dS m–1), and severe drought (30–35% soil moisture content). The transgenic plants retained higher relative water content (RWC), chlorophyll content, K+/Na+ ratio, stomatal conductance, and photosynthetic efficiency compared to the wild-type under these stresses. Positive correlations between trehalose overproduction and high-yield parameters were observed under drought, saline, and sodic conditions. Metabolic profiling using GC-MS indicated that overproduction of trehalose in leaves differently modulated other metabolic switches, leading to significant changes in the levels of sugars, amino acids, and organic acids in transgenic plants under control and stress conditions. Our findings reveal a novel potential technological solution to tackle multiple stresses under changing climatic conditions.

Classification of higher education institutions (HEIs) of a country allows viewing higher education as a differentiated system which respects the diversity of purposes and aspirations of different HEIs. Classification is fundamentally different from ranking, which aims to rank universities in order with higher ranked HEIs being “better” than lower ranked ones. In classification, the universities in a class are grouped by their purpose and mission, and no attempt is made to rank them. Carnegie Classification of universities in the USA is the oldest classification system, which groups universities into a few categories like Research Universities, Masters Universities, Baccalaureate Universities, and Secondary. This classification has been found extremely useful over decades for various purposes including policy making and planning. This has thus motivated similar exercises in many other countries, particularly for research universities. In this paper, we evolve an approach to classify research universities in India, based on the Carnegie Classification approach. We propose a simple basic criterion for identifying research universities, and apply it to the top 100 universities and top 100 engineering institutions in India. Using this criteria, 40 universities and 32 engineering institutions were identified as research HEIs. Based on the data on the level of research activity in these HEIs, we apply a clustering approach similar to the one Carnegie uses to group research HEIs into two sub-categories, viz. “highest research activity” and “moderate research activity”. The clustering approach identified six universities and eight engineering institutions in India to be in the highest research activity category. The level of research activity uses data on the number of full time PhD students, the number of faculty, research grants, and publications.

Glyoxalase pathway is the primary route for metabolism of methylglyoxal (MG), a toxic ubiquitous metabolite that affects redox homeostasis. It neutralizes MG using Glyoxalase I and Glyoxalase II (GLYI and GLYII) enzymes in the presence of reduced glutathione. In addition, there also exists a shorter route for the MG detoxification in the form of Glyoxalase III (GLYIII) enzymes, which can convert MG into D-lactate in a single-step without involving glutathione. GLYIII proteins in different systems demonstrate diverse functional capacities and play a vital role in oxidative stress response. To gain insight into their evolutionary patterns, here we studied the evolution of GLYIII enzymes across prokaryotes and eukaryotes, with special emphasis on plants. GLYIII proteins are characterized by the presence of DJ-1_PfpI domains thereby, belonging to the DJ-1_PfpI protein superfamily. Our analysis delineated evolution of double DJ-1_PfpI domains in plant GLYIII. Based on sequence and structural characteristics, plant GLYIII enzymes could be categorized into three different clusters, which followed different evolutionary trajectories. Importantly, GLYIII proteins from monocots and dicots group separately in each cluster and the each of the two domains of these proteins also cluster differentially. Overall, our findings suggested that GLYIII proteins have undergone significant evolutionary changes in plants, which is likely to confer diversity and flexibility in their functions.

Methylglyoxal (MG) is ubiquitously produced in all living organisms as a byproduct of glycolysis, higher levels of which are cytotoxic, leading to oxidative stress and apoptosis in the living systems. Though its generation is spontaneous but its detoxification involves glyoxalase pathway genes. Based on this understanding, the present study describes the possible role of MG as a novel non-antibiotic-based selection agent in rice. Further, by metabolizing MG, the glyoxalase pathway genes viz. glyoxalase I (GLYI) and glyoxalase II (GLYII), may serve as selection markers. Therefore, herein, transgenic rice harboring GLYI-GLYII genes (as selection markers) were developed and the effect of MG as a selection agent was assessed. The 3 mM MG concentration was observed as optimum for the selection of transformed calli, allowing efficient callus induction and proliferation along with high regeneration frequency (55 ± 2%) of the transgenic calli. Since the transformed calli exhibited constitutively higher activity of GLYI and GLYII enzymes compared to the wild type calli, the rise in MG levels was restricted even upon exogenous addition of MG during the selection process, resulting in efficient selection of the transformed calli. Therefore, MG-based selection method is a useful and efficient system for selection of transformed plants without significantly compromising the transformation efficiency. Further, this MG-based selection system is bio-safe and can pave way towards better public acceptance of transgenic plants.

Rice plants often encounter high temperature stress, but the associated coping strategies are poorly understood. It is known that a prior shorter exposure to high temperature, called thermo-priming, generally results in better adaptation of the plants to subsequent exposure to high temperature stress. High throughput sequencing of transcript and small RNA libraries of rice seedlings primed with short exposure to high temperature followed by high temperature stress and from plants exposed to high temperature without priming was performed. This identified a number of transcripts and microRNAs (miRs) that are induced or down regulated. Among them osa-miR531b, osa-miR5149, osa-miR168a-5p, osa-miR1846d-5p, osa-miR5077, osa-miR156b-3p, osa-miR167e-3p and their respective targets, coding for heat shock activators and repressors, showed differential expression between primed and non-primed plants. These findings were further validated by qRT-PCR. The results indicate that the miR-regulated heat shock proteins (HSPs)/heat shock transcription factors (HSFs) may serve as important regulatory nodes which are induced during thermo-priming for plant survival and development under high temperatures.

Environmental factors play a significant role in controlling growth, development and defense responses of plants. Changes in the abiotic environment not only significantly alter the physiological and molecular pathways in plants, but also result in attracting the insect pests that carry a payload of viruses. Invasion of plants by viruses triggers the RNA silencing based defense mechanism in plants. In counter defense the viruses have gained the ability to suppress the host RNA silencing activities. A new paradigm has emerged, with the recognition that plant viruses also have the intrinsic capacity to modulate host plant response to environmental cues, in an attempt to favour their own survival. Thus, plant-virus interactions provide an excellent system to understand the signals in crosstalk between biotic (virus) and abiotic stresses. In this review, we have summarized the basal plant defense responses to pathogen invasion while emphasizing on the role of RNA silencing as a front line of defense response to virus infection. The emerging knowledge indicates overlap between RNA silencing with the innate immune responses during antiviral defense. The suppressors of RNA silencing serve as Avr proteins, which can be recognized by the host R proteins. The defense signals also function in concert with the phytohormones to influence plant responses to abiotic stresses. The current evidence on the role of virus induced host tolerance to abiotic stresses is also discussed.

Prions can be considered as molecular memory devices, generating reproducible memory of a conformational change. Prion-like proteins (PrLPs) have been demonstrated to be present in plants, but their role in plant stress and memory remains largely unexplored. In this work, we report the widespread presence of PrLPs in plants through a comprehensive analysis of 39 genomes representing major taxonomic groups. We find diverse functional roles associated with plant ‘prionomes’. Investigation of the rice transcriptome further delineated the role of PrLPs in stress and developmental responses, leading us to explore whether and to what extent PrLPs may build stress memory. The rice prionome is significantly enriched for Transposons/Retrotransposons (Ts/RTRs), and we derived transcriptional regulatory inferences from diurnal gene expression revealing a complex regulatory network between PrLPs, transcription factors and genes known to be involved in stress priming, as well as transient and trans-generational plant memory. Overall, our data suggest that plant memory mechanisms may rely upon protein-based signals embedded in PrLPs, in addition to chromatin-based epigenetic signals and provides important insights into the anticipated role of prions in stress and memory.

A plant&rsquo, s response to stress conditions is governed by intricately coordinated gene expression. The microRNAs (miRs) have emerged as relatively new players in the genetic network, regulating gene expression at the transcriptional and post-transcriptional level. In this study, we performed comprehensive profiling of miRs in roots of the naturally salt-tolerant Pokkali rice variety to understand their role in regulating plant physiology in the presence of salt. For comparisons, root miR profiles of the salt-sensitive rice variety Pusa Basmati were generated. It was seen that the expression levels of 65 miRs were similar for roots of Pokkali grown in the absence of salt (PKNR) and Pusa Basmati grown in the presence of salt (PBSR). The salt-induced dis-regulations in expression profiles of miRs showed controlled changes in the roots of Pokkali (PKSR) as compared to larger variations seen in the roots of Pusa Basmati. Target analysis of salt-deregulated miRs identified key transcription factors, ion-transporters, and signaling molecules that act to maintain cellular Ca2+ homeostasis and limit ROS production. These miR:mRNA nodes were mapped to the Quantitative trait loci (QTLs) to identify the correlated root traits for understanding their significance in plant physiology. The results obtained indicate that the adaptability of Pokkali to excess salt may be due to the genetic regulation of different cellular components by a variety of miRs.

Here, we report the 4.34-Mb draft genome assembly of Bacillus marisflavi CK-NBRI-03 (or P3), a Gram-positive bacterium, with an average G+C content of 48.66%. P3 was isolated from agricultural soil from the Badaun (midwestern plain zone) region of Uttar Pradesh, India.

Background The glyoxalase pathway is evolutionarily conserved and involved in the glutathione-dependent detoxification of methylglyoxal (MG), a cytotoxic by-product of glycolysis. It acts via two metallo-enzymes, glyoxalase I (GLYI) and glyoxalase II (GLYII), to convert MG into D-lactate, which is further metabolized to pyruvate by D-lactate dehydrogenases (D-LDH). Since D-lactate formation occurs solely by the action of glyoxalase enzymes, its metabolism may be considered as the ultimate step of MG detoxification. By maintaining steady state levels of MG and other reactive dicarbonyl compounds, the glyoxalase pathway serves as an important line of defence against glycation and oxidative stress in living organisms. Therefore, considering the general role of glyoxalases in stress adaptation and the ability of Sorghum bicolor to withstand prolonged drought, the sorghum glyoxalase pathway warrants an in-depth investigation with regard to the presence, regulation and distribution of glyoxalase and D-LDH genes. Result Through this study, we have identified 15 GLYI and 6 GLYII genes in sorghum. In addition, 4 D-LDH genes were also identified, forming the first ever report on genome-wide identification of any plant D-LDH family. Our in silico analysis indicates homology of putatively active SbGLYI, SbGLYII and SbDLDH proteins to several functionally characterised glyoxalases and D-LDHs from Arabidopsis and rice. Further, these three gene families exhibit development and tissue-specific variations in their expression patterns. Importantly, we could predict the distribution of putatively active SbGLYI, SbGLYII and SbDLDH proteins in at least four different sub-cellular compartments namely, cytoplasm, chloroplast, nucleus and mitochondria. Most of the members of the sorghum glyoxalase and D-LDH gene families are indeed found to be highly stress responsive. Conclusion This study emphasizes the role of glyoxalases as well as that of D-LDH in the complete detoxification of MG in sorghum. In particular, we propose that D-LDH which metabolizes the specific end product of glyoxalases pathway is essential for complete MG detoxification. By proposing a cellular model for detoxification of MG via glyoxalase pathway in sorghum, we suggest that different sub-cellular organelles are actively involved in MG metabolism in plants.

Light and temperature are two major environmental factors influencing plant growth and development. High temperature caused by increasing global warming and greenhouse gases has become a worldwide concern. In plants, it triggers various morphological, biochemical and physiological changes that adversely influence growth and development leading to substantial decrease in crop yield. The declining food production and the requirements of the rapidly growing population pose further threat to global food security. Plants have elaborate genetic and epigenetic regulatory mechanisms to endure through the unfavorable abiotic conditions. The induction of heat stress response at the genetic level involves de-regulation of transcription factors and chaperones which play an indispensable role in acclimation to heat. The epigenetic regulation includes induction or repression of specific microRNAs (miRs) that target a variety of transcripts. This is a signature response by plants that is observed in almost all stresses such as high light intensities, drought and salinity. Since earlier reports have indicated a strong relationship between light and temperature signaling pathways in plants, so their integration in influencing the miR based regulatory networks needs to be elucidated. Here we present the mirador (window designed to command an extensive outlook) on the potential role of miRs in regulating plant growth and development in synergistic response to light and heat networks.

Crop plants face a multitude of diverse abiotic and biotic stresses in the farmers' fields. Although there now exists a considerable knowledge of the underlying mechanisms of response to individual stresses, the crosstalk between response pathways to various abiotic and biotic stresses remains enigmatic. Here, we investigated if the cytotoxic metabolite methylglyoxal (MG), excess of which is generated as a common consequence of many abiotic and biotic stresses, may serve as a key molecule linking responses to diverse stresses. For this, we generated transgenic rice plants overexpressing the entire two-step glyoxalase pathway for MG detoxification. Through assessment of various morphological, physiological and agronomic parameters, we found that glyoxalase-overexpression imparts tolerance towards abiotic stresses like salinity, drought and heat and also provides resistance towards damage caused by the sheath blight fungus (Rhizoctonia solani) toxin phenylacetic acid. We show that the mechanism of observed tolerance of the glyoxalase-overexpressing plants towards these diverse abiotic and biotic stresses involves improved MG detoxification and reduced oxidative damage leading to better protection of chloroplast and mitochondrial ultrastructure and maintained photosynthetic efficiency under stress conditions. Together, our findings indicate that MG may serve as a key link between abiotic and biotic stress response in plants.

Agricultural production worldwide has been severely impacted by cold and freezing stresses. Plant capacity to acclimate to environmental conditions in their immediate vicinity largely control their survival, growth, and productivity. Molecular as well as biochemical mechanisms underpinning plant cold acclimation are very complex and interwoven. The cold-impacted plants try to modulate expression of variety genes controlling cell membrane lipid composition, mitogen-activated protein kinase cascade, total soluble proteins, polyamines, glycinebetaine, proline, reactive oxygen species (ROS) scavengers, cryoprotectants, and a large number of cold responsive factors. To this end, this paper dissects the array of transcriptional factors/genes down- or up-regulated, their identification in different plant species, recognition of cold tolerant/resistant transgenic plants, complexity of the mitogen-activated protein kinase cascade, as well as their cross talk under different stresses and molecular mechanisms. Furthermore, it also comprehensively elucidates physio-biochemical interferences in cold acclimation with a particular emphasis on endogenous content as well as exogenously supplied different types of polyamines, ROS, and osmoprotectants. Overall, low temperature stress tolerance or cold acclimation varies greatly among species depending on the stress intensity and duration and type of plant species.

Pseudomonas sp. strain CK-NBRI-02 is a potential plant growth-promoting Gram-negative rhizobacterium isolated from the rhizosphere of maize plants growing in fields in Srinagar, Jammu, and Kashmir, India. Here, we report a 5.25-Mb draft assembly of the genome sequence of Pseudomonas sp. strain CK-NBRI-02 with an average G+C content of 62.47%.

Ancestors of modern land plants evolved in aquatic environments, with the first land plants appearing around 470–700 million years ago. Terrestrial colonization has been credited to a series of major revolutions in the body plan, anatomy and biochemistry of plants which is required for their survival and reproduction. Plant adaptations to life on land encompassed development of many specialized structures such as water-repellent cuticles, stomata for regulating water evaporation, structures for collecting sunlight, a vascular transport system and many more. In addition, intricate signalling mechanisms regulated by hormones for the perception of the environment have also come into place in higher plants. How these features have evolved in modern-day plants and how these have contributed to diversity are fascinating. In this chapter, we aim to shed light on a few interesting facets of plant functions with a bearing on evolution, which have not only contributed to their establishment on land but also allowed their enormous expansion leading to huge diversity. We believe that plants have a remarkable ability to adapt themselves in the ever-changing environments, despite being rooted to ground.

From the previous chapters in this volume, it is evident that plants have developed very subtle molecular mechanisms to perceive ever-changing environment and respond accordingly to ensure their proper development as engraved in their genome. Plants, thus able to sense and adaptively reciprocate to extraneous signals, anticipate inevitable threats and stresses via the elaborate intercellular systems especially the receptors, microtubules, organ-to-organ communications as well as communicating with both allies and enemies. The appropriate response by plants is needed not only for their own survival but also for reproduction, developing seeds and their dispersal for the continuation of the progeny. There are reports that some form of plant “memory” is used for rapid adaptability of plants to stress and strengthen their defence mechanisms. Exploration of this emerging avenue of research in plant sensory biology is becoming more ascribable with avant-garde breakthroughs via omics approach, high-throughput sequencing technologies and time-lapse as well as Kirlian photography. These new technological interventions would ensure unprecedented deciphering of the secret new-fangled mysteries of the plant world. In many ways, the sensory behaviour of plants seems to be similar to that noticed in the animal world. The question that we have also tried to discuss in this chapter is whether this “intelligent” response of plants falls into the domain of awareness or consciousness as has been proposed by some authors.

Methylglyoxal (MG) is a potent cytotoxin being produced primarily as result of non-enzymatic reactions in the living systems. The toxicity of MG is believed to be due to its ability to carry out glycation of proteins, nucleic acids and phospholipids. MG can readily modify proteins and nucleic acids, forming advanced glycation end-products which results in cellular dysfunction. In order to limit MG levels, several detoxification enzymes exist in the living systems which can metabolize MG. Of these, glyoxalase pathway is considered to be the primary route of MG metabolism. Here, we have discussed the effects of MG on the functioning of different subcellular compartments and the detoxification machineries existing in these organelles to counter excess MG. We describe how MG exerts its toxic effects at the cellular level and how the cell defends itself from MG. In this context, we have highlighted the important role of glyoxalase pathway in plants. Further, we have also discussed how glycolytic products and enzymes control MG levels and how MG regulates the activity of these glycolytic products as a part of substrate and end-product regulatory mechanisms. Overall, we present a comprehensive overview of MG toxicity and detoxification in the cell and its regulation.

In this paper we provide evidence for involvement of chloroplast as alternate organelle for initiating PCD in plants under light and abiotic stress. In animals, mitochondria are the major source of reactive oxygen species (ROS) and key executioner of programmed cell death (PCD). In plants, however, the primary site of generation of ROS is chloroplast and yet its involvement in PCD has not been worked out in details. We found by Evans blue staining that salt (150 mM NaCl)-treated protoplasts obtained from green seedlings had higher rate of cell death than protoplasts obtained from etiolated seedlings. This indicated that cell death induced by NaCl is accentuated by light. Imposition of salt-stress to green protoplasts generated H2O2. Known hallmarks of PCD i.e., blebbing of cell membrane, loabing in nucleus, nick in DNA were observed in light-exposed salt-treated protoplasts and seedlings. TUNEL-FACS assay demonstrate several DNA nicks in the salt-treated green protoplasts exposed to light. Conversely, salt-treated etiolated protoplasts kept in dark had only a few TUNEL-positive nuclei. Similarly, a substantial numbers of TUNEL positive nuclei were observed in green seedlings due to salt treatment in light. However, salt-treated etiolated seedlings kept in dark had very few TUNEL positive nuclei. Addition of Caspase 3 inhibitor (DAVD-CHO) rescued (~50 %) green protoplasts from salt-stress induced cell death suggesting an involvement of apoptosis like PCD (AL-PCD). Ultra structure studies of chloroplast, mitochondria and nucleus from the leaves obtained from salt treated seedlings at the time point that showed PCD signature, resulted to severe granal de-stacking in chloroplasts while structural integrity of mitochondria was maintained. These studies demonstrate the photo-modulation of salinity-induced PCD in photosynthetic tissues is mainly executed by chloroplasts.

Cystathionine β-synthase (CBS) domains have been identified in a wide range of proteins of unrelated functions such as, metabolic enzymes, kinases and channels, and usually occur as tandem re-peats, often in combination with other domains. In plants, CBS Domain-Containing Proteins (CDCPs) form a multi-gene family and only a few are so far been reported to have a role in development via regu-lation of thioredoxin system as well as in abiotic and biotic stress response. However, the function of majority of CDCPs still remains to be elucidated in plants. Here, we report the cloning, characterization and functional validation of a CBS domain containing protein, OsCBSCBSPB4 from rice, which pos-sesses two CBS domains and one PB1 domain. We show that OsCBSCBSPB4 encodes a nucleo-cytoplasmic protein whose expression is induced in response to various abiotic stress conditions in salt-sensitive IR64 and salt-tolerant Pokkali rice cultivars. Further, heterologous expression of OsCBSCB-SPB4 in E. coli and tobacco confers marked tolerance against various abiotic stresses. Transgenic tobac-co seedlings over-expressing OsCBSCBSPB4 were found to exhibit better growth in terms of delayed leaf senescence, profuse root growth and increased biomass in contrast to the wild-type seedlings when subjected to salinity, dehydration, oxidative and extreme temperature treatments. Yeast-two hybrid stud-ies revealed that OsCBSCBSPB4 interacts with various proteins. Of these, some are known to be in-volved in abiotic stress tolerance. Our results suggest that OsCBSCBSPB4 is involved in abiotic stress response and is a potential candidate for raising multiple abiotic stress tolerant plants.

Soil salinity is one of the most devastating factors threatening cultivable land. While low to moderate salt stress affects the plant growth rate and yields, the high-salt stress conditions are detrimental for plant growth. The problem of salinity is often compounded by mineral deficiencies and toxicities. Rice is essentially rated as salt-sensitive crop and its salt susceptibility is dependent on the growth stage. The genetic processes that are involved in coordinating the responses to salinity in rice are not very clearly understood, but can be somewhat explained by mechanisms that regulate ion homeostasis, osmolyte production, scavenging of toxic radicals, water conduction and root–shoot response coordination. This is achieved by modifying the gene expression. The microRNAs (miRs) represent an important class of endogenous small RNAs that have the potential to regulate gene expression at post-transcriptional levels. This review discusses the impact of salinity on rice crop and the current knowledge in the area of miRs with their role in the adaptive response to salt and other abiotic stresses, with a focus on rice.

Anhydrobiosis is an adaptive strategy of certain organisms or specialised propagules to survive in the absence of water while programmed cell death (PCD) is a finely tuned cellular process of the selective elimination of targeted cell during developmental programme and perturbed biotic and abiotic conditions. Particularly during water stress both the strategies serve single purpose i.e., survival indicating PCD may also function as an adaptive process under certain conditions. During stress conditions PCD cause targeted cells death in order to keep the homeostatic balance required for the organism survival, whereas anhydrobiosis suspends cellular metabolic functions mimicking a state similar to death until reestablishment of the favourable conditions. Anhydrobiosis is commonly observed among organisms that have ability to revive their metabolism on rehydration after removal of all or almost all cellular water without damage. This feature is widely represented in terrestrial cyanobacteria and bryophytes where it is very common in both vegetative and reproductive stages of life-cycle. In the course of evolution, with the development of advanced vascular system in higher plants, anhydrobiosis was gradually lost from the vegetative phase of life-cycle. Though it is retained in resurrection plants that primarily belong to thallophytes and a small group of vascular angiosperm, it can be mostly found restricted in orthodox seeds of higher plants. On the contrary, PCD is a common process in all eukaryotes from unicellular to multicellular organisms including higher plants and mammals. In this review we discuss physiological and biochemical commonalities and differences between anhydrobiosis and PCD.

Crop plants face a multitude of diverse abiotic and biotic stresses in the farmers' fields. Although there now exists a considerable knowledge of the underlying mechanisms of response to individual stresses, the crosstalk between response pathways to various abiotic and biotic stresses remains enigmatic. Here, we investigated if the cytotoxic metabolite methylglyoxal (MG), excess of which is generated as a common consequence of many abiotic and biotic stresses, may serve as a key molecule linking responses to diverse stresses. For this, we generated transgenic rice plants overexpressing the entire two-step glyoxalase pathway for MG detoxification. Through assessment of various morphological, physiological and agronomic parameters, we found that glyoxalase-overexpression imparts tolerance towards abiotic stresses like salinity, drought and heat and also provides resistance towards damage caused by the sheath blight fungus (Rhizoctonia solani) toxin phenylacetic acid. We show that the mechanism of observed tolerance of the glyoxalase-overexpressing plants towards these diverse abiotic and biotic stresses involves improved MG detoxification and reduced oxidative damage leading to better protection of chloroplast and mitochondrial ultrastructure and maintained photosynthetic efficiency under stress conditions. Together, our findings indicate that MG may serve as a key link between abiotic and biotic stress response in plants.

Oxidative stress is one of the crucial outcomes of biotic and abiotic stress which leads to the physiological and metabolic alterations in plant system and, therefore, requires a balanced control of reactive oxygen species (ROS) production and its scavenging through antioxidative enzymes and proteins. The enzymatic components of antioxidative defense system consist of several enzymes including catalase (CAT), superoxide dismutase (SOD), and guaiacol peroxidase (GPX) and also the enzymes of ascorbate-glutathione (AsA-GSH) cycle, i.e., ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR), and glutathione reductase (GR). The nonenzymatic components include ascorbate (AsA) and glutathione (GSH) along with carotenoids and tocopherols along with other phenolic compounds. Exploiting the antioxidative behavior of these enzymes, several plants have been modified using transgenic techniques overexpressing the components of antioxidative stress pathways. Moreover, there are several other redox proteins, which have been genetically engineered to help plant survive in adverse conditions. This suggests that the development of transgenic plants overexpressing enzymes and redox-sensitive proteins associated with oxidative stress and antioxidative stress pathways will surely provide an important link of their role in tolerance to oxidative damage in crops. This chapter elucidates the recent advances in the defense system of plants during oxidative stress and also discusses the potential strategies for enhancing tolerance to oxidative stress.

Glyoxalases are known to play a very important role in abiotic stress tolerance. This two-step pathway detoxifies ubiquitously present cytotoxic metabolite methylglyoxal, which otherwise increases to lethal concentrations under various stress conditions. Methylglyoxal initiates stress-induced signaling cascade via reactive oxygen species, resulting in the modifications of proteins involved in various signal transduction pathways, that eventually culminates in cell death or growth arrest. The associated mechanism of tolerance conferred by over-expression of methylglyoxal-detoxifying glyoxalase pathway mainly involves lowering of methylglyoxal levels, thereby reducing subsequently induced cellular toxicity. Apart from abiotic stresses, expression of glyoxalases is affected by a wide variety of other stimuli such as biotic, chemical and hormonal treatments. Additionally, alterations in cellular milieu during plant growth and development also affect expression of glyoxalases. The multiple stress-inducible nat...

†Both authors contributed equally to this work. SUMMARY The glyoxalase system constitutes the major pathway for the detoxification of metabolically produced cytotoxin methylglyoxal (MG) into a non-toxic metabolite D-lactate. Glyoxalase I (GLY I) is an evolutionarily conserved metalloenzyme requiring divalent metal ions for its activity: Zn 2+ in the case of eukaryotes or Ni 2+ for enzymes of prokaryotic origin. Plant GLY I proteins are part of a multimember family; however, not much is known about their physiological function, structure and metal dependency. In this study, we report a unique GLY I (OsGLYI-11.2) from Oryza sativa (rice) that requires Ni 2+ for its activity. Its biochemical, structural and functional characterization revealed it to be a monomeric enzyme, possessing a single Ni 2+ coordination site despite containing two GLY I domains. The requirement of Ni 2+ as a cofactor by an enzyme involved in cellular detoxification suggests an essential role for this otherwise toxic heavy metal in the stress response. Intriguingly, the expression of OsGLYI-11.2 was found to be highly substrate inducible, suggesting an important mode of regulation for its cellular levels. Heterologous expression of OsGLYI-11.2 in Escherichia coli and model plant Nicotiana tabacum (tobacco) resulted in improved adaptation to various abiotic stresses caused by increased scavenging of MG, lower Na + /K + ratio and maintenance of reduced glutathione levels. Together, our results suggest interesting links between MG cellular levels, its detoxification by GLY I, and Ni 2+ – the heavy metal cofactor of OsGLYI-11.2, in relation to stress response and adaptation in plants.

The glyoxalase pathway is required for detoxification of cytotoxic metabolite MG (methylglyoxal) that would otherwise increase to lethal concentrations under adverse environmental conditions. Since its discovery 100 years ago, several roles have been assigned to glyoxalases, but, in plants, their involvement in stress response and tolerance is the most widely accepted role. The plant glyoxalases have emerged as multigene family and this expansion is considered to be important from the perspective of maintaining a robust defence machinery in these sessile species. Glyoxalases are known to be differentially regulated under stress conditions and their overexpression in plants confers tolerance to multiple abiotic stresses. In the present article, we review the importance of glyoxalases in plants, discussing possible roles with emphasis on involvement of the glyoxalase pathway in plant stress tolerance.

ETHYLMALONIC ENCEPHALOPATHY PROTEIN 1 (ETHE1) encodes sulfur dioxygenase (SDO) activity regulating sulfide levels in living organisms. It is an essential gene and mutations in ETHE1 leads to ethylmalonic encephalopathy (EE) in humans and embryo lethality in Arabidopsis. At present, very little is known regarding the role of ETHE1 beyond the context of EE and almost nothing is known about factors affecting its regulation in plant systems. In this study, we have identified, cloned and characterized OsETHE1, a gene encoding ETHE1-like protein from Oryza sativa. ETHE1 proteins in general are most similar to glyoxalase II (GLYII) and hence OsETHE1 has been earlier annotated as OsGLYII1, a putative GLYII gene. Here we show that OsETHE1 lacks GLYII activity and is instead an ETHE1 homolog being localized in mitochondria like its human and Arabidopsis counterparts. We have isolated and analyzed 1618 bp OsETHE1 promoter (pOsETHE1) to examine the factors affecting OsETHE1 expression. For this, transcriptional promoter pOsETHE1: 5-bromo-5-chloro-3-indolyl-β-D-glucuronide (GUS) fusion construct was made and stably transformed into rice. GUS expression pattern of transgenic pOsETHE1:GUS plants reveal a high root-specific expression of OsETHE1. The pOsETHE1 activity was stimulated by Ca(II) and required light for induction. Moreover, pOsETHE1 activity was induced under various abiotic stresses such as heat, salinity and oxidative stress, suggesting a potential role of OsETHE1 in stress response.

The topoisomerase II expression varies as a function of cell proliferation. Maximal topoisomerase II expression was tightly coupled to S phase and G2/M phase via both transcriptional and post-transcriptional regulation. Investigation in meiosis using pollen mother cells also revealed that it is not the major component of meiotic chromosomes, it seems to diffuse out once meiotic chromosomal condensation is completed. Synchronized tobacco BY-2 cell cultures were used to study the role of topoisomerase II in various stages of the cell cycle. Topoisomerase II transcript accumulation was observed during the S- and G2/M- phase of cell cycle. This biphasic expression pattern indicates the active requirement of topoisomerase II during these stages of the cell cycle. Through immuno-localization of topoisomerase II was observed diffusely throughout the nucleoplasm in interphase nuclei, whereas, the nucleolus region exhibited a more prominent immuno-positive staining that correlated with rRNA transcription, as shown by propidium iodide staining and BrUTP incorporation. The immuno-staining analysis also showed that topoisomerase II is the major component of mitotic chromosomes and remain attached to the chromosomes during cell division. The inhibition of topoisomerase II activity using specific inhibitors revealed quite dramatic effect on condensation of chromatin and chromosome individualization from prophase to metaphase transition. Partially condensed chromosomes were not arranged on metaphase plate and chromosomal perturbations were observed when advance to anaphase, suggesting the importance of topoisomerase II activity for proper chromosome condensation and segregation during mitosis. Contrary, topoisomerase II is not the major component of meiotic chromosomes, even though mitosis and meiosis share many processes, including the DNA replication, chromosome condensation and precisely regulated partitioning of chromosomes into daughter cells. Even if topoisomerase II is required for individualization and condensation of meiotic chromosomes, it seems to diffuse out once meiotic chromosomal condensation is completed.

Several ribosomal proteins are down-regulated at the translational, post-transcriptional and transcriptional level in response to abiotic stress, resulting in retardation of growth and productivity in various plants. However, transcriptional mechanisms associated with the stress-mediated down-regulation of such genes have not been well studied. Recently, we reported salt-responsive transcriptional down-regulation of a gene encoding 60S ribosomal protein L32, rpL32_8.1, in rice. In the present work, chromatin remodeling mechanisms associated with down-regulation of this gene under salt stress have been studied. The data obtained from the sodium bisulfite sequencing of a genomic region containing portions of the promoter and 5'-untranslated region of rpL32_8.1 exclude a role for DNA methylation in this down-regulation event. Fine mapping of the first nucleosome after the transcription start site (TSS) revealed its occupancy over the TSS to a greater extent under salt stress than that observed in a controlled environment, possibly causing a hindrance to transcription initiation during its down-regulation. Using chromatin immunoprecipitation, it was observed that histone modifications that are generally associated with highly active genes were markedly reduced under stress conditions at the 5' end region of this gene under salt stress. By applying a modified chromatin conformation capture (mCCC) technique, promoter-terminator interaction was detected for this gene under control conditions. The efficiency of this interaction was diminished under salt stress. This work indicates that the stress-responsive transcriptional down-regulation of rpL32_8.1 is accompanied by alterations in nucleosome positioning, histone modification and gene looping, but not DNA methylation.