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The basic helix-loop-helix (bHLH) is the second-largest TF family in plants that play important roles in plant growth, development, and stress responses. In this study, a total of 100 bHLHs were identified using Hidden Markov Model profiles in the Nicotiana tabacum genome, clustered into 15 major groups (I–XV) based on their conserved domains and phylogenetic relationships. Group VIII genes were found to be the most abundant, with 27 NtbHLH members. The expansion of NtbHLHs in the genome was due to segmental and tandem duplication. The purifying selection was found to have an important role in the evolution of NtHLHs. Subsequent qRT-PCR validation of five selected genes from transcriptome data revealed that NtbHLH3.1, NtbHLH3.2, NtbHLH24, NtbHLH50, and NtbHLH59.2 have higher expressions at 12 and 24 h in comparison to 0 h (control) of chilling stress. The validated results demonstrated that NtbHLH3.2 and NtbHLH24 genes have 3 and fivefold higher expression at 12 h and 2 and threefold higher expression at 24 h than control plant, shows high sensitivity towards chilling stress. Moreover, the co-expression of positively correlated genes of NtbHLH3.2 and NtbHLH24 confirmed their functional significance in chilling stress response. Therefore, suggesting the importance of NtbHLH3.2 and NtbHLH24 genes in exerting control over the chilling stress responses in tobacco.

Solanum viarum serves as a raw material for the steroidal drug industry due to its alkaloid and glycoalkaloid content. Elicitation is well-known for measuring the increase in the yield of bioactive compounds in in vitro cultures. The current study was performed for the accumulation of metabolites viz. solasodine, solanidine, and α-solanine in S. viarum culture using microbial-based elicitors added in 1%, 3%, 5%, and 7% on 25

Zinc (Zn) is a vital micronutrient in both plants and humans for healthy growth and development. The lesser Zn accessibility causes about 20% yield loss along with low Zn content in grains. About 30% of the human population in the world rely on Zn deficient diets. Dietry Zn deficiency causes impairment of physical growth, immune system functioning, reproductive health, and neurobehavioral development in humans. In various physiological processes, Zn plays a key role and serves as a cofactor for various enzymes and proteins in numerous essential biochemical pathways in both plant and animal. Consequently, it is important to increase Zn content of cereal grains such as rice, maize and wheat. Many investigations have been accomplished to improve Zn deficiency tolerance and improving Zn content in grains. In this regard, improving Zn use efficiency is the most meaningful approach that involves modifying root system architecture, solubilization of Zn complex by organic acids, root exudates, and Zn uptake and translocation mechanism in plants. Here we present an outlook of different biotechnological approaches to improve Zn use efficiency and producing cereals with superior grain quality.

One of the most interesting signaling molecules that regulates a wide array of adaptive stress responses in plants are the micro RNAs (miRNAs) that are a unique class of non-coding RNAs constituting novel mechanisms of post-transcriptional gene regulation. Recent studies revealed the role of miRNAs in several biotic and abiotic stresses by regulating various phytohormone signaling pathways as well as by targeting a number of transcription factors (TFs) and defense related genes. Phytohormones are signal molecules modulating the plant growth and developmental processes by regulating gene expression. Studies concerning miRNAs in abiotic stress response also show their vital roles in abiotic stress signaling. Current research indicates that miRNAs may act as possible candidates to create abiotic stress tolerant crop plants by genetic engineering. Yet, the detailed mechanism governing the dynamic expression networks of miRNAs in response to stress tolerance remains unclear. In this review, we provide recent updates on miRNA-mediated regulation of phytohormones combating various stress and its role in adaptive stress response in crop plants.

Soil salinity is one of the critical issue worldwide that adversely affect soil fertility. Salt stress significantly limits crop yield and grain quality; therefore, there is an urgent need to develop a strategy to improve salt stress tolerance. In present study, we reported that rice glutaredoxin (OsGrx_C7) plays a positive response in salt induced stress. Gene expression analysis, silencing, and overexpression of OsGrx_C7 gene were used to discover the role of OsGrx_C7 in response to salt stress. Gene expression analysis suggested that OsGrx_C7 expression was induced under salt stress and ubiquitously expressed in rice including root and shoot. The silencing of osgrx_c7 gene leads to increased sensitivity to salt stress, indicating its importance in salt stress tolerance. A gain-of-function approach showed that OsGrx_C7 may act as an important determinant in salt stress, compared with WT, and revealed higher biomass accumulation, improved root and plant growth under salt stress. Under salt stress condition, OsGrx_C7 overexpressing rice plants showed lower level of lipid peroxidation and Na+/K+ ratio, while proline accumulation, soluble sugar content and GSH/GSSG ratio was higher compared to WT. Furthermore, expression analysis suggested that OsGrx_C7 acted as positive regulator of salt tolerance by reinforcing the expression of transporters (OsHKT2;1, OsHKT1;5 and OsSOS1) engaged in Na+ homeostasis in overexpressing plants. Overall our study revealed that OsGrx_C7 emerged as a key mediator in response to salt stress in rice and could be used for engineering tolerance against salt stress in rice and other crops.

Plant responds to Agrobacterium via three-layered immunity that determines its susceptibility or resistance to Agrobacterium infection. Agrobacterium tumefaciens is a soil-borne Gram-negative bacterium that causes crown gall disease in plants. The remarkable feat of interkingdom gene transfer has been extensively utilised in plant biotechnology to transform plant as well as non-host systems. In the past two decades, the molecular mode of the pathogenesis of A. tumefaciens has been extensively studied. Agrobacterium has also been utilised as a premier model to understand the defence response of plants during plant-Agrobacterium interaction. Nonetheless, the threat of Agrobacterium-mediated crown gall disease persists and is associated with a huge loss of plant vigour in agriculture. Understanding the molecular dialogues between these two interkingdom species might provide a cure for crown gall disease. Plants respond to A. tumefaciens by mounting a three-layered immune response, which is manipulated by Agrobacterium via its virulence effector proteins. Comparative studies on plant defence proteins versus the counter-defence of Agrobacterium have shed light on plant susceptibility and tolerance. It is possible to manipulate a plant's immune system to overcome the crown gall disease and increase its competence via A. tumefaciens-mediated transformation. This review summarises the recent advances in the molecular mode of Agrobacterium pathogenesis as well as the three-layered immune response of plants against Agrobacterium infection.

Excessive levels of heavy metals (HMs) in agricultural soil is a critical concerns for crop production and food safety and pose potential hazards to human and animal health. Anthropogenic sources including agriculture, mining, smelting, electroplating, and other industrial activities have resulted in the deposition of undesirable concentration of metals, such as arsenic (As), cadmium (Cd), chromium (Cr), and lead (Pb) in the soil. Unlike many other pollutants, HMs are difficult to remove from the environment as they cannot be degraded by any method, and are ultimately indestructible. The use of microorganisms and plants for soil remediation of HMs are of great interest because of their high efficiency, ease of use, and cost-effective application. Microorganisms can be used to remediate contaminated soil by detoxification, sequestration, and solubilization of HMs to facilitate their extraction. These microbes may act on HMs by chelation, precipitation, transformation (oxidation-reduction, methylation), biosorption, and accumulation. However, high concentrations of HMs in soil lead to decreased number of soil microbes. These symbiotic rhizospheric microbes depend on plant root exudates for their nutrition, thus to improve the number of microbes, it is also essential to optimize microbial nutrition by optimizing plant-microbe interaction. Different approaches were adopted to address these problems, from enrichment with rhizosphere bacterial consortia resistant to HMs to genetic engineering of plant growth-promoting symbiotic microbes. These genetically engineered (GE) microbes show improved bioremediation potential by enhancing their metal metabolism efficiency as well as increasing soil fertility. In this review, we describe how GE microbes and their association with plants enhance metal tolerance, accumulation, and detoxification in microbes and plants. We also describe the potential of bioremediation using symbiosis between plants and GE microbes.

The adverse effects of arsenic (As) contamination are well known. Rice is a staple food for 50% of the world population but the accumulation of As into rice hampers the food security and safety. Thus the amelioration of As stress and reduction of As levels in rice are needed. In this study, transcriptome (Illumina sequencing) and proteome (2D gel electrophoresis) explored mechanisms of consortium (P. putida+C. vulgaris) mediated growth promotion and As amelioration in rice. The rice seedlings grown hydroponically in the Hewitt nutrient medium and after acclimatization, exposed to 50 µM As alone as well as with microbial consortium to observe the impact at morphological and molecular level. The inoculation of microbial consortium significantly ameliorated the As toxicity, improved growth of root hairs and maintained cellular integrity of the epidermis, exodermis and the stele region during As exposure. Several genes showed differential expression in As and As+P. putida. Down-regulation of As transporters (OsPIP2;2 and OsPIP2;3, OsTIP2;1) and higher expression of WRKY gene (WRKY28) during As+P. putida+C.vulgaris suggested reduction of As uptake in rice. The up-regulation of nutrient elements transporters (OsZIFL9, OsZIFL5, OsZIFL12 and OsZIP2, OsYSL15 and OsCOPT6) in the presence of consortium indicated the improved nutrient status of rice. Higher expression of regulatory elements like auxin/indole 3 acetic acid (AUX/IAA), WRKY and myeloblastosis (MYB) TFs and down-regulation of defense responsive genes such Glutathione-S-transferase, Peroxidase and Glutaredoxinduring As+P. putida+C.vulgaris exposure was also observed. Proteome profiling demonstrated differential abundance of proteins involved in photosynthesis (chlorophyll a/b binding protein, photosystem I Fe-S centre), energy metabolism (ATP synthase subunit beta) transport, signaling (tubulin 1, actin 1), defense (glutathione S-transferase, phenylalanine ammonia lyase) and amino acid metabolism (cysteine synthase, glutamine synthetase), which supported the As ameliorative and growth-promoting potential of microbial consortium during As stress in rice plants. The study gives comprehensive information about gene and protein changes in rice plants in As+consortium exposure.

OsGSTU5 interacts and glutathionylates the VirE2 protein of Agrobacterium and its (OsGSTU5) overexpression and downregulation showed a low and high AMT efficiency in rice, respectively. During Agrobacterium-mediated transformation (AMT), T-DNA along with several virulence proteins such as VirD2, VirE2, VirE3, VirD5, and VirF enter the plant cytoplasm. VirE2 serves as a single-stranded DNA binding (SSB) protein that assists the cytoplasmic trafficking of T-DNA inside the host cell. Though the regulatory roles of VirE2 have been established, the cellular reaction of their host, especially in monocots, has not been characterized in detail. This study identified a cellular interactor of VirE2 from the cDNA library of rice. The identified plant protein encoded by the gene cloned from rice was designated OsGSTU5, it interacted specifically with VirE2 in the host cytoplasm. OsGSTU5 was upregulated during Agrobacterium infection and involved in the post-translational glutathionylation of VirE2 (gVirE2). Interestingly, the in silico analysis showed that the 'gVirE2 + ssDNA' complex was structurally less stable than the 'VirE2 + ssDNA' complex. The gel shift assay also confirmed the attenuated SSB property of gVirE2 over VirE2. Moreover, knock-down and overexpression of OsGSTU5 in rice showed increased and decreased T-DNA expression, respectively after Agrobacterium infection. The present finding establishes the role of OsGSTU5 as an important target for modulation of AMT efficiency in rice.

Nickel (Ni) is an essential element for plant growth and is a constituent of several metalloenzymes, such as urease, Ni-Fe hydrogenase, Ni-superoxide dismutase. However, in high concentrations, Ni is toxic and hazardous to plants, humans and animals. High levels of Ni inhibit plant germination, reduce chlorophyll content, and cause osmotic imbalance and oxidative stress. Sustainable plant-bacterial native associations are formed under Ni-stress, such as Ni hyperaccumulator plants and rhizobacteria showed tolerance to high levels of Ni. Both partners (plants and bacteria) are capable to reduce the Ni toxicity and developed different mechanisms and strategies which they manifest in plant-bacterial associations. In addition to physical barriers, such as plants cell walls, thick cuticles and trichomes, which reduce the elevated levels of Ni entrance, plants are mitigating the Ni toxicity using their own antioxidant defense mechanisms including enzymes and other antioxidants. Bacteria in its turn effectively protect plants from Ni stress and can be used in phytoremediation. PGPR (plant growth promotion rhizobacteria) possess various mechanisms of biological protection of plants at both whole population and single cell levels. In this review, we highlighted the current understanding of the bacterial induced protective mechanisms in plant-bacterial associations under Ni stress.

The present study demonstrates the Ni toxicity-ameliorating and growth-promoting abilities of two different bacterial isolates when applied to wheat (Triticum aestivum L.) as the host plant. Two bacterial strains tolerant to Ni stress were isolated from wheat seeds and selected based on their ability to improve the germination of wheat plants; they were identified as Bacillus megaterium AFI1 and Paenibacillus nicotianae AFI2. The protective effects of these epiphytic bacteria against Ni stress were studied in model experiments with two wheat cultivars: Ni stress-tolerant Leningradskaya 6 and susceptible Chinese spring. When these isolates were used as the inoculants applied to Ni-treated wheat plants, the growth parameters and the levels of photosynthetic pigments of the two wheat cultivars both under normal and Ni-stress conditions were increased, though B. megaterium AFI1 had a more pronounced ameliorative effect on the Ni contents in plant tissues due to its synthesis of siderophores. Over the 10 days of Ni exposure, the plant growth promotion bacteria (PGPB) significantly reduced the lipid peroxidation (LPO), ascorbate peroxidase (APX), superoxide dismutase (SOD) activities and proline content in the leaves of both wheat cultivars. The PGPB also increased peroxidase (POX) activity and the levels of chlorophyll a, chlorophyll b, and carotenoids in the wheat leaves. It was concluded that B. megaterium AFI1 is an ideal candidate for bioremediation and wheat growth promotion against Ni-induced oxidative stress, as it increases photosynthetic pigment contents, induces the antioxidant defense system, and lowers Ni metal uptake.

The Bacillus amyloliquefaciens-SN13 and model crop rice (Oryza sativa) were chosen to understand the complex regulatory networks that govern plant-PGPR interaction under salt stress. During stress, inoculation with SN13 significantly increased biomass, relative water content, proline and total soluble sugar in rice while decreased lipid peroxidation and electrolyte leakage. Extensive alterations in gene expression were also observed in rice root transcriptome under stress in the presence of SN13. Rhizobacteria induced changes in expression of a considerable number of photosynthesis, hormone, and stress-responsive genes, in addition to cell-wall and lipid metabolism-related genes under salt stress as compared to salt stress or SN13 inoculation alone, indicating its potential role in reducing the harmful effects of salinity. To validate RNA-seq data, qRT-PCR was performed for selected differentially expressed genes representing various functional categories including metabolism, regulation, stress-response, and transporters. Results indicate qualitative and quantitative differences between roots responses to SN13 under stressed and unstressed conditions. Functional expressions of OsNAM and OsGRAM in yeast showed enhanced tolerance to various abiotic stresses, indicating crucial SN13-rice interaction in imparting beneficial effects under stress. This is first detailed report on understanding molecular mechanism underlying beneficial plant-microbe interaction in any economically important model crop plant under abiotic stress.

An array of eight CsI(Tl) detectors has been set up to measure the light charged particles in nuclear reactions using heavy ions from the Pelletron Linac Facility, Mumbai. The energy response of CsI(Tl) detector to α -particles from 5 to 40 MeV is measured using radioactive sources and the 12C(12C, α ) reaction populating discrete states in 20Ne. The energy non-linearity and the count rate effect on the pulse shape discrimination property have also been measured and observed the deterioration of pulse shape discrimination with higher count rate.

Arsenic is listed as the most extensive environmental carcinogens. The present report demonstrated that yeast expressing WaarsM gene encoding arsenic methyltransferase from soil fungus Westerdykella aurantiaca removes arsenic from contaminated soil. We investigated the potential use of yeast as a plant growth-promoting agent to stimulate rice plant growth. The genetically engineered (GE) Saccharomyces cerevisiae showed high arsenic methylation and volatilization activity under arsenic stress. Rice seeds bio-primed with GE yeast showed higher seedling vigor index compared to untreated control. Yeast also promotes the growth of rice plants in control condition while GE yeast significantly (P

Dehydrins (DHNs) have been reported to act as chaperones to combat drought stress. While in this study, we explored a novel nuclear complex of Oryza sativa FKBP and YSK2-type dehydrin (OsDhn-Rab16D), associated with ABA signaling to impart drought stress tolerance. The transcript levels of OsDhn-Rab16D in rice seedlings were induced in response to drought, abscisic acid (ABA) and H2O2 exposure. Ectopic expression of OsDhn-Rab16D transgenic lines showed enhanced tolerance to both osmotic stresses caused by PEG and drought. Using the yeast two-hybrid (Y2H) assay, O. sativa FKBP (a prolyl cis-trans isomerase) was identified as an interacting partner of OsDhn-Rab16D. Fluorescence signals between OsDhn-Rab16D and OsFKBP were observed in the nucleus. The pull-down assay confirmed the physical interaction between OsDhn-Rab16D and OsFKBP. qRT-PCR of drought and ABA-responsive gene reveals the higher transcript abundance in PEG-treated transgenic lines. Moreover, under drought conditions, transgenic lines maintain membrane integrity and increase the lignification in adventitious rice roots due to the higher expression level of catalase and lignin biosynthesis enzyme, respectively, as compared with wild-type. Overall, our findings suggest that OsDhn-Rab16D and OsFKBP complex involved in ABA-responsive drought stress signaling in rice and probably act as a positive transcriptional co-regulator, in addition, to act as chaperones.

During Agrobacterium-mediated transformation (AMT), T-DNA along with several virulence proteins like VirD2, VirE2, VirE3, VirD5, and VirF enter into the plant cytoplasm. VirE2 is supposed to serve as single-stranded DNA binding (SSB) protein and assist the cytoplasmic trafficking of T-DNA inside the host cell. In the present study, a rice glutathione-S-transferase (OsGSTU5) that interacts with VirE2 protein in plant cytoplasm has been identified. OsGSTU5 is observed to be involved in post-translational glutathionylation of VirE2 protein (gVirE2). In silico analysis revealed that ‘gVirE2+ssDNA’ complex is structurally less stable than ‘VirE2+ ssDNA’ complex. The gel shift activity confirms the attenuated SSB property of gVirE2 over VirE2 protein under in vitro condition. Moreover, knock-down and overexpression OsGSTU5 phenotypes of rice showed increased and decreased T-DNA expression, respectively after Agrobacterium infection. The present finding convincingly establishes the role of OsGSTU5 as defense protein in rice that can further serve as an important target for modulation of AMT efficiency in rice.

Arsenic (As) considered as one of the hazardous metalloid that hampers various physiological activities in rice. To study the mechanism of As tolerance in rice, one differentially expressed tau class glutathione-S-transferase (OsGSTU5) has been selected and transgenic rice plants with knockdown (KD) and overexpressing (OE) OsGSTU5 were generated. Our results suggested that KD lines became less tolerant to As stress than WT plants, while OE lines showed enhanced tolerance to As. Under As toxicity, OE and KD lines showed enhanced and reduced antioxidant activities such as, SOD, PRX and catalase, respectively indicating its role in ROS homeostasis. In addition, higher malondialdehyde content, poor photosynthetic parameters and higher reactive oxygen species (ROS) in KD plant, suggests that knockdown of OsGSTU5 renders KD plants more susceptible to oxidative damage. Also, the relative expression profile of various transporters such as OsABCC1 (As sequestration), Lsi2 and Lsi6 (As translocaters) and GSH dependent activity of GSTU5 suggests that GSTU5 might help in chelation of As with GSH and sequester it into the root vacuole using OsABCC1 transporter and thus limits the upward translocation of As towards shoot. This study suggests the importance of GSTU5 as a good target to improve the As tolerance in rice.

Authors’ original file for figure 7

Additional file of Transcriptomic and metabolomic shifts in rice roots in response to Cr (VI) stress

Additional File 7:Figure S3 a-e. K-means clustering and GO annotations of genes up-regulated during chromium stress. The clustering was done according to the presence of the motifs identified in this study. a(i) cluster1 a(ii) GO annotation of cluster1. b(i) cluster2 b(ii) GO annotation of cluster2 c(i) cluster3 c(ii) GO annotation of cluster3 d(i) cluster4 d(ii) GO annotation of cluster4 e(i) cluster5 e(ii) GO annotation of cluster5 (PPT 656 KB)

Additional File 5:Table S4. List of root-specific genes down-regulated during Cr (VI) stress in rice. (DOC 66 KB)