Intrinsically disordered protein, DNA binding with one finger transcription factor (OsDOF27) implicates thermotolerance in yeast and rice

Intrinsically disorder regions or proteins (IDRs or IDPs) constitute a large subset of the eukaryotic proteome, which challenges the protein structure–function paradigm. These IDPs lack a stable tertiary structure, yet they play a crucial role in the diverse biological process of plants. This study represents the intrinsically disordered nature of a plant-specific DNA binding with one finger transcription factor (DOF-TF). Here, we have investigated the role of OsDOF27 and characterized it as an intrinsically disordered protein. Furthermore, the molecular role of OsDOF27 in thermal stress tolerance has been elucidated. The qRT-PCR analysis revealed that OsDOF27 was significantly upregulated under different abiotic stress treatments in rice, particularly under heat stress. The stress-responsive transcript induction of OsDOF27 was further correlated with enriched abiotic stress-related cis-regulatory elements present in its promoter region. The in vivo functional analysis of the potential role of OsDOF27 in thermotolerance was further studied in yeast and in planta. Ectopic expression of OsDOF27 in yeast implicates thermotolerance response. Furthermore, the rice transgenic lines with overexpressing OsDOF27 revealed a positive role in mitigating heat stress tolerance. Collectively, our results evidently show the intrinsically disorderedness in OsDOF27 and its role in thermal stress response in rice

Molecular cloning, expression and Insilco analysis of drought stress inducible MYB transcription factor encoding gene from C4 plant Eleusine coracana

Drought is one of the key abiotic stresses that critically influences the crops by restraining their growth and yield potential. Being sessile, plant tackle the detrimental effects of drought stress by modulating the cellular state by changing the gene expression. The transcriptional syndicate essentially drives such alteration of gene expression. Transcription factors (TF) are the key regulatory protein that controls the expression of their target gene by binding to the cis-regulatory elements present in the promoter region. Myb-TF, ubiquitously present in all eukaryotes belong to one of the largest TF family, and play a wide array of biological functions in plants, including anthocyanin biosynthesis, vasculature system, cell signalling, seed maturation and abiotc stress responses. The present performed isolation and molecular cloning of full length Myb TF from Eleusine corocana. The isolated full-length coding sequence has 1053 bp and 350 aa was submitted to NCBI (Accession number MT312253). The transcript level of EcMYB increases under different abiotic stress treatments including dehydration, salinity, and high-temperature stress. The promoter region of EcMyb1 was found to be enriched in stress-responsive cis-regulatory elements such as DRE, HSE, ABRE etc. In phylogenetic analysis, EcMyb1 appeared to have high homology with its monocot orthologs particularly Sateria italica, Hordeum vulgare, Saccharum barberi and Oryza sativa. The three-dimension protein structure was generated based on homology modeling and structural aspects were discussed. Further, Insilco analysis was conducted to explore the physiological properties, subcellular localization, potential posttranslational modification sites (phosphorylation and glycosylation sites), and molecular and biological function of the full-length protein. Overall, the expression profiling and Insilco analysis of EcMyb1 strongly indicated its potential role in abiotic stress response in Eleusine corocana.

Differentially expressed seed aging responsive heat shock protein OsHSP18.2 implicates in seed vigor, longevity and improves germination and seedling establishment under abiotic stress

Small heat shock proteins (sHSP) are a diverse group of proteins and are highly abundant in plant species. Although majority of these sHSPs were shown to express specifically in seed, their potential function in seed physiology remains to be fully explored. Our proteomic analysis revealed that OsHSP18.2, a class II cytosolic HSP is an aging responsive protein as its abundance significantly increased after artificial aging in rice seeds. OsHSP18.2 transcript was found to markedly increase at the late maturation stage being highly abundant in dry seeds and sharply decreased after germination. Our biochemical study clearly demonstrated that OsHSP18.2 forms homooligomeric complex and is dodecameric in nature and functions as a molecular chaperon. OsHSP18.2 displayed chaperone activity as it was effective in preventing thermal inactivation of Citrate Synthase. Further, to analyze the function of this protein in seed physiology, seed specific Arabidopsis overexpression lines for OsHSP18.2 were generated. Our subsequent functional analysis clearly demonstrated that OsHSP18.2 has ability to improve seed vigor and longevity by reducing deleterious ROS accumulation in seeds. In addition, transformed Arabidopsis seeds displayed better performance in germination and cotyledon emergence under adverse conditions as well. Collectively, our work demonstrates that OsHSP18.2 is an aging responsive protein which functions as a molecular chaperon and possibly protect and stabilize the cellular proteins from irreversible damage particularly during maturation drying, desiccation and aging in seeds by restricting ROS accumulation and thereby improves seed vigor, longevity and seedling establishment

Antimicrobial Potential of Essential Oils from Aromatic Plant Ocimum sp.; A Comparative Biochemical Profiling and In-Silico Analysis

Medicinal and aromatic plants (MAPs) are a rich source of bioactive compounds that are immensely important due to their potential use in pharmacological and agricultural applications. Here, we have evaluated the antimicrobial activity of essential oils (EOs) from three different species of Ocimum: O. gratissimum (EO1), O. tenuiflorum (EO2), and O. sanctum (EO3). The EOs were screened for antibacterial activity against pathogenic strains of Escherichia coli, Enterobacter cloacae and methicillin-resistant Staphylococcus aureus (MRSA). The essential oils EO1 and EO3 showed significant growth inhibition of the tested bacteria. Likewise, all EOs exhibited antifungal potential against the broad-spectrum plant fungal pathogen Sclerotinia sclerotiorum that causes white-mould disease in plants. Moreover, the antimicrobial potential of the EOs correlates well with their antioxidant activity determined by DPPH free radical scavenging activity. The biochemical analysis of the EOs employing high-performance thin-layer chromatography, gas chromatography-mass spectrometry, and Fourier transform infrared spectroscopy, revealed the presence of distinct phytoconstituents that might be responsible for their differential bioactivity. Furthermore, an in-silico evaluation of the candidate phytoconstituents using molecular docking analysis suggests their potential for antimicrobial applications. Altogether, our results clearly show that EO1 and EO3 possess promising antimicrobial properties, and therefore could be utilized as a potential antimicrobial agent

Fostering nanoscience’s strategies: A new frontier in sustainable crop improvement for abiotic stress tolerance

Advanced nano-engineering is a convenient technology to attain food security and ensure sustainable agricultural yield and productivity. In addition to addressing the yield barrier, the application of nanoscience emphasizes its potential through innovations such as precision farming, site-targeted delivery of agrochemicals, disease control, and mitigation of environmental stresses in plants. Abiotic stresses negatively influence growth and yield of plants by affecting the physiological, biochemical, and molecular aspects of plants. As seen in recent years, such precedents in plants can be significantly alleviated through the implementation of nanoparticles. The application of nanoparticles helps in understanding the appropriate mechanisms in plants against abiotic stresses and enhances those responses more effectively. Biochemical and physiological adaptations stimulated by nanoparticles include the activation of the antioxidative defense system, stress regulatory gene expressions, stimulation of crucial biochemical pathways, and hormonal regulations. Considering the potential advantages of nanomaterials to date, their full implementation is yet to be a reality in the agricultural sector, largely limited due to concerns regarding the uptake, translocation, bioavailability, and eco-toxicity of nanoparticles. Understanding the underlying mechanisms and responses induced by nanoparticles through molecular approaches is critical in assessing nanomaterials' biological potential. The present review addresses the possible scope of nanotechnology to counter abiotic stress in economically important crops, and their influence on development, growth, absorption, and translocation in plants. Here, an attempt is made to provide an elucidative framework on recent findings related to nanoparticle-induced stress tolerance in plants through a comprehensive insight into molecular mechanisms and biochemical responses that may help to meet the need for adaptive measures in crops during abiotic stress conditions

PUB63, a novel early heat responsive ubiquitin E3 ligase from rice implicates heat stress tolerance

Heat stress has emerged as a major environmental challenge triggering reduced crop yields, compromised growth, and negatively affecting the development of crops. The cellular response to heat stress involves transcriptional and translational reprogramming and regulation of protein homeostasis which is crucial for the heat stress response and leads to the differential expression of many genes associated with protein turnover mechanisms. Plant U-box (PUB) proteins are a class of the Ubiquitin-E3 ligase involved in the Ubiquitin-proteasome pathway. In the present study, we investigated the role of a PUB protein from rice, OsPUB63, involved in heat stress responses in rice. We also identified all PUB coding genes from O. sativa indica and compared with O. sativa japonica. A quantitative real time PCR demonstrated, OsPUB63 as an early heat inducible gene in rice and the subcellular localization study confirmed its presence in the nucleus and cytosol. OsPUB63 was demonstrated as a TPR repeat containing protein with molecular weight of 39.3 KDa. Using a combination of genetic, biochemical, and physiological assays, our work establishes OsPUB63 as crucial in regulating the heat stress response in rice. Specifically, we show that OsPUB63 enhances the heat stress tolerance in transgenic Arabidopsis by promoting the accumulation of proline and reduced MDA content. Our work has identified PUB63 as the first PUB gene family E3 ligase from rice which is implicated in heat stress response and provide a crucial target for improving crop yields under high-temperature conditions in rice

Morpho-physiological and biochemical responses in wheat foliar sprayed with zinc-chitosan-salicylic acid nanoparticles during drought stress

Drought is considered as a significant stress that hampers growth, development as well as productivity of wheat crop around the globe. The present investigation was performed to determine the changes induced by drought on morpho-physiology, antioxidative system, metabolism and yield parameters in wheat crop. The study also focussed on evaluating the effect of zinc-chitosan-salicylic acid (ZCS) nanoparticles in alleviating physiological and biochemical alterations and overcoming yield losses caused by drought. Drought was provided during the vegetative stage on four different varieties (two drought tolerant viz. C-306 and PBW-644 and two drought susceptible viz. HUW-322 and HUW-843) by withholding irrigation and maintaining moisture capacity of soil at 40%. ZCS nanoparticles were foliar sprayed on wheat at concentrations of 100, 200 and 400 mg L−1. Application of ZCS nanoparticles at 100 mg L−1 significantly (p < 0.05) enhanced relative water content (RWC), alleviated levels of antioxidative enzymes like superoxide dismutase, ascorbate peroxidase, glutathione reductase and guaiacol peroxidase and metabolites like proline, ascorbate, malondialdehyde and flavanoid in wheat leaves subjected to drought. Drought recovery was noteworthy in tolerant as well as sensitive varieties. Water stress reduced grain yield / plant by nearly 45% in tolerant varieties and nearly 50% in sensitive varieties. Spray of the nanoparticles on wheat foliage incremented the yield to 63% and 41% in tolerant varieties and 50% and 46% in sensitive varieties. This study suggests an outstanding role of ZCS nanoparticles at a concentration of 100 mg L−1 in mitigation of ill effects of drought. These nanoparticles have the ability to improve osmotic status of plant, enhance synthesis of osmoprotectants, activate ROS scavenging enzymes for maintaining membrane integrity and cellular protection and promote yield increment during drought stress. This implicates its role in ensuring food security and sustainable agriculture with reduction in environmental pollution due to limited use of fertilizers

Efficient Genetic Transformation of Rice for CRISPR/Cas9 Mediated Genome-Editing and Stable Overexpression Studies: A Case Study on Rice Lipase 1 and Galactinol Synthase Encoding Genes

Rice is a staple food crop for almost half of the world&rsquo;s population, especially in the developing countries of Asia and Africa. It is widely grown in different climatic conditions, depending on the quality of the water, soil, and genetic makeup of the rice cultivar. Many (a)biotic stresses severely curtail rice growth and development, with an eventual reduction in crop yield. However, for molecular functional analysis, the availability of an efficient genetic transformation protocol is essential. To ensure food security and safety for the continuously increasing global population, the development of climate-resilient crops is crucial. Here, in this study, the rice transformation protocol has been effectively optimized for the efficient and rapid generation of rice transgenic plants. We also highlighted the critical steps and precautionary measures to be taken while performing the rice transformation. We further assess the efficacy of this protocol by transforming rice with two different transformation constructs for generating galactinol synthase (GolS) overexpression lines and CRISPR/Cas9-mediated edited lines of lipase (Lip) encoding the OsLip1 gene. The putative transformants were subjected to molecular analysis to confirm gene integration/editing, respectively. Collectively, the easy, efficient, and rapid rice transformation protocol used in this present study can be applied as a potential tool for gene(s) function studies in rice and eventually to the rice crop improvement

C terminal consensus sequence present in putative peroxisomal MDAR protein of monocot and dicots.

<p>C terminal consensus sequence present in putative peroxisomal MDAR protein of monocot and dicots.</p

qPCR analysis of <i>Ecmdar</i> expression in wild type and two transgenic <i>Arabidopsis thaliana</i> lines.

<p>Data are presented as Mean±SE. Different letters indicate means that differ significantly (P < 0.05).</p