Publications by authors named "Amit Rai"

112 Publications

Safety and efficacy of Ayurvedic interventions and Yoga on long term effects of COVID-19: A structured summary of a study protocol for a randomized controlled trial.

Trials 2021 Jun 3;22(1):378. Epub 2021 Jun 3.

Central Council for Research in Ayurvedic Sciences Headquarters, 61-65, Institutional Area, Opp. 'D' Block, Janakpuri, New Delhi, 110058, India.

Objectives: Primary Objective • To assess the efficacy of Ayurveda interventions and Yoga in rehabilitation of COVID-19 cases suffering with long term effects of COVID 19 as compared to WHO Rehabilitation Self-Management after COVID-19- Related Illness. Secondary Objective • To assess the safety of Ayurvedic interventions in cases suffering with long term effects of COVID 19 TRIAL DESIGN: Multi-centric, randomized, controlled, parallel group, open-label, exploratory study. The study duration is 9 months and the intervention period is 90 days from the day of enrolment of the participant.

Participants: Patients of either sex between 18 to 60 years, ambulatory, willing to participate, with history (not more than 4 weeks) of positive RT-PCR for COVID-19 or IgM antibodies positivity for SARS CoV-2, but having negative RT-PCR for COVID-19 at the time of screening will be considered eligible for enrolment in the study. Critically ill patients with ARDS (acute respiratory distress syndrome), requiring invasive respiratory support in the intensive care unit, known case of any malignancy, immune-compromised state (e.g. HIV), diabetes mellitus, active pulmonary tuberculosis, past history of any chronic respiratory disease, motor neuron disease, multiple sclerosis, stroke, impaired cognition, atrial fibrillation, acute coronary syndrome, myocardial infarction, severe arrhythmia, concurrent serious hepatic disease or renal disease, pregnant or lactating women, patients on immunosuppressive medications, history of hypersensitivity to the trial drugs or their ingredients, depressive illness (before COVID-19), diagnosed psychotic illnesses, substance dependence or alcoholism will be excluded. The trial will be conducted at two medical colleges in Maharashtra, India.

Intervention And Comparator: Intervention Arm (Group-I): Ayurveda interventions including Agastya Haritaki six gram and Ashwagandha tablet 500 mg twice daily orally after meals with warm water and two sessions of yoga (morning 30 minutes and evening 15 minutes) daily for 90 days, as per the post-COVID-19 care protocol provided in National Clinical Management Protocol based on Ayurveda and Yoga for management of COVID-19 published by Ministry of AYUSH, Government of India. Comparator Arm (Group-II): WHO Rehabilitation Self-Management after COVID-19 related illness for 90 days. The trial drugs are being procured from a GMP certified pharmaceutical company.

Main Outcomes: Primary Outcome: Change in respiratory function to be assessed by San Diego shortness of breath Questionnaire, 6-minutes walk test and pulmonary function test.

Secondary Outcomes: Change in High-resolution Computed Tomography (HRCT) Chest Change in Fatigue score assessed by Modified Fatigue Impact Scale Change in Anxiety score assessed by Hospital Anxiety and Depression Scale Score Change in Sleep Quality assessed by Pittsburgh Sleep Quality Index Change in the quality of life assessed by COV19-QoL scale Safety of the interventions will be assessed by comparing hematological and biochemical investigations before and after the intervention period and Adverse Event/ Adverse drug reaction TIMELINES FOR OUTCOME ASSESSMENT: Subjective parameters and clinical assessment will be assessed at baseline, 15 day, 30 day, 60 day and 90 day. Laboratory parameters (CBC, LFT, KFT, HbA1c, Hs-CRP, D-dimer), Pulmonary function test and HRCT Chest will be done at baseline and after completion of study period i.e. 90 day.

Randomisation: Statistical package for Social Sciences (SPSS) version 15.0 is used to generate the random number sequences. The participants will be randomized to two study groups in the ratio of 1:1.

Blinding (masking): The study is open-label design. However, the outcome assessor will be kept blinded regarding the study group allocation of the participants.

Numbers To Be Randomised (sample Size) Sample Size: The sample size for the study is calculated assuming improvement in 6-minutes walk test by 40 meter in Group I and a change of 10 meter in Group II with a standard deviation of 50 meter based on the results of the previous studies, with 95% Confidence Level (α = 0.05) and 80% power and expecting a dropout rate of 20%. The number of participants to be enrolled in the study should be approximately 55 in each group. Hence, a total of 110 participants will be enrolled in the trial at each study site.

Trial Status: Participants' recruitment started on 1 May 2021. Anticipated end of recruitment is August 2021. Protocol number: CCRAS-01 Protocol version number: 1.1, 13th January 2021.

Trial Registration: The trial is prospectively registered with the Clinical Trial Registry of India (CTRI) on 03 March 2021 [ CTRI/2021/03/031686 ].

Full Protocol: The full protocol is attached as an additional file, accessible from the Journal website (Additional file 1). This communication serves as a summary of the key elements of the full protocol.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s13063-021-05326-1DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8173507PMC
June 2021

A Multifunctional Peptide From Fermented Soybean for Effective Inhibition of SARS-CoV-2 S1 Receptor Binding Domain and Modulation of Toll Like Receptor 4: A Molecular Docking Study.

Front Mol Biosci 2021 31;8:636647. Epub 2021 Mar 31.

Institute of Bioresources and Sustainable Development, Regional Centre, Gangtok, India.

Fermented soybean products are traditionally consumed and popular in many Asian countries and the northeastern part of India. To search for potential agents for the interruption of the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Spike glycoprotein 1 (S1) and human angiotensin-converting enzyme 2 (ACE2) receptor interactions, the antiviral prospective of peptides identified from the proteome of was investigated. Soybean was fermented using KN1G, KN2G and two different strains of (KN2B and KN2M). The peptides were screened for possible antiviral activity using two different web servers (AVPpred and meta-iAVP), and binding interactions of selected 44 peptides were further explored against the receptor-binding domain (RBD) of the S1 protein (PDB ID: 6M0J) by molecular docking using ZDOCK. The results showed that a peptide ALPEEVIQHTFNLKSQ (P13) belonging to KN1G fermented was able to make contacts with the binding motif of RBD by blocking specific residues designated as critical (GLN493, ASN501) in the binding of human angiotensin-converting enzyme 2 (ACE2) cell receptor. The selected peptide was also observed to have a significant affinity towards human toll like receptor 4 (TLR4)/Myeloid Differentiation factor 2 (MD2) (PDB ID: 3FXI) complex known for its essential role in cytokine storm. The energy properties of the docked complexes were analyzed through the Generalized Born model and Solvent Accessibility method (MM/GBSA) using HawkDock server. The results showed peptidyl amino acids GLU5, GLN8, PHE11, and LEU13 contributed most to P13-RBD binding. Similarly, ARG90, PHE121, LEU61, PHE126, and ILE94 were appeared to be significant in P13-TLR4/MD2 complex. The findings of the study suggest that the peptides from fermented soy prepared using KN1G have better potential to be used as antiviral agents. The specific peptide ALPEEVIQHTFNLKSQ could be synthesized and used in combination with experimental studies to validate its effect on SARS-CoV-2-hACE2 interaction and modulation of TLR4 activity. Subsequently, the protein hydrolysate comprising these peptides could be used as prophylaxis against viral diseases, including COVID-19.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fmolb.2021.636647DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8044374PMC
March 2021

Deciphering the role of microRNAs during gene mediated resistance response in rice.

Physiol Mol Biol Plants 2021 Mar 10;27(3):633-647. Epub 2021 Mar 10.

ICAR-National Institute for Plant Biotechnology, Pusa Campus, New Delhi, 110012 India.

The broad-spectrum resistance gene confers resistance to multiple isolates of in rice. In order to decipher the molecular mechanism underlying the mediated resistance in rice line Taipei309 (carrying ), miRNAome study was performed at 24 h post-inoculation (hpi) with . A total of 222 known miRNAs representing 101 miRNA families were found in this study. Of these, 29 and 24 miRNAs were respectively up- and down-regulated in the resistant Taipei309 . Defence response () genes, like, , and , and genes related to transcription factors were up-regulated in Taipei309 line. The vast array of miRNA candidates identified here are miR159c, miR167c, miR2100, miR2118o, miR2118l, miR319a, miR393, miR395l, miR397a, miR397b, miR398, miR439g, miR531b, miR812f, and miR815c, and they manifest their role in balancing the interplay between various genes during mediated resistance. We also validated miRNA/target gene pairs involved in hormone signalling, and cross-talk among hormone pathways regulating the rice immunity. This study suggests that the gene mediated blast resistance is influenced by several microRNAs through PTI and ETI components in the rice line Taipei309 , leading to incompatible host-pathogen interaction.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1007/s12298-021-00960-0DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7981355PMC
March 2021

A novel β-glucosidase from a hot-spring metagenome shows elevated thermal stability and tolerance to glucose and ethanol.

Enzyme Microb Technol 2021 Apr 11;145:109764. Epub 2021 Feb 11.

Center of Innovative and Applied Bioprocessing (DBT-CIAB), S.A.S. Nagar, Sector-81, Knowledge City, Mohali 140 306, India. Electronic address:

β-glucosidase causes hydrolysis of β-1,4-glycosidic bond in glycosides and oligosaccharides. It is an industrially important enzyme owing to its potential in biomass processing applications. In this study, computational screening of an extreme temperature aquatic habitat metagenomic resource was done, leading to the identification of a novel gene, bgl, encoding a β-glucosidase. The comparative protein sequence and homology structure analyses designated it as a GH1 family β-glucosidase. The bgl gene was expressed in a heterologous host, Escherichia coli. The purified protein, Bgl, was biochemically characterized for β-glucosidase activity. Bgl exhibited noteworthy hydrolytic potential towards cellobiose and lactose. Bgl, showed substantial catalytic activity in the pH range of 5.0-7.0 and at the temperature 40 °C-70 °C. The enzyme was found quite stable at 50 °C with a loss of hardly 20% after 40 h of heat exposure. Furthermore, any drastically negative effect was not observed on the enzyme's activity in the presence of metal ions, non-ionic surfactants, metal chelating, and denaturing agents. A significantly high glucose tolerance, retaining 80% relative activity at 1 M, and 40% at 5 M glucose, and ethanol tolerance, exhibiting 80% relative activity in 10% ethanol, enrolled Bgl as a promising enzyme for cellulose saccharification. Furthermore, its ability to catalyze the hydrolysis of daidzin and polydatin ascertained it as an admirably suited biocatalyst for enhancement of nutritional values in soya and wine industries.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.enzmictec.2021.109764DOI Listing
April 2021

A multi-objective hybrid machine learning approach-based optimization for enhanced biomass and bioactive phycobiliproteins production in Nostoc sp. CCC-403.

Bioresour Technol 2021 Jun 26;329:124908. Epub 2021 Feb 26.

Enzyme Technology and Protein Bioinformatics Laboratory, Department of Microbiology, Maharshi Dayanand University, Rohtak 124001, Haryana, India; School of Biotechnology, Institute of Science, Banaras Hindu University, Varanasi 221005, India. Electronic address:

The cyanobacterial phycobiliproteins (PBPs) are an important natural colorant for nutraceutical industries. Here, a multi-objective hybrid machine learning-based optimization approach was used for enhanced cell biomass and PBPs production simultaneously in Nostoc sp. CCC-403. A central composite design (CCD) was employed to design an experimental setup for four input parameters, including three BG-11 medium components and pH. We achieved a 61.76% increase in total PBPs production and an almost 90% increase in cell biomass by our prediction model. We also established a test genome-scale metabolic network (GSMN) for Nostoc sp. and identified potential metabolic fluxes contributing to PBPs enhanced production. This study highlights the advantage of the hybrid machine learning approach and GSMN to achieve optimization for more than one objective and serves as the foundation for future efforts to convert cyanobacteria as an economically viable source for biofuels and natural products.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.biortech.2021.124908DOI Listing
June 2021

Production, characterization and molecular docking of antioxidant peptides from peptidome of kinema fermented with proteolytic Bacillus spp.

Food Res Int 2021 03 19;141:110161. Epub 2021 Jan 19.

Institute of Bioresources and Sustainable Development, Sikkim Centre, Tadong, India. Electronic address:

Kinema is an alkaline traditionally fermented soybean product popularly consumed in Sikkim Himalayan region. Kinema was prepared by soybean fermented with different species of Bacillus and analyzed for peptide content, antioxidant activity and consequence of gastrointestinal enzymes (pepsin and pancreatin) on the antioxidant effect. Antioxidant effect was enhanced during soybean fermentation using different starters, which further increased during gastrointestinal digestion. The peptides formed during soybean fermentation were analyzed using LC-MS/MS. Soybean fermented using different starters resulted in the production of some common peptides and a large number of unique peptides, which may affect the functional property of kinema. Peptides having antioxidative amino acids (histidine, phenylalanine, methionine, tryptophan and tyrosine) and significant GRAVY value were selected for their molecular interaction with myeloperoxidase (MPO), a key enzyme responsible for elevated oxidative stress. A peptide SEDDVFVIPAAYPF produced in kinema fermented using Bacillus licheniformis 1G had interaction with four out of five catalytic residues identified in MPO. Kinema prepared using specific starter can produce unique peptides responsible for specific health benefits.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.foodres.2021.110161DOI Listing
March 2021

Current Status and Potential Therapeutic Strategies for Using Non-coding RNA to Treat Diabetic Cardiomyopathy.

Front Physiol 2020 22;11:612722. Epub 2021 Jan 22.

Department of Emergency Medicine, Institute of Behavioral Medicine and Research, Dorothy M. Davis Heart Lung and Research Institute, The Ohio State University Wexner Medical Center, Columbus, OH, United States.

Diabetic cardiomyopathy (DMCM) is the leading cause of mortality and morbidity among diabetic patients. DMCM is characterized by an increase in oxidative stress with systemic inflammation that leads to cardiac fibrosis, ultimately causing diastolic and systolic dysfunction. Even though DMCM pathophysiology is well studied, the approach to limit this condition is not met with success. This highlights the need for more knowledge of underlying mechanisms and innovative therapies. In this regard, emerging evidence suggests a potential role of non-coding RNAs (ncRNAs), including micro-RNAs (miRNAs), long non-coding RNAs (lncRNAs), and circular RNAs (circRNAs) as novel diagnostics, mechanisms, and therapeutics in the context of DMCM. However, our understanding of ncRNAs' role in diabetic heart disease is still in its infancy. This review provides a comprehensive update on pre-clinical and clinical studies that might develop therapeutic strategies to limit/prevent DMCM.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fphys.2020.612722DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7862744PMC
January 2021

Chromosome-level genome assembly of Ophiorrhiza pumila reveals the evolution of camptothecin biosynthesis.

Nat Commun 2021 01 15;12(1):405. Epub 2021 Jan 15.

Graduate School of Pharmaceutical Sciences, Chiba University, Chiba, Japan.

Plant genomes remain highly fragmented and are often characterized by hundreds to thousands of assembly gaps. Here, we report chromosome-level reference and phased genome assembly of Ophiorrhiza pumila, a camptothecin-producing medicinal plant, through an ordered multi-scaffolding and experimental validation approach. With 21 assembly gaps and a contig N50 of 18.49 Mb, Ophiorrhiza genome is one of the most complete plant genomes assembled to date. We also report 273 nitrogen-containing metabolites, including diverse monoterpene indole alkaloids (MIAs). A comparative genomics approach identifies strictosidine biogenesis as the origin of MIA evolution. The emergence of strictosidine biosynthesis-catalyzing enzymes precede downstream enzymes' evolution post γ whole-genome triplication, which occurred approximately 110 Mya in O. pumila, and before the whole-genome duplication in Camptotheca acuminata identified here. Combining comparative genome analysis, multi-omics analysis, and metabolic gene-cluster analysis, we propose a working model for MIA evolution, and a pangenome for MIA biosynthesis, which will help in establishing a sustainable supply of camptothecin.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41467-020-20508-2DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7810986PMC
January 2021

A Potential Peptide From Soy Cheese Produced Using WS4 for Effective Inhibition of SARS-CoV-2 Main Protease and S1 Glycoprotein.

Front Mol Biosci 2020 11;7:601753. Epub 2020 Dec 11.

Institute of Bioresources and Sustainable Development (DBT-IBSD), Regional Centre, Sikkim, India.

The COVID-19 pandemic caused by novel SARS-CoV-2 has resulted in an unprecedented loss of lives and economy around the world. In this study, search for potential inhibitors against two of the best characterized SARS-CoV-2 drug targets: S1 glycoprotein receptor-binding domain (RBD) and main protease (3CL), was carried out using the soy cheese peptides. A total of 1,420 peptides identified from the cheese peptidome produced using WS4 were screened for antiviral activity by employing the web tools, AVPpred, and meta-iAVP. Molecular docking studies of the selected peptides revealed one potential peptide "KFVPKQPNMIL" that demonstrated strong affinity toward significant amino acid residues responsible for the host cell entry (RBD) and multiplication (3CL) of SARS-CoV-2. The peptide was also assessed for its ability to interact with the critical residues of S1 RBD and 3CL of other β-coronaviruses. High binding affinity was observed toward critical amino acids of both the targeted proteins in SARS-CoV, MERS-CoV, and HCoV-HKU1. The binding energy of KFVPKQPNMIL against RBD and 3CL of the four viruses ranged from -8.45 to -26.8 kcal/mol and -15.22 to -22.85 kcal/mol, respectively. The findings conclude that cheese, produced by using WS4, could be explored as a prophylactic food for SARS-CoV-2 and related viruses. In addition, the multi-target inhibitor peptide, which effectively inhibited both the viral proteins, could further be used as a terminus a quo for the and function against SARS-CoV-2.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fmolb.2020.601753DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7759660PMC
December 2020

Microwave-assisted efficient synthesis of pyrazole-fibrate derivatives as stimulators of glucose uptake in skeletal muscle cells.

Bioorg Med Chem Lett 2021 02 25;34:127760. Epub 2020 Dec 25.

Medicinal and Process Chemistry Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Sitapur Road, Lucknow 226031, India; Academy of Scientific and Innovative Research, Ghaziabad 201002, U.P., India. Electronic address:

The design and synthesis of a series of pyrazolo[3,4-d]pyrimidinones containing fibrate side chains have been accomplished by utilizing the concept of molecular hybridization. All the synthesized compounds were evaluated for the glucose uptake stimulatory effect in L6 rat skeletal muscle cells. Four compounds (3f, 3g, 3j and 3q) were found to show significant stimulation of glucose uptake. Further these four compounds have been examined for their Glut4 translocation stimulatory effect in L6-Glut4myc myotubes. Compound 3q was found to exert maximum increase in GLUT4myc translocation.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.bmcl.2020.127760DOI Listing
February 2021

Biotechnological approaches for the production of designer cheese with improved functionality.

Compr Rev Food Sci Food Saf 2021 Jan 15;20(1):960-979. Epub 2020 Dec 15.

Institute of Bioresources and Sustainable Development, Regional Centre, Tadong, Sikkim, India.

Cheese is a product of ancient biotechnological practices, which has been revolutionized as a functional food product in many parts of the world. Bioactive compounds, such as peptides, polysaccharides, and fatty acids, have been identified in traditional cheese products, which demonstrate functional properties such as antihypertensive, antioxidant, immunomodulation, antidiabetic, and anticancer activities. Besides, cheese-making probiotic lactic acid bacteria (LAB) exert a positive impact on gut health, aiding in digestion, and improved nutrient absorption. Advancement in biotechnological research revealed the potential of metabolite production with prebiotics and bioactive functions in several strains of LAB, yeast, and filamentous fungi. The application of specific biocatalyst producing microbial strains enhances nutraceutical value, resulting in designer cheese products with multifarious health beneficial effects. This review summarizes the biotechnological approaches applied in designing cheese products with improved functional properties.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/1541-4337.12680DOI Listing
January 2021

Comparison of Antimicrobial Efficacy of Four Different Plant Extracts against Cariogenic Bacteria: An Study.

Int J Clin Pediatr Dent 2020 Jul-Aug;13(4):361-367

Department of Pedodontics and Preventive Dentistry, Sardar Patel Postgraduate Institute of Dental and Medical Sciences, Lucknow, Uttar Pradesh, India.

Background: Dental caries is an oral disease which is infectious in nature associated with various pathogenic microorganisms and is prevalent across the world.

Aim: To evaluate and compare the anticariogenic properties of the different plant extracts against various cariogenic microorganisms.

Materials And Methods: In phase I, the anticariogenic efficacy of four different herbal extracts, namely (), (), (), and (Licorice), was evaluated against two strains of bacteria viz. and In the second phase of the study, the overall decay depth was compared.

Results: In phase I, zone of inhibition was significantly higher in (Licorice) followed the descending order by (), (), and (), respectively. In the second phase, it was found that the decay depth was significantly higher in () followed the descending order by (), (), and (Licorice), respectively.

Conclusion: (Licorice) extract had potent antibacterial efficacy against and

Clinical Significance: Synthetic drugs have many adverse effects, so more attention has been paid to natural remedies because they are safe and effective. Today, the main interest is shifted toward the drugs which is derived from plants, which has led to the screening of many herbal plants for their potential antimicrobial activity and the same can be used clinically as an alternative of synthetic drugs used in dentistry.

How To Cite This Article: Rai A, Tripathi AM, Saha S, . Comparison of Antimicrobial Efficacy of Four Different Plant Extracts against Cariogenic Bacteria: An Study. Int J Clin Pediatr Dent 2020;13(4):361-367.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.5005/jp-journals-10005-1796DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7586476PMC
November 2020

Bioconservation of iron and enhancement of antioxidant and antibacterial properties of chicken gizzard protein hydrolysate fermented by Pediococcus acidilactici ATTC 8042.

J Sci Food Agric 2021 May 12;101(7):2718-2726. Epub 2020 Nov 12.

Department of Fermentation Technology, Faculty of Food-Industry, King Mongkut's Institute of Technology Ladkrabang, Bangkok, Thailand.

Background: The poultry industry is one of the fastest growing sectors, and it generates considerable quantities of chicken gizzards (CG) every day. However, due to their hard texture and high microbial load, and due to cultural beliefs, they are not preferred by consumers. Chicken gizzards are a substantial source of proteins, iron, and other nutrients, which can be used effectively to produce nutraceuticals, rich in peptides (antioxidants and antibacterial), bio-iron, essential free amino acids, and fatty acids vital for human health.

Results: Lactic acid fermentation of CG by Pediococcus acidilactici ATTC 8042 increased the antioxidant activity of 2,2-diphenyl-1-picrylhydrazyl (DPPH), azino-bis (3-ethylbenzothiaziline-6-sulphonic acid) (ABTS), and ferric reducing antioxidant power (FRAP) by up to 26 times compared with unfermented CG (P < 0.05). The amount of hydrolysis and solvents (ethanol and water) used for extracting protein hydrolysates significantly affected the antioxidant properties. Moreover, fermented CG showed a negligible reduction in bio-iron (2-3%) compared with heat-processed CG (85 °C for 15 min), in which bio-iron was reduced by up to 20.3% (P < 0.05). The presence of unsaturated fatty acids such as C20:4 and C22:4 n-6 indicated a low level of lipid oxidation.

Conclusion: Fermented CG, with its reasonably high antioxidant and antibacterial activity, together with a substantial amount of bio-iron and other nutritional components can serve as a functional food or feed additive to reduce oxidative stress and to treat iron deficiency. © 2020 Society of Chemical Industry.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/jsfa.10898DOI Listing
May 2021

A novel cold-active type I pullulanase from a hot-spring metagenome for effective debranching and production of resistant starch.

Bioresour Technol 2021 Jan 20;320(Pt A):124288. Epub 2020 Oct 20.

Center of Innovative and Applied Bioprocessing (DBT-CIAB), SAS Nagar, Sector 81, Mohali, India. Electronic address:

Pullulanase is a potent enzyme for starch debranching. In this study, a novel type I pullulanase (Pul) was identified from the metagenome of a thermal aquatic habitat that exhibits optimal activity of debranching at 40 °C temperature and pH 6.0 to 7.0. More than 50% enzymatic activity was detected at the low temperature of 4 °C, determining it a cold-active type I pullulanase. It was able to efficiently catalyze the hydrolysis of α-1,6-glycosidic linkages in pullulan, with a specific activity of 177 U mg. The results determined Pul to be a potential starch debranching biocatalyst, causing a significant increase of about 80% in the apparent amylose content of potato starch. Retrogradation of the debranched starch resulted in the formation of resistant starch 3. The yield of resistant starch was estimated to be about 45%. The resistant starch exhibited higher crystallinity, enhanced heat-stability, and resistance to α-amylase digestion, as compared to native starch.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.biortech.2020.124288DOI Listing
January 2021

ADMET profile and virtual screening of plant and microbial natural metabolites as SARS-CoV-2 S1 glycoprotein receptor binding domain and main protease inhibitors.

Eur J Pharmacol 2021 Jan 16;890:173648. Epub 2020 Oct 16.

Department of Chemical Sciences, University of Naples Federico II, Via Cintia 4, 80126, Naples, Italy. Electronic address:

In an attempt to search for selective inhibitors against the SARS-CoV-2 which caused devastating of lives and livelihoods across the globe, 415 natural metabolites isolated from several plants, fungi and bacteria, belonging to different classes, were investigated. The drug metabolism and safety profiles were computed in silico and the results showed seven compounds namely fusaric acid, jasmonic acid, jasmonic acid methyl ester, putaminoxin, putaminoxin B and D, and stagonolide K were predicted to having considerable absorption, metabolism, distribution and excretion parameters (ADME) and safety indices. Molecular docking against the receptor binding domain (RBD) of spike glycoprotein (S1) and the main protease (M) exposed the compounds having better binding affinity to main protease as compared to the S1 receptor binding domain. The docking results were compared to an antiviral drug penciclovir reportedly of clinical significance in treating the SARS-CoV-2 infected patients. The results demonstrated the test compounds jasmonic acid, putaminoxins B and D bound to the HIS-CYS catalytic dyad as well as to other residues within the M active site with much greater affinity than penciclovir. The findings of the study suggest that these compounds could be explored as potential SARS-CoV-2 inhibitors, and could further be combined with the experimental investigations to develop effective therapeutics to deal with the present pandemic.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.ejphar.2020.173648DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7561576PMC
January 2021

Metagenomics revealing molecular profiling of community structure and metabolic pathways in natural hot springs of the Sikkim Himalaya.

BMC Microbiol 2020 08 10;20(1):246. Epub 2020 Aug 10.

Center of Innovative and Applied Bioprocessing (DBT-CIAB), SAS Nagar, Mohali, India.

Background: Himalaya is an ecologically pristine environment. The geo-tectonic activities have shaped various environmental niches with diverse microbial populations throughout the Himalayan biosphere region. Albeit, limited information is available in terms of molecular insights into the microbiome, including the uncultured microbes, of the Himalayan habitat. Hence, a vast majority of genomic resources are still under-explored from this region. Metagenome analysis has simplified the extensive in-depth exploration of diverse habitats. In the present study, the culture-independent whole metagenome sequencing methodology was employed for microbial diversity exploration and identification of genes involved in various metabolic pathways in two geothermal springs located at different altitudes in the Sikkim Himalaya.

Results: The two hot springs, Polok and Reshi, have distinct abiotic conditions. The average temperature of Polok and Reshi was recorded to be 62 °C and 43 °C, respectively. Both the aquatic habitats have alkaline geochemistry with pH in the range of 7-8. Community profile analysis revealed genomic evidence of plentiful bacteria, with a minute fraction of the archaeal population in hot water reservoirs of Polok and Reshi hot spring. Mesophilic microbes belonging to Proteobacteria and Firmicutes phyla were predominant at both the sites. Polok exhibited an extravagant representation of Chloroflexi, Deinococcus-Thermus, Aquificae, and Thermotogae. Metabolic potential analysis depicted orthologous genes associated with sulfur, nitrogen, and methane metabolism, contributed by the microflora in the hydrothermal system. The genomic information of many novel carbohydrate-transforming enzymes was deciphered in the metagenomic description. Further, the genomic capacity of antimicrobial biomolecules and antibiotic resistance were discerned.

Conclusion: The study provided comprehensive molecular information about the microbial treasury as well as the metabolic features of the two geothermal sites. The thermal aquatic niches were found a potential bioresource of biocatalyst systems for biomass-processing. Overall, this study provides the whole metagenome based insights into the taxonomic and functional profiles of Polok and Reshi hot springs of the Sikkim Himalaya. The study generated a wealth of genomic data that can be explored for the discovery and characterization of novel genes encoding proteins of industrial importance.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1186/s12866-020-01923-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7418396PMC
August 2020

Multiomics-based characterization of specialized metabolites biosynthesis in Cornus Officinalis.

DNA Res 2020 Apr;27(2)

Graduate School of Pharmaceutical Sciences, Chiba University, Chiba 260-8675, Japan.

Cornus officinalis, an important traditional medicinal plant, is used as major constituents of tonics, analgesics, and diuretics. While several studies have focused on its characteristic bioactive compounds, little is known on their biosynthesis. In this study, we performed LC-QTOF-MS-based metabolome and RNA-seq-based transcriptome profiling for seven tissues of C. officinalis. Untargeted metabolome analysis assigned chemical identities to 1,215 metabolites and showed tissue-specific accumulation for specialized metabolites with medicinal properties. De novo transcriptome assembly established for C. officinalis showed 96% of transcriptome completeness. Co-expression analysis identified candidate genes involved in the biosynthesis of iridoids, triterpenoids, and gallotannins, the major group of bioactive metabolites identified in C. officinalis. Integrative omics analysis identified 45 cytochrome P450s genes correlated with iridoids accumulation in C. officinalis. Network-based integration of genes assigned to iridoids biosynthesis pathways with these candidate CYPs further identified seven promising CYPs associated with iridoids' metabolism. This study provides a valuable resource for further investigation of specialized metabolites' biosynthesis in C. officinalis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1093/dnares/dsaa009DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7320821PMC
April 2020

Production and characterisation of lipase for application in detergent industry from a novel Pseudomonas helmanticensis HS6.

Bioresour Technol 2020 Aug 8;309:123352. Epub 2020 Apr 8.

Institute of Bioresources and Sustainable Development, Sikkim Centre, Tadong 737102, Sikkim, India. Electronic address:

The aim of this study was to explore novel source of lipase from biodiversity hot spot region of Sikkim with activity at broad temperature range for application in detergent industry. Among the isolates, Pseudomonas helmanticensis HS6 showed activity at wide range of temperatures was selected for lipase production. Statistical optimisation for enhanced production of lipase resulted in enhancement of lipase activity from 2.3 to 179.3 U/mg. Lipase was purified resulting in 18.78 fold purification, 5.58% yield and high specific activity of 3368 U/mg. The partially purified lipase was found to be active in wide range of temperature (5-80 °C) and pH (6-9), showing optimum activity at 50 °C at pH 7. Peptide sequences on mass spectrometric analysis of purified lipase showed similarity to lipase family protein of three species of Pseudomonas. Both crude and purified lipase retained residual activity of 40-80% after 3 h of incubation with commercial detergents suggesting its application in detergent industry.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.biortech.2020.123352DOI Listing
August 2020

Author Correction: Novel Indole-fused benzo-oxazepines (IFBOs) inhibit invasion of hepatocellular carcinoma by targeting IL-6 mediated JAK2/STAT3 oncogenic signals.

Sci Rep 2020 Feb 6;10(1):2391. Epub 2020 Feb 6.

Department of Pharmaceutical Sciences, Babasaheb Bhimrao Ambedkar University, Vidya Vihar, Raibareli Road, Lucknow, 226025, India.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41598-020-59134-9DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7002741PMC
February 2020

Flavonoids from prevents advanced glycation end products formation and protein oxidation aligned with fructose-induced protein glycation.

Nat Prod Res 2019 Oct 4:1-5. Epub 2019 Oct 4.

Biochemistry Division, CSIR-Central Drug Research Institute , Lucknow , India.

Advanced glycation end products (AGEs) are reactive chemical entities formed by non-enzymatic reaction between reducing sugars and amino group of proteins. Enhanced accumulation of AGEs and associated protein oxidation contribute to pathogenesis of diabetes-associated complications. Here, we evaluated the inhibitory activity of flavonoid compounds isolated from the leaves of on formation of AGEs and protein oxidation. Antiglycation activity was determined by measuring the formation of AGE fluorescence intensity, Nε-(carboxymethyl) lysine, and level of fructosamine. Protein oxidation was examined using levels of protein carbonyls and thiol group. Compounds significantly ( < 0.001) restricted the formation of fluorescent AGEs in fructose- BSA and methylglyoxal-BSA systems. Furthermore, there was a decrease in levels of fructosamine and protein carbonyls, and elevation in level of thiol group in fructose-BSA in presence of flavonoids. In summary, flavonoids from inhibit fructose-mediated protein glycation and oxidation, and can be potential agent for preventing AGE-mediated diabetic complications.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1080/14786419.2019.1672690DOI Listing
October 2019

Metagenomic Insights Into the Taxonomic and Functional Features of , a Traditional Fermented Soybean Product of Sikkim Himalaya.

Front Microbiol 2019 2;10:1744. Epub 2019 Aug 2.

Center of Innovative and Applied Bioprocessing, Mohali, India.

is an ethnic, naturally fermented soybean product consumed in the Sikkim Himalayan region of India. In the present study, the whole metagenome sequencing approach was adopted to examine the microbial diversity and related functional potential of , consumed in different seasons. Firmicutes was the abundant phylum in , ranging from 82.31 to 93.99% in different seasons, followed by Actinobacteria and Proteobacteria. At the species level, the prevalent microorganisms were , , , , , and . The abundance of microbial species varied significantly in different seasons. Further, the genomic presence of some undesirable microbes like , , , and , were also detected in the specific season. The metagenomic analysis also revealed the existence of bacteriophages belonging to the family , , and . Examination of the metabolic potential of the metagenome depicted information about the biocatalysts, presumably involved in the transformation of protein and carbohydrate polymers into bioactive molecules of health-beneficial effects. The genomic resource of several desirable enzymes was identified, such as β-galactosidase, β-glucosidase, β-xylosidase, and glutamate decarboxylase, etc. The catalytic function of a novel glutamate decarboxylase gene was validated for the biosynthesis of γ-aminobutyric acid (GABA). The results of the present study highlight the microbial and genomic resources associated with , and its importance in functional food industry.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.3389/fmicb.2019.01744DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6688588PMC
August 2019

Metabolic diversification of nitrogen-containing metabolites by the expression of a heterologous lysine decarboxylase gene in Arabidopsis.

Plant J 2019 11 27;100(3):505-521. Epub 2019 Aug 27.

Graduate School of Pharmaceutical Sciences, Chiba University, 1-8-1 Inohana, Chuo-ku, Chiba, 260-8675, Japan.

Lysine decarboxylase converts l-lysine to cadaverine as a branching point for the biosynthesis of plant Lys-derived alkaloids. Although cadaverine contributes towards the biosynthesis of Lys-derived alkaloids, its catabolism, including metabolic intermediates and the enzymes involved, is not known. Here, we generated transgenic Arabidopsis lines by expressing an exogenous lysine/ornithine decarboxylase gene from Lupinus angustifolius (La-L/ODC) and identified cadaverine-derived metabolites as the products of the emerged biosynthetic pathway. Through untargeted metabolic profiling, we observed the upregulation of polyamine metabolism, phenylpropanoid biosynthesis and the biosynthesis of several Lys-derived alkaloids in the transgenic lines. Moreover, we found several cadaverine-derived metabolites specifically detected in the transgenic lines compared with the non-transformed control. Among these, three specific metabolites were identified and confirmed as 5-aminopentanal, 5-aminopentanoate and δ-valerolactam. Cadaverine catabolism in a representative transgenic line (DC29) was traced by feeding stable isotope-labeled [α- N]- or [ε- N]-l-lysine. Our results show similar N incorporation ratios from both isotopomers for the specific metabolite features identified, indicating that these metabolites were synthesized via the symmetric structure of cadaverine. We propose biosynthetic pathways for the metabolites on the basis of metabolite chemistry and enzymes known or identified through catalyzing specific biochemical reactions in this study. Our study shows that this pool of enzymes with promiscuous activities is the driving force for metabolite diversification in plants. Thus, this study not only provides valuable information for understanding the catabolic mechanism of cadaverine but also demonstrates that cadaverine accumulation is one of the factors to expand plant chemodiversity, which may lead to the emergence of Lys-derived alkaloid biosynthesis.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1111/tpj.14454DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6899585PMC
November 2019

Prediction of inadvertent internal mammary (thoracic) vein cannulation by CVP monitoring.

J Anaesthesiol Clin Pharmacol 2019 Apr-Jun;35(2):282-283

Department of Anaesthesiology and Critical Care, INHS Asvini, Colaba, Mumbai, Maharashtra, India.

View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.4103/joacp.JOACP_264_18DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6598563PMC
July 2019

Fructose-induced AGEs-RAGE signaling in skeletal muscle contributes to impairment of glucose homeostasis.

J Nutr Biochem 2019 09 8;71:35-44. Epub 2019 Jun 8.

Biochemistry Division, CSIR-Central Drug Research Institute, Lucknow-226031; Academy of Scientific and Innovative Research (AcSIR), New Delhi-201002, India. Electronic address:

Increased fructose intake has been linked to the development of dyslipidemia, obesity and impaired glucose tolerance. Due to its specific metabolic fate, fructose impairs normal lipid and carbohydrate metabolism and facilitates the non-enzymatic glycation reaction leading to enhanced accumulation of advanced glycation end products (AGEs). However, the formation of fructose-AGEs under in vivo setup and its tissue specific accumulation is less explored. Here, we investigated the impact of high fructose on AGEs accumulation in skeletal muscle and its causal role in impaired glucose homeostasis. In L6 rat skeletal muscle cells, chronic exposure to fructose induced AGEs accumulation and the cellular level of the receptor for AGEs (RAGE) and the effect was prevented by pharmacological inhibition of glycation. Under in vivo settings, Sprague Dawley rats exposed to 20% fructose in drinking water for 16 weeks, displayed increased fasting glycemia, impaired glucose tolerance, decreased skeletal muscle Akt (Ser-473) phosphorylation, and enhanced triglyceride levels in serum, liver and gastrocnemius muscle. We also observed a high level of AGEs in serum and gastrocnemius muscle of fructose-supplemented animals, associated with methylglyoxal accumulation and up regulated expression of RAGE in gastrocnemius muscle. Treatment with aminoguanidine inhibited fructose-induced AGEs accumulation and normalized the expression of RAGE and Dolichyl-Diphosphooligosaccharide-Protein Glycosyltransferase (DDOST) in gastrocnemius muscle. Inhibition of AGEs-RAGE axis counteracted fructose-mediated glucose intolerance without affecting energy metabolism. These data reveal diet-derived AGEs accumulation in skeletal muscle and the implication of tissue specific AGEs in metabolic derangement, that may open new perspectives in pathogenic mechanisms and management of metabolic diseases.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jnutbio.2019.05.016DOI Listing
September 2019

Publisher Correction: A cheminformatics approach to characterize metabolomes in stable-isotope-labeled organisms.

Nat Methods 2019 May;16(5):446

RIKEN Center for Sustainable Resource Science, Yokohama, Japan.

In the originally published Supplementary Information for this paper, the files presented as Supplementary Tables 3, 4, and 7 were duplicates of Supplementary Tables 5, 6, and 9, respectively. All Supplementary Table files are now correct online.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41592-019-0423-xDOI Listing
May 2019

Biosynthesis of 2,5-furan dicarboxylic acid by Aspergillus flavus APLS-1: Process optimization and intermediate product analysis.

Bioresour Technol 2019 Jul 21;284:155-160. Epub 2019 Mar 21.

Microbial Processes and Technology Division, CSIR-National Institute for Interdisciplinary Science and Technology, Thiruvananthapuram 695019, Kerala, India. Electronic address:

The aim of the present study was to develop an eco-friendly biological process for the production of 2,5-furan dicarboxylic acid (FDCA) from 5-hydroxy methylfurfuraldehyde (HMF) using microorganisms. Microorganisms were isolated from the soil samples and evaluated for its biotransformation efficiency. Among the isolates, Aspergillus flavus APLS-1 was found to be potent for efficient conversion of HMF to FDCA. The bioconversion parameters were optimized by Box-Behnken design. The optimization resulted in 67% conversion efficiency where 1 g/L HMF (8 mM) was transformed to 0.83 g/L (6.6 mM) FDCA in 14 days at pH6.5 with biomass size of 5.7 g/L and biomass age 60 h. This is the first report on Aspergillus sp., capable of detoxifying HMF and produces FDCA.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.biortech.2019.03.105DOI Listing
July 2019

A cheminformatics approach to characterize metabolomes in stable-isotope-labeled organisms.

Nat Methods 2019 04 28;16(4):295-298. Epub 2019 Mar 28.

RIKEN Center for Sustainable Resource Science, Yokohama, Japan.

We report a computational approach (implemented in MS-DIAL 3.0; http://prime.psc.riken.jp/) for metabolite structure characterization using fully C-labeled and non-labeled plants and LC-MS/MS. Our approach facilitates carbon number determination and metabolite classification for unknown molecules. Applying our method to 31 tissues from 12 plant species, we assigned 1,092 structures and 344 formulae to 3,604 carbon-determined metabolite ions, 69 of which were found to represent structures currently not listed in metabolome databases.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1038/s41592-019-0358-2DOI Listing
April 2019

Bacterial colonization of peripheral intravenous cannulas in a tertiary care hospital: A cross sectional observational study.

Med J Armed Forces India 2019 Jan 6;75(1):65-69. Epub 2018 Jul 6.

Commandant, 174 Military Hospital, C/o 56 APO, India.

Background: The use of intravenous (IV) cannulas is an integral part of patient care in hospitals. These intravenous cannulas are a potential route for microorganisms to enter the blood stream resulting in a variety of local or systemic infections. Studies showing the actual prevalence of colonization of peripheral IV cannulas and its role in BSI are lacking. Hence, this study was aimed to estimate the prevalence of colonization of the injection ports of peripheral IV cannulas.

Methods: This cross sectional study was conducted on patients admitted in ICU and wards in an 800 bedded tertiary care hospital. Swabs were taken from lumens of peripheral IV cannulas and cultured. Patient demographic data and practices followed for maintenance of IV line were noted.

Results: A total of 196 injection port samples were taken, out of which 11 tested positive for microbial growth (5.61%). was the predominant organism contributing 64% of the microbial growth. A significant association was seen between presence of local signs, old age and positive cultures. Flushing IV cannula every 6 h was associated with negative cultures.

Conclusion: Peripheral IV cannulation has significant potential for microbial contamination and is largely ignored. Most of the risk factors associated with growth of microorganisms in the injection ports of peripheral intravenous cannulas (which has a potential to cause catheter-related blood stream infections) can be prevented by improving protocols for management. To prevent infection from occurring, practitioners should be educated and trained about the care and management of IV.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.mjafi.2018.04.004DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6349642PMC
January 2019

Nod1-mediated lipolysis promotes diacylglycerol accumulation and successive inflammation via PKCδ-IRAK axis in adipocytes.

Biochim Biophys Acta Mol Basis Dis 2019 01 2;1865(1):136-146. Epub 2018 Nov 2.

Division of Biochemistry, CSIR-Central Drug Research Institute, Lucknow 226031, India; Academy of Scientific and Innovative Research (AcSIR), New Delhi 201002, India. Electronic address:

Chronic inflammation contributes to obesity mediated metabolic disturbances, including insulin resistance. Obesity is associated with altered microbial load in metabolic tissues that can contribute to metabolic inflammation. Different bacterial components such as, LPS, peptidoglycans have been shown to underpin metabolic disturbances through interaction with host innate immune receptors. Activation of Nucleotide-binding oligomerization domain-containing protein 1 (Nod1) with specific peptidoglycan moieties promotes insulin resistance, inflammation and lipolysis in adipocytes. However, it was not clear how Nod1-mediated lipolysis and inflammation is linked. Here, we tested if Nod1-mediated lipolysis caused accumulation of lipid intermediates and promoted cell autonomous inflammation in adipocytes. We showed that Nod1-mediated lipolysis caused accumulation of diacylglycerol (DAG) and activation of PKCδ in 3T3-L1 adipocytes, which was prevented with a Nod1 inhibitor. Nod1-activated PKCδ caused downstream stimulation of IRAK1/4 and was associated with increased expression of proinflammatory cytokines such as, IL-1β, IL-18, IL-6, TNFα and MCP-1. Pharmacological inhibition or siRNA mediated knockdown of IRAK1/4 attenuated Nod1-mediated activation of NF-κB, JNK, and the expression of proinflammatory cytokines. These results reveal that Nod1-mediated lipolysis promoted accumulation of DAG, which engaged PKCδ and IRAK1/4 to augment inflammation in 3T3-L1 adipocytes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.bbadis.2018.10.036DOI Listing
January 2019

Functional validation of the Pi54 gene by knocking down its expression in a blast-resistant rice line using RNA interference and its effects on other traits.

Funct Plant Biol 2018 11;45(12):1241-1250

Indian Council of Agricultural Research (ICAR) National Research Centre on Plant Biotechnology, New Delhi-110012, India.

Rice blast disease caused by Magnaporthe oryzae is one of the major diseases affecting the rice (Oryza sativa L.) crop. A major blast resistance gene, Pi54, has already been cloned and deployed in different rice varieties. To understand the role of Pi54 in providing rice blast resistance, we used the RNA interferences (RNAi) approach to knock down the expression of this gene. We showed a high frequency of Agrobacterium tumefaciens-mediated transformation of rice line Taipei 309 containing a single gene (Pi54) for blast resistance. Pi54 RNAi leads to a decreased level of Pi54 transcripts, leading to the susceptibility of otherwise M. oryzae-resistant rice lines. However, among the RNAi knockdown plants, the severity of blast disease varied between the lines. Histochemical analysis of the leaves of knockdown plants inoculated with M. oryzae spores also showed typical cell death and blast lesions. Additionally, Pi54 RNAi also showed an effect on the Hda3 gene, a florigen gene playing a role in rice flowering. By using the RNAi technique, for the first time, we showed that the directed degradation of Pi54 transcripts results in a significant reduction in the rice blast resistance response, suggesting that RNAi is a powerful tool for functional validation of genes.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1071/FP18083DOI Listing
November 2018