Evidence for Rho-dependent control of a virulence switch in Acinetobacter baumannii.

IMPORTANCE: Acinetobacter baumannii is a significant cause of infections in the healthcare setting. More recently, A. baumannii has been a leading cause of secondary bacterial pneumonia in patients infected with SARS-CoV-2 and the overall frequency of A. baumannii infection increased 78% during the COVID-19 pandemic. A. baumannii can exist in virulent or avirulent subpopulations and this interconversion is mediated by the expression of a family of TetR-type transcriptional regulators. In this study, we demonstrate that Rho is a key regulatory component in the expression of these TetR regulators. Overall, this study is the first to address a role for Rho in A. baumannii and provides additional evidence for the role of Rho in regulating diversity in bacterial subpopulations.

Indoor air quality guidelines from across the world: An appraisal considering energy saving, health, productivity, and comfort.

Buildings are constructed and operated to satisfy human needs and improve quality of life. Good indoor air quality (IAQ) and thermal comfort are prerequisites for human health and well-being. For their provision, buildings often rely on heating, ventilation, and air conditioning (HVAC) systems, which may lead to higher energy consumption. This directly impacts energy efficiency goals and the linked climate change considerations. The balance between energy use, optimum IAQ and thermal comfort calls for scientifically solid and well-established limit values for exposures experienced by building occupants in indoor spaces, including homes, schools, and offices. The present paper aims to appraise limit values for selected indoor pollutants reported in the scientific literature, and to present how they are handled in international and national guidelines and standards. The pollutants include carbon dioxide (CO2), formaldehyde (CH2O), particulate matter (PM), nitrogen dioxide (NO2), carbon monoxide (CO), and radon (Rn). Furthermore, acknowledging the particularly strong impact on energy use from HVAC, ventilation, indoor temperature (T), and relative humidity (RH) are also included, as they relate to both thermal comfort and the possibilities to avoid moisture related problems, such as mould growth and proliferation of house dust mites. Examples of national regulations for these parameters are presented, both in relation to human requirements in buildings and considering aspects related to energy saving. The work is based on the Indoor Environmental Quality (IEQ) guidelines database, which spans across countries and institutions, and aids in taking steps in the direction towards a more uniform guidance for values of indoor parameters. The database is coordinated by the Scientific and Technical Committee (STC) 34, as part of ISIAQ, the International Society of Indoor Air Quality and Climate.

Compliance With Higher Education-Related Tobacco Control Law Provisions by Institutions of National Importance in India.

Introduction India formulated an anti-tobacco and anti-smoking law in 2003 in response to its resolutions in the United Nations' bodies. This law has been detailed subsequently to make it focussed on educational institutions, which are supposed to perform on-ground action in a decentralized manner. The first step is to put up signboards prohibiting the sale of cigarettes and tobacco products within 100 yards, the second is prohibiting smoking within the campus, and the third is implementing the law related to collecting fines from the offenders as well as the presence of vendors within 100 yards. Additional information on awareness activities was also sought. The focus of this paper is on India's premier educational institutions called the "Institutions of National Importance" by the Indian legislature. These are India's premier public institutions which have the maximum focus of the Indian government by making them of high quality, and non-compliance with the tobacco control law in these institutions should be taken seriously. Methods The paper checked for compliance with the Indian Tobacco Control Law in 79 of these Institutions of National Importance. The requirement for information to be collected from institutions was first derived from the legal act and the rules. Then, the Right to Information Act of 2005, India's transparency law, was used to file applications for information, and certified information from the institutions was collected and reported. Results Only 39.2% of the institutions had the requisite boards prohibiting sales within 100 yards of the institutions. The requirement of having boards prohibiting smoking fared a little better at 73.4% of institutions complying but was not universal. A total of 43% of institutions denied the information pertaining to the collection of fines, either by not providing a requisite reply or stating that this was not part of the record. The information regarding the presence of vendors was not universally supplied with 65.8% of institutions stating the same to not in record or not within the purview. With respect to the awareness activities though, 72.2% of the institutions stated to have some awareness activity for tobacco control and cessation. Conclusion The results show an overall weak compliance with the law. India's health regulators and educational watchdogs must implement anti-smoking and anti-tobacco laws strictly in Indian educational institutions as this is where young people are found. Something as simple as the installation of signboards by educational institutions cannot be overlooked by them. The law must become stricter with deterrence. This must be most intense in the Institutions of National Importance which form the premier institutions in India and become the role model for other institutions in India.

Decentralisation of the Compliance of Anti-tobacco Law in India: The Case of Higher Educational Institutions in New Delhi, India.

Introduction This paper studies the decentralised compliance responsibility of India's tobacco control legislation, called the Cigarettes and Other Tobacco Products (Prohibition of Advertisement and Regulation of Trade and Commerce, Production, Supply and Distribution) Act of 2003, with its rules, which the government has outsourced to higher educational institutions, studied through an example of New Delhi. The study looks into the three most important parameters of decentralised compliance. Two of these require the installation of signboards by higher educational institutions, and the third involves imposing and collecting fines against persons found smoking within the educational institutions. Regarding the boards, the first board is about the warning prohibiting the sale within 100 yards of educational institutions, and the second one prohibits smoking in educational institutions. The study also checks with the educational institutions whether there is a presence of cigarette and tobacco product vendors within 100 yards of the institution, where the sale of such products has been banned by law. The study also found educational activities to create awareness in the institutions for tobacco control and cessation. Methods Thirty-six higher educational institutions, including universities, were randomly selected and studied through a unique methodology using India's transparency law, i.e., the Right to Information Act 2005. The portions of the law, which was direct compliance, or related compliance by the higher educational institution was included in the study. This justified the decentralised responsibility of these higher educational institutions. Applications for information under the transparency law were requested and supplied. Out of the 47 institutions, in which information requests were filed, 36 provided the information under the law. Credible information was provided by the higher educational institutions and this information was collated and interpreted to provide insights into the compliance by the higher educational institutions. Results Only 44.4% of the institutions had a board prohibiting the sale of cigarettes and other tobacco products. This was a non-universal compliance by the higher educational institutions. 88.9 per cent had boards prohibiting smoking in the higher educational institutions. Only one out of the 36 had an instance of smoking recorded and collection of fines. While 47.2% reported to not have any instance of smoking and fine collection. Fifty per cent of the institutions had no record of fine collection amounting to defiance of the law. 75 per cent of institutions did some kind of awareness activities which was not a direct statutory requirement, but a recommended guideline. Conclusion The results show that the intent to decentralise the compliance of the tobacco control law in New Delhi by the higher educational institutions has not been universally successful and requires much effort for its on-ground penetration. Such studies have a policy impact as they serve as an example for the generalisability of such statutes, which only work when there is implementation from the bottom-up and when the deterrent is also strong with incentivisation to the educational institutions to implement tobacco control with vigour.

Disparities in prostate cancer screening, diagnoses, management, and outcomes between Indigenous and non-Indigenous men in a universal health care system.

BACKGROUND: Indigenous Peoples have higher morbidity rates and lower life expectancies than non-Indigenous Canadians. Identification of disparities between Indigenous and non-Indigenous men regarding prostate cancer (PCa) screening, diagnoses, management, and outcomes was sought. METHODS: An observational cohort of men diagnosed with PCa between June 2014 and October 2022 was studied. Men were prospectively enrolled in the province-wide Alberta Prostate Cancer Research Initiative. The primary outcomes were tumor characteristics (stage, grade, and prostate-specific antigen [PSA]) at diagnosis. Secondary outcomes were PSA testing rates, time from diagnosis to treatment, treatment modality, and metastasis-free, cancer-specific, and overall survivals. RESULTS: Examination of 1,444,974 men for whom aggregate PSA testing data were available was performed. Men in Indigenous communities were less likely to have PSA testing performed than men outside of Indigenous communities (32 vs. 46 PSA tests per 100 men [aged 50-70 years] within 1 year; p < .001). Among 6049 men diagnosed with PCa, Indigenous men had higher risk disease characteristics: a higher proportion of Indigenous men had PSA ≥ 10 ng/mL (48% vs. 30%; p < .01), TNM stage ≥ T2 (65% vs. 47%; p < .01), and Gleason grade group ≥ 2 (79% vs. 64%; p < .01) compared to non-Indigenous men. With a median follow-up of 40 months (interquartile range, 25-65 months), Indigenous men were at higher risk of developing PCa metastases (hazard ratio, 2.3; 95% CI, 1.2-4.2; p < .01) than non-Indigenous men. CONCLUSIONS: Despite receiving care in a universal health care system, Indigenous men were less likely to receive PSA testing and more likely to be diagnosed with aggressive tumors and develop PCa metastases than non-Indigenous men.

Does the Presence of Asbestos-Containing Materials in Buildings Post-construction and Before Demolition Have an Impact on the Exposure to Occupants in Non-occupational Settings?

This narrative review aims to determine if asbestos-containing materials in buildings pose a hazard to building occupants in non-occupational settings. This paper is limited to the post-construction and pre-demolition stages of a building. The researchers selected 19 studies from the 126 studies screened, concerning exposure to asbestos fibers in non-occupational building settings, with a focus on post-construction and pre-demolition phases. The literature review found that certain conditions, such as the measurement techniques, standards, and previous data availability, prevent a conclusive answer to the research question. Some studies have pointed towards an effect of asbestos-containing materials on health of occupants in non-occupational settings. But, there are some that do not suggest a positive relationship between non-occupational exposure and the presence of asbestos-containing materials, and therefore these provide scope for further research, as these studies also do not rule out the relationship completely. The present study highlights the gaps in current knowledge and indicates areas for further research. Until conclusive evidence based on revised threshold standards and accurate measurement techniques is available, asbestos-containing materials may be considered unsafe for use in non-occupational settings, especially ones that young people and children occupy.

Pedagogy for Teaching Fire Safety through Design-Based Immersion of the National Building Code 2016 with Feedback from Students of Undergraduate Architecture.

This study is based on the feedback from 3rd-year architectural undergraduate students at a leading architectural education college in India. An undergraduate degree in architecture in India leads to a professional license to practice as an architect in India. Fire safety is also a component of the architectural curriculum, but there are concerns worldwide that architecture colleges may somehow not give the impetus fire safety education may require. A studio-based, immersive pedagogy was created to make fire safety more relevant and easy to grasp for architecture students. This method used integrating the interventions from the country's fire code into the design using students' self-created design problems, with which they were familiar. This design-based immersive integration of the National Building Code 2016 and its fire provisions were tested in this study. The detailed course pedagogical structure has been presented. The study was tested using feedback from the students at the end of the semester using an 11-part questionnaire which 32 students answered in an anonymous mode. The results show an overall positive response where the students prefer a design-based integrated fire safety curriculum which introduces fire codes to the students in the applied format. This study paves the way for more replications of the studio design-based integration of fire codes into the curricula of architecture colleges. Further studies will require this technique to undergo further testing by involving practitioners who have undergone this pedagogy and testing the same in building projects.

Unraveling topoisomerase IA gate dynamics in presence of PPEF and its preclinical evaluation against multidrug-resistant pathogens.

Type IA topoisomerases maintain DNA topology by cleaving ssDNA and relaxing negative supercoils. The inhibition of its activity in bacteria prevents the relaxation of negative supercoils, which in turn impedes DNA metabolic processes leading to cell death. Using this hypothesis, two bisbenzimidazoles, PPEF and BPVF are synthesized, selectively inhibiting bacterial TopoIA and TopoIII. PPEF stabilizes the topoisomerase and topoisomerase-ssDNA complex, acts as an interfacial inhibitor. PPEF display high efficacy against ~455 multi-drug resistant gram positive and negative bacteria. To understand molecular mechanism of inhibition of TopoIA and PPEF, accelerated MD simulation is carried out, and results suggested that PPEF binds, stabilizes the closed conformation of TopoIA with -6Kcal/mol binding energy and destabilizes the binding of ssDNA. The TopoIA gate dynamics model can be used as a tool to screen TopoIA inhibitors as therapeutic candidates. PPEF and BPVF cause cellular filamentation and DNA fragmentation leading to bacterial cell death. PPEF and BPVF show potent efficacy against systemic and neutropenic mouse models harboring E. coli, VRSA, and MRSA infection without cellular toxicity.

Moonlighting proteins: beacon of hope in era of drug resistance in bacteria.

Moonlighting proteins (MLPs) are ubiquitous and provide a unique advantage to bacteria performing multiple functions using the same genomic content. Targeting MLPs can be considered as a futuristic approach in fighting drug resistance problem. This review follows the MLP trail from its inception to the present-day state, describing a few bacterial MLPs, viz., glyceraldehyde 3'-phosphate dehydrogenase, phosphoglucose isomerase glutamate racemase (GR), and DNA gyrase. Here, we carve out that targeting MLPs are the beacon of hope in an era of increasing drug resistance in bacteria. Evolutionary stability, structure-functional relationships, protein diversity, possible drug targets, and identification of new drugs against bacterial MLP are given due consideration. Before the final curtain calls, we provide a comprehensive list of small molecules that inhibit the biochemical activity of MLPs, which can aid the development of novel molecules to target MLPs for therapeutic applications.

Public Health Issue of Indoor Dilution Ventilation for Disease Prevention Versus PM2.5 in Intake Air in Auditoriums of Premier Engineering Institutes in India.

BACKGROUND: Dilution ventilation by enhancing fresh air intake has been prescribed to reduce airborne infection spread during the COVID-19 pandemic. This is all the more important in assembly spaces like auditoriums. Premier technology institutes have large campuses with large auditoriums for academic and cultural events in India. These institutes serve as role models for society, where gatherings are essential, but there is also the possibility of transmission of all airborne respiratory infections, including tuberculosis, into the community. The fresh air taken in should also be filtered for pollution to prevent other lung issues. AIMS: Fresh air intake and filtration have been studied in order to understand whether the outside air supplied indoors is filtered for PM2.5, which is a major ambient polluter in India. Settings and design/methods: In this study, the Right to Information Act of 2005 has been used to obtain first-hand information from the institutes with respect to the heating, ventilation, and air conditioning (HVAC) systems in their auditoriums. Twelve of the 19 institutes fall in cities with non-attainment of ambient air quality standards. RESULTS: Eleven out of all those had recently integrated fresh air supply, and six replied in the negative. Only one out of all of them had appropriate filters. CONCLUSION: This study highlights the need for a possible trade-off between the use of air conditioners for thermal comfort + assumed protection against PM2.5, which is the switching off of air conditioners and manually opening up windows and using fans for ventilation. Indian HVAC design for gathering spaces, especially educational institutes, needs to factor in fresh air for dilution ventilation as well as PM2.5 filtration.

Studying the Double Paradox in Air Conditioning at Indian Airports for Airborne Infection Prevention and Filtration of Harmful Suspended Particulate Matter.

Background Airports are hubs of diverse human interactions. During pandemics, they may serve as centers for the spread of airborne infection. Appropriate methods for the prevention of the spread of airborne infections must be integrated into the air conditioning systems of airports. Along with ultraviolet germicidal irradiation and other sanitization methods, dilution ventilation can be the easiest and most available method for the prevention of airborne infection, which means the intake of outside air into the indoors, which flushes out the aerosolized droplets containing pathogens. Though this process has been adopted by multiple buildings in reaction to the pandemic, it may present the challenge of intake of high concentration of suspended particulate matter in the intake air, a major air pollutant in developing countries, which may enter through the air conditioning systems. Appropriate filtration is necessary so that along with dilution ventilation for airborne disease prevention, the risk of suspended particulate matter of diameter 2.5 micron or PM2.5 induced lung issues is also reduced. Methodology The Right to Information Act, 2005, was used to file applications for information on the details of the air conditioning systems in Indian airports. The 58 airports in the study were also listed according to the list of cities that fall under the criteria for non-attainment of good air quality standards. Results Out of 58 airports considered, 27 fell in the 'non-attainment' of good air quality list. On appraisal of filter systems, it was found that 23 had an intake of fresh air but only five had filters with a minimum efficiency reporting value (MERV) of 10 and above in their air conditioning systems, as is recommended for filtration of suspended particulate matter. Conclusion It can be concluded that most airports did not have the appropriate filter required for filtering PM2.5, which is a major pollutant in Indian cities. In light of coronavirus disease 2019, where dilution ventilation through the intake of outdoor air is suggested, it may also lead to the entry of air with high particulate matter into the indoors.

The Risk Status of Waiting Areas for Airborne Infection Control in Delhi Hospitals.

Background Hospital waiting areas are overlooked from the airborne infection control viewpoint as they are not classified as critical for infection control. This is the area where undiagnosed and potentially infected patients gather with susceptible and vulnerable patients, and there is no mechanism to segregate the two, especially when the potentially infected visitors/patients themselves are unaware of the infection or may be asymptomatic. It is important to know whether hospitals in Delhi, a populated, low-resource setting having community transmission/occurrence of airborne diseases such as tuberculosis, consider waiting areas as critical. Hence, this study aims to determine whether hospitals in Delhi consider waiting areas as critical areas from the airborne infection control viewpoint. Methodology The Right to Information Act, 2005, was used to request information from 11 hospitals included in this study. Results After compiling the results, it was found that five out of the 11 hospitals did not consider waiting areas as critical from the infection spread point of view. Two of the 11 hospitals acknowledged the criticality of waiting areas but did not include the same in the list of critical areas. Only three out of the 11 considered waiting areas as critical and included these in the list of critical areas in a hospital. Conclusions This study provided evidence that most hospitals in Delhi do not include waiting areas in the list of critical areas in a hospital.

Synthesis and Biological Evaluation of Novel 1H-Benzo[d]imidazole Derivatives as Potential Anticancer Agents Targeting Human Topoisomerase I.

Small molecules that modulate biological functions are targets of modern-day drug discovery efforts. A new series of novel 1H-benzo[d]imidazoles (BBZs) were designed and synthesized with different functional groups at the phenyl ring and variable lengths of the alkyl chain at the piperazine end as anticancer agents. We identified human topoisomerase I (Hu Topo I) as a probable target of these molecules through a computational study and DNA relaxation assay, a functional assay of the Hu Topo I enzyme. UV absorption, fluorescence, and circular dichroism spectroscopy were used to study interactions between BBZ and DNA. Out of 16 compounds, 11a, 12a, and 12b showed strong binding affinity and thermal stabilization of AT sequence-specific DNA. BBZs were screened against a panel of 60 human cancer cell lines at National Cancer Institute, USA. Most potent molecules 11a, 12a, and 12b showed 50% growth inhibition (GI50) in a concentration range from 0.16 to 3.6 µM cancer cells. Moreover, 12b showed 50% inhibition of the relaxation of DNA by Hu Topo I at 16 µM. Furthermore, flow cytometry revealed that 11a, 12a, and 12b cause prominent G2M arrest of cancer cells. In view of the above, we propose that 12b deserves to be further evaluated for its therapeutic use as an anticancer agent.

Pyrazoline analogs as potential anticancer agents and their apoptosis, molecular docking, MD simulation, DNA binding and antioxidant studies.

N-formyl pyrazoline derivatives (3a-3l) were designed and synthesized via Michael addition reaction through cyclization of chalcones with hydrazine hydrate in presence of formic acid. The structural elucidation of N-formyl pyrazoline derivatives was carried out by various spectroscopic techniques such as 1H, 13C NMR, FT-IR, UV-visible spectroscopy, mass spectrometry and elemental analysis. Anticancer activity of the pyrazoline derivatives (3a-3l) was evaluated against human lung cancer (A549), fibrosarcoma cell lines (HT1080) and human primary normal lung cells (HFL-1) by MTT assay. The results of anticancer activity showed that potent analogs 3b and 3d exhibited promising activity against A549 (IC50 = 12.47 ± 1.08 and 14.46 ± 2.76 µM) and HT1080 (IC50 = 11.40 ± 0.66 and 23.74 ± 13.30 µM) but low toxic against the HFL-1 (IC50 = 116.47 ± 43.38 and 152.36 ± 22.18 µM). The anticancer activity of potent derivatives (3b and 3d) against A549 cancer cell line was further confirmed by flow cytometry based approach. DNA binding interactions of the pyrazoline derivatives 3b and 3d have been carried out with calf thymus DNA (Ct-DNA) using absorption, fluorescence and viscosity measurements, circular dichroism and cyclic voltammetry. Antioxidant potential of N-formyl pyrazoline derivatives (3a-3l) has been also estimated through DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical and H2O2. Results revealed that all the compounds exhibited significant antioxidant activity. In silico molecular modelling and ADMET properties of pyrazoline derivatives were also studied using PyRx software against topoisomerase II receptor with PDB ID: 1ZXM to explore their best hits. MD simulation of 3b and 3d was also carried out with topoisomerase II for structure-function correlation in a protein. HuTopoII inhibitory activity of the analogs (3a-3l) was examined by relaxation assay at varying concentrations 100-1000 µM.

Gut microbiome-mediated epigenetic regulation of brain disorder and application of machine learning for multi-omics data analysis.

The gut-brain axis (GBA) is a biochemical link that connects the central nervous system (CNS) and enteric nervous system (ENS). Clinical and experimental evidence suggests gut microbiota as a key regulator of the GBA. Microbes living in the gut not only interact locally with intestinal cells and the ENS but have also been found to modulate the CNS through neuroendocrine and metabolic pathways. Studies have also explored the involvement of gut microbiota dysbiosis in depression, anxiety, autism, stroke, and pathophysiology of other neurodegenerative diseases. Recent reports suggest that microbe-derived metabolites can influence host metabolism by acting as epigenetic regulators. Butyrate, an intestinal bacterial metabolite, is a known histone deacetylase inhibitor that has shown to improve learning and memory in animal models. Due to high disease variability amongst the population, a multi-omics approach that utilizes artificial intelligence and machine learning to analyze and integrate omics data is necessary to better understand the role of the GBA in pathogenesis of neurological disorders, to generate predictive models, and to develop precise and personalized therapeutics. This review examines our current understanding of epigenetic regulation of the GBA and proposes a framework to integrate multi-omics data for prediction, prevention, and development of precision health approaches to treat brain disorders.

The impact of epigenetics on cardiovascular disease.

Mortality and morbidity from cardiovascular diseases (CVDs) represents a huge burden to society. It is recognized that environmental factors and individual lifestyles play important roles in disease susceptibility, but the link between these external risk factors and our genetics has been unclear. However, the discovery of sequence-independent heritable DNA changes (epigenetics) have helped us to explain the link between genes and the environment. Multiple diverse epigenetic processes, including DNA methylation, histone modification, and the expression of non-coding RNA molecules affect the expression of genes that produce important changes in cellular differentiation and function, influencing the health and adaptability of the organism. CVDs such as congenital heart disease, cardiomyopathy, heart failure, cardiac fibrosis, hypertension, and atherosclerosis are now being viewed as much more complex and dynamic disorders. The role of epigenetics in these and other CVDs is currently under intense scrutiny, and we can expect important insights to emerge, including novel biomarkers and new approaches to enable precision medicine. This review summarizes the recent advances in our understanding of the role of epigenetics in CVD.

PPEF: A bisbenzimdazole potent antimicrobial agent interacts at acidic triad of catalytic domain of E. coli topoisomerase IA.

BACKGROUND: Topoisomerase is a well known target to develop effective antibacterial agents. In pursuance of searching novel antibacterial agents, we have established a novel bisbenzimidazole (PPEF) as potent E. coli topoisomerase IA poison inhibitor. METHODS: In order to gain insights into the mechanism of action of PPEF and understanding protein-ligand interactions, we have produced wild type EcTopo 67 N-terminal domain (catalytic domain) and its six mutant proteins at acidic triad (D111, D113, E115). The DDE motif is replaced by alanine (A) to create three single mutants: D111A, D113A, E115A and three double mutants: D111A-D113A, D113A-E115A and D111A-E115A. RESULTS: Calorimetric study of PPEF with single mutants showed 10 fold lower affinity than that of wild type EcTopo 67 (7.32 × 106 M-1for wild type, 0.89 × 106 M-1for D111A) and 100 fold lower binding with double mutant D113A-E115A (0.02 × 106 M-1) was observed. The mutated proteins showed different CD signature as compared to wild type protein. CD and fluorescence titrations were done to study the interaction between EcTopo 67 and ligands. Molecular docking study validated that PPEF has decreased binding affinity towards mutated enzymes as compared to wild type. CONCLUSION: The overall study reveals that PPEF binds to D113 and E115 of acidic triad of EcTopo 67. Point mutations decrease binding affinity of PPEF towards DDE motif of topoisomerase. GENERAL SIGNIFICANCE: This study concludes PPEF as poison inhibitor of E. coli Topoisomerase IA, which binds to acidic triad of topoisomerase IA, responsible for its function. PPEF can be considered as therapeutic agent against bacteria.

Correction: Discerning morpho-anatomical, physiological and molecular multiformity in cultivated and wild genotypes of lentil with reconciliation to salinity stress.

[This corrects the article DOI: 10.1371/journal.pone.0177465.].

Discerning morpho-anatomical, physiological and molecular multiformity in cultivated and wild genotypes of lentil with reconciliation to salinity stress.

One hundred and sixty two genotypes of different Lens species were screened for salinity tolerance in hydroponics at 40, 80 and 120 mM sodium chloride (NaCl) for 30 d. The germination, seedling growth, biomass accumulation, seedling survivability, salinity scores, root and shoot anatomy, sodium ion (Na+), chloride ion (Cl-) and potassium ion (K+) concentrations, proline and antioxidant activities were measured to evaluate the performance of all the genotypes. The results were compared in respect of physiological (Na+, K+ and Cl-) and seed yield components obtained from field trials for salinity stress conducted during two years. Expression of salt tolerance in hydroponics was found to be reliable indicator for similarity in salt tolerance between genotypes and was evident in saline soil based comparisons. Impressive genotypic variation for salinity tolerance was observed among the genotypes screened under hydroponic and saline field conditions. Plant concentrations of Na+ and Cl- at 120 mM NaCl were found significantly correlated with germination, root and shoot length, fresh and dry weight of roots and shoots, seedling survivability, salinity scores and K+ under controlled conditions and ranked the genotypes along with their seed yield in the field. Root and shoot anatomy of tolerant line (PDL-1) and wild accession (ILWL-137) showed restricted uptake of Na+ and Cl- due to thick layer of their epidermis and endodermis as compared to sensitive cultigen (L-4076). All the genotypes were scanned using SSR markers for genetic diversity, which generated high polymorphism. On the basis of cluster analysis and population structure the contrasting genotypes were grouped into different classes. These markers may further be tested to explore their potential in marker-assisted selection.