Search results for: NaCl tolerant mutant

Authors: Apurva Gupta, Surendra Singh

Abstract:

Spirulina spp., as a promising source of many commercially valuable products, is grown photo autotrophically in open ponds and raceways on a large scale. However, the economic exploitation in an open system seems to have been limited because of lack of multiple stress-tolerant strains. The present study aims to isolate a stable stress tolerant mutant of Spirulina platensis with improved growth rate and enhanced potential to produce its commercially valuable bioactive compounds. N-methyl-n'-nitro-n-nitrosoguanidine (NTG) at 250 μg/mL (concentration permitted 1% survival) was employed for chemical mutagenesis to generate random mutants and screened against NaCl. In a preliminary experiment, wild type S. platensis was treated with NaCl concentrations from 0.5-1.5 M to calculate its LC₅₀. Mutagenized colonies were then screened for tolerance at 0.8 M NaCl (LC₅₀), and the surviving colonies were designated as NaCl tolerant mutants of S. platensis. The mutant cells exhibited 1.5 times improved growth against NaCl stress as compared to the wild type strain in control conditions. This might be due to the ability of the mutant cells to protect its metabolic machinery against inhibitory effects of salt stress. Salt stress is known to adversely affect the rate of photosynthesis in cyanobacteria by causing degradation of the pigments. Interestingly, the mutant cells were able to protect its photosynthetic machinery and exhibited 4.23 and 1.72 times enhanced accumulation of Chl a and phycobiliproteins, respectively, which resulted in enhanced rate of photosynthesis (2.43 times) and respiration (1.38 times) against salt stress. Phycocyanin production in mutant cells was observed to enhance by 1.63 fold. Nitrogen metabolism plays a vital role in conferring halotolerance to cyanobacterial cells by influx of nitrate and efflux of Na+ ions from the cell. The NaCl tolerant mutant cells took up 2.29 times more nitrate as compared to the wild type and efficiently reduce it. Nitrate reductase and nitrite reductase activity in the mutant cells also improved by 2.45 and 2.31 times, respectively against salt stress. From these preliminary results, it could be deduced that enhanced nitrogen uptake and its efficient reduction might be a reason for adaptive and halotolerant behavior of the S. platensis mutant cells. Also, the NaCl tolerant mutant of S. platensis with significant improved growth and phycocyanin accumulation compared to the wild type can be commercially promising.

Keywords: chemical mutagenesis, NaCl tolerant mutant, nitrogen metabolism, photosynthetic machinery, phycocyanin

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Authors: Faheema Khan

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We have investigated the effects of salt stress on the stability of plant growth, water relations, photosynthetic variables, lipid peroxidation and antioxidant system in salt-tolerant (PK-327) and salt-sensitive (PK-471) soybean genotypes. Ten-day-old salt-tolerant and salt-sensitive soybean plants were subjected to 0-150 mM NaCl for 15 days. While the growth of genotype PK-327 was not affected significantly up to 75 mM NaCl treatment, the growth of the PK-471 was reduced significantly beyond 25 mM NaCl treatments. Salt stress caused severe impairments in photosynthetic variables like photosynthetic rate, chlorophyll fluorescence and chlorophyll content, being more pronounced in salt-sensitive genotype than in salt-tolerant.The activities of antioxidant enzymes (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) were higher in PK-327 than in PK-471 at various levels of salt treatments.It is concluded that tolerance capacity of PK-327 against salinity can be associated with the ability of this genotype in keeping an active photosynthetic system and strong antioxidant defence system.

Keywords: salt stress, soybean, antioxidant, photosynthesis

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Authors: Faheema Khan

Abstract:

To investigate the ability of sensitive and tolerant genotype of Glycine max to adapt to a saline environment in a field, we examined the growth performance, water relation and activities of antioxidant enzymes in relation to photosynthetic rate, chlorophyll a fluorescence, photosynthetic pigment concentration, protein and proline in plants exposed to salt stress. Ten soybean genotypes (Pusa-20, Pusa-40, Pusa-37, Pusa-16, Pusa-24, Pusa-22, BRAGG, PK-416, PK-1042, and DS-9712) were selected and grown hydroponically. After 3 days of proper germination, the seedlings were transferred to Hoagland’s solution (Hoagland and Arnon 1950). The growth chamber was maintained at a photosynthetic photon flux density of 430 μmol m−2 s−1, 14 h of light, 10 h of dark and a relative humidity of 60%. The nutrient solution was bubbled with sterile air and changed on alternate days. Ten-day-old seedlings were given seven levels of salt in the form of NaCl viz., T1 = 0 mM NaCl, T2=25 mM NaCl, T3=50 mM NaCl, T4=75 mM NaCl, T5=100 mM NaCl, T6=125 mM NaCl, T7=150 mM NaCl. The investigation showed that genotype Pusa-24, PK-416 and Pusa-20 appeared to be the most salt-sensitive. genotypes as inferred from their significantly reduced length, fresh weight and dry weight in response to the NaCl exposure. Pusa-37 appeared to be the most tolerant genotype since no significant effect of NaCl treatment on growth was found. We observed a greater decline in the photosynthetic variables like photosynthetic rate, chlorophyll fluorescence and chlorophyll content, in salt-sensitive (Pusa-24) genotype than in salt-tolerant Pusa-37 under high salinity. Numerous primers were verified on ten soybean genotypes obtained from Operon technologies among which 30 RAPD primers shown high polymorphism and genetic variation. The Jaccard’s similarity coefficient values for each pairwise comparison between cultivars were calculated and similarity coefficient matrix was constructed. The closer varieties in the cluster behaved similar in their response to salinity tolerance. Intra-clustering within the two clusters precisely grouped the 10 genotypes in sub-cluster as expected from their physiological findings.Salt tolerant genotype Pusa-37, was further analysed by 2-Dimensional gel electrophoresis to analyse the differential expression of proteins at high salt stress. In the Present study, 173 protein spots were identified. Of these, 40 proteins responsive to salinity were either up- or down-regulated in Pusa-37. Proteomic analysis in salt-tolerant genotype (Pusa-37) led to the detection of proteins involved in a variety of biological processes, such as protein synthesis (12 %), redox regulation (19 %), primary and secondary metabolism (25 %), or disease- and defence-related processes (32 %). In conclusion, the soybean plants in our study responded to salt stress by changing their protein expression pattern. The photosynthetic, biochemical and molecular study showed that there is variability in salt tolerance behaviour in soybean genotypes. Pusa-24 is the salt-sensitive and Pusa-37 is the salt-tolerant genotype. Moreover this study gives new insights into the salt-stress response in soybean and demonstrates the power of genomic and proteomic approach in plant biology studies which finally could help us in identifying the possible regulatory switches (gene/s) controlling the salt tolerant genotype of the crop plants and their possible role in defence mechanism.

Keywords: glycine max, salt stress, RAPD, genomic and proteomic variability

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Authors: Abdul Qayyum, Hafiz Muhammad Saeed, Mamoona Hanif, Etrat Noor, Waqas Malik, Shoaib Liaqat

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Salinity is extremely serious problem that has a drastic effect on maize crop, environment and causes economic losses of country. An advance technique to overcome salinity is to develop salt tolerant geno types which require screening of huge germplasm to start a breeding program. Therefore, present study was undertaken to screen out 25 maize hybrids of different origin for salinity tolerance at seedling stage under three levels of salt stress 250 and 300 mM NaCl including one control. The existence of variation for tolerance to enhanced NaCl salinity levels at seedling stage in maize proved that hybrids had differing ability to grow under saline environment and potential variability within specie. Almost all the twenty five maize hybrids behaved varyingly in response to different salinity levels. However, the maize hybrids H6, H13, H21, H23 and H24 expressed better performance under salt stress in terms of all six characters and proved to be as highly tolerant while H22, H17 H20, H18, H4, H9, and H8 were identified as moderately tolerant. Hybrids H14, H5, H11 and H3 H12, H2, were expressed as most sensitive to salinity suggesting that screening is an effective tool to exploit genetic variation among maize hybrids and salt tolerance in maize can be enhanced through selection and breeding procedure.

Keywords: salinity, hybrids, maize, variation

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Authors: Abdu Qayyum, Hafiz Muhammad Saeed, Mamoona Hanif, Etrat Noor, Waqas Malik, Shoaib Liaqat

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Salinity is extremely serious problem that has a drastic effect on maize crop, environment and causes economic losses of country. An advance technique to overcome salinity is to develop salt tolerant geno types which require screening of huge germ plasm to start a breeding program. Therefore, present study was undertaken to screen out 25 maize hybrids of different origin for salinity tolerance at seedling stage under three levels of salt stress 250 and 300 mM NaCl including one control. The existence of variation for tolerance to enhanced NaCl salinity levels at seedling stage in maize proved that hybrids had differing ability to grow under saline environment and potential variability within specie. Almost all the twenty five maize hybrids behaved varyingly in response to different salinity levels. However, the maize hybrids H6, H13, H21, H23 and H24 expressed better performance under salt stress in terms of all six characters and proved to be as highly tolerant while H22, H17 H20, H18, H4, H9, and H8 were identified as moderately tolerant. Hybrids H14, H5, H11 and H3 H12, H2, were expressed as most sensitive to salinity suggesting that screening is an effective tool to exploit genetic variation among maize hybrids and salt tolerance in maize can be enhanced through selection and breeding procedure.

Keywords: salinity, hybrids, maize, variation

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Authors: S. Noreen, S. Ahmad

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Salinity is one of the most important environmental factors that limit the production of crop plants to the greatest proportion than any other ones. Salt-induced changes in growth, pigment concentration, water status, malondialdehydes (MDA) and H₂O₂ content, enzymatic and non-enzymatic antioxidants, Na⁺, K⁺ content and yield attributes were examined in the glasshouse on ten pea (Pisum Sativum L.) accessions, namely ‘13240’, ‘18302’, ‘19666’, ‘19700’, ‘19776’, ‘19785’, ‘19788’, ‘20153’, ‘20155’, ‘26719’ were subjected to non-stress (0 mM NaCl) and salt stress (100 mM and150 mM NaCl) in pots containing sand medium. The results showed that salt stress at level150 mM substantially reduced biomass production, leaf water status, pigment concentration (chlorophyll ‘a’, ‘b’, ‘carotenoid content’ total chlorophyll), K⁺ content, quantum yield and yield attributes as compared to plants treated with 100 mM NaCl. Antioxidant enzymes, Catalase (CAT), Peroxidase (POD), Superoxide dismutase (SOD) and Ascorbate peroxidase (APX), proline content, total soluble protein, total amino acids, Malondialdehyde content (MDA), Hydrogen peroxide (H₂O₂) content and Na⁺ uptake markedly enhanced due to the influence of salt stress. On the basis of analyses (expressed as percent of control), of 10 accessions of pea plant, two were ranked as salt tolerant namely (‘19666’, ‘20153’), four were moderately tolerant namely (‘19700’, ‘19776’, ‘19785’, ‘20155’), and three were salt sensitive namely (‘13240’, ‘18302’, ‘26719’) at 150 mM NaCl level.

Keywords: antioxidant enzymes, ion uptake, pigment concentration, salt stress, yield attributes

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Authors: Samia Alblwi, Amani Ayad

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Mutation testing is the art of generating syntactic variations of a base program and checking whether a candidate test suite can identify all the mutants that are not semantically equivalent to the base: this technique is widely used by researchers to select quality test suites. One of the main obstacles to the widespread use of mutation testing is cost: even small pro-grams (a few dozen lines of code) can give rise to a large number of mutants (up to hundreds): this has created an incentive to seek to reduce the number of mutants while preserving their collective effectiveness. Two criteria have been used to reduce the size of mutant sets: equiva-lence, which aims to partition the set of mutants into equivalence classes modulo semantic equivalence, and selecting one representative per class; subsumption, which aims to define a partial ordering among mutants that ranks mutants by effectiveness and seeks to select maximal elements in this ordering. In this paper we analyze these two policies using analytical and em-pirical criteria.

Keywords: mutation testing, mutant sets, mutant equivalence, mutant subsumption, mutant set minimization

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Authors: Shang-Han Tsai, Yong-Hong Lin, Chung-Ruey Yen

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Wax apple is an important tropical fruit in Taiwan. The famous producing area is located on the coast in Pingtung county. Land subsidence and climate change will tend to soil alkalization more seriously. This study was to evaluate the effects of NaCl in wax apple seedlings. NaCl salinity reduced wax apple shoot growth, it may due to reducing relative water content in leaf and new shoot. Leaf Cl and Na concentration were increased but K, Ca, and Mg content had no significant difference after irrigated with NaCl for six weeks. In roots, Na and Cl content increase significantly with 90 mM NaCl treatment, but K, Ca, and Mg content was reduced. 30-90 mM Nacl treatment do not affect K/Na, Ca/Na, and Mg/Na ratio, but decrease significantly in 90 mM treatment in roots. The leaf and root electrolyte leakage were significantly affected by 90 mM NaCl treatment. Suggesting 90 mM was optimum concentration for sieve out other tolerance wax apple verities.

Keywords: growth, NaCl stress, nutrient, wax apple

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Authors: Bassam O AlJohny

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The growth of Bacillus aquimaris was inhibited from 6 - 20 % of NaCl but it showed some tolerance when Diatomaceous earth (DE) added from 2 - 12% NaCl. Concerning the effect of NaCl on polyol production, we can conclude that, the test bacterium showed some tolerance to NaCl by producing glycerol up to 8 % of NaCl. Then decreased sharply. The addition of DE decrease the amount of polyol and glycerol remarkably and this due to the productive effect of DE to the bacterial cells. The SEM figures represented the presence of electron dense bodies due to the accumulation of small particles of DE as protective molecules.

Keywords: Bacillus aquimaris, Diatomaceous earth (DE), osmoticstress, sodium chloride

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Authors: Kanlaya Kong-Ngern, Chutima Homwonk, Sittiruk Roytrakul

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In this research, we compared the proteomic profile of two rice leaf sheaths under salt stress, Thai moderately salt tolerant rice (Leaung Anan), and high salt tolerant rice (Pokkali). Seeds were grown in hydroponic culture for 21 days before NaCl was introduced initially at the level of 12 dS m⁻¹ for 10 days. Then the leaf sheath proteomes were analyzed by 1D-SDS-PAGE and NanoLC-MS/MS. In this study, 873 proteins were detected. Among these proteins, 219 proteins were known proteins and the other proteins were unnamed and unknown proteins. By using Mev software, we found that only 31 proteins in treated plants of both rice cultivars significantly expressed, 21 proteins were up-regulated and 10 proteins were down-regulated. Interestingly, the intensity of the 3 proteins in the Leaung Anan more expressed than in the Pokkali. The results indicate that the up-regulated proteins were more expressed in less tolerant rice may play an important role in helping rice to survive under salt stress.

Keywords: mass spectrometry, proteomics, rice leaf sheaths, salt stress

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Authors: Hülya Torun, Ondřej Novák, Jaromír Mikulík, Miroslav Strnad, Faik A. Ayaz

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World climate is changing. Millions of people in the world still face chronic undernourishment for conducting a healthy life and the world’s population is growing steadily. To meet this growing demand, agriculture and food systems must adapt to the adverse effects of climate change and become more resilient, productive and sustainable. From this perspective, to determine tolerant cultivars for undesirable environmental conditions will be necessary food production for sustainable development. Among abiotic stresses, soil salinity is one of the most detrimental global fact restricting plant sources. Development of salt-tolerant lines is required in order to increase the crop productivity and quality in salt-treated lands. Therefore, the objective of this study was to investigate the morphological and physiological responses of barley cultivars accessions to salinity stress by NaCl. For this purpose, it was aimed to determine the crosstalk between some endogenous phytohormones and exogenous salicylic acid (SA) in two different vegetative parts (leaves and roots) of barley (Hordeum vulgare L.; Poaceae; 2n=14; Ince-04) which is detected salt-tolerant. The effects of SA on growth parameters, leaf relative water content (RWC), endogenous phytohormones; including indole-3-acetic acid (IAA), cytokinins (CKs), abscisic acid (ABA), jasmonic acid (JA) and ethylene were investigated in barley cultivars under salinity stress. SA was applied to 17-day-old seedlings of barley in two different ways including before (pre-treated for 24 h) and simultaneously with NaCl stress treatment. NaCl (0, 150, 300 mM) exposure in the hydrophonic system was associated with a rapid decrease in growth parameters and RWC, which is an indicator of plant water status, resulted in a strong up-regulation of ABA as a stress indicator. Roots were more dramatically affected than leaves. Water conservation in 150 mM NaCl treated-barley plants did not change, but decreased in 300 mM NaCl treated plants. Pre- and simultaneously treatment of SA did not significantly alter growth parameters and RWC. ABA, JA and ethylene are known to be related with stress. In the present work, ethylene also increased, similarly to ABA, but not with the same intensity. While ABA and ethylene increased by the increment of salt concentrations, JA levels rapidly decreased especially in roots. Both pre- and simultaneously SA applications alleviated salt-induced decreases in 300 mM NaCl resulted in the increment of ABA levels. CKs and IAA are related to cell growth and development. At high salinity (300 mM NaCl), CKs (cZ+cZR) contents increased in both vegetative organs while IAA levels stayed at the same level with control groups. However, IAA increased and cZ+cZR rapidly decreased in leaves of barley plants with SA treatments before salt applications (in pre- SA treated groups). Simultaneously application of SA decreased CKs levels in both leaves and roots of the cultivar. Due to increasing concentrations of NaCl in association with decreasing ABA, JA and ethylene content and increments in CKs and IAA were recorded with SA treatments. As results of the study, in view of all the phytohormones that we tested, exogenous SA induced greater tolerance to salinity particularly when applied before salinity stress.

Keywords: Barley, Hordeum vulgare, phytohormones, salicylic acid, salinity

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Authors: Azlin S Azmi, Mus’ab Zainal, Sarina Sulaiman, Azura Amid, Zaki Zainudin

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Microalgae lipid is a promising feedstock for biodiesel production. The objective of this work was to study growth factors affecting marine mutant Synechococcus sp. (PCC 7002) for high lipid production. Four growth factors were investigated; nitrogen-phosporus-potassium (NPK) concentration, light intensity, temperature and NaNO3 concentration on mutant strain growth and lipid production were studied. Design Expert v8.0 was used to design the experimental and analyze the data. The experimental design selected was Min-Run Res IV which consists of 12 runs and the response surfaces measured were specific growth rate and lipid concentration. The extraction of lipid was conducted by chloroform/methanol solvents system. Based on the study, mutant Synechococcus sp. PCC 7002 gave the highest specific growth rate of 0.0014 h-1 at 0% NPK, 2500 lux, 40oC and 0% NaNO3. On the other hand, the highest lipid concentration was obtained at 0% NPK, 3500 lux, 30°C and 1% NaNO3.

Keywords: Cyanobacteria, lipid, mutant, marine Synechococcus sp. (PCC 7002), specific growth rate

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Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

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PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provides better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

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Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra

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PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provide better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.

Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity

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Authors: A. Nusrath Unissa, Sameer Hassan, Luke Elizabeth Hanna

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Altered drug binding may be an important factor in isoniazid (INH) resistance, rather than major changes in the enzyme’s activity as a catalase or peroxidase (KatG). The identification of structural or functional defects in the mutant KatGs responsible for INH resistance remains as an area to be explored. In this connection, the differences in the binding affinity between wild-type (WT) and mutants of KatG were investigated, through the generation of three mutants of KatG, Ser315Thr [S315T], Ser315Asn [S315N], Ser315Arg [S315R] and a WT [S315]) with the help of software-MODELLER. The mutants were docked with INH using the software-GOLD. The affinity is lower for WT than mutant, suggesting the tight binding of INH with the mutant protein compared to WT type. These models provide the in silico evidence for the binding interaction of KatG with INH and implicate the basis for rationalization of INH resistance in naturally occurring KatG mutant strains of Mycobacterium tuberculosis.

Keywords: Mycobacterium tuberculosis, KatG, INH resistance, mutants, modelling, docking

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Authors: Imed Mezghani, Yousra Ben Salah, Mohamed Chaieb

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Tricholaena teneriffae L. is a xerophytic perennial herb that belongs to the Poaceae family likely to be used for ecological restoration programs. It's a dominant and economically important species widely distributed in the Bou-Hedma National Park, Tunisia. Reintroduction and expansion of T. teneriffae depend solely on sexual reproduction. This makes the understanding of its germination requirements vital for conservation and management. To provide basic information for its conservation and reintroduction, we studied the influence of environmental factors on seed germination patterns. The germination responses of seeds were determined over a wide range of constant temperatures (15–35°C), polyethylene glycol solutions of different osmotic potentials (0 to −2 MPa) and salt solution (0 to 150 mM of NaCl). Results indicated that the optimum temperature germination was attained at 25°C which corresponds to temperatures prevailing during mid spring season in the Mediterranean area. Seeds germinated in Polyethylene Glycol solutions exhibited significantly lower germination than control especially when water potential fell below -0.6 MPa. Germination percentage and rate decreased with an increase NaCl concentration. Seeds germination was substantially delayed and reduced with an increase in NaCl to levels above 50 mM. T. teneriffae is moderately salt tolerant at germination stage.

Keywords: germination, temperature, Tricholaena teneriffae L., salt stress, water stress, rehabilitation

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Authors: F. Z. Aliyat, M. El Guilli, L. Nassiri, J. Ibijbijen

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Climate change is becoming a crucial factor that can significantly impact all ecosystems. It has a negative impact on the environment in many parts of the planet. Agriculture is the main sector affected by climate change. Particularly, the salinity of agricultural soils is among the problems caused by climate change. The use of phosphate solubilizing bacteria (PSB) as a biofertilizer requires previous research on their tolerance to abiotic stress, specifically saline stress tolerance, before the formation of biofertilizers. In this context, the main goal of this research was to assess the salinity tolerance of four strains: Serratia rubidaea strain JCM1240, Enterobacter bugandensis strain 247BMC, Pantoea agglomerans strain ATCC 27155, Pseudomonas brassicacearum subsp. Neoaurantiaca strain CIP109457, which was isolated from solid phosphate sludge. Additionally, their capacity to solubilize potassium and zinc, as well as their effect on Wheat (Triticum Durum 'Karim') germination. The four PSB strains were tested for their ability to solubilize phosphate in NBRIP medium with tricalcium phosphate (TCP) as the sole source of phosphorus under salt stress. Five concentrations of NaCl were used (0%, 0.5%, 1%, 2.5%, 5%). Their phosphate solubilizing activity was estimated by the vanadate-molybdate method. The potassium and zinc solubilization has been tested qualitatively and separately on solid media with mica and zinc oxide as the only sources of potassium and zinc, respectively. The result showed that the solubilization decreases with the increase in the concentration of NaCl; all the strains solubilize the TCP even with 5% NaCl, with a significant difference among the four strains. The Serratia rubidaea strain was the most tolerant strain. In addition, the four strains solubilize the potassium and the zinc. The Serratia rubidaea strain was the most efficient. Therefore, biofertilization with PSB salt-tolerant strains could be a climate-change-preparedness strategy for agriculture in salt soil.

Keywords: bioavailability of mineral nutrients, phosphate solid sludge; phosphate solubilization, potassium solubilization, salt stress, zinc solubilization.

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Authors: Faycal Boughalleb, Raoudha Abdellaoui

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The effect of NaCl stress on root and leaf anatomy was investigated in three Astragalus species grown in 0-300 mM NaCl for 30 days under greenhouse conditions. Root cross section and cortex thickness was reduced under salt stress in both species while A. tenuifolius showed thinner cortex and the root cross section was unchanged. The epidermis stele thickness was unaffected by salinity in A. armatus and A. tenuifolius and was reduced in A. mareoticus with smaller xylem vessel size. In addition, vessel density and wall thickness of xylem was increased under salt conditions in the studies species. The entire lamina and mesophyll of the three species were thinner in salt-stressed plants. A. armatus and A. tenuifolius showed the higher thickness with increased size of the lower epidermis. NaCl (300 mM) reduced leaf water content by 41.5 % in A. mareoticus while it was unchanged in the other species. The size of the vascular bundle increased under salinity in A. tenuifolius leaves and it was unchanged in the other ones. A longer distance between leaf vascular bundle was occurred in A. mareoticus. The effects of NaCl on root and leaf ultrastructure are discussed in relation to the degree of salt resistance of these species. The unchanged biomass production under salt stress confirmed the higher tolerance oft A. tenuifolius to salinity. A. armatus was moderately salt tolerant with decrease of biomass production by 14.2 % while A. mareoticus was considered as salt sensitive plant when the decrease in biomass production reached 56.8%.

Keywords: Astragalus species, leaf ultrastructure, root anatomy, salt stress

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Authors: R. Loukil, M. Chtourou, T. Damak

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In this work, we use the Fault detection and isolation and the Fault tolerant control based on sliding mode observer in order to introduce the well diagnosis of a nonlinear system. The robustness of the proposed observer for the two techniques is tested through a physical example. The results in this paper show the interaction between the Fault tolerant control and the Diagnosis procedure.

Keywords: fault detection and isolation FDI, fault tolerant control FTC, sliding mode observer, nonlinear system, robustness, stability

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Authors: Azene Tesfaye, Yohannes Petros, Habtamu Zeleke

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To evaluate genetic variation among Ethiopian lentil, laboratory experiment were conducted to screen 12 accessions of lentil (Lens culinaris M.) for salt tolerance. Seeds of 12 Lentil accessions were grown at laboratory (Petri dish) condition with different levels of salinity (0, 2, 4, and 8 dSm-1 NaCl) for 4 weeks. The experimental design was completely randomized design (CRD) in factorial combination with three replications. Data analysis was carried out using SAS software. Average germination time, germination percentage, seedling shoot and root traits, seedling shoot and root weight were evaluated. The two way ANOVA for varieties revealed statistically significant variation among lentil accession, NaCl level and their interactions (p<0.001) with respect to the entire parameters. It was found that salt stress significantly delays germination rate and decreases germination percentage, shoot and root length, seedling shoot and root weight of lentil accessions. The degree of decrement varied with accessions and salinity levels. Accessions 36120, 9235 and 36004 were better salt tolerant than the other accessions. As the result, it is recommended to be used as a genetic resource for the development of lentil accession and other very salt sensitive crop with improved germination under salt stress condition.

Keywords: accession, germination, lentil, NaCl, screening, seedling stage

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Authors: G. P. Abdel Rahim, M. María Guadalupe Moreno Armenta, Jairo Arbey Rodriguez

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We investigated the structure and electronic properties of the compound Mg1-xBixO with varying concentrations of 0, ¼, ½, and ¾ x bismuth in the the NaCl (rock-salt) and WZ (wurtzite) phases. The calculations were performed using the first-principles pseudo-potential method within the framework of spin density functional theory (DFT). Our calculations predict that for Bi concentrations greater than ~70%, the WZ structure is more favorable than the NaCl one and that for x = 0 (pure MgO), x = 0.25 and x = 0.50 of Bi concentration the NaCl structure is more favorable than the WZ one. For x = 0.75 of Bi, a transition from wurtzite towards NaCl is possible, when the pressure is about 22 GPa. Also It has been observed the crystal lattice constant closely follows Vegard’s law, that the bulk modulus and the cohesion energy decrease with the concentration x of Bi.

Keywords: DFT, Mg1-xBixO, pseudo-potential, rock-salt, wurtzite

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Authors: Mohamed M. Bumadian, D. James Gilmour

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Three bacterial species were isolated from the River Wye (Derbyshire, England) and identified using 16S rRNA gene sequencing as Serratia proteamaculans, Aeromonas hydrophila and Klebsiella pneumoniae. Respiration rates of the strains were measured in order to determine the metabolic activity under salt stress. The highest respiration rates of all three strains were found at 0.17 M and 0.5 M NaCl and then the respiration rate decreased with increasing concentrations of NaCl. In addition, the effect of increasing concentrations of NaCl on malate dehydrogenase activity was determined using cell-free extracts of the three strains. Malate dehydrogenase activity was stimulated at NaCl concentrations up to 0.5 M, and a small level of activity remained even at 3.5 M NaCl. The pH optimum of the malate dehydrogenase in cell-free extracts of all strains was higher than pH 7.5.

Keywords: fresh water, halotolerant pathogenic bacteria, 16S rRNA gene, cell-free extracts, respiration rates, malate dehydrogenase

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Authors: Abdul Hameed, Muhammad Zaheer Ahmed, Salman Gulzar, Bilquees Gul, Jan Alam, Ahmad K. Hegazy, Abdel Rehman A. Alatar, M. Ajmal Khan

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Seed germination and recovery from salt stress of an annual halophyte Suaeda heterophylla (Kar. and Kir.) Bunge to different iso-osmotic concentrations (0, -0.46, -0.92, -1.38, -1.84, and -2.30 MPa) of NaCl and PEG-6000 at 15/25, 20/30 and 25/35°C in both 12-h temperature and light regimes and in complete darkness were studied. Maximum number of seeds germinated in distilled water and increase in concentrations of both NaCl and PEG-6000 decreased germination at all temperature regimes, light and dark conditions, with higher inhibition in NaCl than PEG-6000. Recovery of germination and viability of seeds were lower in NaCl than PEG-6000 both in the light and dark. Moderate alternate temperatures (20/30°C) and 12-h photoperiod were found to be the optimal for seed germination and recovery. Better seed germination of S. heterophylla when osmotic potential caused both by NaCl and PEG 6000 is lower, temperature regime of 20/30°C and light regime is for 12 h.

Keywords: seed germination, abiotic stresses, Suaeda heterophylla, molecular biology

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Authors: Min-Shiuan Liou, Yi Shan Yang, Yang-Zhan Huang, Chia-Wen Hsieh

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A high speed growing and butanol-tolerant Clostridium acetobutylicum HOL1 mutant was screened throughout continuous adaption culture with C. acetobutylicum ATCC 824. The HOL1 strain can grow well in 10 g/L butanol contained CGM medium and can produce about 12.8 g /L butanol during 24 hrs. The C. acetobutylicum HOL1 strain was able to produce 166 mM butanol with 21 mM acetone at pH 4.8, resulting in a butanol selectivity (a molar ratio of butanol to total solvents) of 0.79, which is much higher than that (0.6) of the wild-type strain C. acetobutylicum ATCC 824. The acetate and butyrate accumulation were not observed during fermentation of the HOL1 strain. A hyper-butanol producing C. acetobutylicum HOL1 (pBPHS-3), which was created to overexpress the Bacillus psychrosaccharolyticus originated specific heat-shock protein gene, hspX, from a clostridial phosphotransbutyrylase promoter, was studied for its potential to produce a high titer of butanol. Overexpression of hspX resulted in increased final butanol yield 47% and 30% higher than those of the the ATCC824 and the HOL1 strains, respectively. The remarkable high-speed growth and butanol tolerance of strain HOL1 (pBPHS-3) demonstrates that overexpression of heterogeneous stress protein-encoding gene, hspX, could help C. acetobutylicum to effectively produce a high concentration of butanol.

Keywords: Clostridium acetobutylicum, butanol, heat-shock protein, resistance

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Authors: Madhu Jain, Rakesh Kumar Meena

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This paper investigates a finite M/G/1 fault tolerant multi-component machining system. The system incorporates the features such as standby support, threshold recovery and imperfect coverage make the study closer to real time systems. The performance prediction of M/G/1 fault tolerant system is carried out using recursive approach by treating remaining service time as a supplementary variable. The numerical results are presented to illustrate the computational tractability of analytical results by taking three different service time distributions viz. exponential, 3-stage Erlang and deterministic. Moreover, the cost function is constructed to determine the optimal choice of system descriptors to upgrading the system.

Keywords: fault tolerant, machine repair, threshold recovery policy, imperfect coverage, supplementary variable technique

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Authors: T. N. M. El Sebai, A. A. Khattab, Wafaa M. Abd-El Rahim, H. Moawad

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Fungal mutant strains have produced cellulase and xylanase enzymes, and have induced high hydrolysis with enhanced of rice straw. The mutants were obtained by exposing Penicillium strain to UV-light treatments. Screening and selection after treatment with UV-light were carried out using cellulolytic and xylanolytic clear zones method to select the hypercellulolytic and hyperxylanolytic mutants. These mutants were evaluated for their cellulase and xylanase enzyme production as well as their abilities for biodegradation of rice straw. The mutant 12 UV/1 produced 306.21% and 209.91% cellulase and xylanase, respectively, as compared with the original wild type strain. This mutant showed high capacity of rice straw degradation. The effectiveness of tested mutant strain and that of wild strain was compared in relation to enhancing the composting process of rice straw and animal manures mixture. The results obtained showed that the compost product of inoculated mixture with mutant strain (12 UV/1) was the best compared to the wild strain and un-inoculated mixture. Analysis of the composted materials showed that the characteristics of the produced compost were close to those of the high quality standard compost. The results obtained in the present work suggest that the combination between rice straw and animal manure could be used for enhancing the composting process of rice straw and particularly when applied with fungal decomposer accelerating the composting process.

Keywords: rice straw, composting, UV mutants, Penicillium

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Authors: Abderrahmen Bouguerra, Kamel Kara, Djamel Saigaa, Samir Zeghlache, Keltoum Loukal

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In this paper, a Fault-Tolerant control of 2 DOF Helicopter (TRMS System) Based on Fuzzy Gain-Adaptive PID is presented. In particular, the introduction part of the paper presents a Fault-Tolerant Control (FTC), the first part of this paper presents a description of the mathematical model of TRMS, an adaptive PID controller is proposed for fault-tolerant control of a TRMS helicopter system in the presence of actuator faults, A fuzzy inference scheme is used to tune in real-time the controller gains, The proposed adaptive PID controller is compared with the conventional PID. The obtained results show the effectiveness of the proposed method.

Keywords: fuzzy control, gain-adaptive PID, helicopter model, PID control, TRMS system

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Authors: Khaled M. Khleifat

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An extracellular methanol and ethanol tolerant lipase producing bacterial strain K5b4 was isolated from soil samples contaminated with hydrocarbon residues. It was identified by using morphological and biochemical characteristics and 16srRNA technique as Acinetobacter species. The immobilized lipase from Acinetobacter sp. K5b4 retained more than 98% of its residual activity after incubation with pure methanol and ethanol for 24 hours. The highest hydrolytic activity of the immobilized enzyme was obtained in the presence of 75% (v/v) methanol in the assay solution. In contrary, the enzyme was able to maintain its original activity up to only 25% (v/v) ethanol whereas at elevated concentrations of 50 and 75% (v/v) the enzyme activity was reduced to 10 and 40%, respectively. Maximum lipase activity of 31.5 mU/mL was achieved after 48 hr cultivation when the optimized medium (pH 7.0) that composed of 1.0% (w/v) olive oil, 0.2% (w/v) glycerol, 0.15% (w/v) yeast extract, and 0.05% (w/v) NaCl was inoculated with 0.4% (v/v) seed culture and incubated at 30°C and 150 rpm agitation speed. However, the presence of CaCl2 in the growth media did not show any inhibitory or stimulatory effect on the enzyme production as it compared to the control experiment. Meanwhile, the other mineral salts MgCl2, MnCl2, KCl and CoCl2 were negatively affected the production of lipase enzyme. The inhibition of lipase production from Acinetobacter sp. K5b4 in presence of glucose suggesting that lipase gene expression is prone to catabolic repression.

Keywords: K5B4, methanol and ethanol, acinetobacter, morphological

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Authors: Dedy Pratama, Akhmadu Muradi, Hilman Ibrahim, Raden Suhartono, Alexander Jayadi Utama, Patrianef Darwis, S. Dwi Anita, Luluk Yunaini, Kemas Dahlan

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Introduction: Vascular endothelial growth factor (VEGF) gene shows association with various angiogenesis conditions including Diabetic Foot Ulcer (DFU) disease. In this study, we performed this study to examine VEGF gene polymorphism associated with DFU. Methods: Case-control study of polymorphism of VEGF gene +405 C>G and -460 T>C, of diabetes mellitus (DM) type 2 with Diabetic Foot Ulcer (DFU) in Cipto Mangunkusumo National Hospital (RSCM) Jakarta from June to December 2016. Results: There were 203 patients, 102 patients with DFU and 101 patients without DFU. Forty-nine point 8 percent of total samples is male and 50,2% female with mean age 56,06 years. Distribution of the wild-type genotype VEGF +405 C>G wild type CC was found in 6,9% of respondents, the number of mutant heterozygote CG was 69,5% and mutant homozygote GG was 19,7%. Cumulatively, there were 6,9% wild-type and 85,2% mutant and 3,9% of total blood samples could not be detected on PCR-RFLP. Distribution of VEGF allele +405 C>G C alleles were 43% and G alleles were 57%. Distribution of genotype from VEGF gene -460 T>C is wild type TT 42,9%, mutant heterozygote TC 37,9% and mutant homozygote CC 13,3%. Cumulatively, there were 42,9% wild-type and 51% mutant type. Distribution of VEGF -460 T>C were 62% T allele and 38% C allele. Conclusion: In this study we found the distribution of alleles from VEGF +405 C>G is C 43% and G 57% and from VEGF -460 T>C; T 62% and C 38%. We propose that G allele in VEGF +405 C>G can act as a protective allele and on the other hands T allele in VEGF -460 T>C could be acted as a risk factor for DFU in diabetic patients.

Keywords: diabetic foot ulcer, diabetes mellitus, polymorphism, VEGF

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Authors: Raoudha Abdellaoui, Faycal Boughalleb, Zohra Chebil

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In arid and semi-arid regions, salinity represents a major constraint towards plants’ growth. Stipa lagascae, a psammophile grass, is a promised species since its economic and ecological interests. Our study aims to explore the effects of different salt concentrations (0; 100; 200; 300 and 400 mM) on physiological, biochemical and anatomic parameters. Salt stress was applied on S. lagascae plants cultivated under controlled conditions. Results show that salinity reduces biomass production especially when plants are subjected to severe stress (>200 mM NaCl). Concerning the nutritional level, the fact of enriching soil with NaCl, leads to an accumulation of Na+ against other nutritional elements (K+, Ca2+). To maintain tissues hydration, S. lagascae established osmotic adaptation by accumulation of proline and soluble sugars. Salt stress affected significantly root and foliar anatomy. Indeed, plants increased their vessels’ diameter and mesophyll surface. S. lagascae plants reduced also the surface of the belluforme cells to defeat dehydration. According to our results, S. lagascae seems to be a tolerant plant at acceptable concentrations that do not exceed 6g/l.

Keywords: anatomical adaptations, mineral nutrition, plant growth, salt stress, stipa lagascae

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