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  • 21
    Publication Date: 2018-03-05
    Description: Myceliophthora thermophila (ATCC 42464) is a thermophilic fungus that produces cellulolytic enzymes with high thermal stability. Unlike its mesophile counterparts, study on gene expression regulation of cellulolytic enzymes in M. thermophila is inadequate. This work identified the function of MHR1, a putative transcription regulator of cellulolytic enzymes in M. thermophila that was found through RNA-Seq based gene expression profile analysis. RNA interference was used to study the role of MHR1. A recombinant plasmid, pUC19–P pdc – mhr1 –T pdc , which contained the RNAi sequence for mhr1 was constructed and transformed into M. thermophila . One of the transformants, MtR5, in which the RNA interference efficiency was the highest, was used for the following studies. In the mhr1 -silenced strain MtR5, the filter paper hydrolyzing activity was 1.33-fold; β-1, 4-endoglucanase activity was 1.65-fold; and xylanase activity was 1.48-fold higher than those of the parental strain after induction, respectively, by wheat straw powder. qRT-PCR showed that gene expression of cbh1 , cbh2 , egl3 and xyr1 were 9.56-, 37.36-, 56.14- and 28.30-fold higher in MtR5 than in wild type, respectively. Our findings suggest that the transcription factor MHR1 of M. thermophila can repress cellulase and xylanase activities. Silenced mhr1 results in increased expression not only of the main cellulase genes, but also of the positive regulatory gene xyr1 . This work is relevant to the development of M. thermophila as an industrial production host for cellulolytic and hemicellulolytic enzymes, which could be used to degrade a wide range of different biomass, converting lignocellulosic feedstock into sugar precursors for biofuels.
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    Topics: Process Engineering, Biotechnology, Nutrition Technology
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  • 22
    Publication Date: 2018-03-05
    Description: The synthesis and characterization of an aggregate of AgNPs coated with plant extract (PE) from Alphonsea sclerocarpa and its significant antimicrobial activity and inhibition on K562 (blood cancer) cells have been appended in the article. Synthesis of aggregate [(AgNPs)-(PE)] has been followed by a facile eco-friendly approach without using any harmful chemicals. The morphology of an aggregate [(AgNPs)-(PE)] was confirmed by TEM and SEM microscopic characterizations. Properties like solid state, the presence of functional groups, and elemental composition have been characterized through the XRD, FTIR, and EDAX. The biocompatibility of synthesized aggregate of [(AgNPs)-(PE)] was confirmed by the MTT assay. An in vitro cell (HEK293)-based studies were performed for the biocompatibility tests and it is found that the aggregate [(AgNPs)-(PE)] is not harmful to normal/healthy cells. Even though A. sclerocarpa show the antimicrobial (antibacterial and antifungal) activity, it has been further enhanced with the developed aggregate of [(AgNPs)-(PE)]. Furthermore, it has been extended to examine the cellular inhibition on K562 cells and obtained 〉 75% cell inhibition for 24 h treated cells.
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    Topics: Process Engineering, Biotechnology, Nutrition Technology
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  • 23
    Publication Date: 2018-03-05
    Description: Saraca asoca (Roxb.) De Wilde is an important medicinal plant from the Western Ghats of India, traditionally used in treatment of various gynecological disorders. Increasing commercial demand and decreasing numbers has resulted in this plant becoming endangered with crude drug materials being extensively substituted/adulterated with other plant species. The present study was undertaken with the objective of development and evaluation of multivariate cluster analysis of ISSR fingerprints against rbcL -based DNA barcodes as tool to understand the relationships and to differentiate common adulterants and substituents from S. asoca . ISSR-based Hierarchical Cluster Analysis was carried out on 41 samples of S. asoca and 5 each of the 5 common substituent/adulterant plants and the clustering patterns were evaluated against DNA-sequence-based barcoding of rbcL region of their plastids. Factorial analysis and Principal Coordinate Analysis revealed distinct groups of genetic pools of respective taxa thereby confirming the utility of ISSR fingerprinting as a useful tool for differentiation between the genuine and the adulterants/substituents. NCBI-BLAST search on DNA barcode rbcL region confirmed the results of ISSR assays. Therefore, our study demonstrated the utility of simple, cost-effective method of ISSR fingerprinting coupled with rbcL barcoding in differentiating this important medicinal plant from its common adulterants/substituents. Graphical Abstract
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  • 24
    Publication Date: 2018-03-05
    Description: Drought is a major constraint to the productivity of many crops affecting various physiological and biochemical processes. Seventy percent of the peanuts are grown in semiarid tropics that are frequently prone to drought stress. So, we analyzed its effect in 4 cultivars of peanut, with different degrees of drought tolerance, under 10 and 20 days of water stress using two-dimensional gel electrophoresis and mass spectrometry. A total of 189 differentially expressed protein spots were identified in the leaf proteome of all the 4 cultivars using PD Quest Basic software; 74 in ICGV 91114, 41 in ICGS 76, 44 in J 11 and 30 in JL 24. Of these, 30 protein spots were subjected to in-gel trypsin digestion followed by MALDI-TOF that are functionally categorized into 5 groups: molecular chaperones, signal transducers, photosynthetic proteins, defense proteins and detoxification proteins. Of these, 12 proteins were sequenced. Late embryogenesis abundant protein, calcium ion binding protein, sucrose synthase isoform-1, 17.3 kDa heat shock protein and structural maintenance of chromosome proteins were overexpressed only in the 15 and 20 days stressed plants of ICGV 91114 cultivar while cytosolic ascorbate peroxidase was expressed with varying levels in the 10 and 20 days stressed plants of all the 4 cultivars. Signaling protein like 14-3-3 and defense proteins like alpha-methyl-mannoside-specific lectin and mannose/glucose-binding lectins were differentially expressed in the 4 cultivars. Photosynthetic protein like Rubisco was down-regulated in the stressed plants of all 4 cultivars while Photosystem-I reaction center subunit-II of chloroplast precursor protein was overexpressed in only 20 days stressed plants of ICGV 91114, ICGS 76 and J11 cultivars. These differentially expressed proteins could potentially be used as protein markers for screening the peanut germplasm and further crop improvement.
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  • 25
    Publication Date: 2018-03-05
    Description: Plant glutathione peroxidases (GPXs) are non-heme thiol peroxidases that play vital roles in maintaining H 2 O 2 homeostasis and regulating plant response to abiotic stress. Here, we performed a comparative genomic analysis of the GPX gene family in cucumber ( Cucumis sativus ). As a result, a total of 6 CsGPX genes were identified, which were unevenly located in four out of the seven chromosomes in cucumber genome. Based on the phylogenetic analysis, the GPX genes of cucumber, Arabidopsis and rice could be classified into five groups. Analysis of the distribution of conserved domains of GPX proteins showed that all these proteins contain three highly conserved motifs, as well as other conserved sequences and residues. Gene structure analysis revealed a conserved exon–intron organization pattern of these genes. Through analyzing the promoter regions of CsGPX genes, many hormone-, stress-, and development-responsive cis -elements were identified. Moreover, we also investigated their expression patterns in different tissues and developmental stages as well as in response to abiotic stress and x acid (ABA) treatments. The qRT-PCR results showed that the transcripts of CsGPX genes varied largely under abiotic stress and ABA treatments at different time points. These results demonstrate that cucumber GPX gene family may function in tissue development and plant stress responses.
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  • 26
    Publication Date: 2018-03-05
    Description: In this study, we reported the potential of various methanol fractions of onion (MFO) and its components quercetin and quercetin glucosides on platelet aggregation and antioxidant activity. The onion extracts were separated into several fractions using methanol and water. Further, these extracts were analyzed using simple high-performance liquid chromatography–diode array detector method to separate quercetin (Q), quercetin-4′- O -monoglucoside and quercetin-3, 4′- O -diglucoside from onion sample. It was observed that the bioactive compounds were accumulated in the non-polar portions rather than in the polar one. MFO and flavonol glucosides inhibited collagen-induced platelet aggregation, and the anti-aggregatory effects were comparatively studied using rat PRP (platelet-rich plasma). Among the extracted compounds, quercetin was found as a potent inhibitor of platelet aggregation compared to quercetin glucosides, whereas quercetin glucosides showed high antioxidant activities. These results suggest that MFO, quercetin and quercetin glucosides have powerful antiplatelet and antioxidant activities. These studies provide possible information leading to the intake of onions rich in these flavonols as a dietary supplement and functional food ingredient to prevent thrombosis and cardiovascular and oxidative stress-related diseases and might be utilized as a real source of valuable phytochemicals for the pharmaceutical and food industries for the development of antioxidant, anticoagulant and antiplatelet agents.
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  • 27
    Publication Date: 2018-03-05
    Description: In this study, we have described three steps to produce ethanol from Pogonatherum crinitum , which was derived after the treatment of textile wastewater. (a) Production of biomass: biomass samples collected from a hydroponic P. crinitum phytoreactor treating dye textile effluents and augmented with Ca-alginate immobilized growth-promoting bacterium, Bacillus pumilus strain PgJ (consortium phytoreactor), and waste sorghum husks were collected and dried. Compositional analysis of biomass (consortium phytoreactor) showed that the concentration of cellulose, hemicelluloses and lignin was 42, 30 and 17%, respectively, whereas the biomass samples without the growth-promoting bacterium (normal phytoreactor) was slightly lower, 40, 29 and 16%, respectively. (b) Hydrolysate (sugar) production: a crude sample of the fungus, Phanerochaete chrysosporium containing hydrolytic enzymes such as endoglucanase (53.25 U/ml), exoglucanase (8.38 U/ml), glucoamylase (115.04 U/ml), xylanase (83.88 U/ml), LiP (0.972 U/ml) and MnP (0.459 U/ml) was obtained, and added to consortium, normal and control phytoreactor derived biomass supplemented with Tween-20 (0.2% v/v). The hydrolysate of biomass from consortium phytoreactor produced maximum reducing sugar (0.93 g/l) than hydrolysates of normal phytoreactor biomass (0.82 g/l) and control phytoreactor biomass (0.79 g/l). FTIR and XRD analysis confirmed structural changes in treated biomass. (c) Ethanol production: the bioethanol produced from enzymatic hydrolysates of waste biomass of consortium and normal phytoreactor using Saccharomyces cerevisiae (KCTC 7296) was 42.2 and 39.4 g/l, respectively, while control phytoreactor biomass hydrolysate showed only 25.5 g/l. Thus, the amalgamation of phytoremediation and bioethanol production can be the truly environment-friendly way to eliminate the problem of textile dye along with bioenergy generation.
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  • 28
    Publication Date: 2018-03-05
    Description: The present study was aimed to isolate indigenous plant growth-promoting bacteria (PGPB) from Nanmangalam reserve forest, India and to analyze their positive impact on nursery plant species. In total, 160 isolates were obtained from different nitrogen-free Media (LGI, JMV, NFB). Amongst these, 12 isolates were shown positive for 5–8% of ammonia production nif H positive and then isolates were further tested for their plant growth-promoting (PGP) activity. Based on their PGP activity, nine isolates were selected, and applied in nurseries of twelve native plant species, along with organic manure and inorganic fertilizer. All the isolates were shown positive effects when compared to control. In that, five of these bacterial isolates, Paenibacillus sp. RRB2, Azospirillum brasilense RRAK5, Bacillus subtilis subsp. subtilis RRD8, Burholdria kururiensis RRAK1, and Pseudomonas stutzeri RRAN2, enhanced biomass production in several trees.
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  • 29
    Publication Date: 2018-03-05
    Description: Biomass from wetland aquatic grass and buffalo grass can be exploited for biogas production, because this substrate is plenteous and does not compete with food production. In this study, the grass substrate was physically pretreated by boiling with different retention time to increase its biodegradability and was examined in batch mode. Boiling pretreatment suggested that 100 °C with 2 h retention time was the best condition. The results showed that the optimum grass concentration in the 1:1 ratio of co-digestion mixture with manure produced the highest methane yield. The results suggested that co-digestion of buffalo grass and buffalo dung was a promising approach for improving biogas production. This study was achieved the upgraded biogas through biological purification contained 90.42% CH 4 8.04% CO 2 1.43% O 2 and 0.11% other trace gases—a remarkable performance based on an efficiency criteria. Furthermore, the digestate has high nutrient concentrations that can potentially use as fertilizer.
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  • 30
    Publication Date: 2018-03-05
    Description: The repertoire and functions of MADS-box family transcription factors (TFs) largely remains unexplored with respect to floral organogenesis of Momordica dioica Roxb. Degenerative PCR followed by rapid amplification of cDNA ends was employed in the present study to clone and characterize 17 MADS-box genes (designated as MdMADS01 to MdMADS17 ) from the floral buds of M. dioica. The cloned genes were clustered into three subgroups (11 MIKC C , 4 MIKC* and 2 Mα) based on phylogenetic relationships with the MADS-box genes from Cucumis sativus, Cucumis melo and Arabidopsis thaliana . Southern hybridization showed that all the isolated genes were represented by single copy locus in the M. dioica genome. Gene structure analysis revealed 1–8 exons in MdMADS -box genes with the number of exons in MIKC greatly exceeding from that in M-type genes. Motif elicitation of the MdMADS -box genes indicated the presence of additional domains with MIKC type, suggesting that they had more complex structures. Expression analysis of MdMADS genes in six M. dioica transcriptome suggested that, 11 MIKC C —type genes are associated with floral homeotic functions, 4 MIKC*-type genes ( MdMADS12 to MdMADS15 ) controlled the growth of male gametophyte, while the two M -type genes ( MdMADS16 and MdMADS17 ) played significant role in female gametogenesis and seed development. Overall, these are the first set of MADS-box genes from M. dioica exhibiting a differential expression pattern during floral development. The results from this study will provide valuable information for further functional studies of candidate MADS-box genes in the sexual dimorphism of this economically important dioecious cucurbit.
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