MicroRNAs (miRNAs) are an emerging class of small non-coding RNAs that exhibit important role in regulation of gene expression, mostly through the mechanism of cleavage and/or inhibition of translation of target mRNAs during or after transcription. Although much has been unravelled about the role of miRNAs in diverse biological processes like maintenance of functional integrity of genes and genome, growth and development, metabolism, and adaptive responses towards biotic and abiotic stresses in plants, not much is known on their specific roles in majority of cash crops - an area of investigation with potentially significant and gainful economic implications. Tea (Camellia sinensis) is globally the second most consumed beverage after water and its cultivation has major agro-economic and social ramifications. In recent years, global tea production has been greatly challenged by many biotic and abiotic stress factors and a deeper understanding of molecular processes regulating stress adaptation in this largely under investigated crop stands to significantly facilitate potential crop improvement strategies towards durable stress tolerance. This review endeavours to highlight recent advances in our understanding of the role of miRNAs in regulating stress tolerance traits in tea plant with additional focus on their role in determining tea quality attributes.

Semi-continuous production of xanthan gum using self-immobilized Xanthomonas campestris cells in biofilm reactors was studied. Fermentation was carried out using two different designs of biofilm reactor equipped with a) stainless-steel support (SSS) and b) polyethylene support (PES). Fermentation was performed in three cycles with refreshing the media at the beginning of each: cycle 1, 0-27 h; cycle 2, 27-54 h; and cycle 3, 54-78.5 h. Results showed that the glucose consumption and the pH reduction in the PES biofilm reactor was faster compared to the SSS biofilm reactor. Scanning electron microscopy showed that the SSS was capable to immobilize more cells during the growth of X. campestris. The maximum concentration of xanthan gum in the SSS biofilm reactor obtained after 27 h (3.47 ± 0.71 g/L), while the maximum concentration of xanthan in the PES biofilm reactor obtained after 78.5 h (3.21 ± 0.68 g/L). Thermal stability analysis of xanthan using differential scanning calorimetry showed the presence of two fractures attributed to dehydration and degradation of polymer. The thermogram represented both endothermal and exothermal behaviour of xanthan polymer. Furthermore, the functional groups and molecular structure of the xanthan produced in this study was evaluated using Fourier transform infrared spectrometry and also proton nuclear magnetic resonance. in addition, the surface tension of (0.2 %, w/v) xanthan gum solution was in a range of 52.16-56.5 mN/m. Rheological analysis of xanthan showed that the G' values were higher than the G″ in all frequencies demonstrating a relatively high elasticity of the produced xanthan gum.

Algae are omnipresent in all seas and oceans, which make thema target for many applications such as bio-fertilizers, fish feeding and removal of heavy metals. In the present study, different algal species were examined as sustainable alternatives substrates for PHA production by Halomonas sp. Several media simulations were utilized to achieve high polymer productivity. The maximum poly(3-hydroxybutyrate) (PHB) concentrations were determined by using Corallina mediterranea hydrolysates as a carbon and nitrogen source. The isolates Halomonas pacifica ASL10 and Halomonas salifodiane ASL11 were found to be able to produce PHA by 67 % wt and 63 % wt CDW, respectively. PHB nanoparticles (NPs) had high zeta potential values and small particle sizes. These properties make it suitable for several drug delivery and pharmaceutical applications. Interestingly, NPs showed a potent antibacterial activity against several reference strains. The antibacterial efficacy of PHA-NPs has not been previously studied, thus this study opens a promising use of PHA-NPs.

Phototrophic biofilms produce a matrix of extracellular polymeric substances (EPS), which holds the cells together and functions inter alia as nutrient storage and protection layer. EPS mainly consist of water, polysaccharides, proteins, lipids and nucleic acids as well as lysis and hydrolysis products which makes the composition very complex. Thus, rough simplifications are used and commonly one or at most two components of the EPS are examined. In this work a new procedure for separation and analysis of EPS in the main components (i) polysaccharides, (ii) proteins and (iii) lipids is presented with recovery rates of nearly 100 %. The method was established with synthetic EPS, which based on the composition of real EPS described in literature. Afterwards, the method was transferred to real EPS samples allowing a deeper insight in the composition of EPS from only one sample. The composition of EPS-extracts from Nostoc spec, cultivated under heterotrophic and mixotrophic batch and fed-batch conditions, was analysed during a cultivation period of 14 days. It was observed that mixotrophic cultivation led to higher amounts of carbohydrates, lipids and proteins than heterotrophic cultivation respectively, regardless of batch or fed-batch culture. While the amount of proteins in the EPS increased during the cultivation period, carbohydrates and lipids were dominant in the beginning and decreased afterwards.

Basmati and non-basmati rice varieties are commercially important. Aromatic rice varieties are low yielding and recently depletion in aroma is observed due to the shift towards modern agriculture. Therefore, it is necessary to restore the aroma and increase the yield through sustainable agriculture. The use of microbial bioinoculants is one of the promising ways to achieve these targets. With these objectives, rhizospheric bacterial strains Enterobacter hormaechei (AM122) and Lysinibacillus xylanilyticus (DB25) having the property of synthesizing 2-acetyl-1-pyrroline (2AP) were isolated from the rhizosphere of two aromatic rice varieties, Ambemohar-157 and Dehradun Basmati respectively and their effect on plant growth, aroma and yield enhancement under mono-inoculation and consortium conditions was analyzed. The bacterial inoculum in consortium resulted in significant improvement in vegetative growth, yield and 2AP content over mono inoculation and control. The study highlights the potential of E. hormaechei and L. xylanilyticus in plant growth, yield and aroma enhancement in basmati and non-basmati rice varieties. These strains can be taken up further for developing a commercial bioformulation.

Bis(2,4-di-tert-butylphenyl)Phosphate (bDtBPP) leaches out of polyethylene films used by the biopharmaceutical industry in single-use systems (SUS) for the culturing of drug producing cell lines. Previous studies found bDtBPP (0.025 - 0.110 mg/L) negatively affects Chinese hamster ovary (CHO) cell growth and productivity. Less information is known about the potential early stages of subtle pre-lethal cytotoxic effects of bDtBPP. This study aimed to investigate the pre-lethal cytotoxic effects in CHO-K1 cells of bDtBPP (0.005 - 0.25 μg/ml) at process relevant concentrations following 2, 24 and 48 h exposure, using high content analysis to investigate multiple pre-lethal cytotoxicity markers. After 48 h exposure, bDtBPP (0.005 - 0.25 μg/ml; P ≤ 0.001) increased nuclear intensity. A dose- and time-dependent reduction in mitochondrial mass was seen after exposure to bDtBPP. Reactive oxygen species increased after 2 h exposure to 0.25 μg/ml bDtBPP, 24 and 48 h exposure to 0.05 - 0.25 μg/ml bDtBPP (P ≤ 0.01 and P ≤ 0.001). BDtBPP induced subtle pre-lethal cytotoxic effects on CHO-K1 cellular health. This study highlights the cellular health benefits of the biopharmaceutical industry switching to alternative SUS plastics which do not leach bDtBPP, which may enhance CHO-K1 cell productivity.

A novel glucose oxidase (GOD)-perhydrolase-in situ chemical oxidation (ISCO) cascade reaction system was designed, optimized, and verified the operation feasibility in this research. Among the determined four perhydrolases, acyltransferase from Mycobacterium smegmatis (MsAcT) displayed the highest specific activity for perhydrolysis reaction (76.4 U/mg) and the lowest Km value to hydrogen peroxide (13.9 mmol/L). GOD-MsAcT cascade reaction system also displayed high catalytic efficiency. Under the optimal parameters (50:1 activity unit ratio of GOD to MsAcT, pH 8.0, 50 mmol/L of β-d-glucose, and 15 mmol/L of glyceryl triacetate), the melanin decolorization rate using GOD-MsAcT-ISCO cascade reaction system reached 86.8 %. Kinetics of GOD-MsAcT-ISCO cascade reaction system for melanin decolorization fitted the kinetic model of Boltzmann sigmoid. As a substitutive skin whitening technology, GOD-MsAcT-ISCO cascade reaction system displayed an excellent application prospect.

Curcumin has ignited global interest as an elite drugable molecule, owing to its time-honoured pharmacological activities against diverse human ailments. Limited natural accessibility and poor oral bioavailability caused major hurdles in the curcumin-based drug development process. We report the first successful testimony of curcumin and its glucoside synthesis in Atropa belladonna hairy roots (HR) through metabolic engineering. Re-routing the inherent biosynthetic precursors of the phenylpropanoid pathway of A. belladonna by heterologous expression of key curcumin biosynthetic pathway genes (i.e., Diketide-CoA synthase-DCS and Curcumin synthase-CURS3) and glucosyltransferase gene (CaUGT2) resulted in the production of curcumin and its glucoside in HR clones. Under shake-flask cultivation, the PGD2-HR1clone bearing DCS/ CURS3 genes showed the maximum curcumin yield (180.62 ± 4.7 μg/g DW), while the highest content of curcumin monoglucoside (32.63 ± 2.27 μg/g DW) along with curcumin (67.89 ± 2.56 μg/g DW) were noted in the PGD3-HR3 clone co-expressing DCS/CURS3 and CaUGT2 genes. Bioreactor up-scaling showed yield improvements in the PGD2-HR1 (2.3 fold curcumin) and the PGD3-HR3 clone (0.9 and 1.65 folds of curcumin-monoglucoside and curcumin respectively). These findings proved the advantageous use of HR cultures as the production source for curcumin and its glucoside, which remained unexplored so far.

Unicellular green algae, a promising source for renewable biofuels, produce lipid-rich biomass from light and CO2. Productivity in photo-bioreactors is affected by inhomogeneous light distribution from high cell pigment causing heat dissipation of light energy absorbed in excess and shading of the deep layers. Contrasting reports have been published on the relation between photoprotective energy dissipation and productivity. Here, we have re-investigated the relation between energy quenching (qE) activity, photodamage and light use efficiency by comparing WT and two Chlamydomonas reinhardtii strains differing for their complement in LHCSR proteins, which catalyse dissipation of excitation energy in excess (qE). Strains were analysed for ROS production, protein composition, rate of photodamage and productivity assessed under wide light and CO2 conditions. The strain lacking LHCSR1 and knocked down in LHCSR3, thus depleted in qE, produced O2 at significantly higher rate under high light, accompanied by enhanced singlet oxygen release and PSII photodamage. However, biomass productivity of WT was delayed in respect for mutant strains under intermittent light conditions only, implying that PSII activity was not the limiting factor under excess light. Contrary to previous proposals, domestication of Chlamydomonas for carbon assimilation rate in photo-bioreactors by down-regulation of photoprotective energy dissipation was ineffective in increasing algal biomass productivity.

Ganoderic acids (GAs) produced by Ganoderma are a type of lanostane-type triterpenoids with anticancer and antimetastatic activities; however, low production of GAs limits its wide application. In this study, a novel strategy by promoting sporulation of Ganoderma was developed to increase GA production. First, a high-spore producing Ganoderma strain G. 260125 was obtained from dikaryotic strain CGMCC 5.0026, and the sporulation-specific gene of this strain exhibits a higher transcription level than CGMCC 5.0026. Then, the effect of promoting sporulation on GA content was investigated. The maximum ganoderic acid (GA)-T, GA-Mk, and GA-Me contents in G. 260125 in shake flasks were 358.97, 78.32, and 12.75 μg/100 mg dry weight, respectively, which were 3.42, 2.91, and 1.73 times higher than those obtained in CGMCC 5.0026. Moreover, total and individual GA contents in spores were significantly higher than those in liquid static culture. Both concentrations of intermediates and transcription levels of GA biosynthetic genes also improved in G. 260125 during fermentation compared with those in CGMCC 5.0026. For scaling-up experiments, GA-T, GA-Me, and GA-Mk production in G. 260125 improved by 2.2-, 2.6-, and 2.1-fold compared with those in CGMCC 5.0026. In addition, the effectiveness of the developed strategy was also confirmed in three different Ganoderma strains. This work illustrated that promoting sporulation efficiently improves GA production in liquid static cultures of Ganoderma.

A biosorbent composed of non-viable Pseudomonas putida trapped in agar beads was able to remove Cu2+ and Zn2+ from solutions containing one or both metals. The process in batch followed pseudo second-order kinetics, with adsorption capacities of 0.255 mg Cu2+/g and 0.170 mg Zn2+/g according to the Langmuir isotherm. These values were up to ten times lower for beads without biomass. The metals became bound to OH, CH2, CO, COC and COP groups, with the last three being provided by the biomass, which highlights its importance. Adsorption values for single-metal solutions filtered in a fixed-bed column were 0.152 mg Cu2+/g and 0.117 mg Zn2+/g, but decreased to 0.075 and 0.058, respectively, with mixed-metal solutions (1:1 ratio). In 10:1-ratio solutions, the metal in greater proportion was better adsorbed. Under all conditions, removal percentage was ∼60 %. The column could be reused throughout ten absorption/desorption cycles without significant alterations in adsorption capacity.

Aromatic nitro compounds are key building blocks for many industrial syntheses and are also components of explosives, drugs and pesticides. Due to the environmentally unfriendly experimental conditions involved in their chemical syntheses, industrial processes would benefit from the use of biocatalysts. Among potentially useful enzymes, N-oxygenases, whose role is to oxygenate primary amines, are becoming relevant. These enzymes are involved in different secondary metabolic pathways in Streptomyces and in few other bacteria, forming part of the enzyme pools implicated in antibiotic synthesis. In this work, a group of Streptomyces strains, whose biomass was obtained from simple and novel culture media, were identified as new sources of N-oxygenase activity. Furthermore, the use of unspecific metabolic stimulation strategies allowed substantial improvements in the activity of whole cells as biocatalysts. It is remarkable the 6 to 50-fold increase in nitro compound yields compared to the biotransformation under standard conditions when Streptomyces griseus was the biocatalyst. In addition, biocatalyst substrate acceptance was studied in order to determine the biocatalytic potential of this enzyme.

In-depth understanding of microbial growth is crucial for the development of new advances in biotechnology and for combating microbial pathogens. Condition-specific proteome expression is central to microbial physiology and growth. A multitude of processes are dependent on the protein expression, thus, whole-cell analysis of microbial metabolism using genome-scale metabolic models is an attractive toolset to investigate the behaviour of microorganisms and their communities. However, genome-scale models that incorporate macromolecular expression are still inhibitory complex: the conceptual and computational complexity of these models severely limits their potential applications. In the need for alternatives, here we revisit some of the previous attempts to create genome-scale models of metabolism and macromolecular expression to develop a novel framework for integrating protein abundance and turnover costs to conventional genome-scale models. We show that such a model of Escherichia coli successfully reproduces experimentally determined adaptations of metabolism in a growth condition-dependent manner. Moreover, the model can be used as means of investigating underutilization of the protein machinery among different growth settings. Notably, we obtained strongly improved predictions of flux distributions, considering the costs of protein translation explicitly. This finding in turn suggests protein translation being the main regulation hub for cellular growth.

Continuous improvements of cell culture media are required in order to ensure high yield and product quality. However, some components can be instable and lead to detrimental effects on bioprocess performances. l-cysteine is an essential amino acid commonly used in cell culture media. Despite its beneficial effect on recombinant protein production, in some cases, this component can be responsible for product microheterogeneity. In this context, alternative components have to be found in order to reduce product variants while maintaining high productivity. In this study, we have assessed the performance of different cysteine and cystine analogs : N-acetyl-cysteine, s-sulfocysteine, N,N'-diacetyl-l-cystine and the N,N'-diacetyl-l-cystine dimethylester (DACDM). Replacement of cysteine by cystine analogs, and especially DACDM, has shown positive impact on charge variants level and recombinant protein coloration level. Moreover, this molecule contributed to the increase of the intracellular glutathione pool, which suggests a close relationship with the oxidative stress regulation.

Rapid development of effective biotherapeutics has been a concern during the last couple decades. In our work we designed two novel peptide tags, GMOP and mGMOP, derived from the N-terminal region of human granulocyte and macrophage colony stimulating factor (hGM-CSF), which contain four and six potential O-glycosylation sites, respectively. These peptide tags were fused to the N-terminus of human interferon-α2b (hIFN-α2b), a therapeutic antiviral and antiproliferative protein rapidly cleared from circulation. Two new molecules were obtained which, consistently with the presence of O-glycans, showed higher molecular masses, more negatively charged isoforms, and higher sialic acid content compared to wild-type IFN. In vitro bioactivity of purified chimeras revealed a similar antiviral specific biological activity (SBA) compared to unmodified IFN. A reduction of antiproliferative SBA was only observed for mGMOP-IFN. Pharmacokinetic studies in rats showed a notable improvement in terminal half-life (t1/2elim) (3.3 and 2.8 times-longer) and a marked reduction of the apparent clearance (CLapp, 3.7 and 4.1-fold lower for GMOP-IFN and mGMOP-IFN in comparison with native IFN, respectively). Furthermore, the in vitro thermal and plasma stability of both proteins was improved. Finally, a monoclonal antibody (mAb) that recognizes an N-terminal GM-CSF epitope was able to bind both chimeras in western blots and ELISAs. This demonstrates the potential of both peptides to behave as bifunctional tags to create novel long-acting biotherapeutics and to facilitate detection and purification.

In this study, single-stranded DNA aptamers that switch structural conformation upon binding to the salivary peptide histatin 3 have been reported for the first time. Histatin 3 is an antimicrobial peptide that possesses the capability of being a therapeutic agent against oral candidiasis and has recently been linked as a novel biomarker for acute stress. The aptamers were identified through a library immobilization version of an iterative in vitro process known as the Systematic Evolution of Ligands by EXponential enrichment (SELEX). Through the SELEX process, four unique aptamer candidates sharing a consensus sequence were identified. These selected sequences exhibited binding affinity and specificity to histatin 3 and in order to further characterize these aptamers, a direct format enzyme-linked aptamer sorbent assay (ELASA) was developed. The best performing candidate demonstrated an equilibrium dissociation constant (Kd) value of 1.97 ± 0.48 μM. These novel aptamers have the potential to lead to the further development of refined sensing assays and platforms for the detection and quantification of histatin 3 in human saliva and other biological media.

Small millets' is a generic term that includes all the millets except pearl millet and sorghum. These small or minor millets constitute eleven species that are marginally cultivated and consumed worldwide. These small millets possess excellent agronomic-, climate-resilient, and nutritional traits, although they lack popularity. Small millets withstand a broad spectrum of environmental stresses and possess better water-use and nitrogen-use efficiencies. Of note, small millets are five- to seven-fold nutritionally rich in terms of protein, bioactive compounds, micro- and macro-nutrients as compared to major cereals. Irrespective of these merits, small millets have received little research attention compared to major millets and cereals. However, the knowledge generated from such studies is significant for the improvement of millets per se and for translating the information to improve major cereals through breeding and transgene-based approaches. Given this, the review enumerates the efforts invested in dissecting the climate-resilient traits in small millets and provides a roadmap for deploying the information in crop improvement of millets as well as cereals in the scenario of climate change.

Biomass and lipid production by the marine diatom Chaetoceros affinis were characterized under continuous light with aeration. Media based on palm oil mill effluent (POME; 10, 20 and 30 % v/v in distilled water) were used together with a standard control medium. The maximum biomass concentration on day 12 of batch cultures in control medium was 821 ± 71 mg L-1. Under identical conditions, in the best POME medium (20 % POME v/v in distilled water with other inorganic components), the biomass concentration was reduced by ∼11 % to 734 ± 66 mg L-1. The lipid content of the biomass grown in the control medium was 50.8 ± 4.5 % by dry weight, but was a little lower (48.9 ± 4.1 % by dry wt) in the above specified best POME medium. In the best POME medium, oleic acid was the major fatty acid (72.3 ± 5.2 % by weight) in the total lipids extracted from the biomass and monounsaturated fatty acids were the main type of fatty acids (74.6 ± 5.2 %). POME levels of >20 % in the medium suppressed both biomass and lipid production relative to the medium with 20 % POME.

During the Agaricus bisporus canning processes, a large number of water-soluble elements were dissolved into the processing hot water. This study was conducted to use the industrial wastewater of A. bisporus to prepare agricultural microbial fertilizer. In the work, the influence of 6 different liquid fermentation factors on the total biomass of living Bacillus cereus was evaluated with the one-factor-at-a-time method and the Plackett-Burman design. The total biomass of living B. cereus was most influenced by fermentation temperature, shaking speed, and inoculation volume, which were identified as the most critical independent variables for the B. cereus biomass. The approximate ranges of optimal fermentation conditions for the three key factors were identified by the path of steepest ascent. The center point of these factors were 24 ℃ of temperature, 250 rpm of shaking speed and 12 % inoculum amount, respectively. The Box-Behnken design was applied to derive a statistical model for optimizing the three fermentation factors for B. cereus biomass. After further optimizations based on statistical predictions, the optimum fermentation parameters for B. cereus cultured in the A. bisporus industrial wastewater were fermentation temperature of 24.8 °C, shaking speed of 234 rpm, inoculum dose of 12.2 % (v:v, %), industrial wastewater concentration of 4%, initial pH values of 6.5, loading liquid of 60 mL/250 mL, and culture time of 24 h. Culturing with the optimal fermentation conditions resulted in the biomass of B. cereus of 1.35 ± 0.02 × 109 Obj/mL (N = 3), which was consistent with the predicted values (1.32 × 109 Obj/mL) predicted by the corresponding regression models (p < 0.05), and more, was also far higher than that of the standard of agricultural bacterial fertilizers in People's Republic of China. Further, the results of field trial indicated that the of B. cereus liquid fertilizer can remarkably enhance the yield of Brassica chinensis L. It is practicable to make use of the industrial wastewater of A. bisporus to prepare the microbial fertilizer.

Soil salinity is one of the major environmental factors, influencing agricultural productivity of crops. As a non-edible and ideal oilseed crop, castor (Ricinus communis L.) has great industrial value in biofuel, but molecular mechanisms of salt stress regulation are still unknown. In this study, the differentially expressed genes (DEGs) for differential salt tolerance in two castor cultivar (wild castor : Y, cultivated castor 'Tongbi 5': Z) were identified. 12 libraries were sampled for Illumina high-throughput sequencing to consider 132,426 nonredundant unigenes and 31,221 gene loci. Multiple phytohormones and transcription factors (TFs) were correlated with salt-tolerance and differently enriched in these two genotypes. The type 2C protein phosphatases (PP2C) homologs were all upregulated under salt stress. Importantly, IAA (1), DELLA (1) and Jasmonate zim domain (JAZ) (1) were also identified and found to be differentially expressed. Based on the co-expressed module by regulatory networks and heatmap analysis, ERF/AP2, WRKY and bHLH families were prominently participate in high salt stress response of wild and cultivated castor. Finally, these results highlight that the hub DEGs and families were more accumulated in cultivated castor than those in wild castor, providing novel insights into the salinity adaptive mechanisms and genetic improvement in castor.

Ficus carica L. is an important source of phenolic and flavonoid compounds with valuable pharmaceutical application across various diseases. The current study was carried out to investigate the influence of Piriformospora indica elicitation on growth, production of phenolic compounds, antioxidant capacity, and expression level of flavonoid biosynthetic pathway genes in hairy root (HR) cultures of F. carica. The maximum improvement in accumulation of phenolic compounds was observed when HR culture of Ficus carica L. was exposed to 2% culture filtrate of P. indica for 72 h: gallic acid (80.5- fold), caffeic acid (26.2-fold), coumaric acid (4.5-fold), and cinnamic acid (60.1-fold), apigenin (27.6-fold) and rutin (5.7-fold). While the highest levels of chlorogenic acid (4.9-fold) and quercetin flavonoid (8.8-fold) were obtained after 48 h elicitation with culture filtrate and cell extract of P. indica at 6% (v/v), respectively. The analysis of biosynthetic genes revealed that the exposure to fungal elicitors resulted in up-regulation of phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT) and MYB3 transcription factor. This study shows the potential of P. indica as an efficacious elicitor for enhancing the secondary metabolites production by F. carica HRs.

The Bacillus subtilis group comprises species known for their ability to produce a wide variety of antimicrobial peptides. This work focuses on bacilysin, a broad-spectrum active dipeptide, and its prevalence in the B. subtilis group. In silico genome analysis of strains from Bacillus amyloliquefaciens, Bacillus velezensis, Bacillus licheniformis, Bacillus pumilus and B. subtilis subspecies inaquosorum, spizizenii and subtilis revealed that the bacilysin gene cluster is present in all species except for B. licheniformis. This observation was corroborated by PCR detection of the bacilysin genetic determinants on a collection of 168 food and environmental strains from the B. subtilis group. Phylogenetic analyses also demonstrated that the bacilysin gene cluster sequence showed more than 80 % identity within each species of the B. subtilis group. An in vitro screening of the strain collection was performed against foodborne pathogens. Twenty-three strains were selected for their ability of their Cell-Free Supernatant to inhibit foodborne pathogens. After an ammonium sulphate precipitation of their supernatant, eight strains, all belonging to B. velezensis, exhibited antimicrobial activity against Gram-negative pathogens. Using Ultra High Performance Liquid Chromatography - Mass Spectrometry, the presence of bacilysin was confirmed in these eight precipitates. These findings provide evidence that bacilysin is a major player in the antagonistic activity of B. velezensis against Gram-negative foodborne pathogens.

The mini-chromosome maintenance (MCM) family, a large and functionally diverse protein family belonging to the AAA+ superfamily, is essential for DNA replication in all eukaryotic organisms. The MCM 2-7 form a hetero-hexameric complex which serves as licensing factor necessary to ensure the proper genomic DNA replication during the S phase of cell cycle. MCM 8-10 are also associated with the DNA replication process though their roles are particularly unclear. In this study, we report an extensive in silico analysis of MCM gene family (MCM 2-10) in Arabidopsis and rice. Comparative analysis of genomic distribution across eukaryotes revealed conservation of core MCMs 2-7 while MCMs 8-10 are absent in some taxa. Domain architecture analysis underlined MCM 2-10 subfamily specific features. Phylogenetic analyses clustered MCMs into 9 clades as per their subfamily. Duplication events are prominent in plant MCM family, however no duplications are observed in Arabidopsis and rice MCMs. Synteny analysis among Arabidopsis thaliana, Oryza sativa, Glycine max and Zea mays MCMs demonstrated orthologous relationships and duplication events. Further, estimation of synonymous and non-synonymous substitution rates illustrated evolution of MCM family under strong constraints. Expression profiling using available microarray data and qRT-PCR revealed differential expression under various stress conditions, hinting at their potential use to develop stress resilient crops. Homology modeling of Arabidopsis and rice MCM 2-7 and detailed comparison with yeast MCMs identified conservation of eukaryotic specific insertions and extensions as compared to archeal MCMs. Protein-protein interaction analysis revealed an extensive network of putative interacting partners mainly involved in DNA replication and repair. The present study provides novel insights into the MCM family in Arabidopsis and rice and identifies unique features, thus opening new perspectives for further targeted analyses.

Potato (Solanum tuberosum L.) is considered to be frost-susceptible as short spells of frost can reduce the tuber yield and quality. Ice recrystallization inhibition (IRI) protein helps prevent growth of ice crystals in the cell apoplast during frost and help prevent damage associated with freezing stress. In this study, we investigated the in planta potential of Lolium perenne derived IRI3 transgene in improving the tolerance of transgenic potato lines for freezing stress. The codon optimized IRI3 transgene was introduced into potato cultivar Diamant through Agrobacterium mediated transformation. Three transgenic potato lines were successfully generated which were confirmed for transgene insertion and genomic integration by polymerase chain reaction and Southern blot. It was evident that the IRI3 transcript decreased in initial 24 h of cold stress treatment while the IRI3 mRNA expression up regulated in subsequent hours of cold treatment with maximum increase to 20 folds at 96 h post stress. A similar trend was also revealed in ion-leakage assay which showed that during cold stress, the transgenic potato lines depicted reduced ion leakage of 14-22% as compared to non-transgenic control plants. Further, the generated transgenic potato lines were tolerant to the frost spell in quarantine field conditions as compared to the non-transgenic potato lines. Additionally, the transgenic lines exhibited efficient recovery post frost injury in field conditions. The biochemical profiles of chlorophyll, proline and higher levels of antioxidant enzyme (superoxide dismutase, Catalase) activity and malondialdehyde levels showed that despite the phenotypic impact of low temperature, the transgenic potato lines quickly adjusted to maintain their cellular homeostasis post freezing stress by increasing the antioxidant defenses. This study suggests that up regulation of IRI3 transcript and regulatory network of cold stress response in transgenic potato lines improve frost tolerance and help stabilize yield in cultivated potato.