RESUMEN
The efficacy, targeting ability, and biocompatibility of plant-based nanoparticles can be exploited in fields such as agriculture and medicine. This study highlights the use of plant-based ginger nanoparticles as an effective and promising strategy against cancer and for the treatment and prevention of bacterial infections and related disorders. Ginger is a well-known spice with significant medicinal value due to its phytochemical constituents including gingerols, shogaols, zingerones, and paradols. The silver nanoparticles (AgNPs) derived from ginger extracts could be an important non-toxic and eco-friendly nanomaterial for widespread use in medicine. In this study, AgNPs were biosynthesized using an ethanolic extract of ginger rhizome and their phytochemical, antioxidant, antibacterial, and cytotoxic properties were evaluated. UV-visible spectral analysis confirmed the formation of spherical AgNPs. FTIR analysis revealed that the NPs were associated with various functional biomolecules that were associated with the NPs during stabilization. The particle size and SEM analyses revealed that the AgNPs were in the size range of 80-100 nm, with a polydispersity index (PDI) of 0.510, and a zeta potential of -17.1 mV. The purity and crystalline nature of the AgNPs were confirmed by X-ray diffraction analysis. The simple and repeatable phyto-fabrication method reported here may be used for scaling up for large-scale production of ginger-derived NPs. A phytochemical analysis of the ginger extract revealed the presence of alkaloids, glycosides, flavonoids, phenolics, tannins, saponins, and terpenoids, which can serve as active biocatalysts and natural stabilizers of metallic NPs. The ginger extracts at low concentrations demonstrated promising cytotoxicity against Vero cell lines with a 50% reduction in cell viability at 0.6-6 µg/mL. When evaluated for biological activity, the AgNPs exhibited significant antioxidant and antibacterial activity on several Gram-positive and Gram-negative bacterial species, including Escherichia coli, Bacillus subtilis, Pseudomonas aeruginosa, and Staphylococcus aureus. This suggests that the AgNPs may be used against multi-drug-resistant bacteria. Ginger-derived AgNPs have a considerable potential for use in the development of broad-spectrum antimicrobial and anticancer medications, and an optimistic perspective for their use in medicine and pharmaceutical industry.
RESUMEN
MAIN CONCLUSION: The present investigation profoundly asserted the catalytic potential of plant-based aldo-ketoreductase, postulating its role in polyketide biosynthesis and providing new insights for tailored biosynthesis of vital plant polyketides for therapeutics. Plants hold great potential as a future source of innovative biocatalysts, expanding the possibilities within chemical reactions and generating a variety of benefits. The aldo-keto reductase (AKR) superfamily includes a huge collection of NAD(P)H-dependent oxidoreductases that carry out a variety of redox reactions essential for biosynthesis, detoxification, and intermediary metabolism. The present study involved the isolation, cloning, and purification of a novel aldo-ketoreductase (AvAKR) from the leaves of Aloe vera (Aloe barbadensis Miller) by heterologous gene expression in Escherichia coli based on the unigene sequences of putative ketoreductase and cDNA library screening by oligonucleotide hybridization. The in-silico structural analysis, phylogenetic relationship, and molecular modeling were outranged to approach the novelty of the sequence. Additionally, agroinfiltration of the candidate gene tagged with a green fluorescent protein (GFP) was employed for transient expression in the Nicotiana benthamiana to evaluate the sub-cellular localization of the candidate gene. The AvAKR preferred cytoplasmic localization and shared similarities with the known plant AKRs, keeping the majority of the conserved active-site residues in the AKR superfamily enzymes. The enzyme facilitated the NADPH-dependent reduction of various carbonyl substrates, including benzaldehyde and sugars, proclaiming a broad spectrum range. Our study successfully isolated and characterized a novel aldo-ketoreductase (AvAKR) from Aloe vera, highlighting its versatile NADPH-dependent carbonyl reduction proficiency therewith showcasing its potential as a versatile biocatalyst in diverse redox reactions.
Asunto(s)
Aldehído Reductasa , Aloe , Aldo-Ceto Reductasas/genética , Aldehído Reductasa/genética , Aldehído Reductasa/química , Aldehído Reductasa/metabolismo , Oxidorreductasas de Alcohol/genética , Oxidorreductasas de Alcohol/metabolismo , Aloe/genética , Aloe/metabolismo , Filogenia , NADP/genética , Plantas/metabolismoRESUMEN
Asparagus racemosus is known for its diverse content of secondary metabolites, i.e., saponins, alkaloids, and a wide range of flavonoids. Flavonoids, including phenols and polyphenols, have a significant role in plant physiology and are synthesized in several tissues. Despite the diverse role of flavonoids, genetic information is limited for flavonoid biosynthesis pathways in A. racemosus. The current study explores full-scale functional genomics information of A. racemosus by de novo transcriptome sequencing using Illumina paired-end sequencing technology to elucidate the genes involved in flavonoid biosynthesis pathways. The de novo assembly of high-quality paired-end reads resulted in â¼2.3 million high-quality reads with a pooled transcript of 45,647 comprising â¼76 Mb transcriptome with a mean length (bp) of 1,674 and N50 of 1,868bp. Furthermore, the coding sequence (CDS) prediction analysis from 45,647 pooled transcripts resulted in 45,444 CDS with a total length and mean length of 76,398,686 and 1,674, respectively. The Gene Ontology (GO) analysis resulted in a high number of CDSs assigned to 25,342 GO terms, which grouped the predicted CDS into three main domains, i.e., Biological Process (19,550), Molecular Function (19,873), and Cellular Component (14,577). The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database was used to categorize 6,353 CDS into 25 distinct biological pathway categories, in which the majority of mapped CDS were shown to be related to translation (645), followed by signal transduction (532), carbohydrate metabolism (524), folding, sorting, and degradation (522). Among these, only â¼64 and 14 CDSs were found to be involved in the phenylpropanoid and flavonoid biosynthesis pathways, respectively. Quantitative Real-time PCR was used to check the expression profile of fourteen potential flavonoid biosynthesis pathway genes. The qRT-PCR analysis result matches the transcriptome sequence data validating the Illumina sequence results. Moreover, a large number of genes associated with the flavonoids biosynthesis pathway were found to be upregulated under the induction of methyl jasmonate. The present-day study on transcriptome sequence data of A. racemosus can be utilized for characterizing genes involved in flavonoid biosynthesis pathways and for functional genomics analysis in A. racemosus using the reverse genetics approach (CRISPR/Cas9 technology).
RESUMEN
Polyhydroxybutyrate (PHB) has significant potential for replacing non-biodegradable traditional plastic, which is responsible for several global environmental issues. The main problem with switching to bio-based alternatives for petrochemical plastics is the large price gap on the market. To overcome this problem, the present research was focused on the utilization of inexpensive substrates i.e. agricultural residues for cost-effective PHB production by endospore-forming bacteria Bacillus badius MTCC 13004. For efficient PHB production, Box-Behnken Design (BBD) was selected for media optimization and to observe the interactive effects of four variables i.e. pH, Na acetate, Banana peel, and mustard cake. PHB yield of 2.11 g/L was attained under optimized conditions compared to non-optimized conditions (0.72 g/L). FTIR spectra analysis of PHB extracted from Bacillus badius was found to be similar to commercial PHB. NMR data was also matched with the chemical shift signals CH, CH2, and CH3 of PHB. The melting temperature (Tm) and glass transition temperature (Tg) of PHB from Bacillus badius was found to be 165.14 and 2.68 °C, respectively. Further, PCR protocol was also designed to amplify key enzymes of the PHB synthesis pathway i.e. PHB synthase (phb C gene).
Asunto(s)
Bacillus , Análisis Costo-Beneficio , Bacillus/metabolismo , Plásticos/metabolismo , Hidroxibutiratos/química , Poliésteres/químicaRESUMEN
Thirty-five distinct endophytic fungi were isolated from the roots of Asparagus racemosus. Five out of 35 isolates were found to be efficient saponins producers and they were identified as Aspergillus terreus (E.F-1), Aspergillus flavus (E.F-7), Penicillium sp. (E.F-12), Talaromyces pinophilus(S-26), and Aspergillus terreus (Y-2) based on 18 sr RNA sequencing. The crude extracts of endophytic fungi were screened using High-performance liquid chromatography (HPLC) for quantitative analysis of saponin. The crude extracts of endophytic fungi were also characterised using FT-IR spectroscopy and mass spectrometry. The IR spectra of all five endophytic fungi crude extracts revealed the presence of -OH,-CH Alkyl,-CH3,-C-O-C,-C=C,-C=O stretching, which indicated the presence of saponin. Eight types of saponins recognised by mass spectrometry were Cyclamine saponin, Aspoligonin A, Sarsapogenin, Asparacosin A, Schidigera saponinD5, Aspargoside A, Dioscin, and Protodioscin. Endophytic fungi extracts also exhibited antimicrobial activity and antioxidant activity.
RESUMEN
Despite extensive works on miRNA's role during plant-oomycete interaction, its role in Capsicum annuum-Phytophthora capsici pathosystem is not fully explored. Therefore, the present study was designed to identify known and novel miRNAs along with their target genes in two contrasting chili peppers genotypes, i.e., GojamMecha_9086 (resistant) and Dabat_80045 (susceptible) under P. capsici infection associated with modulating the defense response during disease pathogenesis. The result demonstrated 79 known miRNAs corresponding to 24 miRNAs families and 477 novel miRNAs along with 22,895 potential targets, including 30 defense-related target genes against P. capsici infection. The expression analysis of 29 known and 157 novel miRNAs in resistant and 30 known and 177 novel miRNAs in susceptible genotypes revealed differential accumulation patterns. qRT-PCR analysis of 8 defense-related miRNAs representing 4 novels (Pz-novel-miR428-1, Pz-novel-miR160-1, Pz-novel-miR1028-1, Pz-novel-miR204-1) and 4 known miRNAs (Pz-known-miR803-1, Pz-known-miR2059-1, Pz-known-miR2560-1, Pz-known-miR1872-1) revealed differential accumulation pattern in both resistant and susceptible genotypes. Additionally, validation of eight target genes of miRNAs using regional amplification quantitative RT-PCR (RA-PCR), a superior technique to 5'-RNA Ligase-Mediated-rapid amplification of cDNA ends (5' RLM-RACE), revealed expression of six target genes positively correlated with their corresponding miRNAs in RC versus RI leaf, while five target genes observed an inverse correlation with their corresponding miRNAs in SC versus SI leaf, suggesting their key role during disease response. The Pz-known-miR1872-PODs pair showed perfect inverse relation in all four samples. The significant findings of the current study provide comprehensive genome-wide information about the repertoire of miRNAs and their target genes expressed in resistant and susceptible chili pepper genotypes, which can serve as a valuable resource for better understanding the post-transcriptional regulatory mechanism during C. annuum-P. capsici pathosystem.
Asunto(s)
Capsicum , MicroARNs , Phytophthora , Enfermedades de las Plantas , Capsicum/genética , Capsicum/microbiología , Resistencia a la Enfermedad/genética , Genotipo , MicroARNs/genética , Phytophthora/patogenicidad , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/microbiologíaRESUMEN
In the recent past, cross-kingdom movement of miRNAs, small (20-25 bases), and endogenous regulatory RNA molecules has emerged as one of the major research areas to understand the potential implications in modulating the plant's biotic stress response. The current review discussed the recent developments in the mechanism of cross-kingdom movement (long and short distance) and critical cross-talk between host's miRNAs in regulating gene function in bacteria, fungi, viruses, insects, and nematodes, and vice-versa during host-pathogen interaction and their potential implications in crop protection. Moreover, cross-kingdom movement during symbiotic interaction, the emerging role of plant's miRNAs in modulating animal's gene function, and feasibility of spray-induced gene silencing (SIGS) in combating biotic stresses in plants are also critically evaluated. The current review article analysed the horizontal transfer of miRNAs among plants, animals, and microbes that regulates gene expression in the host or pathogenic organisms, contributing to crop protection. Further, it highlighted the challenges and opportunities to harness the full potential of this emerging approach to mitigate biotic stress efficiently.
Asunto(s)
MicroARNs , Animales , Hongos/genética , Silenciador del Gen , MicroARNs/genética , MicroARNs/metabolismo , Plantas/metabolismo , Estrés Fisiológico/genéticaRESUMEN
Chili pepper (Capsicum annuum L.) is economically one of the most important spice. But, it's productivity is highly affected by the pathogen, Phytophthora capsici L. Our current understanding of the molecular mechanisms associated with the defence response in C. annuum-P. capsici pathosystem is limited. The current study used RNA-seq technology to dissect the genes associated with defence response against P. capsici infection in two contrasting landraces, i.e. GojamMecha_9086 (Resistant) and Dabat_80045 (Susceptible) exposed to P. capsici infection. The transcriptomes from four leaf samples (RC, RI, SC and SI) of chili pepper resulted in a total of 118,879 assembled transcripts along with 52,384 pooled unigenes. The enrichment analysis of the transcripts indicated 23 different KEGG pathways under five main categories. Out of 774 and 484 differentially expressed genes (DEGs) of two landraces (under study), respectively, 57 and 29 DEGs were observed as associated with defence responses against P. capsici infection in RC vs. RI and SC vs. SI leaf samples, respectively. qRT-PCR analysis of six randomly selected genes validated the results of Illumina NextSeq500 sequencing. A total of 58 transcription factor families (bHLH most abundant) and 2095 protein families (Protein kinase most abundant) were observed across all the samples with maximum hits in RI and SI samples. Expression analysis revealed differential regulation of genes associated with defence and signalling response with shared coordination of molecular function, cellular component and biological processing. The results presented here would enhance our present understanding of the defence response in chili pepper against P. capsici infection, which the molecular breeders could utilize to develop resistant chili genotypes. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01122-y.
RESUMEN
BACKGROUND: Aloe vera (Aloe barbadensis Miller), commonly known as Ghritkumari/Gwarpatha, is a member of the Liliaceae family, used in the traditional medicine system for ages. Aloe vera has made its importance as a therapeutic agent, acting as a cure for various diseases such as skin problems, lungs, and heart disorders, diabetes, ulcers, various microbial infections, and asthma. Despite its tremendous health benefits, the dark side of the plant is a reason of concern as there are several active compounds present in the plant, raising questions on its safe oral consumption and application. METHODS AND RESULTS: The literature review was compiled from information resourced from various national and international journals available at Google Scholar and curated with Mendeley. The data mining was carried out during the period of January to May 2021. This study explored and summarized the dark side of Aloe vera, subjected to various secondary metabolites present in it. Aloin, the most active compound of Aloe vera, is a type of anthraquinone metabolized by human gut microflora, resulting in the formation of aloe-emodin anthraquinone, later being associated with several harmful effects such as carcinogenicity, genotoxicity, nephrotoxicity, and purgative. Besides this, several alkaloids and polysaccharides present in the plant are reported to cause hepatotoxicity and male infertility, respectively. CONCLUSIONS: The harmful effects of the plants are not adequately discovered yet; hence there is a need to come up with some mechanism to understand and suppress the formation of such toxic compounds completely. This review examined the botany, active compounds, and adverse clinical effects in the range of metabolites associated with this herb - "Aloe vera".
Asunto(s)
Aloe , Antraquinonas , Daño del ADN , Humanos , Masculino , Extractos Vegetales/efectos adversos , PolisacáridosRESUMEN
The study focused on the medicinal properties of citrus fruits and their ability to synthesize silver nanoparticles. As the resistance against the modern antibiotic agents is on increase, finding new and effective natural antibiotic agents is the need of the modern era. Similarly, bio-synthesis of nanoparticles is also being encouraged for eco-friendly reasons. Due to remarkable medicinal and industrial applications of silver nanoparticles (AgNPs), citrus fruit juice is used to reduce silver ions for the green synthesis of AgNPs. Phytochemical analysis revealed the presence of various constituents which impart antibacterial property to citrus fruits, analyzed against four pathogenic bacteria. Also, citrus fruit juice exhibits radical scavenging activity because of these constituents. Further, the AgNPs synthesized using citrus fruits were characterized using Ultra-Violet Visible (UV-VIS) spectroscopy, Fourier Transform Infrared (FTIR) spectroscopy and Transmission Electron Microscopy (TEM) to study the shape and size of the AgNPs. Anticancer activity of AgNPs was also evaluated against Colo-205 cell lines and found to inhibit 37.9% growth of cell lines at the concentration of 10 µg/ml. Hence, synthesized AgNPs can be used effectively against cancer cell lines in combination with other anti-cancer agents.
Asunto(s)
Antineoplásicos/farmacología , Citrus , Nanopartículas del Metal , Antibacterianos/farmacología , Línea Celular Tumoral , Frutas , Tecnología Química Verde , Humanos , Pruebas de Sensibilidad Microbiana , Extractos Vegetales/farmacología , Plata/farmacología , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Industrial wastes, for instance, tannery wastes are rich soups of resistant and bioremediation-potent bacteria. In the present work, Chromium (Cr) and tannic acid (TA) resistance bacterial strains were isolated from tannery effluent and identified as Bacillus subtilis (MCC 3275) and Bacillus safensis (MCC 3283) based on its 16S Ribosomal RNA homology. Hexavalent Cr is highly toxic and mutagenic due to its high mobility and reactivity. Whereas, TA is known to inhibit enzyme activity, substrate deprivation, and interaction with membranes and matrix-metal ions. The developed In vitro co-cultured microcosm of B. subtilis and B. safensis was able to remove Cr(VI) up to 95% and TA up to 23%. The bacteria cultures separately were able to degrade Cr(VI) to 88% by B. subtilis and 91% by B. safensis and TA up to 27%. Plackett Burman design (PBD) followed by Response surface methodology (RSM) was applied for the optimization of physio-chemical parameters. The optimized conditions for co-culture development were recorded as K2HPO4 = 0.2 g/L, MgSO4 = 0.2 g/L, NH4Cl = 0.5 g/L, glucose - 0.2 g/L, TA - 5%, Cr = 200 ppm, incubation period of 96 h, agitation speed of 110 rpm, pH = 5.0, temperature= 30 °C and inoculum size = 3%. Scanning electron microscopy (SEM) and Fourier transform infrared (FTIR) revealed the thorough mechanism of cellular uptake followed by degradation of Cr(VI) and TA. The efficiency of co-culture for other heavy metals was observed as follows: Zn 65%, Pb 63%, Cd 65%, and Ni 65%. Bioremediation using bacteria is an economical and environmentally better alternative to conventional remediation methods. The isolated bacteria are useful in the effluent treatment of tannery or related industries and in metal recovery in mining processes.
Asunto(s)
Bacillus subtilis/metabolismo , Bacillus/metabolismo , Cromo/metabolismo , Taninos/metabolismo , Contaminantes Químicos del Agua/metabolismo , Contaminación Química del Agua/prevención & control , Biodegradación Ambiental , Residuos Industriales/análisisRESUMEN
Wheat (Triticum aestivum L.) crop health assumes unprecedented significance in being the second most important staple crop of the world. It is host to an array of fungal pathogens attacking the plant at different developmental stages and accrues various degrees of yield losses owing to these. Tilletia indica that causes Karnal bunt (KB) disease in wheat is one such fungal pathogen of high quarantine importance restricting the free global trade of wheat besides the loss of grain yield as well as quality. With global climate change, the disease appears to be shifting from its traditional areas of occurrence with reports of increased vulnerabilities of new areas across the continents. This KB vulnerability of new geographies is of serious concern because once established, the disease is extremely difficult to eradicate and no known instance of its complete eradication using any management strategy has been reported yet. The host resistance to KB is the most successful as well as preferred strategy for its mitigation and control. However, breeding of KB resistant wheat cultivars has proven to be not so easy, and the low success rate owes to the scarcity of resistance sources, extremely laborious and regulated field screening protocols delaying identification/validation of putative resistance sources, and complex quantitative nature of resistance with multiple genes conferring only partial resistance. Moreover, given a lack of comprehensive understanding of the KB disease epidemiology, host-pathogen interaction, and pathogen evolution. Here, in this review, we attempt to summarize the progress made and efforts underway toward a holistic understanding of the disease itself with a specific focus on the host-pathogen interaction between T. indica and wheat as key elements in the development of resistant germplasm. In this context, we emphasize the tools and techniques being utilized in development of KB resistant germplasm by illuminating upon the genetics concerning the host responses to the KB pathogen including a future course. As such, this article could act as a one stop information primer on this economically important and re-emerging old foe threatening to cause devastating impacts on food security and well-being of communities that rely on wheat.
RESUMEN
Among several important wheat foliar diseases, Stripe rust (YR), Leaf rust (LR), and Stem rust (SR) have always been an issue of concern to the farmers and wheat breeders. Evolution of virulent pathotypes of these rusts has posed frequent threats to an epidemic. Pyramiding rust-resistant genes are the most economical and environment-friendly approach in postponing this inevitable threat. To achieve durable long term resistance against the three rusts, an attempt in this study was made searching for novel sources of resistant alleles in a panel of 483 spring wheat genotypes. This is a unique and comprehensive study where evaluation of a diverse panel comprising wheat germplasm from various categories and adapted to different wheat agro-climatic zones was challenged with 18 pathotypes of the three rusts with simultaneous screening in field conditions. The panel was genotyped using 35K SNP array and evaluated for each rust at two locations for two consecutive crop seasons. High heritability estimates of disease response were observed between environments for each rust type. A significant effect of population structure in the panel was visible in the disease response. Using a compressed mixed linear model approach, 25 genomic regions were found associated with resistance for at least two rusts. Out of these, seven were associated with all the three rusts on chromosome groups 1 and 6 along with 2B. For resistance against YR, LR, and SR, there were 16, 18, and 27 QTL (quantitative trait loci) identified respectively, associated at least in two out of four environments. Several of these regions got annotated with resistance associated genes viz. NB-LRR, E3-ubiquitin protein ligase, ABC transporter protein, etc. Alien introgressed (on 1B and 3D) and pleiotropic (on 7D) resistance genes were captured in seedling and adult plant disease responses, respectively. The present study demonstrates the use of genome-wide association for identification of a large number of favorable alleles for leaf, stripe, and stem rust resistance for broadening the genetic base. Quick conversion of these QTL into user-friendly markers will accelerate the deployment of these resistance loci in wheat breeding programs.
RESUMEN
Genetic diversity is crucial for successful adaptation and sustained improvement in crops. India is bestowed with diverse agro-climatic conditions which makes it rich in wheat germplasm adapted to various niches. Germplasm repository consists of local landraces, trait specific genetic stocks including introgressions from wild relatives, exotic collections, released varieties, and improved germplasm. Characterization of genetic diversity is done using morpho-physiological characters as well as by analyzing variations at DNA level. However, there are not many reports on array based high throughput SNP markers having characteristics of genome wide coverage employed in Indian spring wheat germplasm. Amongst wheat SNP arrays, 35K Axiom Wheat Breeder's Array has the highest SNP polymorphism efficiency suitable for genetic mapping and genetic diversity characterization. Therefore, genotyping was done using 35K in 483 wheat genotypes resulting in 14,650 quality filtered SNPs, that were distributed across the B (~ 50%), A (~ 39%), and D (~ 10%) genomes. The total genetic distance coverage was 4477.85 cM with 3.27 SNP/cM and 0.49 cM/SNP as average marker density and average inter-marker distance, respectively. The PIC ranged from 0.09 to 0.38 with an average of 0.29 across genomes. Population structure and Principal Coordinate Analysis resulted in two subpopulations (SP1 and SP2). The analysis of molecular variance revealed the genetic variation of 2% among and 98% within subpopulations indicating high gene flow between SP1 and SP2. The subpopulation SP2 showed high level of genetic diversity based on genetic diversity indices viz. Shannon's information index (I) = 0.648, expected heterozygosity (He) = 0.456 and unbiased expected heterozygosity (uHe) = 0.456. To the best of our knowledge, this study is the first to include the largest set of Indian wheat genotypes studied exclusively for genetic diversity. These findings may serve as a potential source for the identification of uncharacterized QTL/gene using genome wide association studies and marker assisted selection in wheat breeding programs.
Asunto(s)
Triticum/genética , Triticum/metabolismo , Mapeo Cromosómico/métodos , Cromosomas de las Plantas/genética , Grano Comestible/genética , Variación Genética/genética , Genoma de Planta/genética , Estudio de Asociación del Genoma Completo/métodos , Genotipo , Fenotipo , Fitomejoramiento/métodos , Poaceae/genética , Polimorfismo de Nucleótido Simple/genéticaRESUMEN
BACKGROUND: Aloe vera is a perennial, succulent, drought-resistant plant that exhibits many pharmacological characteristics such as wound healing ability against skin burns, anti-ulcer, anti-inflammatory, anti-tumor, anti-viral, anti-hypercholesterolemic, anti-hyperglycemic, anti-asthmatic and much more. Despite great medicinal worth, little genomic information is available on Aloe vera. This study is an initiative to explore the full-scale functional genomics of Aloe vera by generating whole transcriptome sequence database, using Illumina HiSeq technology and its progressive annotation specifically with respect to the metabolic specificity of the plant. RESULTS: Transcriptome sequencing of root and leaf tissue of Aloe vera was performed using Illumina paired-end sequencing technology. De novo assembly of high quality paired-end reads, resulted into 1,61,733 and 2,21,792 transcripts with mean length of 709 and 714 nucleotides for root and leaf respectively. The non-redundant transcripts were clustered using CD-HIT-EST, yielding a total of 1,13,063 and 1,41,310 unigenes for root and leaf respectively. A total of 6114 and 6527 CDS for root and leaf tissue were enriched into 24 different biological pathway categories using KEGG pathway database. DGE profile prepared by calculating FPKM values was analyzed for differential expression of specific gene encoding enzymes involved in secondary metabolite biosynthesis. Sixteen putative genes related to saponin, lignin, anthraquinone, and carotenoid biosynthesis were selected for quantitative expression by real-time PCR. DGE as well as qRT PCR expression analysis represented up-regulation of secondary metabolic genes in root as compared to leaf. Furthermore maximum number of genes was found to be up-regulated after the induction of methyl jasmonate, which stipulates the association of secondary metabolite synthesis with the plant's defense mechanism during stress. Various transcription factors including bHLH, NAC, MYB were identified by searching predicted CDS against PlantTFdb. CONCLUSIONS: This is the first transcriptome database of Aloe vera and can be potentially utilized to characterize the genes involved in the biosynthesis of important secondary metabolites, metabolic regulation, signal transduction mechanism, understanding function of a particular gene in the biology and physiology of plant of this species as well as other species of Aloe genus.
Asunto(s)
Aloe/genética , Aloe/metabolismo , Antraquinonas/metabolismo , Perfilación de la Expresión Génica , Genes de Plantas/genética , Saponinas/metabolismo , Análisis de Secuencia , Genómica , Anotación de Secuencia Molecular , Factores de Transcripción/metabolismoRESUMEN
This study was carried out to analyze the heavy metals biosorption potential of bacteria isolated from soil contaminated with electroplating industrial effluents. Bacterial isolates were screened for their multi-metal biosorption potential against copper, nickel, lead, and chromium. Bacterial isolate CU4A showed the maximum uptake of copper, nickel, lead, and chromium in aqueous solution, with a biosorption efficiency of 87.16 %, 79.62%, 84.92%, and 68.12%, respectively. The bacterial strain CU4A was identified as Bacillus cereus, following 16S rRNA gene sequence analysis. The surface chemical functional groups of bacterial biomass were identified by Fourier transform infrared (FTIR) spectroscopy as hydroxyl, carboxyl, amine, and halide, which may be involved in the biosorption of heavy metals. Analysis with scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) confirmed the adsorption of metals on the bacterial cell mass. The results of this study are significant and could be further investigated for the removal of heavy metals from contaminated environments.
Asunto(s)
Bacillus cereus/metabolismo , Contaminación Ambiental/prevención & control , Metales Pesados/metabolismo , Microbiología del Suelo , Contaminantes del Suelo/metabolismo , Adsorción , Restauración y Remediación Ambiental , Microscopía Electrónica de Rastreo , Filogenia , ARN Bacteriano/genética , ARN Ribosómico 16S/genética , Análisis de Secuencia de ARN , Espectrometría por Rayos X , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
Mutations in gyrA are the primary cause of quinolone resistance encountered in gram-negative clinical isolates. The prospect of this work was to analyze the role of gyrA mutations in eliciting high quinolone resistance in uropathogenic E.coli (UPEC) through molecular docking studies. Quinolone susceptibility testing of 18 E.coli strains isolated from UTI patients revealed unusually high resistance level to all the quinolones used; especially norfloxacin and ciprofloxacin. The QRDR of gyrA was amplified and sequenced. Mutations identified in gyrA of E.coli included Ser83Leu, Asp87Asn and Ala93Gly/Glu. Contrasting previous reports, we found Ser83Leu substitution in sensitive strains. Strains with S83L, D87N and A93E (A15 and A26) demonstrated norfloxacin MICs ≥1024mg/L which could be proof that Asp87Asn is necessary for resistance phenotype. Resistance to levofloxacin was comparatively lower in all the isolates. Docking of 4 quinolones (ciprofloxacin, ofloxacin, levofloxacin and norfloxacin) to normal and mutated E.coli gyrase A protein demonstrated lower binding energies for the latter, with significant displacement of norfloxacin in the mutated GyrA complex and least displacement in case of levofloxacin.
Asunto(s)
Antibacterianos/farmacología , Girasa de ADN/genética , Farmacorresistencia Bacteriana/genética , Proteínas de Escherichia coli/genética , Quinolonas/farmacología , Escherichia coli Uropatógena/genética , Girasa de ADN/metabolismo , Infecciones por Escherichia coli/microbiología , Proteínas de Escherichia coli/metabolismo , Humanos , Pruebas de Sensibilidad Microbiana , Simulación del Acoplamiento Molecular , Mutación , Homología de Secuencia de Ácido Nucleico , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/efectos de los fármacos , Escherichia coli Uropatógena/metabolismoRESUMEN
A chromium and tannic acid resistance fungal strain was isolated from tannery effluent, and identified as Aspergillus niveus MCC 1318 based on its rDNA gene sequence. The MIC (minimum inhibitory concentration) of the isolate against chromium and tannic acid was found to be 200 ppm and 5% respectively. Optimization of physiochemical parameters for biosorption of chromium and tannic acid degradation was carried out by Plackett-Burman design followed by response surface methodology (RSM). The maximum chromium removal and tannic acid degradation was found to be 92 and 68% respectively by A. niveus. Chromium removal and tannic acid degradation was increased up to 11 and 6% respectively after optimization. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) was used to investigate biosorption phenomena.
RESUMEN
Tannin acyl hydrolase (E.C. 3.1.1.20) commonly referred as tannase, is a hydrolytic enzyme that catalyses the hydrolysis of ester bonds present in gallotannins, ellagitannins, complex tannins and gallic acid esters. Tannases are the important group of botechnologically relevant enzymes distributed throughout the animal, plant and microbial kingdoms. However, microbial tannases are currently receiving a great deal of attention. Tannases are extensively used in food, feed, pharmaceutical, beverage, brewing and chemical industries. Owing to its diverse area of applications, a number of patents have been appeared in the recent past. The present review pretends to present the advances and perspectives in the industrial application of tannase with special emphasis on patents.
Asunto(s)
Hidrolasas de Éster Carboxílico/metabolismo , Alimentación Animal/análisis , Cerveza/análisis , Bebidas/análisis , Catálisis , Ácido Gálico/química , Hidrólisis , Patentes como Asunto , Taninos/química , Té/química , Vino/análisisRESUMEN
Diacylglycerol O-acyltransferase 1 (DGAT1) is a microsomal enzyme that catalyzes the final step of triglyceride synthesis. The DGAT1 gene is a strong functional candidate for determining milk fat content in cattle. In this work, we used PCR-SSCP (polymerase chain reaction-single-strand conformation polymorphism) and DNA sequencing to examine polymorphism in the region spanning exon 7 to exon 9 of the DGAT1 gene in Murrah and Pandharpuri buffaloes. Three alleles (A, B and C) and four novel single-nucleotide polymorphisms were identified in the buffalo DGAT1 gene. The frequencies of the alleles differed between the two buffalo breeds, with allele C being present in Murrah but not in Pandharpuri buffalo. The allele variation detected in this work may influence DGAT1 expression and function. The results described here could be useful in examining the association between the DGAT1 gene and milk traits in buffalo.