RESUMO
Coriander is a herbaceous spice and condiment crop also known for its medicinal properties. The present study identified two putative novel deltapartitiviruses and an enamovirus tentatively named as Coriandrum sativum deltapartitivirus 1, 2 (CsDPV1, 2) and Coriandrum sativum enamovirus (CsEV) in the publicly available transcriptome-assembled contigs derived from coriander grown in India. CsDPV1 and 2 contained tripartite and bipartite genomes, respectively, with each genome segment encoding a single open reading frame (ORF). CsEV contained five ORFs encoding proteins P0, P1, P2, P3 and P5. Phylogenetic analysis revealed three distinct subgroups of deltapartitiviruses wherein CsDPV1 and 2 grouped in subgroup 3 and 1, respectively, whilst CsEV formed a distinct sub-clade within enamoviruses. Further, the presence of CsDPV2 in fruit samples of one of the cultivars from where the virus was identified was confirmed through RT-PCR assay and Sanger sequencing. The study highlights the need for further studies on understanding the importance and the biological properties of identified novel viruses.
Assuntos
Coriandrum , Luteoviridae , Filogenia , Transcriptoma , FrutasRESUMO
Knowledgebase for rice sheath blight information (KRiShI) is a manually curated user-friendly knowledgebase for rice sheath blight (SB) disease that allows users to efficiently mine, visualize, search, benchmark, download, and update meaningful data and information related to SB using its easy and interactive interface. KRiShI collects and integrates widely scattered and unstructured information from various scientific literatures, stores it under a single window, and makes it available to the community in a user-friendly manner. From basic information, best management practices, host resistance, differentially expressed genes, proteins, metabolites, resistance genes, pathways, and OMICS scale experiments, KRiShI presents these in the form of easy and comprehensive tables, diagrams, and pictures. The "Search" tab allows users to verify if their input rice gene id(s) are Rhizoctonia solani (R. solani) responsive and/or resistant. KRiShI will serve as a valuable resource for easy and quick access to data and information related to rice SB disease for both the researchers and the farmers. To encourage community curation a submission facility is made available. KRiShI can be found at http://www.tezu.ernet.in/krishi .
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Oryza , Oryza/genética , Doenças das Plantas/genética , Bases de ConhecimentoRESUMO
Coriander (Coriandrum sativum L.) is well known vegetable and spice crop grown globally for its leaves and seeds. Stem gall (Protomyces macrosporus L.) is a fungal disease affecting its quality and yield. However, no information is available on SSR markers linked to disease resistance in coriander. Hence, development of co-dominant genetic markers is prerequisite for disease investigations in coriander. In-house stem gall resistance and susceptible cultivars transcriptome data were utilized. Totally, 59,933 and 56,861 transcripts were examined, 9141 and 8346 Simple Sequence Repeats (SSR) were identified and the most abundant type was the tri, followed by di, tetra, penta and hexa nucleotide repeats. A total of ten selected SSR-Functional Domain Markers (FDM) were developed based on functional annotation terms associated with pathogen response and validated among ten coriander cultivars and their transferability was examined in five fennel (Foeniculum vulgare L.) cultivars. Nine primer pairs resulted from amplified bands. Marker ACorSGD-1 shown monomorphic bands among coriander genotypes except Acr-1 showed heterologouse and multiple bands in fennel cultivars. Markers ACorSGD-4, 5, 7 and 9 shown presence in resistant cultivars and absence of bands among susceptible cultivars of coriander and thus, considered to be the candidate markers for disease screening. Marker ACorSGD-6 shown monomorphic bands among coriander. Markers ACorSGD-1, 2, 3, and 5 shown transferability among fennel cultivars. A total of 136 alleles in coriander and fennel were produced. Using UPGMA clustering method a dendrogram was generated and cultivars were grouped into two separate clusters with coriander and fennel. Identified and developed SSR-FDM markers are useful for linkage mapping for disease resistant in coriander.
Assuntos
Coriandrum/genética , Coriandrum/metabolismo , Repetições de Microssatélites/genética , Ascomicetos/patogenicidade , Mapeamento Cromossômico/métodos , Resistência à Doença/genética , Expressão Gênica/genética , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas/genética , Marcadores Genéticos/genética , Genótipo , Micoses/genética , Filogenia , Folhas de Planta/genética , Polimorfismo Genético/genética , Transcriptoma/genéticaRESUMO
Moringa oleifera is a plant well-known for its nutrition value, drought resistance and medicinal properties. cDNA libraries from five different tissues (leaf, root, stem, seed and flower) of M.â¯oleifera cultivar Bhagya were generated and sequenced. We developed a bioinformatics pipeline to assemble transcriptome, along with the previously published M.â¯oleifera genome, to predict 17,148 gene models. Few candidate genes related to biosynthesis of secondary metabolites, vitamins and ion transporters were identified. Expressions were further confirmed by real-time quantitative PCR experiments for few promising leads. Quantitative estimation of metabolites, as well as elemental analysis, was also carried out to support our observations. Enzymes in the biosynthesis of vitamins and metabolites like quercetin and kaempferol are highly expressed in leaves, flowers and seeds. The expression of iron transporters and calcium storage proteins were observed in root and leaves. In general, leaves retain the highest amount of small molecules of interest.
Assuntos
Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/fisiologia , Moringa oleifera , Metabolismo Secundário/fisiologia , Transcriptoma/fisiologia , Biblioteca Gênica , Moringa oleifera/genética , Moringa oleifera/metabolismoRESUMO
Coriander (Coriandrum sativum L.) is a well-known spice and aromatic crop cultivated globally. Stem gall disease is one of the major constraints for its leaf and seed quality used for consumption and also affecting the yield. The identification of resistance genes and further characterization of such genes could help to understand the molecular basis of resistance and lay a solid ground for cloning of stem gall resistance genes in coriander. To evaluate the genetic expression of disease resistance-relevant genes in popularly grown coriander cultivars in India such as Pant Haritma, Hisar Sugandh, Hisar Surabhi, Hisar Anand, Rajendra Swathi, ACr-1, ACr-2, AgCr-1, CO-2 and CS-6 were used for LRR, GDSL, USP, ANK and PDR gene expression using Real Time PCR along with 18S housekeeping gene as internal control for the normalization. Result revealed the different expression pattern of genes among the cultivars tested. Highest expression was shown in cultivar AgCr-1 followed by Pant Haritma, Hisar Sugandh and ACr-1, and least expression in Hisar Anand, ACr-2, CO-2, Rajendra Swathi and CS-6. Domain analysis revealed the conserved domain relevance of the genes. This is the first report on stem gall resistance gene expression in coriander. The identified genes have a potential role in coriander and further utilize in crop improvement program. We hypothesize that contrasting cultivars can be a good source for candidate gene evaluation and further to use them as potential markers and used in hybridization program focus on incorporating and develop durable disease-resistance into the adapted cultivars of the region.
Assuntos
Coriandrum/genética , Resistência à Doença/genética , Doenças das Plantas/genética , Expressão Gênica/genética , Regulação da Expressão Gênica de Plantas/genética , Índia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Caules de Planta/genética , Caules de Planta/metabolismo , Transcriptoma/genéticaRESUMO
BACKGROUND: Krishna Tulsi, a member of Lamiaceae family, is a herb well known for its spiritual, religious and medicinal importance in India. The common name of this plant is 'Tulsi' (or 'Tulasi' or 'Thulasi') and is considered sacred by Hindus. We present the draft genome of Ocimum tenuiflurum L (subtype Krishna Tulsi) in this report. The paired-end and mate-pair sequence libraries were generated for the whole genome sequenced with the Illumina Hiseq 1000, resulting in an assembled genome of 374 Mb, with a genome coverage of 61 % (612 Mb estimated genome size). We have also studied transcriptomes (RNA-Seq) of two subtypes of O. tenuiflorum, Krishna and Rama Tulsi and report the relative expression of genes in both the varieties. RESULTS: The pathways leading to the production of medicinally-important specialized metabolites have been studied in detail, in relation to similar pathways in Arabidopsis thaliana and other plants. Expression levels of anthocyanin biosynthesis-related genes in leaf samples of Krishna Tulsi were observed to be relatively high, explaining the purple colouration of Krishna Tulsi leaves. The expression of six important genes identified from genome data were validated by performing q-RT-PCR in different tissues of five different species, which shows the high extent of urosolic acid-producing genes in young leaves of the Rama subtype. In addition, the presence of eugenol and ursolic acid, implied as potential drugs in the cure of many diseases including cancer was confirmed using mass spectrometry. CONCLUSIONS: The availability of the whole genome of O.tenuiflorum and our sequence analysis suggests that small amino acid changes at the functional sites of genes involved in metabolite synthesis pathways confer special medicinal properties to this herb.
Assuntos
Regulação da Expressão Gênica de Plantas , Genoma de Planta , Ocimum/genética , Índia , Ocimum/metabolismo , Folhas de Planta/metabolismo , Plantas Medicinais/genética , Plantas Medicinais/metabolismoRESUMO
Understanding the principles of abiotic and biotic stress responses, tolerance and adaptation remains important in plant physiology research to develop better varieties of crop plants. Better understanding of plant stress response mechanisms and application of knowledge derived from integrated experimental and bioinformatics approaches are gaining importance. Earlier, we showed that compiling a database of stress-responsive transcription factors and their corresponding target binding sites in the form of Hidden Markov models at promoter, untranslated and upstream regions of stress-up-regulated genes from expression analysis can help in elucidating various aspects of the stress response in Arabidopsis. In addition to the extensive content in the first version, STIFDB2 is now updated with 15 stress signals, 31 transcription factors and 5,984 stress-responsive genes from three species (Arabidopsis thaliana, Oryza sativa subsp. japonica and Oryza sativa subsp. indica). We have employed an integrated biocuration and genomic data mining approach to characterize the data set of transcription factors and consensus binding sites from literature mining and stress-responsive genes from the Gene Expression Omnibus. STIFDB2 currently has 38,798 associations of stress signals, stress-responsive genes and transcription factor binding sites predicted using the Stress-responsive Transcription Factor (STIF) algorithm, along with various functional annotation data. As a unique plant stress regulatory genomics data platform, STIFDB2 can be utilized for targeted as well as high-throughput experimental and computational studies to unravel principles of the stress regulome in dicots and gramineae. STIFDB2 is available from the URL: http://caps.ncbs.res.in/stifdb2.
Assuntos
Arabidopsis/genética , Bases de Dados Genéticas , Genes de Plantas , Oryza/genética , Software , Estresse Fisiológico , Fatores de Transcrição/genética , Adaptação Biológica , Algoritmos , Sítios de Ligação , Internet , Anotação de Sequência Molecular , Transdução de Sinais , Transcrição GênicaRESUMO
Fenugreek (Trigonella foenum-graecum L.) is a self-pollinated leguminous crop belonging to the Fabaceae family. It is a multipurpose crop used as herb, spice, vegetable and forage. It is a traditional medicinal plant in India attributed with several nutritional and medicinal properties including antidiabetic and anticancer. We have performed a combined transcriptome assembly from RNA sequencing data derived from leaf, stem and root tissues. Around 209,831 transcripts were deciphered from the assembly of 92% completeness and an N50 of 1382 bases. Whilst secondary metabolites of medicinal value, such as trigonelline, diosgenin, 4-hydroxyisoleucine and quercetin, are distributed in several tissues, we report transcripts that bear sequence signatures of enzymes involved in the biosynthesis of such metabolites and are highly expressed in leaves, stem and roots. One of the antidiabetic alkaloid, trigonelline and its biosynthesising enzyme, is highly abundant in leaves. These findings are of value to nutritional and the pharmaceutical industry.
Assuntos
Diosgenina , Plantas Medicinais , Trigonella , Diosgenina/metabolismo , Hipoglicemiantes/metabolismo , Extratos Vegetais/metabolismo , Plantas Medicinais/genética , Plantas Medicinais/metabolismo , Transcriptoma , Trigonella/genética , Trigonella/metabolismoRESUMO
Oil palm (Elaeis guineensis Jacq.) is a perennial vegetable and a high oil-yielding crop (4-6 t/ha). There is a large scope for increasing the oil yield by selecting elite planting material for breeding programme in germplasm evaluation, characterization and utilization. In the present study, a diverse range of 150 oil palm genotypes were characterized using 12 quantitative variables with 54genomic microsatellite markers. A wide variation was observed in the morphological traits among indigenous populations. Highly significantand positive correlations were observed between vegetative dry matter (VDM) and total dry matter (TDM) (0.862), and height and height increment (0.838). The first two principal component analyses explained 67.7% of total variation among morphological traits. The genotypes IC0610001-59 (Pune-2) and IC0610001-60 (Pune-2) were found highly promising based on less height increment, more TDMwith high yield. For the mapping study, general linear model (GLM) approach, quantitative-trait loci (QTL) for annual height increment, number of bunches, bunch yield and bunch index were linked to simple-sequence repeat (SSR) loci mEgCIR3649 with phenotypic variance of 15.08, 10.43, 11.74, 15.39. TDM and VDM were linked to mEgCIR0192 (27.34 and 24.19%), mEgCIR3684 (16.84 and 18.30%), SPSC00163 (18.8 and 15.39%) and mEgCIR0555 (16.47 and 18.81%), with at a significant threshold (P) level of B0.001 and by mixed linear model (MLM) approach. TDM was linked to mEgCIR0555 with phenotypic variance of 20.72%, bunch yield and bunch index were linked to mEgCIR2813 at phenotypic variance of 17.11% and 12.88%, respectively, at a significant threshold (P) level of B0.01.
Assuntos
Arecaceae/genética , DNA de Plantas/genética , Repetições de Microssatélites , Locos de Características Quantitativas , Arecaceae/crescimento & desenvolvimento , Mapeamento Cromossômico , Cromossomos de Plantas , Estudos de Associação Genética , Genótipo , Índia , Óleo de Palmeira , Fenótipo , Melhoramento Vegetal , Análise de Sequência de DNARESUMO
In this paper, we present the data acquired during transcriptome analysis of the plant Moringa oleifera [1] from five different tissues (root, stem, leaf, flower and seed) by RNA sequencing. A total of 271 million reads were assembled with an N50 of 2094â¯bp. The combined transcriptome was assessed for transcript abundance across five tissues. The protein coding genes identified from the transcripts were annotated and used for orthology analysis. Further, enzymes involved in the biosynthesis of select medicinally important secondary metabolites, vitamins and ion transporters were identified and their expression levels across tissues were examined. The data generated by RNA sequencing has been deposited to NCBI public repository under the accession number PRJNA394193 (https://www.ncbi.nlm.nih.gov/bioproject/PRJNA394193).
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This protocol describes a stepwise process to identify proteins of interest from a query proteome derived from NGS data. We implemented this protocol on Moringa oleifera transcriptome to identify proteins involved in secondary metabolite and vitamin biosynthesis and ion transport. This knowledge-driven protocol identifies proteins using an integrated approach involving sensitive sequence search and evolutionary relationships. We make use of functionally important residues (FIR) specific for the query protein family identified through its homologous sequences and literature. We screen protein hits based on the clustering with true homologues through phylogenetic tree reconstruction complemented with the FIR mapping. The protocol was validated for the protein hits through qRT-PCR and transcriptome quantification. Our protocol demonstrated a higher specificity as compared to other methods, particularly in distinguishing cross-family hits. This protocol was effective in transcriptome data analysis of M. oleifera as described in Pasha et al.â¢Knowledge-driven protocol to identify secondary metabolite synthesizing protein in a highly specific manner.â¢Use of functionally important residues for screening of true hits.â¢Beneficial for metabolite pathway reconstruction in any (species, metagenomics) NGS data.
RESUMO
Plants are essential facilitators of human life on planet earth. Plants play a critical functional role in mediating the quality of air, availability of food and the sustainability of agricultural resources. However, plants are in constant interaction with its environment and often hampered by various types of stresses like biotic and abiotic ones. Biotic stress is a significant reason for crop-loss and causes yield loss in the range of 31-42%, post-harvest loss due to biotic stress is in the range of 6-20%, and abiotic stress causes 6-20% of the crop damage. Recognizing the molecular factors driving plant stress-related events, and developing molecular strategies to aid plants to tolerate, resist or adapt to biotic and abiotic stress are critical for sustainable agriculture practice. In this review, we discuss how recent advances in bioinformatics, plant genomics, and data science could help to improve our understanding of plant stress biology and improve the scale of global food production. We present various areas of scientific and technological advances, such as increased availability of genomics data through whole genome sequencing that require attention. We also discuss emerging techniques including CRISPR-Cas9 based genome engineering systems to develop plant varieties that can handle combinatorial stress signals. Growing trend of converging multiple omics technologies and availability of accurate, multi-scale models of plant stress through the study of orthologs and synteny studies, would improve our knowledge of how plants perceive, respond, and manage stress to thrive as resilient crop species and thus help to reduce global food crisis.
Assuntos
Modelos Biológicos , Plantas/metabolismo , Estresse Fisiológico , Agricultura , Abastecimento de Alimentos , Regulação da Expressão Gênica de Plantas/genética , Engenharia Genética , Genoma de Planta , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Redes e Vias Metabólicas/genética , Plantas/genética , Biologia de SistemasRESUMO
Stem gall (Protomyces macrosporus Unger), a serious disease that affects leaves, petioles, stems and fruits of coriander (Coriandrum sativum L.) causing heavy loss in yield. Genetic improvement of coriander for stem gall disease is indispensable. Coriander cultivars of stem gall resistance (ACr-1) and susceptible (CS-6) leaf samples were utilized and transcriptome sequenced using Illumina NextSeq500 platform. After trimming low-quality reads and adapter sequences, a total of 49,163,108 and 43,746,120 high-quality reads were retained and further assembly resulted validated transcripts of 59,933 and 56,861. We have predicted 52,506 and 48,858 coding sequences (CDS) ofwhich 50,506 and 46,945 were annotated using NCBInr database. Gene ontology analysis annotated 19,099 and 17,625 terms; pathway analysis obtained 24 different functional pathway categories; signal transduction, transport, catabolism, translation and carbohydrate metabolism pathways etc. were dominated. Differentially expressed genes analysis predicted 13,123 CDS commonly expressed of which 431 and 400 genes were significantly upregulated and downregulated, respectively, in which Rgenes, stress inducible transcription factors such as ERF, NAC, bZIP, MYB, DREB and WRKY and antifungal related genes were predicted. The real-time PCR analysis of HSP20 gene expression in resistance showed upregulation by 10-fold over susceptible sample and 18s used as a housekeeping gene for normalization. The present results provide an insights into various aspects underlying the development of resistance to stem gall in coriander.
Assuntos
Coriandrum/genética , Resistência à Doença/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Doenças das Plantas/genética , Folhas de Planta/genética , Proteínas de Plantas/genética , Coriandrum/microbiologia , Ontologia Genética , Interações Hospedeiro-Patógeno , Anotação de Sequência Molecular , Doenças das Plantas/microbiologia , Folhas de Planta/microbiologia , TranscriptomaRESUMO
Finger millet (Eleusine coracana (L.) Gaertn.), an important C4 species is known for its stress hardiness and nutritional significance. To identify novel drought responsive mechanisms, we generated transcriptome data from leaf tissue of finger millet, variety GPU-28, exposed to gravimetrically imposed drought stress so as to simulate field stress conditions. De novo assembly based approach yielded 80,777 and 90,830 transcripts from well-irrigated (control) and drought-stressed samples, respectively. A total of 1790 transcripts were differentially expressed between the control and drought-stress treatments. Functional annotation and pathway analysis indicated activation of diverse drought-stress signalling cascade genes such as serine threonine protein phosphatase 2A (PP2A), calcineurin B-like interacting protein kinase31 (CIPK31), farnesyl pyrophosphate synthase (FPS), signal recognition particle receptor α (SRPR α) etc. The basal regulatory genes such as TATA-binding protein (TBP)-associated factors (TAFs) werefound to be drought responsive, indicating that genes associated with housekeeping or basal regulatory processes are activated underdrought in finger millet. A significant portion of the expressed genes was uncharacterized, belonging to the category of proteins of unknown functions (PUFs). Among the differentially expressed PUFs, we attempted to assign putative function for a few, using anovel annotation tool, Proteins of Unknown Function Annotation Server. Analysis of PUFs led to the discovery of novel drought responsive genes such as pentatricopeptide repeat proteins and tetratricopeptide repeat proteins that serve as interaction modules in multiprotein interactions. The transcriptome data generated can be utilized for comparative analysis, and functional validation of the genes identified would be useful to understand the drought adaptive mechanisms operating under field conditions in finger millet, as has been already attempted for a few candidates such as CIPK31 and TAF6. Such an attempt is needed to enhance the productivity of finger millet under water-limited conditions, and/or to adopt the implicated mechanisms in other related crops.
Assuntos
Adaptação Biológica/genética , Secas , Eleusine/genética , Perfilação da Expressão Gênica , Estresse Fisiológico/genética , Transcriptoma , Biologia Computacional/métodos , Regulação da Expressão Gênica de Plantas , Genômica/métodos , Anotação de Sequência MolecularRESUMO
BACKGROUND: The midgut of silkworm (Bombyx mori L.) plays an important role as a natural barrier and source of innate immunity. We had purified the novel red fluorescent protein (RFP) from the midgut of the silkworm Bombyx mori L. and bioassay studies confirmed RFPs possess antiviral, antifungal and antibacterial properties. N-terminal sequence of RFP analysis predicted chbp gene and it belongs to lipocalin gene family and is known to involve in anti-pathogenic activities. OBJECTIVE: The main objective of this study was to purify RFP from the midgut of Kolar Gold silkworm and confirm its antimicrobial activity. METHODS: For isolation of RFP, midgut juice was collected by brief exposure to chloroform vapours to fifth instar Kolar Gold silkworm larvae. Juice was purified by 40 % ammonium sulfate precipitation and purified by gel filtration chromatography (GFC) and fractions with fluorescence red under Ultra violet (UV) were collected. Molecular weight and purity of RFP was identified using PAGE, MALDI-TOF and HPLC. Antimicrobial property of purified RFP against BmNPV, Escherichia coli, Klebsiella pneumonia, Bacillus subtilis and Phytophthora meadii was performed. N-terminal sequencing of RFP was performed using Edman degradation method. Using ten amino acid sequence, using default parameter BLAST search was performerd. From the fifth day old fifth instar silkworm midgut mRNA was isolated and cDNA was synthesized using oligo-dt primer and amplification of ChBP gene was carried out by using cDNA as the template and ChBP gene specific primers. chbp protein sequence as a input built the homology model by using SWISS-MODEL. RESULTS: RFP was purified by 40 % ammonium sulfate precipitation and gel filtration chromatography (GFC) and fractions with fluorescence red under Ultra violet (UV) were collected and SDS - PAGE revealed a size of 40 kDa. RFP purified by GFC was further reconfirmed by HPLC with a single peak with a retention time of 8.755 min. MALDI-TOF produced a peak at a molecular mass of 40 kDa. RFP from the midgut juice showed antiviral activity against the silkworm virus BmNPV, antibacterial activity against Escherichia coli, Klebsiella pneumonia, Bacillus subtilis and Phytophthora meadii. N-terminal sequencing of RFP by Edman degradation method sequenced TQTIETDYWV amino acids and BLAST analysis predicted the Chlorophyllide-a Binding Protein (chbp) with B. mori. PCR product was sequenced and obtained 911bp nucleotides encoding 302 amino acid residues and deposited with the accession number KX186723 in NCBI. Sequence analysis revealed Chbp belongs to lipocalin gene family and known to involve in antiviral, antifungal and anti-bacterial properties. Chbp gene homology model was predicted using crystal structure of insecticyanin A from the tobacco hornworm as a template. CONCLUSION: Our results indicated RFP present in midgut juice of 5th instar larvae of kolar gold silkworm. We have purified novel RFP with molecular mass of 40 kDa and showed its antipathogenic activities. Chbp gene synthesises RFP and further it could be utilized for agriculture and pharmaceutical industry.
Assuntos
Antibacterianos/química , Antifúngicos/química , Antivirais/química , Bombyx/química , Sistema Digestório/química , Proteínas Luminescentes/química , Animais , Antibacterianos/isolamento & purificação , Antibacterianos/farmacologia , Antifúngicos/isolamento & purificação , Antifúngicos/farmacologia , Antivirais/isolamento & purificação , Bombyx/metabolismo , Simulação por Computador , Sistema Digestório/metabolismo , Larva/química , Larva/metabolismo , Luz , Proteínas Luminescentes/isolamento & purificação , Proteínas Luminescentes/farmacologia , Conformação Proteica , Homologia de Sequência de Aminoácidos , Proteína Vermelha FluorescenteRESUMO
The modern sequencing technologies are generating large volumes of information at the transcriptome and genome level. Translation of this information into a biological meaning is far behind the race due to which a significant portion of proteins discovered remain as proteins of unknown function (PUFs). Attempts to uncover the functional significance of PUFs are limited due to lack of easy and high throughput functional annotation tools. Here, we report an approach to assign putative functions to PUFs, identified in the transcriptome of mulberry, a perennial tree commonly cultivated as host of silkworm. We utilized the mulberry PUFs generated from leaf tissues exposed to drought stress at whole plant level. A sequence and structure based computational analysis predicted the probable function of the PUFs. For rapid and easy annotation of PUFs, we developed an automated pipeline by integrating diverse bioinformatics tools, designated as PUFs Annotation Server (PUFAS), which also provides a web service API (Application Programming Interface) for a large-scale analysis up to a genome. The expression analysis of three selected PUFs annotated by the pipeline revealed abiotic stress responsiveness of the genes, and hence their potential role in stress acclimation pathways. The automated pipeline developed here could be extended to assign functions to PUFs from any organism in general. PUFAS web server is available at http://caps.ncbs.res.in/pufas/ and the web service is accessible at http://capservices.ncbs.res.in/help/pufas.
Assuntos
Morus/genética , Proteínas de Plantas/genética , Transcriptoma , Sequência de Aminoácidos , Dados de Sequência Molecular , Morus/classificação , Filogenia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Ligação Proteica , Homologia de Sequência de AminoácidosRESUMO
Biological enrichment analysis using gene ontology (GO) provides a global overview of the functional role of genes or proteins identified from large-scale genomic or proteomic experiments. Phenomic enrichment analysis of gene lists can provide an important layer of information as well as cellular components, molecular functions, and biological processes associated with gene lists. Plant phenomic enrichment analysis will be useful for performing new experiments to better understand plant systems and for the interpretation of gene or proteins identified from high-throughput experiments. Plant ontology (PO) is a compendium of terms to define the diverse phenotypic characteristics of plant species, including plant anatomy, morphology, and development stages. Adoption of this highly useful ontology is limited, when compared to GO, because of the lack of user-friendly tools that enable the use of PO for statistical enrichment analysis. To address this challenge, we introduce Plant Ontology Enrichment Analysis Server (POEAS) in the public domain. POEAS uses a simple list of genes as input data and performs enrichment analysis using Ontologizer 2.0 to provide results in two levels, enrichment results and visualization utilities, to generate ontological graphs that are of publication quality. POEAS also offers interactive options to identify user-defined background population sets, various multiple-testing correction methods, different enrichment calculation methods, and resampling tests to improve statistical significance. The availability of such a tool to perform phenomic enrichment analyses using plant genes as a complementary resource will permit the adoption of PO-based phenomic analysis as part of analytical workflows. POEAS can be accessed using the URL http://caps.ncbs.res.in/poeas.
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Biotic and abiotic stresses adversely affect agriculture by reducing crop growth and productivity worldwide. To investigate the abiotic stress-responsive genes in Arabidopsis thaliana, we compiled a dataset of stress signals and differentially upregulated genes (>= 2.5 fold change) from Stress-responsive transcription Factors DataBase (STIFDB) with additional set of stress signals and genes curated from PubMed and Gene Expression Omnibus. A dataset of 3091 genes differentially upregulated due to 14 different stress signals (abscisic acid, aluminum, cold, cold-drought-salt, dehydration, drought, heat, iron, light, NaCl, osmotic stress, oxidative stress, UV-B and wounding) were curated and used for the analysis. Details about stress-responsive enriched genes and their association with stress signals can be obtained from STIFDB2 database . The gene-stress-signal data were analyzed using an enrichment-based meta-analysis framework consisting of two different ontologies (Gene Ontology and Plant Ontology), biological pathway and functional domain annotations. We found several shared and distinct biological processes, cellular components and molecular functions associated with stress-responsive genes. Pathway analysis revealed that stress-responsive genes perturbed the pathways under the "Metabolic pathways" category. We also found several shared and stress-signal specific protein domains, suggesting functional mechanisms regulating stress-response. Phenomic characteristics of abiotic stress-responsive genes were ascertained for several stresses and found to be shared by multiple stresses in both anatomy and temporal categories of Plant Ontology. We found several constitutive stress-responsive genes that are differentially upregulated due to perturbation of different stress signals, for example a gene (AT1G68440) involved in phenylpropanoid metabolism and polyamine catabolism as responsive to seven different stress signals. We also performed structure-function prediction of five genes associated responsive to multiple abiotic stress signals. We envisage that results from our analysis that provide insight into functional repertoire, metabolic pathways and phenomic characteristics common and specifically associated with stress signals would help to understand abiotic stress regulome in Arabidopsis thaliana and may also help to develop an improved plant variety using molecular breeding and genetic engineering techniques that are rapidly stress-responsive and tolerant.