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1.
Br J Nutr ; 131(4): 686-697, 2024 02 28.
Artigo em Inglês | MEDLINE | ID: mdl-37781761

RESUMO

The human gut microbiota can biosynthesize essential micronutrients such as B-vitamins and is also known for its metabolic cooperative behaviour. The present study characterises such B-vitamin biosynthesizers, their biosynthetic pathways, explores their prevalence and abundance, examines how lifestyle or diet affects them in multiple Indian cohorts and compares it with the Chinese cohort. To achieve this, publicly available faecal metagenome data of healthy individuals from multiple Indian (two urban and three tribal populations) and a Chinese cohort were analysed. The distribution of prevalence and abundance of B-vitamin biosynthesizers showed similar profiles to that of the entire gut community of the Indian cohort, and there were 28 B-vitamin biosynthesizers that had modest or higher prevalence and abundance. The omnivorous diet affected only the prevalence of a few B-vitamin biosynthesizers; however, lifestyle and/or location affected both prevalence and abundance. A comparison with the Chinese cohort showed that fourteen B-vitamin biosynthesizers were significantly more prevalent and abundant in Chinese as compared with Indian samples (False Discovery Rate (FDR) <= 0·05). The metabolic potential of the entire gut community for B-vitamin production showed that within India, the tribal cohort has a higher abundance of B-vitamin biosynthesis pathways as compared with two urban cohorts namely, Bhopal and Kasargod, and comparison with the Chinese cohort revealed a higher abundance in the latter group. Potential metabolic cooperative behaviour of the Indian gut microbiome for biosynthesis of the B-vitamins showed multiple pairs of species showed theoretical complementarity for complete biosynthetic pathways genes of thiamine, riboflavin, niacin and pantothenate.


Assuntos
Microbioma Gastrointestinal , Complexo Vitamínico B , Humanos , Microbioma Gastrointestinal/genética , Tiamina , Riboflavina/análise , China
2.
Plant Cell Rep ; 42(6): 1059-1069, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37074465

RESUMO

KEY MESSAGE: Genome-wide structural variants we identified and new NOR-linked markers we developed would be useful for future genome-wide association studies (GWAS), and for new gene/trait mapping purposes. Bioinformatic alignment of the assembled genomes of Col-0 and Sha ecotypes of Arabidopsis thaliana revealed ~ 13,000 genome-wide structural variants involving simple insertions or deletions and repeat contractions or expansions. Using some of these structural variants, we developed new, rapid, and low-cost PCR-based molecular markers that are genetically linked to the nucleolus organizer regions (NORs). A. thaliana has two NORs, one each on chromosome 2 (NOR2) and chromosome 4 (NOR4). Both NORs are ~ 4 Mb each, and hundreds of 45S ribosomal RNA (rRNA) genes are tandemly arrayed at these loci. Using previously characterized recombinant inbred lines (RILs) derived from Sha x Col-0 crosses, we validated the utility of the newly developed NOR-linked markers in genetically mapping rRNA genes and the associated telomeres to either NOR2 or NOR4. Lastly, we sequenced Sha genome using Oxford Nanopore Technology (ONT) and used the data to obtain sequences of NOR-telomere junctions, and with the help of RILs, we mapped them as new genetic markers to their respective NORs (NOR2-TEL2N and NOR4-TEL4N). The structural variants obtained from this study would serve as valuable data for genome-wide association studies (GWAS), and to rapidly design more genome-wide genetic (molecular) markers for new gene/trait mapping purposes.


Assuntos
Arabidopsis , Arabidopsis/genética , Estudo de Associação Genômica Ampla , Mapeamento Cromossômico , Sequência de Bases , Telômero
3.
Parasitol Res ; 122(1): 285-298, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36399171

RESUMO

During amoebiasis, colonization of the gut by Entamoeba histolytica can lead to alterations of the host microbiota. In this study, we have compared the gut microbiota of patients of amoebiasis with healthy controls using 16S rRNA gene variable regions, (V1-V3) and (V3-V5), of the bacterial genome. From this 16S rRNA gene amplicon data, one paired-end and two single-end datasets were selected and compared by the number of OTUs obtained, sequence count, and diversity analysis. Our results showed that the V1-V3-paired-end dataset gave the maximum number of OTUs in comparison to the two single-end datasets studied. The amoebiasis samples showed a significant drop in richness in the alpha diversity measurements and lower intra group similarity compared to the healthy controls. Bacteria of genus Prevotella, Sutterella, and Collinsella were more abundant in healthy controls whereas Escherichia, Klebsiella, and Ruminococcus were more abundant in the E. histolytica-positive patients. All the healthy controls harbored bacteria belonging to Faecalibacterium, Prevotella, Ruminococcus, Subdoligranulum, and Escherichia genera while all the E. histolytica-positive patient samples contained genus Enterobacter. The compositional changes in the gut microbiome observed in our study indicated a higher prevalence of pathogenic bacteria along with a depletion of beneficial bacteria in E. histolytica-infected individuals when compared with healthy controls. These results underline the interplay between E. histolytica and the human gut microbiome, giving important inputs for future studies and treatments.


Assuntos
Entamebíase , Microbioma Gastrointestinal , Humanos , Microbioma Gastrointestinal/genética , RNA Ribossômico 16S/genética , Bactérias/genética , Diarreia , Índia , Fezes/microbiologia
4.
J Obstet Gynaecol Res ; 49(1): 194-200, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36647324

RESUMO

BACKGROUND: Intrapartum care is of paramount importance in overall respectful maternity care (RMC). Uterine fundal pressure maneuver (UFPM) is the most controversial obstetric maneuver considered obsolete in many countries. UFPM is associated with adverse life threatening maternal and fetal effects. The baseline prevalence of UFPM in our tertiary care teaching hospital with a heavy patient load was 78.4% which was quite high. Our aim was to reduce the prevalence of UFPM by 75% from baseline over a period of 10 months. METHODS: After root cause analysis, we formulated the departmental Standard Operating Procedures on safe vaginal delivery and birth practices and initiated the Labour Room Quality Improvement Initiative. We sensitized and created awareness among the resident doctors and nursing staff regarding the high prevalence of UFPM through dedicated lectures, intercommunication via WhatsApp groups and strict vigilance in the labor wards. The point of care quality improvement (QI) methodology was used. The primary outcome was decrease in the prevalence of UFPM. RESULTS: The prevalence of UFPM reduced from the baseline value of 78.4%-4% over the period of 7 months. Post intervention it gradually increased and sustained at 21.2% over a follow-up period of 3 months. CONCLUSION: QI methods can effectively and rapidly improve the acceptance and adherence to newer initiatives in a busy tertiary care health facility to reduce the prevalence of UFPM and eventually improve the overall RMC.


Assuntos
Serviços de Saúde Materna , Melhoria de Qualidade , Gravidez , Feminino , Humanos , Prevalência , Países em Desenvolvimento , Parto Obstétrico/métodos , Segunda Fase do Trabalho de Parto , Qualidade da Assistência à Saúde
5.
Planta ; 253(2): 59, 2021 Feb 04.
Artigo em Inglês | MEDLINE | ID: mdl-33538916

RESUMO

MAIN CONCLUSION: Comparative analysis of genome-wide miRNAs and their gene targets between cytoplasmic male sterile (CMS) and fertile lines of pigeonpea suggests a possible role of miRNA-regulated pathways in reproductive development. Exploitation of hybrid vigor using CMS technology has delivered nearly 50% yield gain in pigeonpea. Among various sterility-inducing cytoplasms (A1-A9) reported so far in pigeonpea, A2 and A4 are the two major sources that facilitate hybrid seed production. Recent evidence suggests involvement of micro RNA in vast array of biological processes including plant reproductive development. In pigeonpea, information about the miRNAs is insufficient. In view of this, we sequenced six small RNA libraries of CMS line UPAS 120A and isogenic fertile line UPAS 120B using Illumina technology. Results revealed 316 miRNAs including 248 known and 68 novel types. A total of 637 gene targets were predicted for known miRNAs, while 324 genes were associated with novel miRNAs. Degradome analysis revealed 77 gene targets of predicted miRNAs, which included a variety of transcription factors playing key roles in plant reproduction such as F-box family proteins, apetala 2, auxin response factors, ethylene-responsive factors, homeodomain-leucine zipper proteins etc. Differential expression of both known and novel miRNAs implied roles for both conserved as well as species-specific players. We also obtained several miRNA families such as miR156, miR159, miR167 that are known to influence crucial aspects of plant fertility. Gene ontology and pathway level analyses of the target genes showed their possible implications for crucial events during male reproductive development such as tapetal degeneration, pollen wall formation, retrograde signaling etc. To the best of our knowledge, present study is first to combine deep sequencing of small RNA and degradome for elucidating the role of miRNAs in flower and male reproductive development in pigeonpea.


Assuntos
Cajanus/genética , MicroRNAs , Infertilidade das Plantas/genética , RNA de Plantas/genética , Cajanus/fisiologia , Citoplasma , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Sequenciamento de Nucleotídeos em Larga Escala , MicroRNAs/genética
6.
J Exp Bot ; 72(8): 3122-3136, 2021 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-33528493

RESUMO

In C4 species, ß-carbonic anhydrase (CA), localized to the cytosol of the mesophyll cells, accelerates the interconversion of CO2 to HCO3-, the substrate used by phosphoenolpyruvate carboxylase (PEPC) in the first step of C4 photosynthesis. Here we describe the identification and characterization of low CO2-responsive mutant 1 (lcr1) isolated from an N-nitroso-N-methylurea- (NMU) treated Setaria viridis mutant population. Forward genetic investigation revealed that the mutated gene Sevir.5G247800 of lcr1 possessed a single nucleotide transition from cytosine to thymine in a ß-CA gene causing an amino acid change from leucine to phenylalanine. This resulted in severe reduction in growth and photosynthesis in the mutant. Both the CO2 compensation point and carbon isotope discrimination values of the mutant were significantly increased. Growth of the mutants was stunted when grown under ambient pCO2 but recovered at elevated pCO2. Further bioinformatics analyses revealed that the mutation has led to functional changes in one of the conserved residues of the protein, situated near the catalytic site. CA transcript accumulation in the mutant was 80% lower, CA protein accumulation 30% lower, and CA activity ~98% lower compared with the wild type. Changes in the abundance of other primary C4 pathway enzymes were observed; accumulation of PEPC protein was significantly increased and accumulation of malate dehydrogenase and malic enzyme decreased. The reduction of CA protein activity and abundance in lcr1 restricts the supply of bicarbonate to PEPC, limiting C4 photosynthesis and growth. This study establishes Sevir.5G247800 as the major CA allele in Setaria for C4 photosynthesis and provides important insights into the function of CA in C4 photosynthesis that would be required to generate a rice plant with a functional C4 biochemical pathway.


Assuntos
Anidrases Carbônicas , Fotossíntese , Proteínas de Plantas , Setaria (Planta) , Dióxido de Carbono , Anidrases Carbônicas/genética , Anidrases Carbônicas/metabolismo , Células do Mesofilo/metabolismo , Setaria (Planta)/enzimologia , Setaria (Planta)/genética
7.
Med J Armed Forces India ; 77(1): 28-31, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33487862

RESUMO

BACKGROUND: The purpose of the present study was to determine the airway changes in skeletal class II division 1 malocclusion patients with mandibular retrognathism, treated with Twin-Block (TB) appliance. METHODS: Airway assessment was carried for twelve patients (mean age 11.7 ± 1.1 years) who underwent myofunctional therapy using TB appliance for correction of skeletal class II division 1 malocclusion with mandibular retrognathism. Acoustic pharyngometry (AP) was used to assess and quantify the comparative changes in the upper airway, pretreatment and posttreatment. RESULTS: Data acquired was subjected to appropriate statistical analysis. The paired 't' test was used to compare pre-treatment (T0) and after the positive pterygoid response (T1). TB appliance increased mean minimum airway area by 0.28 ± 0.25 cm2 and mean airway by 0.47 ± 0.44 cm2 with 95% CI. Posttreatment minimum airway and mean area changes were found to be statistically significant (P-value<0.01). CONCLUSION: TB appliance therapy has a positive effect on upper airway and is beneficial for the treatment of sleep-related disorders associated with Class II division 1 malocclusion for achieving positive functional changes, esthetics, and healthier quality of life.

8.
BMC Genomics ; 20(1): 206, 2019 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-30866809

RESUMO

BACKGROUND: Promoter motifs in Entamoeba histolytica were earlier analysed using microarray data with lower dynamic range of gene expression. Additionally, previous transcriptomic studies did not provide information on the nature of highly transcribed genes, and downstream promoter motifs important for gene expression. To address these issues we generated RNA-Seq data and identified the high and low expressing genes, especially with respect to virulence potential. We analysed sequences both upstream and downstream of start site for important motifs. RESULTS: We used RNA-Seq data to classify genes according to expression levels, which ranged six orders of magnitude. Data were validated by reporter gene expression. Virulence-related genes (except AIG1) were amongst the highly expressed, while some kinases and BspA family genes were poorly expressed. We looked for conserved motifs in sequences upstream and downstream of the initiation codon. Following enrichment by AME we found seven motifs significantly enriched in high expression- and three in low expression-classes. Two of these motifs (M4 and M6) were located downstream of AUG, were exclusively enriched in high expression class, and were mostly found in ribosomal protein, and translation-related genes. Motif deletion resulted in drastic down regulation of reporter gene expression, showing functional relevance. Distribution of core promoter motifs (TATA, GAAC, and Inr) in all genes revealed that genes with downstream motifs were not preferentially associated with TATA-less promoters. We looked at gene expression changes in cells subjected to growth stress by serum starvation, and experimentally validated the data. Genes showing maximum up regulation belonged to the low or medium expression class, and included genes in signalling pathways, lipid metabolism, DNA repair, Myb transcription factors, BspA, and heat shock. Genes showing maximum down regulation belonged to the high or medium expression class. They included genes for signalling factors, actin, Ariel family, and ribosome biogenesis factors. CONCLUSION: Our analysis has added important new information about the E. histolytica transcriptome. We report for the first time two downstream motifs required for gene expression, which could be used for over expression of E. histolytica genes. Most of the virulence-related genes in this parasite are highly expressed in culture.


Assuntos
Entamoeba histolytica/patogenicidade , Perfilação da Expressão Gênica/métodos , Fatores de Virulência/genética , Entamoeba histolytica/genética , Regulação Neoplásica da Expressão Gênica , Redes Reguladoras de Genes , Regiões Promotoras Genéticas , Análise de Sequência de RNA , Sequenciamento Completo do Genoma
9.
Plant Physiol ; 170(3): 1655-74, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26813793

RESUMO

Leaves are derived from heterotrophic meristem tissue that, at some point, must make the transition to autotrophy via the initiation of photosynthesis. However, the timing and spatial coordination of the molecular and cellular processes underpinning this switch are poorly characterized. Here, we report on the identification of a specific stage in rice (Oryza sativa) leaf development (P3/P4 transition) when photosynthetic competence is first established. Using a combined physiological and molecular approach, we show that elements of stomatal and vascular differentiation are coordinated with the onset of measurable light absorption for photosynthesis. Moreover, by exploring the response of the system to environmental perturbation, we show that the earliest stages of rice leaf development have significant plasticity with respect to elements of cellular differentiation of relevance for mature leaf photosynthetic performance. Finally, by performing an RNA sequencing analysis targeted at the early stages of rice leaf development, we uncover a palette of genes whose expression likely underpins the acquisition of photosynthetic capability. Our results identify the P3/P4 transition as a highly dynamic stage in rice leaf development when several processes for the initiation of photosynthetic competence are coordinated. As well as identifying gene targets for future manipulation of rice leaf structure/function, our data highlight a developmental window during which such manipulations are likely to be most effective.


Assuntos
Clorofila/metabolismo , Perfilação da Expressão Gênica/métodos , Oryza/genética , Fotossíntese/genética , Folhas de Planta/genética , Clorofila/química , Fluorescência , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Luz , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Oryza/crescimento & desenvolvimento , Oryza/metabolismo , Fotossíntese/efeitos da radiação , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Estômatos de Plantas/genética , Estômatos de Plantas/metabolismo , Estômatos de Plantas/ultraestrutura , Feixe Vascular de Plantas/genética , Feixe Vascular de Plantas/crescimento & desenvolvimento , Feixe Vascular de Plantas/metabolismo , Plastídeos/genética , Plastídeos/metabolismo , Plastídeos/ultraestrutura , Fatores de Tempo
11.
Plant J ; 84(2): 257-66, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26333774

RESUMO

The specification of vascular patterning in plants has interested plant biologists for many years. In the last decade a new context has emerged for this interest. Specifically, recent proposals to engineer C(4) traits into C(3) plants such as rice require an understanding of how the distinctive venation pattern in the leaves of C(4) plants is determined. High vein density with Kranz anatomy, whereby photosynthetic cells are arranged in encircling layers around vascular bundles, is one of the major traits that differentiate C(4) species from C(3) species. To identify genetic factors that specify C(4) leaf anatomy, we generated ethyl methanesulfonate- and γ-ray-mutagenized populations of the C(4) species sorghum (Sorghum bicolor), and screened for lines with reduced vein density. Two mutations were identified that conferred low vein density. Both mutations segregated in backcrossed F(2) populations as homozygous recessive alleles. Bulk segregant analysis using next-generation sequencing revealed that, in both cases, the mutant phenotype was associated with mutations in the CYP90D2 gene, which encodes an enzyme in the brassinosteroid biosynthesis pathway. Lack of complementation in allelism tests confirmed this result. These data indicate that the brassinosteroid pathway promotes high vein density in the sorghum leaf, and suggest that differences between C(4) and C(3) leaf anatomy may arise in part through differential activity of this pathway in the two leaf types.


Assuntos
Brassinosteroides/metabolismo , Sistema Enzimático do Citocromo P-450/genética , Folhas de Planta/genética , Proteínas de Plantas/genética , Sorghum/genética , Sistema Enzimático do Citocromo P-450/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Sorghum/metabolismo
12.
J Conserv Dent Endod ; 27(2): 214-218, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38463468

RESUMO

Apexification is a technique used to create a mineralized barrier in a root having an open apex or to continue the apical growth of a root that is still not fully developed in teeth with necrotic pulps. Biodentine is a versatile material which can help in achieving apical closure as well as reinforcement of thin dentinal walls, thus improving the overall prognosis and preserving the natural dentition. In the present case, the radiographic evaluation showed thinner dentinal walls and an open apex with periapical radiolucency in relation to the upper right central incisor. The large open root apex and thin dentin walls of immature permanent teeth render them challenging to treat with root canal therapy. This case report describes a modified single-step apexification procedure and strengthening of the dentinal walls of the root canal using Biodentine.

14.
Life (Basel) ; 13(8)2023 Jul 31.
Artigo em Inglês | MEDLINE | ID: mdl-37629524

RESUMO

Sequencing technologies have rapidly evolved over the past two decades, and new technologies are being continually developed and commercialized. The emerging sequencing technologies target generating more data with fewer inputs and at lower costs. This has also translated to an increase in the number and type of corresponding applications in genomics besides enhanced computational capacities (both hardware and software). Alongside the evolving DNA sequencing landscape, bioinformatics research teams have also evolved to accommodate the increasingly demanding techniques used to combine and interpret data, leading to many researchers moving from the lab to the computer. The rich history of DNA sequencing has paved the way for new insights and the development of new analysis methods. Understanding and learning from past technologies can help with the progress of future applications. This review focuses on the evolution of sequencing technologies, their significant enabling role in generating plant genome assemblies and downstream applications, and the parallel development of bioinformatics tools and skills, filling the gap in data analysis techniques.

15.
Am J Bot ; 99(2): 186-92, 2012 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-22301893

RESUMO

PREMISE OF THE STUDY: Next-generation sequencing (NGS) technologies are frequently used for resequencing and mining of single nucleotide polymorphisms (SNPs) by comparison to a reference genome. In crop species such as chickpea (Cicer arietinum) that lack a reference genome sequence, NGS-based SNP discovery is a challenge. Therefore, unlike probability-based statistical approaches for consensus calling and by comparison with a reference sequence, a coverage-based consensus calling (CbCC) approach was applied and two genotypes were compared for SNP identification. METHODS: A CbCC approach is used in this study with four commonly used short read alignment tools (Maq, Bowtie, Novoalign, and SOAP2) and 15.7 and 22.1 million Illumina reads for chickpea genotypes ICC4958 and ICC1882, together with the chickpea trancriptome assembly (CaTA). KEY RESULTS: A nonredundant set of 4543 SNPs was identified between two chickpea genotypes. Experimental validation of 224 randomly selected SNPs showed superiority of Maq among individual tools, as 50.0% of SNPs predicted by Maq were true SNPs. For combinations of two tools, greatest accuracy (55.7%) was reported for Maq and Bowtie, with a combination of Bowtie, Maq, and Novoalign identifying 61.5% true SNPs. SNP prediction accuracy generally increased with increasing reads depth. CONCLUSIONS: This study provides a benchmark comparison of tools as well as read depths for four commonly used tools for NGS SNP discovery in a crop species without a reference genome sequence. In addition, a large number of SNPs have been identified in chickpea that would be useful for molecular breeding.


Assuntos
Cicer/genética , Sequência Consenso , Produtos Agrícolas/genética , Genoma de Planta , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA/métodos , Sequência de Bases , Mapeamento Cromossômico/métodos , Biologia Computacional/métodos , DNA de Plantas/genética , Variação Genética , Genótipo , Padrões de Referência , Reprodutibilidade dos Testes , Alinhamento de Sequência/métodos , Transcriptoma
16.
BMC Genomics ; 12: 108, 2011 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-21324149

RESUMO

BACKGROUND: Several tools are available to identify miRNAs from deep-sequencing data, however, only a few of them, like miRDeep, can identify novel miRNAs and are also available as a standalone application. Given the difference between plant and animal miRNAs, particularly in terms of distribution of hairpin length and the nature of complementarity with its duplex partner (or miRNA star), the underlying (statistical) features of miRDeep and other tools, using similar features, are likely to get affected. RESULTS: The potential effects on features, such as minimum free energy, stability of secondary structures, excision length, etc., were examined, and the parameters of those displaying sizable changes were estimated for plant specific miRNAs. We found most of these features acquired a new set of values or distributions for plant specific miRNAs. While the length of conserved positions (nucleus) in mature miRNAs were relatively longer in plants, the difference in distribution of minimum free energy, between real and background hairpins, was marginal. However, the choice of source (species) of background sequences was found to affect both the minimum free energy and miRNA hairpin stability. The new parameters were tested on an Illumina dataset from maize seedlings, and the results were compared with those obtained using default parameters. The newly parameterized model was found to have much improved specificity and sensitivity over its default counterpart. CONCLUSIONS: In summary, the present study reports behavior of few general and tool-specific statistical features for improving the prediction accuracy of plant miRNAs from deep-sequencing data.


Assuntos
Biologia Computacional/métodos , MicroRNAs/química , Plantas/genética , RNA de Plantas/química , Interpretação Estatística de Dados , Conformação de Ácido Nucleico
17.
Funct Integr Genomics ; 11(4): 651-7, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-21547435

RESUMO

Single feature polymorphisms (SFPs) are microarray-based molecular markers that are detected by hybridization of DNA or cRNA to oligonucleotide probes. With an objective to identify the potential polymorphic markers for drought tolerance in pigeonpea [Cajanus cajan (L.) Millspaugh], an important legume crop for the semi-arid tropics but deficient in genomic resources, Affymetrix Genome Arrays of soybean (Glycine max), a closely related species of pigeonpea were used on cRNA of six parental genotypes of three mapping populations of pigeonpea segregating for agronomic traits like drought tolerance and pod borer (Helicoverpa armigiera) resistance. By using robustified projection pursuit method on 15 pair-wise comparisons for the six parental genotypes, 5,692 SFPs were identified. Number of SFPs varied from 780 (ICPL 8755 × ICPL 227) to 854 (ICPL 151 × ICPL 87) per parental combination of the mapping populations. Randomly selected 179 SFPs were used for validation by Sanger sequencing and good quality sequence data were obtained for 99 genes of which 75 genes showed sequence polymorphisms. While associating the sequence polymorphisms with SFPs detected, true positives were observed for 52.6% SFPs detected. In terms of parental combinations of the mapping populations, occurrence of true positives was 34.48% for ICPL 151 × ICPL 87, 41.86% for ICPL 8755 × ICPL 227, and 81.58% for ICP 28 × ICPW 94. In addition, a set of 139 candidate genes that may be associated with drought tolerance has been identified based on gene ontology analysis of the homologous pigeonpea genes to the soybean genes that detected SFPs between the parents of the mapping populations segregating for drought tolerance.


Assuntos
Cajanus/genética , Desidratação/genética , Polimorfismo Genético , Cajanus/fisiologia , Secas , Perfilação da Expressão Gênica , Marcadores Genéticos , Genoma de Planta , Genótipo , Análise de Sequência com Séries de Oligonucleotídeos , Estresse Fisiológico/genética
18.
Artigo em Inglês | MEDLINE | ID: mdl-33848588

RESUMO

Development of insect resistance to biopesticides is a current and pertinent global issue. Earlier, it was established that lepidopteran larvae can recover from Bt intoxication via a midgut regenerative response and subsequently generate resistance. Molecular aspects of restoration of the midgut integrity following toxin exposure are emerging recently. In the present study, we bring out the pivotal role of gut arylphorin in mediating the midgut regenerative response following sublethal Bt exposure in Achaea janata. Bt-induced midgut damage was characterized by microscopic analysis using differential interference contrast (DIC) and immunofluorescence (IF). Extensive disruption of brush-border membrane, associated with the underlying cytoskeletal alterations including F-actin, α-actin and ß-tubulin was observed. Single-photon fluorescence microscopy combined with fluorescence lifetime imaging (FLIM) established the metabolic state associated with enhanced stem cell proliferation and migration from the basal side towards the luminal side following the damage. In-silico analysis revealed the phylogenetic relationship of gut arylphorin with closely related insect species and indicated the presence of two different subunits. Transient RNAi knockdown of the arylphorin resulted in diminished expression of mitotic Cyclin B mRNA levels. Human monoclonal Cyclin B antibody cross-reactivity with the Cyclin B located in the stem cells further validate the role of arylphorin as the mitogenic factor responsible for stem cell proliferation and epithelial regeneration. An in-depth understanding of resistance mechanisms will aid in the design of new strategies for the long-term usage and efficacy of Bt technology against pest control.


Assuntos
Toxinas de Bacillus thuringiensis/toxicidade , Proteínas de Insetos/metabolismo , Intestinos , Mariposas/metabolismo , Animais , Bacillus thuringiensis
19.
Commun Biol ; 4(1): 254, 2021 02 26.
Artigo em Inglês | MEDLINE | ID: mdl-33637850

RESUMO

C4 photosynthesis provides an effective solution for overcoming the catalytic inefficiency of Rubisco. The pathway is characterised by a biochemical CO2 concentrating mechanism that operates across mesophyll and bundle sheath (BS) cells and relies on a gas tight BS compartment. A screen of a mutant population of Setaria viridis, an NADP-malic enzyme type C4 monocot, generated using N-nitroso-N-methylurea identified a mutant with an amino acid change in the gene coding region of the ABCG transporter, a step in the suberin synthesis pathway. Here, Nile red staining, TEM, and GC/MS confirmed the alteration in suberin deposition in the BS cell wall of the mutant. We show that this has disrupted the suberin lamellae of BS cell wall and increased BS conductance to CO2 diffusion more than two-fold in the mutant. Consequently, BS CO2 partial pressure is reduced and CO2 assimilation was impaired in the mutant. Our findings provide experimental evidence that a functional suberin lamellae is an essential anatomical feature for efficient C4 photosynthesis in NADP-ME plants like S. viridis and have implications for engineering strategies to ensure future food security.


Assuntos
Subfamília G de Transportadores de Cassetes de Ligação de ATP/metabolismo , Dióxido de Carbono/metabolismo , Lipídeos/biossíntese , Mutação , Fotossíntese , Feixe Vascular de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Setaria (Planta)/metabolismo , Subfamília G de Transportadores de Cassetes de Ligação de ATP/genética , Difusão , Cromatografia Gasosa-Espectrometria de Massas , Regulação da Expressão Gênica de Plantas , Microscopia Eletrônica de Transmissão , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Folhas de Planta/ultraestrutura , Feixe Vascular de Plantas/genética , Feixe Vascular de Plantas/crescimento & desenvolvimento , Feixe Vascular de Plantas/ultraestrutura , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/ultraestrutura , Setaria (Planta)/genética , Setaria (Planta)/crescimento & desenvolvimento , Setaria (Planta)/ultraestrutura
20.
Sci Rep ; 10(1): 10294, 2020 06 24.
Artigo em Inglês | MEDLINE | ID: mdl-32581303

RESUMO

The genome sequences of 16 Streptomyces strains, showing potential for plant growth-promotion (PGP) activities in rice, sorghum, chickpea and pigeonpea, isolated from herbal vermicompost, have been decoded. The genome assemblies of the 16 Streptomyces strains ranged from 6.8 Mb to 8.31 Mb, with a GC content of 72 to 73%. The extent of sequence similarity (in terms of shared ortholog) in 16 Streptomyces strains showed 70 to 85% common genes to the closest publicly available Streptomyces genomes. It was possible to identify ~1,850 molecular functions across these 16 strains, of which close to 50% were conserved across the genomes of Streptomyces strains, whereas, ~10% were strain specific and the rest were present in various combinations. Genome assemblies of the 16 Streptomyces strains have also provided genes involved in key pathways related to PGP and biocontrol traits such as siderophores, auxin, hydrocyanic acid, chitinase and cellulase. Further, the genome assemblies provided better understanding of genetic similarity among target strains and with the publically available Streptomyces strains.


Assuntos
Desenvolvimento Vegetal , Rizosfera , Microbiologia do Solo , Streptomyces/genética , Cajanus/crescimento & desenvolvimento , Cajanus/microbiologia , Cicer/crescimento & desenvolvimento , Cicer/microbiologia , DNA Bacteriano/isolamento & purificação , Genoma Bacteriano , Redes e Vias Metabólicas/genética , Oryza/crescimento & desenvolvimento , Oryza/microbiologia , Filogenia , Reguladores de Crescimento de Plantas/metabolismo , Sorghum/crescimento & desenvolvimento , Sorghum/microbiologia , Streptomyces/isolamento & purificação , Streptomyces/metabolismo , Sequenciamento Completo do Genoma
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