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1.
Semin Cancer Biol ; 86(Pt 3): 666-681, 2022 11.
Artigo em Inglês | MEDLINE | ID: mdl-34216789

RESUMO

An overwhelming number of research articles have reported a strong relationship of the microbiome with cancer. Microbes have been observed more commonly in the body fluids like urine, stool, mucus of people with cancer compared to the healthy controls. The microbiota is responsible for both progression and suppression activities of various diseases. Thus, to maintain healthy human physiology, host and microbiota relationship should be in a balanced state. Any disturbance in this equilibrium, referred as microbiome dysbiosis becomes a prime cause for the human body to become more prone to immunodeficiency and cancer. It is well established that some of these microbes are the causative agents, whereas others may encourage the formation of tumours, but very little is known about how these microbial communications causing change at gene and epigenome level and trigger as well as encourage the tumour growth. Various studies have reported that microbes in the gut influence DNA methylation, DNA repair and DNA damage. The genes and pathways that are altered by gut microbes are also associated with cancer advancement, predominantly those implicated in cell growth and cell signalling pathways. This study exhaustively reviews the current research advancements in understanding of dysbiosis linked with colon, lung, ovarian, breast cancers and insights into the potential molecular targets of the microbiome promoting carcinogenesis, the epigenetic alterations of various potential targets by altered microbiota, as well as the role of various chemopreventive agents for timely prevention and customized treatment against various types of cancers.


Assuntos
Microbiota , Neoplasias , Humanos , Disbiose/complicações , Disbiose/genética , Epigenômica , Epigênese Genética , Neoplasias/genética
2.
Genomics ; 112(1): 99-107, 2020 01.
Artigo em Inglês | MEDLINE | ID: mdl-31356969

RESUMO

Snow Mountain Garlic grows in the high altitudes of the Himalayas under low temperature conditions. It contains various bioactive compounds whose metabolic pathways have not been worked out at genomic level. The present work is the first report on the transcriptome sequencing of this plant. >43 million paired-end reads (301 × 2) were generated using Illumina Miseq sequencing technology. Assembling of the sequencing data resulted in 326,785 transcripts. Differentially expressed genes between the clove and leaf tissues were identified and characterized. Besides, greater emphasis was laid on the genes, which were highly expressed in clove since the latter is assumed to contain high content of the bioactive compounds. Further analysis led to the identification of the genes plausibly involved in the organosulfur metabolism. We also identified several simple sequence repeats and single nucleotide polymorphism. These constitute valuable genetic resource for research and further genetic improvement of the plant.


Assuntos
Alho/genética , Compostos de Enxofre/metabolismo , Transcriptoma , Alho/metabolismo , Perfilação da Expressão Gênica , Ontologia Genética , Genes de Plantas , Marcadores Genéticos , Sequenciamento de Nucleotídeos em Larga Escala , Mutação INDEL , Redes e Vias Metabólicas/genética , Repetições de Microssatélites , Folhas de Planta/genética , Folhas de Planta/metabolismo , Polimorfismo de Nucleotídeo Único , Domínios Proteicos
3.
Mycoses ; 61(8): 534-542, 2018 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-29500851

RESUMO

Dermatophytes are considered as the main pathogens responsible for onychomycosis, but recently successive isolations of yeast-like fungi from the infected nails has led to consider these also as primary agents of nail infections. Trichosporon species which are non-candidal, basidiomycetous, yeast-like, anamorphic fungi are commonly isolated from soil but they are also emerging as important etiological agents of onychomycosis. Three species of Trichosporon viz., T. asahii, T. asteroides and T. faecale were isolated from the infected nails of three female members of a family from district Doda of Jammu and Kashmir State. Among the isolated species of Trichosporon, T. asahii was recovered from the nail samples of all the three members, thus confirming its recognition as a main pathogenic species of onychomycosis. So far, there is no report of T. asteroides and T. faecale causing onychomycosis and hence they constitute new additions to the list of onychomycotic fungi. Some of the predisposing factors like low socio-economic condition, poor hygiene, frequent exposure of finger nails to water and dirt, climatic conditions and nail trauma were observed to be the main causes of nail infection in these patients. However, a link between the pathogenic genus and the genetic makeup of the patients is also probable.


Assuntos
Saúde da Família , Unhas/patologia , Onicomicose/diagnóstico , Onicomicose/patologia , Trichosporon/isolamento & purificação , Tricosporonose/diagnóstico , Tricosporonose/patologia , Adolescente , Adulto , Análise por Conglomerados , DNA Fúngico/química , DNA Fúngico/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Feminino , Genes de RNAr , Humanos , Índia , Técnicas Microbiológicas , Microscopia , Unhas/microbiologia , Filogenia , RNA Fúngico/genética , RNA Ribossômico 18S/genética , RNA Ribossômico 28S/genética , Análise de Sequência de DNA , Trichosporon/classificação , Trichosporon/genética
4.
Plant Cell Rep ; 36(1): 203-217, 2017 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-27844102

RESUMO

KEY MESSAGE: Under severe drought conditions, Brassica juncea shows differential methylation and demethylation events, such that certain epialleles are silenced and some are activated. The plant employed avoidance strategy by delaying apoptosis through the activation of several genes. Harsh environmental conditions pose serious threat to normal growth and development of crops, sometimes leading to their death. However, plants have developed an essential mechanism of modulation of gene activities by epigenetic modifications. Brassica juncea is an important oilseed crop contributing effectively to the economy of India. In the present investigation, we studied the changes in the methylation level of various stress-responsive genes of B. juncea variety RH30 by methylation-dependent immune-precipitation-chip in response to severe drought. On the basis of changes in the number of differential methylation regions in response to drought, the promoter regions were designated as hypermethylated and hypomethylated. Gene body methylation increased in all the genes, whereas promoter methylation was dependent on the function of the gene. Overall, the genes responsible for delaying apoptosis were hypomethylated and many genes responsible for normal routine activities were hypermethylated at promoter regions, thereby suggesting that these may be suspending the activities under harsh conditions.


Assuntos
Alelos , Secas , Genes de Plantas , Mostardeira/genética , Mostardeira/fisiologia , Estresse Fisiológico/genética , Arabidopsis/genética , Metilação de DNA/genética , Ontologia Genética , Análise de Sequência com Séries de Oligonucleotídeos , Regiões Promotoras Genéticas , Transdução de Sinais/genética
5.
Phytother Res ; 29(4): 617-27, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25756352

RESUMO

The present study involves evaluation of antioxidant potential of Crocus sativus and its main constituents, safranal (SFN) and crocin (CRO), in bronchial epithelial cells, followed antiinflammatory potential of the active constituent safranal, in a murine model of asthma. To investigate the antioxidizing potential of Crocus sativus and its main constituents in bronchial epithelial cells, the stress was induced in these cells by a combination of different cytokines that resulted in an increase in nitric oxide production (NO), induced nitric oxide synthase (iNOS) levels, peroxynitrite ion generation, and cytochrome c release. Treatment with saffron and its constituents safranal and crocin resulted in a decrease of NO, iNOS levels, peroxynitrite ion generation, and prevented cytochrome c release. However, safranal significantly reduced oxidative stress in bronchial epithelial cells via iNOS reduction besides preventing apoptosis in these cells. In the murine model of asthma study, antiinflammatory role of safranal was characterized by increased airway hyper-responsiveness, airway cellular infiltration, and epithelial cell injury. Safranal pretreatment to these allergically inflamed mice lead to a significant decrease in airway hyper-responsiveness and airway cellular infiltration to the lungs. It also reduced iNOS production, bronchial epithelial cell apoptosis, and Th2 type cytokine production in the lungs.


Assuntos
Asma/tratamento farmacológico , Crocus/química , Cicloexenos/farmacologia , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Terpenos/farmacologia , Animais , Anti-Inflamatórios/farmacologia , Antioxidantes/farmacologia , Células Cultivadas , Citocromos c/metabolismo , Citocinas/metabolismo , Modelos Animais de Doenças , Humanos , Pulmão/efeitos dos fármacos , Camundongos , Camundongos Endogâmicos BALB C , Óxido Nítrico/metabolismo , Estresse Oxidativo/efeitos dos fármacos , Ácido Peroxinitroso/metabolismo , Extratos Vegetais/farmacologia
6.
ScientificWorldJournal ; 2015: 387367, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25685838

RESUMO

The present investigation was aimed at understanding the molecular mechanism of gene amplification. Interplay of fragile sites in promoting gene amplification was also elucidated. The amplification promoting sequences were chosen from the Saccharomyces cerevisiae ARS, 5S rRNA regions of Plantago ovata and P. lagopus, proposed sites of replication pausing at Ste20 gene locus of S. cerevisiae, and the bend DNA sequences within fragile site FRA11A in humans. The gene amplification assays showed that plasmid bearing APS from yeast and human beings led to enhanced protein concentration as compared to the wild type. Both the in silico and in vitro analyses were pointed out at the strong bending potential of these APS. In addition, high mitotic stability and presence of TTTT repeats and SAR amongst these sequences encourage gene amplification. Phylogenetic analysis of S. cerevisiae ARS was also conducted. The combinatorial power of different aspects of APS analyzed in the present investigation was harnessed to reach a consensus about the factors which stimulate gene expression, in presence of these sequences. It was concluded that the mechanism of gene amplification was that AT rich tracts present in fragile sites of yeast serve as binding sites for MAR/SAR and DNA unwinding elements. The DNA protein interactions necessary for ORC activation are facilitated by DNA bending. These specific bindings at ORC promote repeated rounds of DNA replication leading to gene amplification.


Assuntos
Replicação do DNA/genética , Amplificação de Genes/fisiologia , Saccharomyces cerevisiae/genética , Simulação por Computador , Replicação do DNA/fisiologia , Genes Fúngicos/genética , Genes Fúngicos/fisiologia , Filogenia , Plantago/genética , Plantago/fisiologia , Reação em Cadeia da Polimerase , RNA Ribossômico 5S/genética , RNA Ribossômico 5S/fisiologia , Saccharomyces cerevisiae/fisiologia
7.
Protoplasma ; 261(4): 749-769, 2024 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-38340171

RESUMO

Crocus sativus is a valuable plant due to the presence of apocarotenoids in its stigma. Considerable work has been done in the past to understand the apocarotenoid biosynthetic pathway in saffron. However, the reports on understanding the regulation of flowering at the post-transcriptional level are meagre. The study aimed to discover the candidate miRNAs, target genes, transcription factors (TFs), and apocarotenoid biosynthetic pathway genes associated with the regulation and transition of flowering in C. sativus. In the present investigation, miRNA profiling was performed in flowering and non-flowering corms of saffron, along with expression analysis of apocarotenoid genes and transcription factors involved in the synthesis of secondary metabolites. Significant modulation in the expression of miR156, miR159, miR166, miR172, miR395, miR396, miR399, and miR408 gene families was observed. We obtained 36 known miRNAs (26 in flowering and 10 in non-flowering) and 64 novel miRNAs (40 in flowering and 24 in non-flowering) unique to specific tissues in our analysis. TFs, including CsMADS and CsMYb, showed significant modulation in expression in flowering tissue, followed by CsHB. Additionally, the miRNAs were predicted to be involved in carbohydrate metabolism, phytohormone signalling, regulation of flower development, and response to stress, cold, and defence. The comprehensive study has enhanced our understanding of the regulatory machinery comprising factors like phytohormones, abiotic stress, apocarotenoid genes, transcription factors, and miRNAs responsible for the synthesis of apocarotenoids and developmental processes during and after flowering.


Assuntos
Crocus , Flores , Regulação da Expressão Gênica de Plantas , MicroRNAs , MicroRNAs/genética , MicroRNAs/metabolismo , Flores/genética , Flores/crescimento & desenvolvimento , Flores/metabolismo , Crocus/genética , Perfilação da Expressão Gênica , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
8.
Nat Prod Res ; 38(4): 696-700, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-36896764

RESUMO

Fungal endophytes are valued for biosynthesizing chemically diverse metabolic cascade with interesting biological activities. In the current investigation, two compounds were isolated from Penicillium polonicum, an endophyte of Zingiber officinale. The active moieties, glaucanic acid (1) and dihydrocompactin acid (2) were isolated from the ethyl acetate extract of P. polonicum and characterized by NMR and mass spectroscopy. Further, bioactive potential of the isolated compounds was evaluated by antimicrobial, antioxidant and cytotoxicity assays. Compounds 1 and 2 displayed antifungal activity against phytopathogen Colletotrichum gloeosporioides with more than 50% reduction in its growth. Both the compounds exhibited antioxidant activity against free radicals (DPPH and ABTS) and cytotoxicity activity against cancer cell lines respectively. The compounds, glaucanic acid and dihydrocompactin acid are being reported for the first time from an endophytic fungus. This is the first report on the biological activities of Dihydrocompactin acid produced by endophytic fungal strain.


Assuntos
Lovastatina/análogos & derivados , Penicillium , Zingiber officinale , Penicillium/química , Fungos , Antioxidantes/farmacologia , Antioxidantes/metabolismo , Endófitos/química
9.
Methods Mol Biol ; 2575: 153-179, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-36301475

RESUMO

Plants possess a plethora of important secondary metabolites, which are unique sources of natural pigments, pharmaceutical compounds, food additives, natural pesticides, and other industrial components. The commercial significance of such metabolites/compounds has directed the research toward their production and exploration of methods for enhancement of production. Biotechnological tools are critical in selecting, integrating, multiplying, improving, and analyzing medicinal plants for secondary metabolite production. Out of many techniques that are being explored to enhance secondary metabolite production, "plant cell transfection" is the latest tool to achieve maximum output from the plant source. It is based upon the introduction of foreign DNA into the plant cell relying on physical treatment such as electroporation, cell squeezing, sonoporation, optical transfection nanoparticles, magnetofection, and chemical treatment or biological treatment that depends upon carrier. One of the promising tools that have been exploited is CRISPR-Cas9. Overall, the abovementioned tools focus on the stable transfection of desired gene transcripts. Since the integration and continuous expression of transfected gene of particular trait represents stable transfection of host cell genome, resulting from transfer of required trait to daughter cells ultimately leading to enhanced production of secondary metabolites of interest. This chapter will review a set of biotechnological tools that are candidates for achieving the enhanced bioactive compound production indicated here to be used for drug discovery.


Assuntos
Células Vegetais , Plantas Medicinais , Transfecção , Plantas Medicinais/metabolismo , Biotecnologia , Eletroporação
10.
Heliyon ; 9(9): e19487, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37662754

RESUMO

Over the past half century, limited use of synthetic fertilizers, pesticides, and conservation of the environment and natural resources have become the interdependent goals of sustainable agriculture. These practices support agriculture sustainability with less environmental and climatic impacts. Therefore, there is an upsurge in the need to introduce compatible booster methods for maximizing net production. The best straightforward strategy is to explore and utilize plant-associated beneficial microorganisms and their products. Bioinoculants are bioformulations consisting of selected microbial strains on a suitable carrier used in the enhancement of crop production. Fungal endophytes used as bioinoculants confer various benefits to the host, such as protection against pathogens by eliciting immune response, mineralization of essential nutrients, and promoting plant growth. Besides, they also produce various bioactive metabolites, phytohormones, and volatile organic compounds. To design various bioformulations, transdisciplinary approaches like genomics, transcriptomics, metabolomics, proteomics, and microbiome modulation strategies like gene editing and metabolic reconstruction have been explored. These studies will refine the existing knowledge on the diversity, phylogeny and beneficial traits of the microbes. This will also help in synthesizing microbial consortia by evaluating the role of structural and functional elements of communities in a controlled manner. The present review summarizes the beneficial aspects associated with fungal endophytes for capitalizing agricultural outputs, enlists various multi-omics techniques for understanding and modulating the mechanism involved in endophytism and the generation of new bioformulations for providing novel solutions for the enhancement of crop production.

11.
Pathogens ; 12(1)2022 Dec 31.
Artigo em Inglês | MEDLINE | ID: mdl-36678413

RESUMO

Apple scab, caused by Venturia inaequalis, is one of the world's most commercially significant apple diseases. The fungi have a catastrophic impact on apples, causing considerable losses in fruit quality and productivity in many apple-growing locations despite numerous control agents. Fungi secrete various effectors and other virulence-associated proteins that suppress or alter the host's immune system, and several such proteins were discovered in this work. Using state-of-the-art bioinformatics techniques, we examined the V. inaequalis reference genome (EU-B04), resulting in the identification of 647 secreted proteins, of which 328 were classified as small secreted proteins (SSPs), with 76.52% of SSPs identified as anticipated effector proteins. The more prevalent CAZyme proteins were the enzymes engaged in plant cell wall disintegration (targeting pectin and xylanase), adhesion and penetration (Cutinases/acetyl xylan esterase), and reactive oxygen species formation (multicopper oxidases). Furthermore, members of the S9 prolyl oligopeptidase family were identified as the most abundant host defense peptidases. Several known effector proteins were discovered to be expressed during the V. inaequalis infection process on apple leaves. The present study provides valuable data that can be used to develop new strategies for controlling apple scab.

12.
Plant Cell Rep ; 30(5): 799-806, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21249368

RESUMO

Plant Biotechnology involves manipulation of genetic material to develop better crops. Keeping in view the challenges being faced by humanity in terms of shortage of food and other resources, we need to continuously upgrade the genomic technologies and fine tune the existing methods. For efficient genetic transformation, Agrobacterium-mediated as well as direct delivery methods have been used successfully. However, these methods suffer from many disadvantages especially in terms of transfer of large genes, gene complexes and gene silencing. To overcome these problems, recently, some efforts have been made to develop genetic transformation systems based on engineered plant chromosomes called minichromosomes or plant artificial chromosomes. Two approaches namely, "top-down" or "bottom-up" have been used for minichromosomes. The former involves engineering of the existing chromosomes within a cell and the latter de novo assembling of chromosomes from the basic constituents. While some success has been achieved using these chromosomes as vectors for genetic transformation in maize, however, more studies are needed to extend this technology to crop plants. The present review attempts to trace the genesis of minichromosomes and discusses their potential of development into plant artificial chromosome vectors. The use of these vectors in genetic transformation will greatly ameliorate the food problem and help to achieve the UN Millennium development goals.


Assuntos
Cromossomos Artificiais/genética , Cromossomos de Plantas/genética , Produtos Agrícolas/genética , Engenharia Genética/métodos , Plantas Geneticamente Modificadas/genética , Biotecnologia/métodos , Centrômero , Produtos Agrícolas/normas , Técnicas de Transferência de Genes , Vetores Genéticos , Telômero , Transformação Genética , Zea mays/genética
13.
Front Bioeng Biotechnol ; 9: 664705, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34222213

RESUMO

Catalysis is a process carried out in the presence of a heterogenous catalyst for accelerating the rate of a chemical reaction. It plays a pivotal role in transition from take, make, and dispose technology to sustainable technology via chemo- and biocatalytic processes. However, chemocatalyzed reactions are usually associated with copious amounts of perilous/hazardous environmental footprints. Therefore, whole-cell biotransformations or enzyme cocktails serve as cleaner biocatalytic alternatives in replacing the classical chemical procedures. These benchmark bioconversion reactions serve as important key technology in achieving the goals of green chemistry by eliminating waste generation at source. For this, nature has always been a driving force in fuelling natural product discovery and related applications. The fungal endophytic community, in particular, has undergone co-evolution with their host plant and has emerged as a powerful tool of genetic diversity. They can serve as a treasure trove of biocatalysts, catalyzing organic transformations of a wide range of substances into enantiopure compounds with biotechnological relevance. Additionally, the biocatalytic potential of endophytic fungi as whole-intact organisms/isolated enzyme systems has been greatly expanded beyond the existing boundaries with the advancement in high-throughput screening, molecular biology techniques, metabolic engineering, and protein engineering. Therefore, the present review illustrates the promising applications of endophytic fungi as biocatalysts for the synthesis of new structural analogs and pharmaceutical intermediates and refinement of existing proteins for novel chemistries.

14.
Front Microbiol ; 12: 635917, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34122359

RESUMO

Dynamic consortium of microbial communities (bacteria, fungi, protists, viruses, and nematodes) colonizing multiple tissue types and coevolving conclusively with the host plant is designated as a plant microbiome. The interplay between plant and its microbial mutualists supports several agronomic functions, establishing its crucial role in plant beneficial activities. Deeper functional and mechanistic understanding of plant-microbial ecosystems will render many "ecosystem services" by emulating symbiotic interactions between plants, soil, and microbes for enhanced productivity and sustainability. Therefore, microbiome engineering represents an emerging biotechnological tool to directly add, remove, or modify properties of microbial communities for higher specificity and efficacy. The main goal of microbiome engineering is enhancement of plant functions such as biotic/abiotic stresses, plant fitness and productivities, etc. Various ecological-, biochemical-, and molecular-based approaches have come up as a new paradigm for disentangling many microbiome-based agromanagement hurdles. Furthermore, multidisciplinary approaches provide a predictive framework in achieving a reliable and sustainably engineered plant-microbiome for stress physiology, nutrient recycling, and high-yielding disease-resistant genotypes.

15.
Protoplasma ; 257(4): 1093-1108, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32152722

RESUMO

Tomato is an excellent model for studying fruit development, ripening, and other secondary metabolic pathways such as carotenoid biosynthetic pathway, flavonoid pathway, and many more. Tomato fruit development and ripening occurs under tight genetic control and involves the expression of thousands of genes affecting fruit quality and accumulation of pigments and metabolites. Here, we have described the development of a microarray platform that has allowed establishment of a framework for quantification of the expression of large number of genes and transcription factors possibly regulating various secondary metabolic pathways in tomato. To unravel the molecular mechanisms of fruit development and ripening, a tomato 60-mer oligonucleotide 44 K microarray along with the custom array for many genes and transcription factors was designed and validated in the fruit and leaf tissues. Comparative profiling of gene expression studies has allowed us to identify a large number of differentially expressed genes and transcription factors. Gene ontology revealed the involvement of these genes in various biological, cellular, and molecular processes like isoprenoid, terpenoid, pigment, ethylene biosynthesis, phytohormone signaling, and fruit ripening. Further, correlation, as well as differential expression studies, has revealed that several transcription factors like RIN, AGAMOUS, TAGL1, MYB, MADS-box etc. could be the possible regulators of various secondary metabolic pathways. The present study has identified various metabolites, their biosynthetic pathways and genes which may possibly be controlled by different transcription factors. The present findings have laid a base for understanding the transcriptional and metabolic shifts which occur in parallel during programmed fruit ripening and developmental processes in tomato.


Assuntos
Frutas/química , Regulação da Expressão Gênica de Plantas/genética , Solanum lycopersicum/química , Fatores de Transcrição/metabolismo
16.
Plant Methods ; 16: 47, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32280363

RESUMO

BACKGROUND: Crocus sativus is a recalcitrant plant for genetic transformation and genetic improvement, largely due to difficulties in Agrobacterium mediated transformation and vegetative reproduction. Effective genome editing requires proficient callus production and an efficient method to deliver Cas9 and sgRNAs into the plant. Here, we demonstrate Agrobacterium-mediated transformation of saffron. Further, we developed a CRISPR-Cas9 based system in this plant, for efficient gene knockout or edits in future. RESULTS: Efficient callus production and regeneration confers important benefits in developing competent transformation system in plants. More than 70% multiplication rate of callus initiation was achieved from corm slices of saffron subjected to a two-step sterilization procedure and grown on complete MS medium supplemented with 2,4-D (0.5 mg/L), BAP (1 mg/L), IAA (1 mg/L), photoperiod of 16/8 h and 45% relative humidity at 20 ± 2 °C. In vitro cormlet generation was accomplished in 8 weeks by using mature somatic embryos on MS medium supplemented with TDZ (0.5 mg/L) + IAA (1 mg/L) + Activated charcoal (0.1 g/L) at 15 ± 2 °C. The attempt of using Agrobacterium-mediated transformation resulted in successful integration of the binary vector into the somatic embryos of saffron with a transformation efficiency of 4%. PCR and Southern blot analysis confirmed the integration of Cas9 into saffron. CONCLUSION: The protocol for callus production, somatic embryogenesis and regeneration was standardised. Successful demonstration of integrated Cas9 in this study constitutes first step in developing strategies for genetic manipulation of saffron, which has so far been considered recalcitrant. Furthering the development of this technology holds significant potential for advancing genetic research in saffron by integrating multigene targeting and/or use of recyclable cassettes.

17.
Brief Funct Genomics ; 19(1): 1-9, 2020 01 22.
Artigo em Inglês | MEDLINE | ID: mdl-31875900

RESUMO

A plant communicates within itself and with the outside world by deploying an array of agents that include several attractants by virtue of their color and smell. In this category, the contribution of 'carotenoids and apocarotenoids' is very significant. Apocarotenoids, the carotenoid-derived compounds, show wide representation among organisms. Their biosynthesis occurs by oxidative cleavage of carotenoids, a high-value reaction, mediated by carotenoid cleavage oxygenases or carotenoid cleavage dioxygenases (CCDs)-a family of non-heme iron enzymes. Structurally, this protein family displays wide diversity but is limited in its distribution among plants. Functionally, this protein family has been recognized to offer a role in phytohormones, volatiles and signal production. Further, their wide presence and clade-specific functional disparity demands a comprehensive account. This review focuses on the critical assessment of CCDs of higher plants, describing recent progress in their functional aspects and regulatory mechanisms, domain architecture, classification and localization. The work also highlights the relevant discussion for further exploration of this multi-prospective protein family for the betterment of its functional understanding and improvement of crops.


Assuntos
Carotenoides/metabolismo , Regulação da Expressão Gênica de Plantas , Oxigenases/química , Oxigenases/metabolismo , Desenvolvimento Vegetal , Plantas/metabolismo , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Relação Estrutura-Atividade
18.
Plant Cell Rep ; 28(12): 1857-67, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19847437

RESUMO

Gene amplification is prevalent in many eukaryotes and has been found linked to various phenomena such as ontogenesis, carcinogenesis, in vitro culturing, neoplasia and drug resistance. Earlier, we reported a novel B chromosome in Plantago lagopus L., which was found to have arisen as a result of massive amplification of 5S rDNA. In addition, the chromosome is also composed of 45S rDNA and transposable elements. While the importance of gene amplification cannot be underestimated, its mechanism of origin is still unclear. Therefore, the aim of the present study was to determine whether amplification can be reactivated in the novel B chromosome. For this purpose, in vitro culture was used as stress. Three modes of tissue culture, i.e., direct, indirect and somatic embryogenesis were used for raising in vitro cultures. The variations due to genetic and epigenetic mechanisms were assessed in regenerants using molecular techniques, namely, PCR-RFLP, SSAP and MSAP. The retrotransposon-based molecular markers were applied to detect the polymorphism within transposable elements of in vitro regenerated and mother plants. We detected the variations that may be due to genetic changes either because of element recombination or activation of transposable elements which can lead to increase in the copy number. MSAP analysis revealed the differences in the DNA methylation pattern of the regenerants derived from novel chromosome bearing mother plants. Some regenerated plants were associated with increase and decrease in DNA methylation of both internal and external cytosine of the CCGG sequence.


Assuntos
Cromossomos de Plantas/genética , Epigênese Genética , Amplificação de Genes/genética , Instabilidade Genômica/genética , Plantago/genética , Técnicas de Cultura de Tecidos/métodos , Sequência de Bases , Meios de Cultura , Metilação de DNA/efeitos dos fármacos , Metilação de DNA/genética , DNA Intergênico/genética , DNA Ribossômico/genética , Epigênese Genética/efeitos dos fármacos , Instabilidade Genômica/efeitos dos fármacos , Reguladores de Crescimento de Plantas/farmacologia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/crescimento & desenvolvimento , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Plantago/efeitos dos fármacos , Plantago/fisiologia , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Regeneração/efeitos dos fármacos , Regeneração/genética
19.
Protoplasma ; 256(1): 249-260, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30078109

RESUMO

The dried stigmas of saffron constitute the world's costliest spice. Saffron has many therapeutic applications due to the presence of apocarotenoids. The latter are synthesized at different stages of development, and the biosynthetic pathway involves several genes encoding different enzymes. In order to understand the differential expression of various genes of the pathway, eight distinct developmental stages (S1-early to S8-late) were identified. The corms were assorted into three groups (I, II, and III) based on corm weight. Expression profiles of 12 carotenoid/apocarotenoid genes were studied. The expression of all genes was minimum/least in groups I and II corms during bud development. Lowest expression of carotenogenic genes (CsPSY, CsPDS, CsZDS, CsCRTISO, CsLYC-ß1, CsLYC-ε, CsBCH2, and CsNCED) was observed during early stages (S1-S3) of corm growth (dormant period). In group III corms, increased expression of apocarotenoid genes (CsZCO, CsCCD2, CsUGT, and CsALDH) was observed during S4 to S8 stages (reproductive period, floral differentiation). Besides, expression profiles of genes in apical and axillary buds were also examined. Of all the genes studied, apocarotenoid biosynthesis genes (CsBCH2, CsZCO, CsCCD2, CsALDH, and CsUGT) were found to be upregulated in apical bud than in the axillary bud. The results indicated that interaction of phytohormones and sugars, mother corm reserves and the influence of internal and external factors may be contributing to the growth of saffron corm/bud. The study has laid a foundation for further research on the molecular mechanisms underlying bud dormancy/growth in saffron.


Assuntos
Carotenoides/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Proteínas de Plantas/genética , Crocus , Ativação Transcricional
20.
Genes (Basel) ; 10(2)2019 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-30781667

RESUMO

B chromosomes have been reported in many eukaryotic organisms. These chromosomes occur in addition to the standard complement of a species. Bs do not pair with any of the A chromosomes and they have generally been considered to be non-essential and genetically inert. However, due to tremendous advancements in the technologies, the molecular composition of B chromosomes has been determined. The sequencing data has revealed that B chromosomes have originated from A chromosomes and they are rich in repetitive elements. In our laboratory, a novel B chromosome was discovered in Plantago lagopus. Using molecular cytogenetic techniques, the B chromosome was found to be composed of ribosomal DNA sequences. However, further characterization of the chromosome using next generation sequencing (NGS) etc. revealed that the B chromosome is a mosaic of sequences derived from A chromosomes, 5S ribosomal DNA (rDNA), 45S rDNA, and various types of repetitive elements. The transmission of B chromosome through the female sex track did not follow the Mendelian principles. The chromosome was found to have drive due to which it was perpetuating in populations. The present paper attempts to summarize the information on nature, transmission, and origin of B chromosomes, particularly the current status of our knowledge in P. lagopus.


Assuntos
Cromossomos de Plantas/genética , Evolução Molecular , Plantago/genética , Mapeamento Cromossômico , Plantago/crescimento & desenvolvimento , Sequências Repetitivas de Ácido Nucleico/genética
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