A novel assay for genetic and environmental changes in the architecture of intact root systems of plants grown in vitro

1991 ◽  
Vol 27 (2) ◽  
pp. 129-139 ◽  
Author(s):  
Gert Ooms ◽  
Karen Moore
Keyword(s):  
Environmental Changes ◽  
Root Systems ◽  
Intact Root

scholarly journals In Vitro Compression Model for Orthodontic Tooth Movement Modulates Human Periodontal Ligament Fibroblast Proliferation, Apoptosis and Cell Cycle

Biomolecules ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 932
Author(s):  
Julia Brockhaus ◽  
Rogerio B. Craveiro ◽  
Irma Azraq ◽  
Christian Niederau ◽  
Sarah K. Schröder ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Death ◽  
Periodontal Ligament ◽  
Environmental Changes ◽  
Tooth Movement ◽  
Orthodontic Tooth Movement ◽  
Compressive Force ◽  
Periodontal Ligament Fibroblasts ◽  
Human Periodontal Ligament Fibroblasts

Human Periodontal Ligament Fibroblasts (hPDLF), as part of the periodontal apparatus, modulate inflammation, regeneration and bone remodeling. Interferences are clinically manifested as attachment loss, tooth loosening and root resorption. During orthodontic tooth movement (OTM), remodeling and adaptation of the periodontium is required in order to enable tooth movement. hPDLF involvement in the early phase-OTM compression side was investigated for a 72-h period through a well-studied in vitro model. Changes in the morphology, cell proliferation and cell death were analyzed. Specific markers of the cell cycle were investigated by RT-qPCR and Western blot. The study showed that the morphology of hPDLF changes towards more unstructured, unsorted filaments under mechanical compression. The total cell numbers were significantly reduced with a higher cell death rate over the whole observation period. hPDLF started to recover to pretreatment conditions after 48 h. Furthermore, key molecules involved in the cell cycle were significantly reduced under compressive force at the gene expression and protein levels. These findings revealed important information for a better understanding of the preservation and remodeling processes within the periodontium through Periodontal Ligament Fibroblasts during orthodontic tooth movement. OTM initially decelerates the hPDLF cell cycle and proliferation. After adapting to environmental changes, human Periodontal Ligament Fibroblasts can regain homeostasis of the periodontium, affecting its reorganization.

scholarly journals Identification of Putative Virulence Genes by DNA Methylation Studies in the Cereal Pathogen Fusarium graminearum

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1192
Author(s):  
Francesco Tini ◽  
Giovanni Beccari ◽  
Gianpiero Marconi ◽  
Andrea Porceddu ◽  
Micheal Sulyok ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Fusarium Graminearum ◽  
Environmental Changes ◽  
Virulent Strain ◽  
Fungal Virulence ◽  
Wheat Seedlings ◽  
Wide Range ◽  
Conidia Production ◽  
Natural Hosts

DNA methylation mediates organisms’ adaptations to environmental changes in a wide range of species. We investigated if a such a strategy is also adopted by Fusarium graminearum in regulating virulence toward its natural hosts. A virulent strain of this fungus was consecutively sub-cultured for 50 times (once a week) on potato dextrose agar. To assess the effect of subculturing on virulence, wheat seedlings and heads (cv. A416) were inoculated with subcultures (SC) 1, 23, and 50. SC50 was also used to re-infect (three times) wheat heads (SC50×3) to restore virulence. In vitro conidia production, colonies growth and secondary metabolites production were also determined for SC1, SC23, SC50, and SC50×3. Seedling stem base and head assays revealed a virulence decline of all subcultures, whereas virulence was restored in SC50×3. The same trend was observed in conidia production. The DNA isolated from SC50 and SC50×3 was subject to a methylation content-sensitive enzyme and double-digest, restriction-site-associated DNA technique (ddRAD-MCSeEd). DNA methylation analysis indicated 1024 genes, whose methylation levels changed in response to the inoculation on a healthy host after subculturing. Several of these genes are already known to be involved in virulence by functional analysis. These results demonstrate that the physiological shifts following sub-culturing have an impact on genomic DNA methylation levels and suggest that the ddRAD-MCSeEd approach can be an important tool for detecting genes potentially related to fungal virulence.

scholarly journals Study of the Effect of Nutrient Environment on the Biological Characteristics of Fusarium solani Clinical Corneal Isolates

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Dalan Jing ◽  
Yingyu Li ◽  
Ziyuan Liu ◽  
Chen Huang ◽  
Pei Zhang ◽  
...  
Keyword(s):  
Fusarium Solani ◽  
Glucose Concentration ◽  
Environmental Changes ◽  
Biological Characteristics ◽  
Clinical Isolates ◽  
Reproductive Capacity ◽  
Diabetic Patients ◽  
Liquid Media ◽  
Nutritional Conditions

Purpose. The biological characteristics of Fusarium solani clinical corneal isolates growing in different nutritional conditions in vitro were studied in order to find out the key point of pathogenicity. Methods. Five kinds of media with different glucose and nitrogen concentrations were prepared as the liquid and solid forms. The clinical isolates were as follows: 2 Fusarium solani strains. The clinical corneal isolates and the standard strains were inoculated in the solid and liquid media. They were all incubated at 296 for 96 h and observed at defined time points. The optical density was recorded to generate the growth curves in liquid media. Morphologic changes of colonies in the solid media were determined under the light microscope. Results. The clinical isolates of Fusarium solani showed stronger reproductive capacity in the abominable nutritional condition. Besides, when the glucose concentration in the medium was consistent with the glucose concentration of aqueous in diabetic patients, the clinical isolates would show the biological features of quicker growth rate and stronger reproductive capacity. Conclusions. Nitrogen source is essential for fungus reproduction. The clinical isolates showed stronger environmental adaptability under different nutritional conditions and more sensitive to environmental changes.

Conversaziones 1974

1975 ◽  
Vol 29 (2) ◽  
pp. 163-171
Keyword(s):  
Electron Microscopy ◽  
Fine Structure ◽  
Cell Walls ◽  
Structural Changes ◽  
Vascular Tissue ◽  
Root Systems ◽  
Plant Roots ◽  
Tracer Studies ◽  
Water Move ◽  
Intact Root

CONVERSAZIONES were held this year on 9 May and 27 June. At the first conversazione twenty-seven exhibits and two films were shown. The fine structure of plant roots in relation to transport of nutrient ions and water was demonstrated by Dr D. T. Clarkson of the A.R.C. Letcombe Laboratory, Wantage and Dr A. W. Robards of the Department of Biology, University of York. Two major pathways by which nutrients and water move radially across the cortex towards the central vascular tissue have been distinguished by the use of tracer studies of adsorption by different zones of intact root systems, microautoradiography and electron microscopy. Movement can be apoplastic through cell walls, or symplastic between cells joined by plasmodesmata. As the root ages, structural changes in the endodermis reduce movement in the former pathway but the symplast is not interrupted by the elaboration of endodermal walls because plasmodesmatal connexions remain intact. These observations help explain the contrasting extent to which different ions and water reach the shoot from young and mature parts of root systems.

3 OXIDATIVE STRESS INDUCED CHANGES IN EPIGENETIC MODIFYING GENE mRNA IN PRE-IMPLANTATION IN VITRO BOVINE EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 149 ◽  
Author(s):  
C. A. Burroughs ◽  
G. L. Williamson ◽  
M. C. Golding ◽  
C. R. Long
Keyword(s):  
Oxidative Stress ◽  
Quantitative Pcr ◽  
Environmental Changes ◽  
Geometric Mean ◽  
Hypoxia Inducible Factor ◽  
Sirtuin 1 ◽  
Transcript Levels ◽  
Development Rates ◽  
High O2

In vitro embryo culture systems introduce a myriad of environmental changes to embryos when compared with in vivo development. Accordingly, in vitro culture has been associated with decreased embryo development rates and alterations to the fetal epigenome. We hypothesized that an interaction exists between oxidative stress and the expression of key epigenetic modifying enzymes. To test this hypothesis, we used quantitative PCR to measure transcript levels of several genes known to be associated with oxidative stress and epigenetic modifications during bovine in vitro pre-implantation development. Epigenetic modifiers examined included histone deacetylases (SIRT1, SIRT6), whose actions result in gene repression; a histone demethylase (JMJD1A) associated with gene activation; and DNA hydroxylases (TET1-3), which have been associated with conversion of 5-methylcytosine to 5-hydroxy-methylcytosine. Bovine ova were matured, fertilized, and cultured in vitro according to standard laboratory procedures. At 18 h post-fertilization, cumulus cells were removed and presumptive zygotes were cultured in 5% CO2 and air (high O2), or 5% CO2, 5% O2 and 90% N (low O2). Groups of 16-cell- (n = 20) and blastocyst- (n = 15) stage embryos were collected, RNA was extracted using an RNeasy Mini Kit, and cDNA was generated using qScript reagents. Quantitative PCR was performed using Perfecta SYBR to determine transcript levels of superoxide dismutase 2, mitochondrial (SOD2), lactate dehydrogenase A (LDHA), hypoxia-inducible factor 1 alpha (HIF1A), hypoxia-inducible factor 2 alpha (HIF2A), histone deacetylase 1 (HDAC1), jumonji domain containing 1A (JMJD1A), sirtuin 1 (SIRT1), sirtuin 6 (SIRT6), tet oncogene 1 (TET1), tet oncogene 2 (TET2), and tet oncogene 3 (TET3) in 3 replicates (each measured in triplicate). Data were analyzed using the comparative Ct method normalized to the geometric mean of 2 endogenous control genes [glyceraldehyde-3-phosphate dehydrogenase (GAPDH), succinate dehydrogenase (SDHA)]. Statistical analyses were performed using a paired t-test comparing the relative expression of each gene in high O2 with low O2 within each individual embryo stage. Blastocyst development rates were 35% (n = 710 oocytes) for low O2 and 19% for high O2 (n = 1221 oocytes; P < 0.001). The HIF2A transcript levels were elevated at the 16-cell stage in high-O2 embryos (4.26 ± 0.55 v. 1.22 ± 0.22; P < 0.01), but were not different at the blastocyst stage. Elevated HIF2A was likely in response to oxidative stress and was associated with elevated levels of JMJD1A (1.73 ± 0.18 v. 1.15 ± 0.19), SIRT1 (1.68 ± 0.20 v. 1.09 ± 0.16), SIRT6 (2.30 ± 0.34 v. 1.07 ± 0.13), and TET2 (3.44 ± 0.89 v. 1.23 ± 0.19; P < 0.05) in high-O2 16-cell embryos. The JMJD1A (1.31 ± 0.17 v. 1.05 ± 0.10), SIRT1 (1.45 ± 0.24 v. 1.05 ± 0.11), and TET1 (1.45 ± 0.19 v. 1.02 ± 0.07) transcripts were also elevated in high-O2 blastocysts, whereas TET3 transcript levels were lower (0.69 ± 0.12) than in low O2 (1.07 ± 0.14; P < 0.05). Genes associated with epigenome modification and with oxidative stress were altered in response to the high oxygen tension. The lower blastocyst rate observed in the high-O2 group may be due to the damaging effects of oxidative stress and this stress alters transcript levels associated with the epigenome. These data suggest that embryos surviving oxidative stress may exhibit epigenetic changes that could influence subsequent development.

scholarly journals Effects of Environmental Changes on Expression of the Oligopeptide Permease (opp) Genes of Borrelia burgdorferi

2002 ◽  
Vol 184 (22) ◽  
pp. 6198-6206 ◽  
Author(s):  
Xing-Guo Wang ◽  
Bo Lin ◽  
J. Michael Kidder ◽  
Samuel Telford ◽  
Linden T. Hu
Keyword(s):  
Borrelia Burgdorferi ◽  
Promoter Region ◽  
Nitrogen Availability ◽  
Environmental Changes ◽  
Substrate Binding ◽  
Environmental Response ◽  
Promoter Regions ◽  
Promoter Sequences ◽  
Oligopeptide Permease

ABSTRACT We analyzed expression of a putative oligopeptide permease (Opp) of Borrelia burgdorferi. Unlike the opp operons of other bacteria for which there is a single substrate binding protein, B. burgdorferi codes for three substrate binding proteins (OppA-I to -III) in its opp operon and an additional two homologs on plasmids (OppA-IV and -V). Instead of a single promoter region regulating transcription of the entire operon, as seen in other bacterial opp operons, it appears that among oppA-I, -II, and -III, as well as oppA-IV and -V, each has a potential upstream promoter region. We tested the function of these putative promoter sequences by fusion to a promoterless β-galactosidase reporter gene in pCB182. Each of the promoter regions was found to be active. The level of activity in the reporter constructs closely paralleled the level of expression of each gene in in vitro-grown B. burgdorferi. Changes in carbon and nitrogen availability differentially affected individual promoters, but no changes in promoter activity were seen when Escherichia coli bacteria (with the promoter constructs) were grown in various concentrations of phosphate and leucine and changes in pH. Expression of specific oppA genes with B. burgdorferi varied significantly between its mouse and fed and unfed tick hosts. Differences in regulation of opp gene expression suggest a potential role in environmental response by the organism.

scholarly journals Some physiological and biochemical changes of micropropagated Rhynchostylis gigantea during ex vitro acclimatization and greenhouse stages

2020 ◽  
Vol 4 (4) ◽  
pp. First
Author(s):  
Cao Phi Bang
Keyword(s):  
Water Content ◽  
Environmental Changes ◽  
Economic Value ◽  
Matter Content ◽  
Leaf Water ◽  
Leaf Water Content ◽  
Dry Matter Content ◽  
Ex Vitro ◽  
Fresh Leaf

The ex vitro acclimatization and greenhouse periods play a significant role for the in vitro originated plantlets. In these stages, the micropropagated plantlets have to rapidly adapt to environmental changes. Rhynchostylis gigantea is widely in vitro produced due to highly aesthetic and economic value. The aim of this work was to update the physiological changes of micropropagated R. gigantea plantlets during ex vitro acclimatization and greenhouse stages. The analysis results showed that leaf water content was significantly decreased at day 14 (90.36%) and day 28 (90.17%) stages but increased at day 84 (92.52%) and day 140 (92.34%) stages in compared to in vitro stages, day 0 (92.7%). Dry matter content was changing in the opposite direction to the leaf water content with the highest values at day 14 (9.63%) and day 28 (9.83%), respectively. The leaf transpiration rate was the highest at day zero (0.146 g/dm2/h) in compared to all other studied points. Oppositively, GPX activity was the lowest in plantlets at day zero (13.2 UI/g fresh leaf ) and the highest in planlets at day 14 (36,4 UI/g fresh leaf ). The leaf proline content was higher at day 7 and day 14 stages (132.3 and 150.8 m g/g fresh leaf, respectively) but lower at day 84 and day 140 stages (44.3 and 53.3 microgram/g fresh leaf, respectively) than at day zero (73.7 microgram/g fresh leaf ).

scholarly journals Laboratory Exercise Demonstrating the Importance of Leaves in the Rooting of Herbaceous Stem Cuttings

HortScience ◽  
1990 ◽  
Vol 25 (11) ◽  
pp. 1441-1442
Author(s):  
Michael Marcotrigiano ◽  
Susan P. McGlew
Keyword(s):  
Root Systems ◽  
Stem Cuttings ◽  
Laboratory Exercise ◽  
The Poor ◽  
Rooting Response

A teaching exercise that rapidly and inexpensively demonstrates the effects of leaf reduction on the rooting of stem cuttings is described. Coleus stem cuttings with whole leaves, half leaves, or no leaves are placed in sand on either misted or nonmisted greenhouse benches. Rooting response is recorded by ranking root systems by comparison to a diagram. A demonstration is also described that uses leafless cuttings cultured in vitro on solidified media containing a carbohydrate and/or auxin source. This demonstration gives students information that will help them speculate on the physiological reasons for the poor rooting response of leafless herbaceous cuttings.

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