scholarly journals Functional Characterization of9-/13-LOXsin Rice and Silencing Their Expressions to Improve Grain Qualities

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Moytri RoyChowdhury ◽  
Xiaobai Li ◽  
Hangying Qi ◽  
Wenxu Li ◽  
Jian Sun ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Functional Characterization ◽  
Negative Regulation ◽  
Human Consumption ◽  
Direct Products ◽  
Oxidative Rancidity ◽  
Storage Quality ◽  
Specific Expression ◽  
Transgenic Lines ◽  
Regenerated Plant

Lipoxygenases (LOXs) are involved in oxidative rancidity and render rice unsuitable for human consumption. Here, RNA interference- (RNAi-) induced gene expression inhibition was used to analyze the functions of the bran/seed-specific LOXs in rice.r9-LOX1andL-2(9-LOX category) were the candidate genes expressing a bran/seed-specific LOX, whileRCI-1was (13-LOX category) a plastid-specific LOX. Real-time PCR showed that three LOXs were cultivar/tissue specific expression on a certain level.r9-LOX1andL-2were generally much higher in active bran/seed than in stabilized bran, mature seed, and regenerated plant.RCI-1was barely expressed in seed. In transgenic lines,r9-LOX1, as well asL-2, expression was dramatically downregulated, compared to the nontransgenic controls. SPME/GC-MS analysis ofr9-LOX1RNAi transgenic lines showed 74.33% decrease in nonanal content (formed during oxidation of linoleic acid by lipoxygenase), but 388.24% increase in acetic acid and 184.84% hexanal (direct products of 13-LOX). These results indicate thatr9-LOX1positively regulates the amount of nonanal but negatively regulates acetic acid and hexanal. The negative regulation may be due to a mechanism of negative feedback between LOX family members. The information will help comprehensively understand the function of the bran/seed-specific LOXs,r9-LOX1, and improve the storage quality in the future.

scholarly journals Function of Circular RNAs in Fish and Their Potential Application as Biomarkers

2021 ◽  
Vol 22 (13) ◽  
pp. 7119
Author(s):  
Golam Rbbani ◽  
Artem Nedoluzhko ◽  
Jorge Galindo-Villegas ◽  
Jorge M. O. Fernandes
Keyword(s):  
Growth Regulation ◽  
Functional Characterization ◽  
Circular Rnas ◽  
Farmed Fish ◽  
Developmentally Regulated ◽  
Specific Expression ◽  
Cell Type Specific Expression ◽  
Cell Type Specific ◽  
Taxonomic Groups ◽  
And Function

Circular RNAs (circRNAs) are an emerging class of regulatory RNAs with a covalently closed-loop structure formed during pre-mRNA splicing. Recent advances in high-throughput RNA sequencing and circRNA-specific computational tools have driven the development of novel approaches to their identification and functional characterization. CircRNAs are stable, developmentally regulated, and show tissue- and cell-type-specific expression across different taxonomic groups. They play a crucial role in regulating various biological processes at post-transcriptional and translational levels. However, the involvement of circRNAs in fish immunity has only recently been recognized. There is also broad evidence in mammals that the timely expression of circRNAs in muscle plays an essential role in growth regulation but our understanding of their expression and function in teleosts is still very limited. Here, we discuss the available knowledge about circRNAs and their role in growth and immunity in vertebrates from a comparative perspective, with emphasis on cultured teleost fish. We expect that the interest in teleost circRNAs will increase substantially soon, and we propose that they may be used as biomarkers for selective breeding of farmed fish, thus contributing to the sustainability of the aquaculture sector.

scholarly journals A combination of MEF3 and NFI proteins activates transcription in a subset of fast-twitch muscles.

1997 ◽  
Vol 17 (2) ◽  
pp. 656-666 ◽  
Author(s):  
F Spitz ◽  
M Salminen ◽  
J Demignon ◽  
A Kahn ◽  
D Daegelen ◽  
...  
Keyword(s):  
Regulatory Elements ◽  
Specific Expression ◽  
Transgenic Lines ◽  
Slow Twitch ◽  
Myogenic Factors ◽  
Fast Twitch ◽  
Restricted Pattern ◽  
Aldolase A ◽  
Drive Reporter Gene Expression

The human aldolase A pM promoter is active in fast-twitch muscles. To understand the role of the different transcription factors which bind to this promoter and determine which ones are responsible for its restricted pattern of expression, we analyzed several transgenic lines harboring different combinations of pM regulatory elements. We show that muscle-specific expression can be achieved without any binding sites for the myogenic factors MyoD and MEF2 and that a 64-bp fragment comprising a MEF3 motif and an NFI binding site is sufficient to drive reporter gene expression in some but, interestingly, not all fast-twitch muscles. A result related to this pattern of expression is that some isoforms of NFI proteins accumulate differentially in fast- and slow-twitch muscles and in distinct fast-twitch muscles. We propose that these isoforms of NFI proteins might provide a molecular basis for skeletal muscle diversity.

scholarly journals Identification and Functional Characterization of the CVOMT and EOMT Genes Promoters from Ocimum Basilicum L

2021 ◽  
Author(s):  
Fatemeh Khakdan ◽  
Zahra Shirazi ◽  
Mojtaba Ranjbar
Keyword(s):  
Water Deficit ◽  
Transcriptional Control ◽  
Functional Characterization ◽  
Pcr Analysis ◽  
Water Deficit Stress ◽  
Specific Expression ◽  
Stress Dependent ◽  
First Time ◽  
Dependent Mechanism

Abstract Methyl chavicol and methyl eugenol are important phenylpropanoid compounds previously purified from basil. These compounds are significantly enhanced by the water deficit stress-dependent mechanism. Here, for the first time, pObCVOMT and pObEOMT promoters were extracted by the genome walking method. They were then cloned into the upstream of the β-glucuronidase (GUS) reporter gene to identify the pattern of GUS water deficit stress-specific expression. Histochemical GUS assays showed in transgenic tobacco lines bearing the GUS gene driven by pObCVOMT and pObEOMT promoters, GUS was strongly expressed under water deficit stress. qRT-PCR analysis of pObCVOMT and pObEOMT transgenic plants confirmed the histochemical assays, indicating that the GUS expression is also significantly induced and up-regulated by increasing density of water deficit stress. This indicates these promoters are able to drive inducible expression. The cis-acting elements analysis showed that the pObCVOMT and pObEOMT promoters contained dehydration or water deficit-related transcriptional control elements.

scholarly journals Modulation of Auxin Levels in Pollen Grains Affects Stamen Development and Anther Dehiscence in Arabidopsis

2018 ◽  
Vol 19 (9) ◽  
pp. 2480 ◽  
Author(s):  
Hernán Salinas-Grenet ◽  
Ariel Herrera-Vásquez ◽  
Samuel Parra ◽  
Allan Cortez ◽  
Lilian Gutiérrez ◽  
...  
Keyword(s):  
Acetic Acid ◽  
Apical Meristem ◽  
Seed Set ◽  
Transgenic Arabidopsis ◽  
Pollen Grains ◽  
Anther Development ◽  
Anther Dehiscence ◽  
Transgenic Lines ◽  
Indole 3 Acetic Acid ◽  
Auxin Accumulation

Auxin regulates diverse aspects of flower development in plants, such as differentiation of the apical meristem, elongation of the stamen, and maturation of anthers and pollen. It is known that auxin accumulates in pollen, but little information regarding the biological relevance of auxin in this tissue at different times of development is available. In this work, we manipulated the amount of free auxin specifically in developing pollen, using transgenic Arabidopsis lines that express the bacterial indole-3-acetic acid-lysine synthetase (iaaL) gene driven by a collection of pollen-specific promoters. The iaaL gene codes for an indole-3-acetic acid-lysine synthetase that catalyzes the conversion of free auxin into inactive indole-3-acetyl-l-lysine. The transgenic lines showed several abnormalities, including the absence of short stamina, a diminished seed set, aberrant pollen tubes, and perturbations in the synchronization of anther dehiscence and stamina development. This article describes the importance of auxin accumulation in pollen and its role in stamina and anther development.

Beta-MHC and SMLC1 transgene induction in overloaded skeletal muscle of transgenic mice

1996 ◽  
Vol 270 (4) ◽  
pp. C1111-C1121 ◽  
Author(s):  
J. L. Wiedenman ◽  
I. Rivera-Rivera ◽  
D. Vyas ◽  
G. Tsika ◽  
L. Gao ◽  
...  
Keyword(s):  
Transgenic Mice ◽  
Troponin C ◽  
Type I ◽  
Initial Attempt ◽  
Specific Expression ◽  
Slow Type ◽  
Transgenic Lines ◽  
Fast Twitch Muscle ◽  
Mechanical Overload

The hypertrophic responses of white fast-twitch muscle to mechanical overload has been investigated using transgenic mice. After 7 wk of overload, endogenous beta-myosin heavy chain (MHC) and slow myosin light chain 1 and 2 (SMLC1, SMLC2) protein were increased in the overloaded plantaris (OP) muscle compared with sham-operated control plantaris (CP)muscle. Concurrently, the levels of endogenous beta-MHC, SMLC1, SMLC2, and cardiac/slow troponin C (CTnC) mRNA transcripts were significantly increased in OP muscles, whereas skeletal troponin C (sTnC) mRNA transcript levels decreased. As an initial attempt to locate DNA sequence(s) that governs beta-MHC induction in response to mechanical overload, multiple independent transgenic lines harboring four different human beta-MHC transgenes (beta 1286, beta 988, beta 450, beta 141) were generated. Except for transgene beta 141, muscle-specific expression and induction (3- to 22-fold) in OP muscles were observed by measuring chloramphenicol acetyltransferase activity (CAT assay). Induction of a SMLC1 transgene (3920SMLC1) in OP muscles was also observed. Collectively, these in vivo data provide evidence that 1) a mechanical overload inducible element(s) is located between nucleotides -450 and +120 of the human beta-MHC transgene, 2) 3,900 bp of 5' sequence is sufficient to confer mechanical overload induction of a SMLC1 transgene, and 3) the increased expression of slow/type I isomyosin (beta-MHC, SMLC1, SMLC2) in response to mechanical overload is regulated, in part, transcriptionally.

scholarly journals Rice OsHSFA3 Gene Improves Drought Tolerance by Modulating Polyamine Biosynthesis Depending on Abscisic Acid and ROS Levels

2020 ◽  
Vol 21 (5) ◽  
pp. 1857 ◽  
Author(s):  
Ming-Dong Zhu ◽  
Meng Zhang ◽  
Du-Juan Gao ◽  
Kun Zhou ◽  
Shan-Jun Tang ◽  
...  
Keyword(s):  
Abscisic Acid ◽  
Drought Tolerance ◽  
Functional Characterization ◽  
Activity Levels ◽  
Enzymatic Antioxidants ◽  
Heat Shock Factors ◽  
Polyamine Biosynthesis ◽  
Polyamine Content ◽  
Transgenic Lines

Drought is a serious problem, which causes heavy yield losses for rice. Heat-shock factors (HSFs) had been implicated in tolerance to drought and high temperature. However, there has not been much functional characterization and mechanism clarification in rice. Previously, we found an HSF gene, OsHSFA3, was highly related with drought tolerance after screening from 10,000 different samples. Herein, we cloned the OsHSFA3 from rice and overexpressed it in Arabidopsis thaliana to study its regulatory mechanism of drought tolerance. Phenotypic and physiological assays of the transgenic Arabidopsis lines showed that overexpression of OsHSFA3 confers drought tolerance by reducing water loss and reactive oxygen species (ROS) levels, whereas it increases abscisic acid (ABA) levels. However, enzymatic antioxidants such as activity levels of superoxide dismutase, peroxidase and catalase were not significantly different between wild type and transgenic lines. Instead, we observed a significant increase in polyamine content, which was correlated with increased AtADC1, AtADC2, SPDS1 and SPMS expression levels. In silico and in vivo analyses confirmed that OsHSFA3 is a nuclear-localized gene. In addition, OsHSFA3 can bind to the promoter of AtADC1 and OsADC via a yeast one-hybrid assay. Overall, this study reveals that OsHSFA3 improves drought tolerance in Arabidopsis not only by increasing ABA levels, but also by modulating polyamine levels to maintain ROS homeostasis, therefore it could be a strong candidate to develop drought-tolerant rice cultivars.

scholarly journals Isolation and Functional Characterization of cDNA of Serum Amyloid A-Activating Factor That Binds to the Serum Amyloid A Promoter

1998 ◽  
Vol 18 (12) ◽  
pp. 7327-7335 ◽  
Author(s):  
Alpana Ray ◽  
Bimal K. Ray
Keyword(s):  
Transcription Factors ◽  
Dna Binding ◽  
Zinc Finger ◽  
Okadaic Acid ◽  
Serum Amyloid A ◽  
Kinase Inhibitor ◽  
Functional Characterization ◽  
Serum Amyloid ◽  
Specific Expression ◽  
Amyloid A

ABSTRACT Serum amyloid A (SAA), a plasma protein inducible in response to many inflammatory conditions, is associated with the pathogenesis of several diseases including reactive amyloidosis, rheumatoid arthritis, and atherosclerosis. We have previously reported an element of the SAA promoter, designated SAA-activating sequence (SAS), that is involved in the inflammation-induced SAA expression, and a nuclear factor, SAS-binding factor (SAF), that interacts with the SAS element has been identified previously (A. Ray and B. K. Ray, Mol. Cell. Biol. 16:1584–1594, 1996). To evaluate how SAF is involved in SAA promoter activation, we have investigated structural features and functional characteristics of this transcription factor. Our studies indicate that SAF belongs to a family of transcription factors characterized by the presence of multiple zinc finger motifs of the Cys2-His2 type at the carboxyl end. Of the three cloned SAF cDNAs (SAF-1, SAF-5, and SAF-8), SAF-1 isoform showed a high degree of homology to MAZ/ZF87/Pur-1 protein while SAF-5 and SAF-8 isoforms are unique and are related to SAF-1/MAZ/ZF87/Pur-1 at the zinc finger domains but different elsewhere. Although structurally distinct, all members are capable of activating SAS element-mediated expression and display virtually identical sequence specificities. However, varying levels of expression of members of this gene family were observed in different tissues. Functional activity of SAF is regulated by a posttranslational event as SAF DNA-binding and transactivation abilities are increased by a protein phosphatase inhibitor, okadaic acid, and inhibited by a protein kinase inhibitor, H7. Consistent with this observation, increased DNA binding of the cloned SAF and its hyperphosphorylation, in response to okadaic acid treatment of the transfected cells, were observed. Taken together, our results suggest that, in addition to tissue-specific expression, SAFs, a family of zinc finger transcription factors, undergo a modification by a posttranslational event that confers their SAA promoter-binding activity and transactivation potential.

Hematopoietic Stem and Progenitor Cell Biology in the Zebrafish.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4160-4160
Author(s):  
David Traver ◽  
Julien Bertrand ◽  
Albert Kim ◽  
Jennifer Cisson ◽  
Emily Violette
Keyword(s):  
Cell Biology ◽  
Progenitor Cell ◽  
Zebrafish Embryo ◽  
Functional Characterization ◽  
Hematopoietic Stem ◽  
Blood Island ◽  
The Past ◽  
Transgenic Lines ◽  
Zebrafish Embryogenesis

Abstract Over the past decade, the development of forward genetic approaches in the zebrafish system has provided unprecedented power in understanding the molecular basis of vertebrate blood development. Establishment of cellular and hematological approaches to better understand the biology of resulting blood mutants, however, has lagged behind these efforts. We have recently developed the means to identify zebrafish hematopoietic stem cells (HSCs), transgenic lines to mark hematopoietic precursors and their progeny, and the assays to test these populations functionally. Like other vertebrates, zebrafish demonstrate differential waves of hematopoiesis during embryogenesis. These waves can be visualized directly by fluorescent transgenesis in living embryos. The earliest blood-forming cells in the zebrafish embryo express the scl and lmo2 genes. By directing expression of GFP to early blood precursors using the lmo2 promoter, we have isolated early hematopoietic cells by flow cytometry and tested them functionally by transplantation. Transplantation of lmo2::GFP+ cells isolated from embryos at 14 hours post-fertilization (hpf) resulted in only transient reconstitution of erythrocytes, suggesting that the earliest identifiable blood-forming cells are committed to the erythroid lineage. Later in embryogenesis, lmo2:GFP+ GATA-1:dsRED+ cells are found in the posterior blood island (PBI) from approximately 30–60 hpf. Molecular and functional characterization of these cells suggests that they possess limited myeloid and erythroid, but not lymphoid differentiation potentials. This suggests that committed progenitors with definitive hematopoietic potential arise in the embryo before HSCs can be identified. Additional studies have suggested that the first multipotent HSCs are born later in the zebrafish aorta/gonad/mesonephros (AGM) region. We have visualized putative HSCs in the AGM by their expression of the lmo2 and cd41 transgenes. Using confocal timelapse imaging in living embryos, lmo2::GFP+ cells have been observed to emigrate from the AGM region into circulation. Transplantation studies are underway to test putative HSC populations for repopulation activity. Taken together, our findings suggest that at least three independent waves of blood cell precursors are formed during zebrafish embryogenesis.

Functional characterization of orchardgrass cytosolic Hsp70 (DgHsp70) and the negative regulation by Ca2+/AtCaM2 binding

2012 ◽  
Vol 58 ◽  
pp. 29-36 ◽  
Author(s):  
Joon-Yung Cha ◽  
Mukhamad Su'udi ◽  
Woe-Yeon Kim ◽  
Deok Ryong Kim ◽  
Youn-Sig Kwak ◽  
...  
Keyword(s):  
Functional Characterization ◽  
Negative Regulation

Meat composition and quality of young growing Belgian Blue bulls offered a fattening diet with selenium enriched cereals

2015 ◽  
Vol 95 (3) ◽  
pp. 465-473 ◽  
Author(s):  
Youcef Mehdi ◽  
Antoine Clinquart ◽  
Jean-Luc Hornick ◽  
Jean-François Cabaraux ◽  
Louis Istasse ◽  
...  
Keyword(s):  
Chemical Composition ◽  
Meat Quality ◽  
Dry Matter ◽  
Control Diet ◽  
Human Consumption ◽  
Oxidative Rancidity ◽  
Water Losses ◽  
Meat Composition ◽  
Basal Concentration

Mehdi, Y., Clinquart, A., Hornick, J.-L., Cabaraux, J.-F., Istasse, L. and Dufrasne, I. 2015. Meat composition and quality of young growing Belgian Blue bulls offered a fattening diet with selenium enriched cereals. Can. J. Anim. Sci. 95: 465–473. The objective of this study was to evaluate the effects of selenium (Se) enrichment of cereals on the performance of Belgian Blue bulls, meat quality and chemical composition. Twenty-three bulls were used in the present study. Twelve bulls were offered a control diet containing Se at a basal concentration of 58 µg kg−1 of dry matter (DM) and the other 11 bulls were given a diet containing 173 µg kg−1 DM of Se by means of Se-enriched spelt and barley. The Se enrichment of the diet did not affect the growth performance, the slaughter data or meat quality (P > 0.05). There were no effects of Se on tenderness, oxidative rancidity and water losses. However, there were some effects of Se enrichment on the meat chemical composition. The ether extract was decreased from 2.1 to 1.7% DM (P<0.05). There was also significant Se enrichment (P<0.001) in the longissimus thoracis muscle (177 vs. 477 ng g−1 DM) and organs: liver (474 vs. 1126 ng g−1 DM) and kidney (4956 vs. 5655 ng g−1 DM), Under such conditions, the human consumption of a piece of such meat or liver can provide a large part of the recommended daily Se intake, estimated between 30 and 57%.

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