scholarly journals TRANSIENT EXPRESSION OF REPORTER GENES IN CULTIVARS OF Amaranthus caudatus L.

2021 ◽  
Vol 14 (4) ◽  
pp. 53-63
Author(s):  
O. M. Yaroshko ◽  
Keyword(s):  
Transient Expression ◽  
Plant Material ◽  
Food Industry ◽  
Target Genes ◽  
Plant Biomass ◽  
Reporter Genes ◽  
Optimal Conditions ◽  
Uida Gene ◽  
Amaranthus Caudatus ◽  
Series Of Experiments

Local cultivars of A. caudatus: Helios and Karmin were used as plant material. Amaranth is a new pseudocereal introduced in Ukraine. The plant biomass of amaranth is used in medicine, food industry and cosmetology industry. Aim. The purpose of the work was to identify the optimal conditions for the transient expression of reporter genes in Amaranthus caudatus cultivars. Methods. Biochemical and microscopy methods were used in the following work. Seedlings and adult plants of different age were infiltrated with agrobacterial suspensions separately (genetic vector pCBV19 with a uidA gene and genetic vector pNMD2501 with a gfp gene in Agrobacterium tumefaciens GV3101 strain). Results. Transient expression of the uidA and gfp genes was obtained in amaranth plants after conduction series of experiments. The most intensive transient expression of gfp and uidA genes was observed in seedlings infiltrated at the age of 1 day. The maximum fluorescence of the GFP protein was observed on 5th–6th days. Conclusions. It was shown that the cultivar Helios was more susceptible to agrobacterial infection than the cultivar Karmin. The effectiveness of Agrobacterium mediated transformation was from 16% to 95% for the Helios cultivar and from 12% to 93% for the Karmin cultivar. The obtained results indicate that the studied amaranth cultivars can potentially be used for obtaining transient expression of target genes and synthesizing target proteins in their tissues in the future.

scholarly journals The Process of Producing Bioethanol from Delignified Cellulose Isolated from Plants of the Miscanthus Genus

2020 ◽  
Vol 7 (2) ◽  
pp. 61
Author(s):  
Olga Kriger ◽  
Ekaterina Budenkova ◽  
Olga Babich ◽  
Stanislav Suhih ◽  
Nikolay Patyukov ◽  
...  
Keyword(s):  
Plant Material ◽  
Plant Biomass ◽  
Raw Materials ◽  
Cellulose Content ◽  
Optimal Conditions ◽  
Pachysolen Tannophilus ◽  
Initial Amount ◽  
High Degree ◽  
High Cellulose ◽  
Unique Index

Plants of the Miscanthus genus (Miscanthus Anderss.) have a unique index of biomass production in relation to the occupied area. Miscanthus plants can be attributed to promising second-generation raw materials for the production of bioethanol and biofuel. Miscanthus plants are characterized by a high cellulose content. Herein, we report the results of a study on the obtained delignified cellulose with subsequent processing into bioethanol using microbial communities. In the course of the study, the optimal conditions for the delignification of the initial plant material for cellulose were selected. Ethanol with a high degree of conversion was successfully obtained from the isolated delignified cellulose. The article describes the pilot technological scheme for the conversion of Miscanthus plant biomass to bioethanol involving the delignification stages, followed by the conversion of the resulting cellulose into bioethanol by a consortium of microorganisms. As a result of the study, it was found that delignification using trifluoroacetic acid leads to the production of cellulose of high purity. Bioethanol with a yield of 3.1% to 3.4% in terms of the initial amount of biomass was successfully obtained by a microorganism consortium of Saccharomyces cerevisiae M Y-4242/Pachysolen tannophilus Y-3269, and Scheffersomyces stipitis Y-3264.

Eco-Friendly Dye-free Coloration Technology for Wool Based on Fenton Reagent Pretreatment

2011 ◽  
Vol 239-242 ◽  
pp. 3327-3330 ◽  
Author(s):  
Yan Ling Sui ◽  
Yong Zhu Cui
Keyword(s):  
Nitric Acid ◽  
Emission Reduction ◽  
Optimal Conditions ◽  
Fenton Reagent ◽  
Saving Energy ◽  
Before And After ◽  
Series Of Experiments

The wool was pretreated with Fenton reagent in this paper, on this basis, 4-hydroxy benzaldehyde and concentrated nitric acid were used to discuss the dye-free coloration deeply. It was analyzed comparatively through a series of experiments on the wool before and after Fenton reagent pretreatment, and the effects of concentrations, temperature and time on the coloration were further discussed. The experiment indicated that, compared with untreated wool, the color of wool with Fenton reagent pretreatment was deeper and the coloration rate was quicker. It realized good coloration at lower temperatures, which achieved the goal of saving energy and emission reduction. The optimal conditions were that concentrations of 4-hydroxy benzaldehyde and concentrated nitric acid were 2.5% and 3% respectively, reacting time was 90min, and reacting temperature was 70°C.

scholarly journals Melanin synthesis by black yeast-like fungi Psedonadsoniella brunnea: dependence of L-tyrosine quantity in the cultural medium

2019 ◽  
Vol 26 (1) ◽  
pp. 41-46 ◽  
Author(s):  
T. Kondratiuk ◽  
T. Beregova ◽  
T. Akulenko ◽  
Ie. Torgalo ◽  
V. Vereschaka
Keyword(s):  
Nutrient Medium ◽  
Culture Medium ◽  
Statistical Processing ◽  
Black Yeast ◽  
Malt Extract ◽  
Melanin Synthesis ◽  
Optimal Conditions ◽  
Cultural Medium ◽  
Yeast Fungi ◽  
Series Of Experiments

To determine the optimal conditions for the synthesis of melanin by black yeast fungi Pseudonadsoniella brunnea (Basidiomycota, Agaricomycotina, Agaricomycetes, Polyporales, Meripilaceae), depending on the amount of L-tyrosine in the culture medium was the purpose of the work. The standard Malt Extract Broth (MEB) liquid nutrient medium was used within this study. L-tyrosine was added to the culture medium in a quantity of 0.01, 0.025 and 0.05%.To obtain the melanin the cultivation of Pseudonadsoniella brunnea was carried out at pH 1-1.5, temperature + 21 ± 1 ° C during 7 days. Statistical processing of the results was carried out using generally accepted methods of variation statistics. It has been established that the level of melanin synthesis by black yeast-like fungi Pseudonadsoniella brunnea depends on the amount of L-tyrosine introduced into the culture medium. The MEB nutrient medium containing 0.05% L-tyrosine in this series of experiments found to be the best composition for obtaining melanin by the strain-producer Pseudonadsoniella brunnea. Compared to control (MEB without L-tyrosine), the amount of melanin synthesized by Ps. brunnea in these conditions increased by 2.5 times. The further research into the optimal conditions for the cultivation of black yeast-like fungi Pseudonadsoniella brunnea in order to obtain melanin is relevant and promising.

scholarly journals Complementary Contribution of Fungi and Bacteria to Lignocellulose Digestion in the Food Stored by a Neotropical Higher Termite

2021 ◽  
Vol 9 ◽  
Author(s):  
Edimar A. Moreira ◽  
Gabriela F. Persinoti ◽  
Letícia R. Menezes ◽  
Douglas A. A. Paixão ◽  
Thabata M. Alvarez ◽  
...  
Keyword(s):  
Plant Material ◽  
Plant Biomass ◽  
Carbohydrate Active Enzymes ◽  
Expression Of Genes ◽  
Its Sequence Analysis ◽  
Gut Symbionts ◽  
Cazy Genes ◽  
Complementary Set ◽  
New Evidence ◽  
Differential Expression Of Genes

Lignocellulose digestion in termites is achieved through the functional synergy between gut symbionts and host enzymes. However, some species have evolved additional associations with nest microorganisms that collaborate in the decomposition of plant biomass. In a previous study, we determined that plant material packed with feces inside the nests of Cornitermes cumulans (Syntermitinae) harbors a distinct microbial assemblage. These food nodules also showed a high hemicellulolytic activity, possibly acting as an external place for complementary lignocellulose digestion. In this study, we used a combination of ITS sequence analysis, metagenomics, and metatranscriptomics to investigate the presence and differential expression of genes coding for carbohydrate-active enzymes (CAZy) in the food nodules and the gut of workers and soldiers. Our results confirm that food nodules express a distinct set of CAZy genes suggesting that stored plant material is initially decomposed by enzymes that target the lignin and complex polysaccharides from fungi and bacteria before the passage through the gut, where it is further targeted by a complementary set of cellulases, xylanases, and esterases produced by the gut microbiota and the termite host. We also showed that the expression of CAZy transcripts associated to endoglucanases and xylanases was higher in the gut of termites than in the food nodules. An additional finding in this study was the presence of fungi in the termite gut that expressed CAZy genes. This study highlights the importance of externalization of digestion by nest microbes and provides new evidence of complementary digestion in the context of higher termite evolution.

scholarly journals Large distances separate coregulated genes in living Drosophila embryos

2019 ◽  
Vol 116 (30) ◽  
pp. 15062-15067 ◽  
Author(s):  
Tyler Heist ◽  
Takashi Fukaya ◽  
Michael Levine
Keyword(s):  
Rna Polymerase Ii ◽  
Transcriptional Activity ◽  
Target Genes ◽  
Reporter Genes ◽  
Transcriptional Enhancers ◽  
Close Proximity ◽  
Resolution Imaging ◽  
In Cis ◽  
Target Promoters ◽  
Drosophila Embryos

Transcriptional enhancers are short segments of DNA that switch genes on and off in response to a variety of cellular signals. Many enhancers map quite far from their target genes, on the order of tens or even hundreds of kilobases. There is extensive evidence that remote enhancers are brought into proximity with their target promoters via long-range looping interactions. However, the exact physical distances of these enhancer–promoter interactions remain uncertain. Here, we employ high-resolution imaging of living Drosophila embryos to visualize the distances separating linked genes that are coregulated by a shared enhancer. Cotransvection assays (linked genes on separate homologs) suggest a surprisingly large distance during transcriptional activity: at least 100–200 nm. Similar distances were observed when a shared enhancer was placed into close proximity with linked reporter genes in cis. These observations are consistent with the occurrence of “transcription hubs,” whereby clusters (or condensates) of multiple RNA polymerase II complexes and associated cofactors are periodically recruited to active promoters. The dynamics of this process might be responsible for rapid fluctuations in the distances separating the transcription of coregulated reporter genes during transvection. We propose that enhancer-promoter communication depends on a combination of classical looping and linking models.

Co-operation between protein-acetylating and protein-methylating co-activators in transcriptional activation

2000 ◽  
Vol 28 (4) ◽  
pp. 415-418 ◽  
Author(s):  
M. R. Stallcup ◽  
D. Chen ◽  
S. S. Koh ◽  
H. Ma ◽  
Y.-H. Lee ◽  
...  
Keyword(s):  
Transcriptional Activation ◽  
Transcription Initiation ◽  
Protein Modification ◽  
Hormone Receptors ◽  
Target Genes ◽  
Reporter Genes ◽  
Specific Protein ◽  
Synergistic Enhancement ◽  
Arginine Residues

Nuclear hormone receptors (NRs) activate transcription by binding to specific enhancer elements associated with target genes. Transcriptional activation is accomplished with the help of complexes of co-activator proteins that bind to NRs. p160 co-activators, a family of three related 160 kDa proteins, serve as primary co-activators by binding directly to NRs and recruiting additional secondary co-activators. Some of these (CBP/p300 and p/CAF) can acetylate histones and other proteins in the transcription complex, thus helping to modify chromatin structure and form an active transcription initiation complex. We recently discovered co-activator-associated arginine methyltransferase 1 (CARM1), which binds to p160 co-activators and thereby enhances transcriptional activation by NRs on transiently transfected reporter genes. CARM1 also methylates specific arginine residues in the N-terminal tail of histone H3 in vitro. A related arginine-specific protein methyltransferase, PRMT1, also binds p160 co-activators and enhances NR function. PRMT1 methylates histone H4 in vitro. The enhancement of NR function by CARM1, PRMT1 and p300 depends on their interactions with p160 co-activators. In the presence of p160 co-activators, some pairs of these three secondary co-activators provide a highly synergistic enhancement of NR function on transiently transfected reporter genes. We have also observed an enhancement of NR function on stably integrated reporter genes by these co-activators. We propose that the synergy of co-activator function between p300, CARM1 and PRMT1 is due to their different but complementary protein modification activities.

scholarly journals Differential Morphophysiological and Biochemical Responses of Cotton Genotypes Under Various Salinity Stress Levels During Early Growth Stage

2021 ◽  
Vol 12 ◽  
Author(s):  
Wajeeha Munawar ◽  
Amjad Hameed ◽  
Muhammad Khashif Riaz Khan
Keyword(s):  
Salt Tolerance ◽  
Salinity Stress ◽  
Photosynthetic Rate ◽  
Plant Biomass ◽  
Fiber Quality ◽  
Enzymatic Antioxidants ◽  
Cotton Production ◽  
Cotton Genotypes ◽  
Mda Content ◽  
Series Of Experiments

Cotton is a primary agriculture product important for fiber use in textiles and the second major oil seed crop. Cotton is considered as moderately tolerant to salt stress with salinity threshold of 7.7 dS/m at seedling stage. Salinity causes reduction in the growth of seedlings and cotton production that limits fiber quality and cotton yield. In this study, initially, 22 cotton genotypes were screened for relative salt tolerance using germination test in Petri plates (growth chamber). Selected 11 genotypes were further tested in pot experiment (sand) with 0, 15, and 20 dS/m NaCl treatments under glass house conditions. At four-leaves stage, different morphological and physiological traits were measured for all genotypes while biochemical analysis was performed on selected seven highly tolerant and sensitive genotypes. NaCl treatment significantly reduced plant biomass in two genotypes IR-NIBGE-13 and BS-2018, while NIAB-135, NIAB-512, and GH-HADI had least difference in fresh weight between the control and NaCl-treated plants. Photosynthetic rate was maintained in all the genotypes with the exception of SITARA-16. In two sensitive genotypes (IR-NIBGE-13 and 6071/16), Na+ ion accumulated more in leaves as compared to K+ ion under stress conditions, and an increase in Na+/K+ ratio was also observed. The lesser accumulation of malondialdehyde (MDA) content and higher activity of enzymatic antioxidants such as superoxide dismutase (SOD), peroxidase (POD), and ascorbate peroxidase (APX) in stressed plants of NIAB-135, NIAB-512, and FH-152 indicated that these genotypes had adaption capacity for salinity stress in comparison with sensitive genotypes, i.e., IR-NIBGE-13 and 6071/16. The observed salt tolerance was corelated with plant biomass maintenance (morphological), photosynthetic rate, and ionic homeostasis (K+/Na+ ratio, physiological) and biochemical stress marker regulations. After a series of experiments, it was concluded that NIAB-135, NIAB-512, and FH-152 could be utilized in breeding programs aimed at improving salinity tolerance in cotton and can expand cotton cultivation in saline area.

scholarly journals Integrated sRNAome and RNA-Seq Analysis Reveals miRNA Effects on Betalain Biosynthesis in Pitaya

2020 ◽  
Author(s):  
Canbin Chen ◽  
Fangfang Xie ◽  
Qingzhu Hua ◽  
Noemi Tel Zur ◽  
Lulu Zhang ◽  
...  
Keyword(s):  
Transient Expression ◽  
Plant Species ◽  
Negative Correlation ◽  
Target Genes ◽  
Regulatory Mechanism ◽  
Expression System ◽  
Expression Patterns ◽  
Rna Seq ◽  
Chlorophyll Accumulation ◽  
Regulatory Functions

Abstract Background: MicroRNAs (miRNAs) and their regulatory functions in anthocyanin, carotenoid, and chlorophyll accumulation have been extensively characterized in many plant species. However, the miRNA regulatory mechanism in betalain biosynthesis remains mostly unknown. Results: In this study, 126 conserved miRNAs and 41 novel miRNAs were first isolated from Hylocereus monacanthus, among which 95 conserved miRNAs belonged to 53 miRNA families. 34 candidate miRNAs related to betalain biosynthesis were found to be differentially expressed. The expression patterns of those differential expressed miRNAs were analyzed in various tissues of the pitaya by RT-qPCR. A significantly negative correlation was detected between the expression levels of half those miRNAs and corresponding target genes. Target genes of miRNAs i.e. aly-miR157d-5p_L+1_1ss4AC-comp25631_c0, aau-miR160_L-4R+ 1-comp36993_c0_seq3, nta-miR6020b-comp234190_c0, PC-5p-192_7269-comp29967_c0, PC-5p-23845_39-comp28219_c0, mdm-miR828a_1ss22AT-comp24967_c0, mdm-miR858- comp15143_c0, mdm-miR858-comp24362_c0 and mdm-miR858-comp403340_c0 were verified by 5′RACE and transient expression system in tobacco.Conclusions: aly-miR157d-5p_L+1_1ss4AC, aau-miR160_L-4R+1, nta-miR6020b PC-5p-192_7269, PC-5p-23845_39, mdm-miR828a_1ss22AT and mdm-miR858 may play important roles in pitaya fruit coloration and betalain accumulation. Our findings provide insights into the roles of miRNAs and their target genes of regulatory functions involved in betalain biosynthesis of pitaya.

Analysis of competence and of Brachyury autoinduction by use of hormone-inducible Xbra

Development ◽  
1997 ◽  
Vol 124 (11) ◽  
pp. 2225-2234 ◽  
Author(s):  
M. Tada ◽  
M.A. O'Reilly ◽  
J.C. Smith
Keyword(s):  
Glucocorticoid Receptor ◽  
Target Genes ◽  
Early Embryo ◽  
Fate Map ◽  
Animal Pole ◽  
Reading Frame ◽  
Xenopus Development ◽  
Series Of Experiments ◽  
Inducing Factors ◽  
Mesoderm Inducing Factors

Analysis of gene function in Xenopus development frequently involves over-expression experiments, in which RNA encoding the protein of interest is microinjected into the early embryo. By taking advantage of the fate map of Xenopus, it is possible to direct expression of the protein to particular regions of the embryo, but it has not been possible to exert control over the timing of expression; the protein is translated immediately after injection. To overcome this problem in our analysis of the role of Brachyury in Xenopus development, we have, like Kolm and Sive (1995; Dev. Biol. 171, 267–272), explored the use of hormone-inducible constructs. Animal pole regions derived from embryos expressing a fusion protein (Xbra-GR) in which the Xbra open reading frame is fused to the ligand-binding domain of the human glucocorticoid receptor develop as atypical epidermis, presumably because Xbra is sequestered by the heat-shock apparatus of the cell. Addition of dexamethasone, which binds to the glucocorticoid receptor and releases Xbra, causes formation of mesoderm. We have used this approach to investigate the competence of animal pole explants to respond to Xbra-GR, and have found that competence persists until late gastrula stages, even though by this time animal caps have lost the ability to respond to mesoderm-inducing factors such as activin and FGF. In a second series of experiments, we demonstrate that Xbra is capable of inducing its own expression, but that this auto-induction requires intercellular signals and FGF signalling. Finally, we suggest that the use of inducible constructs may assist in the search for target genes of Brachyury.

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