The regulation of post-germinative transition from the cotyledon- to vegetative-leaf stages by microRNA-targeted SQUAMOSA PROMOTER-BINDING PROTEIN LIKE13 in Arabidopsis

2010 ◽  
Vol 20 (2) ◽  
pp. 89-96 ◽  
Author(s):  
Ruth C. Martin ◽  
Masashi Asahina ◽  
Po-Pu Liu ◽  
Jessica R. Kristof ◽  
Jennifer L. Coppersmith ◽  
...  
Keyword(s):  
Binding Protein ◽  
Leaf Primordium ◽  
Mutant Form ◽  
Wild Type ◽  
Cotyledon Stage ◽  
Squamosa Promoter Binding Protein ◽  
Early Seedling ◽  
Early Seedling Development ◽  
Vegetative Leaf

AbstractGermination and early seedling development are critical for successful stand establishment of plants. Following germination, the cotyledons, which are derived from embryonic tissue, emerge from the seed. Arabidopsis seedlings at post-germinative stages are supported mainly by the supply of nutrition from the cotyledons until vegetative leaves emerge and initiate photosynthesis. The switch to autotrophic growth is a significant transition at the post-germinative stage. Here, we provide evidence that down-regulation of SQUAMOSA PROMOTER-BINDING PROTEIN LIKE13 (SPL13) by microRNA156 (miR156) plays an important role in the regulation of the post-germinative switch from the cotyledon stage to the vegetative-leaf stage. Silent mutations created in the SPL13 sequence in the region that is complementary to the miR156 sequence caused the deregulation of the mutant form of SPL13 (mSPL13) mRNA from miR156. Mutant seedlings over-accumulated miRNA-resistant messages and exhibited a delay in the emergence of vegetative leaves compared to wild-type seedlings. The delay was not observed in control transgenic plants expressing non-mutated SPL13, indicating that the phenotype was caused specifically by the silent mutations and deregulation of SPL13 from miR156. Characterization of the SPL13 promoter indicated that this gene is expressed mainly in the hypocotyl and affects leaf primordium development. These results suggest that the repression of SPL13 by miR156 is essential for normal post-germinative growth in Arabidopsis.

scholarly journals Designing and studying a mutant form of the ice-binding protein from Choristoneura fumiferana

2020 ◽  
Author(s):  
Ksenia A. Glukhova ◽  
Julia D. Okulova ◽  
Bogdan S. Melnik
Keyword(s):  
Binding Protein ◽  
Choristoneura Fumiferana ◽  
Freezing Temperature ◽  
Mutant Form ◽  
Wild Type ◽  
Isolation And Purification ◽  
Eastern Spruce Budworm ◽  
Ice Surface ◽  
Ice Binding ◽  
Ice Binding Protein

AbstractIce-binding proteins are expressed in the cells of some organisms, helping them to survive extremely low temperatures. One of the problems in study of such proteins is the difficulty of isolation and purification. For example, eight cysteine residues in cfAFP from Choristoneura fumiferana (the eastern spruce budworm) form intermolecular bridges during the overexpression of this protein. This impedes the process of the protein purification dramatically.In this work we designed a mutant form of ice-binding protein cfAFP, which is much more easy to isolate that the wild-type protein. The mutant form named mIBP83 did not lose the ability to bind to ice surface. Besides, observation of the processes of freezing and melting of ice in presence of mIBP83 showed that this protein affects the process of ice melting, increasing its melting temperature, and at least does not decrease the freezing temperature.

scholarly journals Ectopic Expression of Glycine max GmNAC109 Enhances Drought Tolerance and ABA Sensitivity in Arabidopsis

Biomolecules ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 714 ◽  
Author(s):  
Nguyen ◽  
Hoang ◽  
Nguyen ◽  
Binh ◽  
Watanabe ◽  
...  
Keyword(s):  
Glycine Max ◽  
Drought Tolerance ◽  
Water Deficit ◽  
Ectopic Expression ◽  
Wild Type ◽  
Aba Sensitivity ◽  
Leaf Tissues ◽  
Early Seedling ◽  
Early Seedling Development ◽  
Higher Sensitivity

The NAC (NAM, ATAF1/2, CUC2) transcription factors are widely known for their various functions in plant development and stress tolerance. Previous studies have demonstrated that genetic engineering can be applied to enhance drought tolerance via overexpression/ectopic expression of NAC genes. In the present study, the dehydration- and drought-inducible GmNAC109 from Glycine max was ectopically expressed in Arabidopsis (GmNAC109-EX) plants to study its biological functions in mediating plant adaptation to water deficit conditions. Results revealed an improved drought tolerance in the transgenic plants, which displayed greater recovery rates by 20% to 54% than did the wild-type plants. In support of this finding, GmNAC109-EX plants exhibited lower water loss rates and decreased endogenous hydrogen peroxide production in leaf tissues under drought, as well as higher sensitivity to exogenous abscisic acid (ABA) treatment at germination and early seedling development stages. In addition, analyses of antioxidant enzymes indicated that GmNAC109-EX plants possessed stronger activities of superoxide dismutase and catalase under drought stress. These results together demonstrated that GmNAC109 acts as a positive transcriptional regulator in the ABA-signaling pathway, enabling plants to cope with adverse water deficit conditions.

scholarly journals Magnitude of the CREB-Dependent Transcriptional Response Is Determined by the Strength of the Interaction between the Kinase-Inducible Domain of CREB and the KIX Domain of CREB-Binding Protein

2000 ◽  
Vol 20 (24) ◽  
pp. 9409-9422 ◽  
Author(s):  
Adam J. Shaywitz ◽  
Simon L. Dove ◽  
Jon M. Kornhauser ◽  
Ann Hochschild ◽  
Michael E. Greenberg
Keyword(s):  
Amino Acid ◽  
Amino Acid Substitution ◽  
Protein Interactions ◽  
Mammalian Cells ◽  
Binding Protein ◽  
Full Length ◽  
Mutant Form ◽  
Creb Binding Protein ◽  
Wild Type ◽  
Kix Domain

ABSTRACT The activity of the transcription factor CREB is regulated by extracellular stimuli that result in its phosphorylation at a critical serine residue, Ser133. Phosphorylation of Ser133 is believed to promote CREB-dependent transcription by allowing CREB to interact with the transcriptional coactivator CREB-binding protein (CBP). Previous studies have established that the domain encompassing Ser133 on CREB, known as the kinase-inducible domain (KID), interacts specifically with a short domain in CBP termed the KIX domain and that this interaction depends on the phosphorylation of Ser133. In this study, we adapted a recently described Escherichia coli-based two-hybrid system for the examination of phosphorylation-dependent protein-protein interactions, and we used this system to study the kinase-induced interaction between the KID and the KIX domain. We identified residues of the KID and the KIX domain that are critical for their interaction as well as two pairs of oppositely charged residues that apparently interact at the KID-KIX interface. We then isolated a mutant form of the KIX domain that interacts more tightly with wild-type and mutant forms of the KID than does the wild-type KIX domain. We show that in the context of full-length CBP, the corresponding amino acid substitution resulted in an enhanced ability of CBP to stimulate CREB-dependent transcription in mammalian cells. Conversely, an amino acid substitution in the KIX domain that weakens its interaction with the KID resulted in a decreased ability of full-length CBP to stimulate CREB-dependent transcription. These findings demonstrate that the magnitude of CREB-dependent transcription in mammalian cells depends on the strength of the KID-KIX interaction and suggest that the level of transcription induced by coactivator-dependent transcriptional activators can be specified by the strength of the activator-coactivator interaction.

scholarly journals Characterization of dacC, Which Encodes a New Low-Molecular-Weight Penicillin-Binding Protein in Bacillus subtilis

1998 ◽  
Vol 180 (18) ◽  
pp. 4967-4973 ◽  
Author(s):  
Lotte B. Pedersen ◽  
Thomas Murray ◽  
David L. Popham ◽  
Peter Setlow
Keyword(s):  
Molecular Weight ◽  
Bacillus Subtilis ◽  
Binding Protein ◽  
Low Molecular Weight ◽  
Penicillin Binding Protein ◽  
Wild Type ◽  
Genome Sequencing Project ◽  
Penicillin Binding Proteins ◽  
Sequencing Project

ABSTRACT The pbp gene (renamed dacC), identified by the Bacillus subtilis genome sequencing project, encodes a putative 491-residue protein with sequence homology to low-molecular-weight penicillin-binding proteins. Use of a transcriptional dacC-lacZ fusion revealed thatdacC expression (i) is initiated at the end of stationary phase; (ii) depends strongly on transcription factor ςH; and (iii) appears to be initiated from a promoter located immediately upstream of yoxA, a gene of unknown function located upstream of dacC on the B. subtilis chromosome. A B. subtilis dacCinsertional mutant grew and sporulated identically to wild-type cells, and dacC and wild-type spores had the same heat resistance, cortex structure, and germination and outgrowth kinetics. Expression ofdacC in Escherichia coli showed that this gene encodes an ∼59-kDa membrane-associated penicillin-binding protein which is highly toxic when overexpressed.

scholarly journals Morphological characterization of fruit, seed and seedling and germination of Hymenaea courbaril L. (Fabaceae) ('Jatobá')

2016 ◽  
Vol 38 (3) ◽  
pp. 204-211 ◽  
Author(s):  
Manoela Mendes Duarte ◽  
Sheilly Raquelly Prado de Paula ◽  
Flaick Rodrigo de Lima Ferreira ◽  
Antonio Carlos Nogueira
Keyword(s):  
Germination Percentage ◽  
Morphological Characterization ◽  
Morphological Description ◽  
Hymenaea Courbaril ◽  
Randomized Design ◽  
Early Seedling ◽  
Mean Germination Time ◽  
Completely Randomized Design ◽  
Early Seedling Development

Abstract This study has aimed to characterize the morphology of fruit, seed and seedling and evaluate the germination of Hymenaea courbaril L., under different substrates and temperatures. The physical characteristics were determined, as well as the morphological description of the internal and external parts of fruits and seeds, stages of germination and early seedling development. Germination tests were conducted in a completely randomized design, in a 3x2 factorial scheme, with three temperatures (20 °C, 25 °C and 30 °C) and two substrates (sand and vermiculite), with six plots of 30 seeds. The germination percentage, germination speed index (GSI) and mean germination time (MGT) were calculated. The fruits are dry and woody and the seeds are exalbuminous, with an axial embryo. The germination is epigeal and the seedling is phanerocotylar. The combination of vermiculite or sand associated to temperatures of 25 °C or 30 °C is effective for conducting the germination tests.

scholarly journals The CRISPR/Cas9-Mediated Modulation of SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE 8 in Alfalfa Leads to Distinct Phenotypic Outcomes

2022 ◽  
Vol 12 ◽  
Author(s):  
Stacy D. Singer ◽  
Kimberley Burton Hughes ◽  
Udaya Subedi ◽  
Gaganpreet Kaur Dhariwal ◽  
Kazi Kader ◽  
...  
Keyword(s):  
Drought Tolerance ◽  
First Generation ◽  
Binding Protein ◽  
Leaf Size ◽  
Growth Conditions ◽  
Wild Type ◽  
Induced Mutations ◽  
Forage Crop ◽  
Morphological Alterations ◽  
Squamosa Promoter Binding Protein

Alfalfa (Medicago sativa L.) is the most widely grown perennial leguminous forage and is an essential component of the livestock industry. Previously, the RNAi-mediated down-regulation of alfalfa SQUAMOSA PROMOTER-BINDING PROTEIN-LIKE 8 (MsSPL8) was found to lead to increased branching, regrowth and biomass, as well as enhanced drought tolerance. In this study, we aimed to further characterize the function of MsSPL8 in alfalfa using CRISPR/Cas9-induced mutations in this gene. We successfully generated alfalfa genotypes with small insertions/deletions (indels) at the target site in up to three of four MsSPL8 alleles in the first generation. The efficiency of editing appeared to be tightly linked to the particular gRNA used. The resulting genotypes displayed consistent morphological alterations, even with the presence of up to two wild-type MsSPL8 alleles, including reduced leaf size and early flowering. Other phenotypic effects appeared to be dependent upon mutational dosage, with those plants with the highest number of mutated MsSPL8 alleles also exhibiting significant decreases in internode length, plant height, shoot and root biomass, and root length. Furthermore, MsSPL8 mutants displayed improvements in their ability to withstand water-deficit compared to empty vector control genotypes. Taken together, our findings suggest that allelic mutational dosage can elicit phenotypic gradients in alfalfa, and discrepancies may exist in terms of MsSPL8 function between alfalfa genotypes, growth conditions, or specific alleles. In addition, our results provide the foundation for further research exploring drought tolerance mechanisms in a forage crop.

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