scholarly journals Overexpression of ANAC046 Promotes Suberin Biosynthesis in Roots of Arabidopsis thaliana

2019 ◽  
Vol 20 (24) ◽  
pp. 6117 ◽  
Author(s):  
Kashif Mahmood ◽  
Viktoria Valeska Zeisler-Diehl ◽  
Lukas Schreiber ◽  
Yong-Mei Bi ◽  
Steven J. Rothstein ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Pcr Analysis ◽  
Transcription Activator ◽  
Cuticular Waxes ◽  
Plant Parts ◽  
Wide Range ◽  
Transgenic Lines ◽  
Blue Stain ◽  
Root Endodermis

NAC (NAM (no apical meristem), ATAF1/2, and CUC2 (cup-shaped cotyledon)) proteins are one of the largest families of plant-specific transcription factors, and this family is present in a wide range of land plants. Here, we have investigated the role of ANAC046 in the regulation of suberin biosynthesis and deposition in Arabidopsis. Subcellular localization and transcriptional activity assays showed that ANAC046 localizes in the nucleus, where it functions as a transcription activator. Analysis of the PANAC046:GUS lines revealed that ANAC046 is mainly expressed in the root endodermis and periderm, and is also induced in leaves by wounding. The transgenic lines overexpressing ANAC046 exhibited defective surfaces on the aerial plant parts compared to the wild-type (WT) as characterized by increased permeability for Toluidine blue stain and greater chlorophyll leaching. Quantitative RT-PCR analysis showed that the expression of suberin biosynthesis genes was significantly higher in the roots and leaves of overexpression lines compared to the WT. The biochemical analysis of leaf cuticular waxes showed that the overexpression lines accumulated 30% more waxes than the WT. Concurrently, overexpression lines also deposited almost twice the amount of suberin content in their roots compared with the WT. Taken together, these results showed that ANAC046 is an important transcription factor that promotes suberin biosynthesis in Arabidopsis thaliana roots.

scholarly journals Conserved Cu-MicroRNAs in Arabidopsis thaliana Function in Copper Economy under Deficiency

Plants ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 141 ◽  
Author(s):  
Muhammad Shahbaz ◽  
Marinus Pilon
Keyword(s):  
Arabidopsis Thaliana ◽  
Small Rnas ◽  
Wild Type ◽  
Microrna Target ◽  
Electron Carrier ◽  
Cu Deficiency ◽  
Regulatory Circuits ◽  
Transgenic Lines ◽  
Cu Homeostasis

Copper (Cu) is a micronutrient for plants. Three small RNAs, which are up-regulated by Cu deficiency and target transcripts for Cu proteins, are among the most conserved microRNAs in plants. It was hypothesized that these Cu-microRNAs help save Cu for the most essential Cu-proteins under deficiency. Testing this hypothesis has been a challenge due to the redundancy of the Cu microRNAs and the properties of the regulatory circuits that control Cu homeostasis. In order to investigate the role of Cu-microRNAs in Cu homeostasis during vegetative growth, we used a tandem target mimicry strategy to simultaneously inhibit the function of three conserved Cu-microRNAs in Arabidopsis thaliana. When compared to wild-type, transgenic lines that express the tandem target mimicry construct showed reduced Cu-microRNA accumulation and increased accumulation of transcripts that encode Cu proteins. As a result, these mimicry lines showed impaired photosynthesis and growth compared to wild type on low Cu, which could be ascribed to a defect in accumulation of plastocyanin, a Cu-containing photosynthetic electron carrier, which is itself not a Cu-microRNA target. These data provide experimental support for a Cu economy model where the Cu-microRNAs together function to allow maturation of essential Cu proteins under impending deficiency.

scholarly journals Intersection of the tocopherol and plastoquinol metabolic pathways at the plastoglobule

2009 ◽  
Vol 425 (2) ◽  
pp. 389-399 ◽  
Author(s):  
Anna Maria Zbierzak ◽  
Marion Kanwischer ◽  
Christina Wille ◽  
Pierre-Alexandre Vidi ◽  
Patrick Giavalisco ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Photosystem Ii ◽  
Metabolic Pathways ◽  
Polar Lipids ◽  
Protein Bodies ◽  
Side Chain ◽  
Considerable Proportion ◽  
Transgenic Lines ◽  
Tocopherol Cyclase

Plastoglobules, lipid–protein bodies in the stroma of plant chloroplasts, are enriched in non-polar lipids, in particular prenyl quinols. In the present study we show that, in addition to the thylakoids, plastoglobules also contain a considerable proportion of the plastidial PQ-9 (plastoquinol-9), the redox component of photosystem II, and of the cyclized product of PQ-9, PC-8 (plastochromanol-8), a tocochromanol with a structure similar to γ-tocopherol and γ-tocotrienol, but with a C-40 prenyl side chain. PC-8 formation was abolished in the Arabidopsis thaliana tocopherol cyclase mutant vte1, but accumulated in VTE1-overexpressing plants, in agreement with a role of tocopherol cyclase (VTE1) in PC-8 synthesis. VTE1 overexpression resulted in the proliferation of the number of plastoglobules which occurred in the form of clusters in the transgenic lines. Simultaneous overexpression of VTE1 and of the methyltransferase VTE4 resulted in the accumulation of a compound tentatively identified as 5-methyl-PC-8, the methylated form of PC-8. The results of the present study suggest that the existence of a plastoglobular pool of PQ-9, along with the partial conversion of PQ-9 into PC-8, might represent a mechanism for the regulation of the antioxidant content in thylakoids and of the PQ-9 pool that is available for photosynthesis.

scholarly journals Increased Cuticle Waxes by Overexpression of WSD1 Improves Osmotic Stress Tolerance in Arabidopsis thaliana and Camelina sativa

2021 ◽  
Vol 22 (10) ◽  
pp. 5173
Author(s):  
Hesham M. Abdullah ◽  
Jessica Rodriguez ◽  
Jeffrey M. Salacup ◽  
Isla S. Castañeda ◽  
Danny J. Schnell ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Critical Role ◽  
Plant Productivity ◽  
Camelina Sativa ◽  
Wax Ester ◽  
Pcr Analysis ◽  
Oilseed Crop ◽  
Cuticular Waxes ◽  
Changing Climate ◽  
Wax Synthase

To ensure global food security under the changing climate, there is a strong need for developing ‘climate resilient crops’ that can thrive and produce better yields under extreme environmental conditions such as drought, salinity, and high temperature. To enhance plant productivity under the adverse conditions, we constitutively overexpressed a bifunctional wax synthase/acyl-CoA:diacylglycerol acyltransferase (WSD1) gene, which plays a critical role in wax ester synthesis in Arabidopsis stem and leaf tissues. The qRT-PCR analysis showed a strong upregulation of WSD1 transcripts by mannitol, NaCl, and abscisic acid (ABA) treatments, particularly in Arabidopsis thaliana shoots. Gas chromatography and electron microscopy analyses of Arabidopsis seedlings overexpressing WSD1 showed higher deposition of epicuticular wax crystals and increased leaf and stem wax loading in WSD1 transgenics compared to wildtype (WT) plants. WSD1 transgenics exhibited enhanced tolerance to ABA, mannitol, drought and salinity, which suggested new physiological roles for WSD1 in stress response aside from its wax synthase activity. Transgenic plants were able to recover from drought and salinity better than the WT plants. Furthermore, transgenics showed reduced cuticular transpirational rates and cuticle permeability, as well as less chlorophyll leaching than the WT. The knowledge from Arabidopsis was translated to the oilseed crop Camelina sativa (L.) Crantz. Similar to Arabidopsis, transgenic Camelina lines overexpressing WSD1 also showed enhanced tolerance to drought stress. Our results clearly show that the manipulation of cuticular waxes will be advantageous for enhancing plant productivity under a changing climate.

scholarly journals Jasmonates-Mediated Rewiring of Central Metabolism Regulates Adaptive Responses

2019 ◽  
Vol 60 (12) ◽  
pp. 2613-2620 ◽  
Author(s):  
Tatyana V Savchenko ◽  
Hardy Rolletschek ◽  
Katayoon Dehesh
Keyword(s):  
Central Metabolism ◽  
Adaptive Responses ◽  
Active Growth ◽  
Environmental Challenges ◽  
Collective Data ◽  
Specific Distribution ◽  
Wide Range ◽  
Transgenic Lines ◽  
Branched Chain

Abstract The lipid-derived hormones jasmonates (JAs) play key functions in a wide range of physiological and developmental processes that regulate growth, secondary metabolism and defense against biotic and abiotic stresses. In this connection, biosynthesis, tissue-specific distribution, metabolism, perception, signaling of JAs have been the target of extensive studies. In recent years, the involvement of JAs signaling pathway in the regulation of growth and adaptive responses to environmental challenges has been further examined. However, JAs-mediated mechanisms underlying the transition from ‘growth mode’ to ‘adaptive mode’ remain ambiguous. Combined analysis of transgenic lines deficient in JAs signaling in conjunction with the data from JAs-treated plants revealed the function of these hormones in rewiring of central metabolism. The collective data illustrate JAs-mediated decrease in the levels of metabolites associated with active growth such as sucrose, raffinose, orotate, citrate, malate, and an increase in phosphorylated hexoses, responsible for the suppression of growth and photosynthesis, concurrent with the induction of protective metabolites, such as aromatic and branched-chain amino acids, and aspartate family of metabolites. This finding provides an insight into the function of JAs in shifting the central metabolism from the production of growth-promoting metabolites to protective compounds and expands our understanding of the role of JAs in resource allocation in response to environmental challenges.

Tolerance to, and Uptake and Degradation of 2,4,6-Trinitrotoluene (TNT) are Enhanced by the Expression of a Bacterial Nitroreductase Gene in Arabidopsis thaliana

2005 ◽  
Vol 60 (3-4) ◽  
pp. 272-278 ◽  
Author(s):  
Mami Kurumata ◽  
Misa Takahashi ◽  
Atsushi Sakamoto ◽  
Juan L. Ramos ◽  
Ales Nepovim ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Transgenic Plants ◽  
Degradation Products ◽  
Plant Genome ◽  
Pcr Analysis ◽  
Wild Type ◽  
In Planta ◽  
Gene Encoding ◽  
Plant Body ◽  
Wide Range

Abstract Arabidopsis thaliana was transformed with a gene encoding a nitroreductase (NTR, E.C. 1.6.99.7) with activity against a wide range of nitroaromatic compounds. The gene was transferred from Escherichia coli by an Agrobacterium-mediated in planta method. The ob­tained seeds were sowed to produce T1 plants, and they were assayed for the integration of the transgene in the plant genome. Transgenic plants that were positive with the PCR analysis were self-pollinated to produce T2 generation plants. Seven lines obtained were assayed for the NTR activity. While the noil-transformed wild-type plants showed no detectable NTR activity, the enzyme activity of the transgenic plant lines was approx. 20 times higher. Using the line with the highest NTR activity, the phytoremediation characteristics of plants against 2,4,6-trinitrotoluene (TNT) was investigated. While the wild-type plants did not grow in the presence of 0.1 mᴍ TNT, the transgenic plants grew almost normally in this condition. The uptake of TNT by seedlings of transgenic plants increased by 7 to 8 times when they were floated on TNT solution. HPLC analysis showed that the peak due to TNT taken up into plant body was much smaller in the transgenic plants as compared with that of the wild type, and that a number of peaks attributable to the degradation products of TNT, including 4-amino-2,6-dinitrotoluene, were detected in the extract from the transgenic plants. This indi­cates that the expression of bacterial NTR improved the capability of plants to degrade TNT.

scholarly journals Ectopic Expression of Fiber Related Gbwri1 Complements Seed Phenotype in Arabidopsis thaliana

2021 ◽  
Vol 25 (01) ◽  
pp. 123-130
Author(s):  
Mahnoor Imran
Keyword(s):  
Arabidopsis Thaliana ◽  
Fatty Acid ◽  
Fatty Acid Biosynthesis ◽  
Ectopic Expression ◽  
Acid Biosynthesis ◽  
Functional Homology ◽  
Normal Seedling ◽  
Transgenic Lines ◽  
Protein Isoform

WRINKLED1 belongs to AP2/EREB family of transcription factors whose role has been well established in seed oil biosynthesis. The objective of the study was to trace the role of fiber related Gbwri1 in seed development and fatty acid biosynthesis. In this study, we isolated a transcript from elite fiber producing cotton (Gossypium barbadense), which is over-expressed in G. barbadense fibers as compared to G. hirsutum and G. arboreum. The putative protein encoded by this transcript exhibited homology in specific domains and protein structure with WRINKLED1 of Arabidopsis thaliana and was thus designated as Gbwri1. In this study, we investigated the functional homology of fiber elongation related Gbwri1 with fatty acid biosynthesis regulator Atwri1. Ectopic expression of Gbwri1 in wri1-3 mutant of A. thaliana was analyzed. In the transgenic lines of A. thaliana, Gbwri1 resumed the seed weight, seed area, and surface morphology to the wild type. Gbwri1 transformation rescued the wrinkled phenotype of wri1-3 mutants by resuming the expression of fatty acid biosynthesis genes biotin carboxyl carrier protein isoform 2 (bccp2) and keto-ACP synthase 1 (kas1). Moreover, the seedling development of transgenic lines on non-sucrose medium demonstrated that the Gbwri1 was able to regulate the supply of sucrose for normal seedling establishment. Our results showed that the transformation of Gbwri1 in A. thaliana wri1-3 mutant was able to complement wri1-3 impaired phenotype. Thus, Gbwri1 is involved in cotton fiber development and fatty acid biosynthesis in seeds. © 2021 Friends Science Publishers

scholarly journals Expression profiling of kinesins, involved in the development of autophagy in Arabidopsis thaliana, and the role of tubulin acetylation in the interaction of Atg8 protein with microtubules

2018 ◽  
Vol 22 ◽  
pp. 162-168
Author(s):  
V. D. Olenieva ◽  
D. I. Lytvyn ◽  
A. I. Yemets ◽  
Ya. B. Blume
Keyword(s):  
Arabidopsis Thaliana ◽  
Structural Biology ◽  
Abiotic Stresses ◽  
Transcriptional Activity ◽  
Pcr Analysis ◽  
Expression Levels ◽  
Tubulin Acetylation ◽  
Atg8 Protein ◽  
Dynamics Simulations

Aim. To investigate the interrelation between changes in the expression levels of kinesin genes that are potentially involved in the development of stress-induced autophagy in Arabidopsis thaliana by means of microtubules, and the structural biology analysis of the role of α-tubulin acetylation in the regulation of interaction of α-tubulin with Atg8. Methods. The simulation of the influence of abiotic stresses. PCR analysis of changes in expression levels of kinesin genes. The molecular dynamics simulations of α-tubulin and Atg8 complexes were performed using the GROMACS 4.5.5 program. Results. It was shown that the changes in expression levels were caused by the influence of stressful stimuli. A significant increase in the transcriptional activity of the KIN5B, KIN12B, KIN12F genes after UV-B irradiation, the KIN6, KIN7O, KIN7D, KIN12B genes under osmotic-, and KIN6, KIN12B under salt stress was detected. By means of bioinformatics it was demonstrated that α-tubulin acetylation provides an enhanced interaction of α-tubulin and Atg8 protein. Conclusions. Obtained data point out the important role of kinesins and α-tubulin acetylation in realization of microtubules’ partaking in the development of stress-induced autophagy in plants. Keywords: microtubules, α-tubulin, kinesins, Atg8 protein, stress-induced autophagy.

scholarly journals Molecular identification of three Arabidopsis thaliana mitochondrial dicarboxylate carrier isoforms: organ distribution, bacterial expression, reconstitution into liposomes and functional characterization

2008 ◽  
Vol 410 (3) ◽  
pp. 621-629 ◽  
Author(s):  
Luigi Palmieri ◽  
Nathalie Picault ◽  
Roberto Arrigoni ◽  
Evelyne Besin ◽  
Ferdinando Palmieri ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Functional Characterization ◽  
Dicarboxylic Acids ◽  
Uncoupling Proteins ◽  
Organ Distribution ◽  
Mitochondrial Carrier ◽  
Wide Range ◽  
Mitochondrial Carrier Family ◽  
Arabidopsis Thaliana Genome

Screening of the Arabidopsis thaliana genome revealed three potential homologues of mammalian and yeast mitochondrial DICs (dicarboxylate carriers) designated as DIC1, DIC2 and DIC3, each belonging to the mitochondrial carrier protein family. DIC1 and DIC2 are broadly expressed at comparable levels in all the tissues investigated. DIC1–DIC3 have been reported previously as uncoupling proteins, but direct transport assays with recombinant and reconstituted DIC proteins clearly demonstrate that their substrate specificity is unique to plants, showing the combined characteristics of the DIC and oxaloacetate carrier in yeast. Indeed, the Arabidopsis DICs transported a wide range of dicarboxylic acids including malate, oxaloacetate and succinate as well as phosphate, sulfate and thiosulfate at high rates, whereas 2-oxoglutarate was revealed to be a very poor substrate. The role of these plant mitochondrial DICs is discussed with respect to other known mitochondrial carrier family members including uncoupling proteins. It is proposed that plant DICs constitute the membrane component of several metabolic processes including the malate–oxaloacetate shuttle, the most important redox connection between the mitochondria and the cytosol.

scholarly journals Overexpression of PtrMYB121 Positively Regulates the Formation of Secondary Cell Wall in Arabidopsis thaliana

2020 ◽  
Vol 21 (20) ◽  
pp. 7734
Author(s):  
Ying Liu ◽  
Jiayin Man ◽  
Yinghao Wang ◽  
Chao Yuan ◽  
Yuyu Shi ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Cell Wall ◽  
Transcription Factors ◽  
Secondary Cell Wall ◽  
Tissue Expression ◽  
Wild Type ◽  
Tissue Expression Analysis ◽  
Wide Range ◽  
Transgenic Lines ◽  
Key Enzyme

MYB transcription factors have a wide range of functions in plant growth, hormone signaling, salt, and drought tolerances. In this study, two homologous transcription factors, PtrMYB55 and PtrMYB121, were isolated and their functions were elucidated. Tissue expression analysis revealed that PtrMYB55 and PtrMYB121 had a similar expression pattern, which had the highest expression in stems. Their expression continuously increased with the growth of poplar, and the expression of PtrMYB121 was significantly upregulated in the process. The full length of PtrMYB121 was 1395 bp, and encoded protein contained 464 amino acids including conserved R2 and R3 MYB domains. We overexpressed PtrMYB121 in Arabidopsis thaliana, and the transgenic lines had the wider xylem as compared with wild-type Arabidopsis. The contents of cellulose and lignin were obviously higher than those in wild-type materials, but there was no significant change in hemicellulose. Quantitative real-time PCR demonstrated that the key enzyme genes regulating the synthesis of lignin and cellulose were significantly upregulated in the transgenic lines. Furthermore, the effector-reporter experiment confirmed that PtrMYB121 bound directly to the promoters of genes relating to the synthesis of lignin and cellulose. These results suggest that PtrMYB121 may positively regulate the formation of secondary cell wall by promoting the synthesis of lignin and cellulose.

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