Stable genetic transformation of Arabidopsis thaliana by Agrobacterium inoculation in planta

1994 ◽  
Vol 5 (4) ◽  
pp. 551-558 ◽  
Author(s):  
Seok So Chang ◽  
Soon Ki Park ◽  
Byung Chul Kim ◽  
Bong Joong Kang ◽  
Dal Ung Kim ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Genetic Transformation ◽  
In Planta

scholarly journals Ca2+-Dependent Protein Kinase 6 Enhances KAT2 Shaker Channel Activity in Arabidopsis thaliana

2021 ◽  
Vol 22 (4) ◽  
pp. 1596
Author(s):  
Elsa Ronzier ◽  
Claire Corratgé-Faillie ◽  
Frédéric Sanchez ◽  
Christian Brière ◽  
Tou Cheu Xiong
Keyword(s):  
Arabidopsis Thaliana ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Physical Interaction ◽  
Fluorescence Lifetime Imaging ◽  
Dependent Manner ◽  
In Planta ◽  
Knock Out ◽  
Calcium Dependent ◽  
Dependent Protein

Post-translational regulations of Shaker-like voltage-gated K+ channels were reported to be essential for rapid responses to environmental stresses in plants. In particular, it has been shown that calcium-dependent protein kinases (CPKs) regulate Shaker channels in plants. Here, the focus was on KAT2, a Shaker channel cloned in the model plant Arabidopsis thaliana, where is it expressed namely in the vascular tissues of leaves. After co-expression of KAT2 with AtCPK6 in Xenopuslaevis oocytes, voltage-clamp recordings demonstrated that AtCPK6 stimulates the activity of KAT2 in a calcium-dependent manner. A physical interaction between these two proteins has also been shown by Förster resonance energy transfer by fluorescence lifetime imaging (FRET-FLIM). Peptide array assays support that AtCPK6 phosphorylates KAT2 at several positions, also in a calcium-dependent manner. Finally, K+ fluorescence imaging in planta suggests that K+ distribution is impaired in kat2 knock-out mutant leaves. We propose that the AtCPK6/KAT2 couple plays a role in the homeostasis of K+ distribution in leaves.

scholarly journals Evidence that the Arabidopsis thaliana 3-hydroxy-3-methylglutaryl-CoA reductase 1 is phosphorylated at Ser577 in planta

2018 ◽  
Vol 35 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Jekson Robertlee ◽  
Keiko Kobayashi ◽  
Jianwei Tang ◽  
Masashi Suzuki ◽  
Toshiya Muranaka
Keyword(s):  
Arabidopsis Thaliana ◽  
In Planta ◽  
Coa Reductase

scholarly journals Characterization of the early response of Arabidopsis thaliana to Dickeya dadantii infection using expression profiling

2018 ◽  
Author(s):  
Frédérique Van Gijsegem ◽  
Frédérique Bitton ◽  
Anne-Laure Laborie ◽  
Yvan Kraepiel ◽  
Jacques Pédron
Keyword(s):  
Arabidopsis Thaliana ◽  
Early Stage ◽  
Plant Defence ◽  
Plant Responses ◽  
Type I ◽  
In Planta ◽  
Secretion Systems ◽  
Dickeya Dadantii ◽  
A Genome

AbstractTo draw a global view of plant responses to interactions with the phytopathogenic enterobacterale Dickeya dadantii, a causal agent of soft rot diseases on many plant species, we analysed the early Arabidopsis responses to D. dadantii infection. We performed a genome-wide analysis of the Arabidopsis thaliana transcriptome during D. dadantii infection and conducted a genetic study of identified responses.A limited set of genes related to plant defence or interactions with the environment were induced at an early stage of infection, with an over-representation of genes involved in both the metabolism of indole glucosinolates (IGs) and the jasmonate (JA) defence pathway. Bacterial type I and type II secretion systems are required to trigger the induction of IG and JA-related genes while the type III secretion system appears to partially inhibit these defence pathways. Using Arabidopsis mutants impaired in JA biosynthesis or perception, we showed that induction of some IG metabolism genes was COI1-dependent but, surprisingly, JA-independent. Moreover, characterisation of D. dadantii disease progression in Arabidopsis mutants impaired in JA or IG pathways showed that JA triggers an efficient plant defence response that does not involve IGs.The induction of the IG pathway by bacterial pathogens has been reported several times in vitro. This study shows for the first time, that this induction does indeed occur in planta, but also that this line of defence is ineffective against D. dadantii infection, in contrast to its role to counteract herbivorous or fungal pathogen attacks.

scholarly journals In Planta Genetic Transformation to Produce CRISPRed High-Oleic Peanut

2021 ◽  
Author(s):  
Han Hong Wei ◽  
Shu Tao Yu ◽  
Zhi Wei Wang ◽  
Zhen Yang ◽  
Guo Sheng Song ◽  
...  
Keyword(s):  
Oleic Acid ◽  
Genetic Transformation ◽  
Genome Editing ◽  
Loss Of Function ◽  
In Planta ◽  
Easy Method ◽  
High Oleic ◽  
Keeping Quality ◽  
Varietal Improvement ◽  
Multiple Health

Abstract In contrast to its normal-oleic counterpart, high-oleic peanut has better keeping quality and multiple health benefits. Breeding high-oleic peanut through conventional means is a tedious process generally requiring several years. Genome editing may shorten the duration. In this study, node injection method was used to transform normal-oleic Huayu 23, a popular peanut cultivar having loss-of-function FAD2A, with CRISPR/Cas9 construct targeting FAD2B, and two peanut mutants with over 80% oleic acid and 442A insertion in FAD2B were obtained. As a genotype-independent, simple and easy method for peanut genetic transformation, node injection has great potential in factional analysis of genes and in peanut varietal improvement.

Identification of Two Arabidopsis thaliana Plasma Membrane Transporters Able to Transport Glucose 1-Phosphate

2019 ◽  
Vol 61 (2) ◽  
pp. 381-392
Author(s):  
Irina Malinova ◽  
Stella Kössler ◽  
Tom Orawetz ◽  
Ulrike Matthes ◽  
Slawomir Orzechowski ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Plasma Membrane ◽  
Carbohydrate Metabolism ◽  
Transport Processes ◽  
Membrane Transporters ◽  
In Planta ◽  
Sugar Derivative ◽  
Sugar Metabolites

Abstract Primary carbohydrate metabolism in plants includes several sugar and sugar-derivative transport processes. Over recent years, evidences have shown that in starch-related transport processes, in addition to glucose 6-phosphate, maltose, glucose and triose-phosphates, glucose 1-phosphate also plays a role and thereby increases the possible fluxes of sugar metabolites in planta. In this study, we report the characterization of two highly similar transporters, At1g34020 and At4g09810, in Arabidopsis thaliana, which allow the import of glucose 1-phosphate through the plasma membrane. Both transporters were expressed in yeast and were biochemically analyzed to reveal an antiport of glucose 1-phosphate/phosphate. Furthermore, we showed that the apoplast of Arabidopsis leaves contained glucose 1-phosphate and that the corresponding mutant of these transporters had higher glucose 1-phosphate amounts in the apoplast and alterations in starch and starch-related metabolism.

scholarly journals Identification of client iron–sulfur proteins of the chloroplastic NFU2 transfer protein in Arabidopsis thaliana

2020 ◽  
Vol 71 (14) ◽  
pp. 4171-4187 ◽  
Author(s):  
Nathalie Berger ◽  
Florence Vignols ◽  
Jonathan Przybyla-Toscano ◽  
Mélanie Roland ◽  
Valérie Rofidal ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
De Novo ◽  
Bimolecular Fluorescence Complementation ◽  
In Planta ◽  
Chlorophyll Metabolism ◽  
Transfer Protein ◽  
Iron Sulfur ◽  
Bimolecular Fluorescence Complementation Assay ◽  
Client Proteins ◽  
S Proteins

Abstract Iron–sulfur (Fe-S) proteins have critical functions in plastids, notably participating in photosynthetic electron transfer, sulfur and nitrogen assimilation, chlorophyll metabolism, and vitamin or amino acid biosynthesis. Their maturation relies on the so-called SUF (sulfur mobilization) assembly machinery. Fe-S clusters are synthesized de novo on a scaffold protein complex and then delivered to client proteins via several transfer proteins. However, the maturation pathways of most client proteins and their specificities for transfer proteins are mostly unknown. In order to decipher the proteins interacting with the Fe-S cluster transfer protein NFU2, one of the three plastidial representatives found in Arabidopsis thaliana, we performed a quantitative proteomic analysis of shoots, roots, and seedlings of nfu2 plants, combined with NFU2 co-immunoprecipitation and binary yeast two-hybrid experiments. We identified 14 new targets, among which nine were validated in planta using a binary bimolecular fluorescence complementation assay. These analyses also revealed a possible role for NFU2 in the plant response to desiccation. Altogether, this study better delineates the maturation pathways of many chloroplast Fe-S proteins, considerably extending the number of NFU2 clients. It also helps to clarify the respective roles of the three NFU paralogs NFU1, NFU2, and NFU3.

scholarly journals Biochemical characteristics of AtFAR2, a fatty acid reductase from Arabidopsis thaliana that reduces fatty acyl-CoA and -ACP substrates into fatty alcohols

2016 ◽  
Vol 63 (3) ◽  
Author(s):  
Thuy T. P. Doan ◽  
Anders S. Carlsson ◽  
Sten Stymne ◽  
Per Hofvander
Keyword(s):  
Arabidopsis Thaliana ◽  
Fatty Acid ◽  
Fatty Acyl ◽  
Fatty Alcohols ◽  
Abnormal Development ◽  
In Planta ◽  
Alcohol Production ◽  
Genes Encoding ◽  
In Vitro Data

Fatty alcohols and derivatives are important for proper deposition of a functional pollen wall. Mutations in specific genes encoding fatty acid reductases (FAR) responsible for fatty alcohol production cause abnormal development of pollen. A disrupted AtFAR2 (MS2) gene in Arabidopsis thaliana results in pollen developing an abnormal exine layer and a reduced fertility phenotype. AtFAR2 has been shown to be targeted to chloroplasts and in a purified form to be specific for acyl-ACP substrates. Here, we present data on the in vitro and in planta characterizations of AtFAR2 from A. thaliana and show that this enzyme has the ability to use both, C16:0-ACP and C16:0-CoA, as substrates to produce C16:0-alcohol. Our results further show that AtFAR2 is highly similar in properties and substrate specificity to AtFAR6 for which in vitro data has been published, and which is also a chloroplast localized enzyme. This suggests that although AtFAR2 is the major enzyme responsible for exine layer functionality, AtFAR6 might provide functional redundancy to AtFAR2.

scholarly journals Distinct Peculiarities of In Planta Synthesis of Isoprenoid and Aromatic Cytokinins

Biomolecules ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 86
Author(s):  
Vladimir Oslovsky ◽  
Ekaterina Savelieva ◽  
Mikhail Drenichev ◽  
Georgy Romanov ◽  
Sergey Mikhailov
Keyword(s):  
Arabidopsis Thaliana ◽  
Plant Cells ◽  
Cytokinin Activity ◽  
Hormonal Activity ◽  
In Planta ◽  
Significant Difference ◽  
Aromatic Cytokinins ◽  
Derivatives Of ◽  
Ribose Moiety

The biosynthesis of aromatic cytokinins in planta, unlike isoprenoid cytokinins, is still unknown. To compare the final steps of biosynthesis pathways of aromatic and isoprenoid cytokinins, we synthesized a series of nucleoside derivatives of natural cytokinins starting from acyl-protected ribofuranosyl-, 2′-deoxyribofuranosyl- and 5′-deoxyribofuranosyladenine derivatives using stereoselective alkylation with further deblocking. Their cytokinin activity was determined in two bioassays based on model plants Arabidopsis thaliana and Amaranthus caudatus. Unlike active cytokinins-bases, cytokinin nucleosides lack the hormonal activity until the ribose moiety is removed. According to our experiments, ribo-, 2′-deoxyribo- and 5′-deoxyribo-derivatives of isoprenoid cytokinin N6-isopentenyladenine turned in planta into active cytokinins with clear hormonal activity. As for aromatic cytokinins, both 2′-deoxyribo- and 5′-deoxyribo-derivatives did not exhibit analogous activity in Arabidopsis. The 5′-deoxyribo-derivatives cannot be phosphorylated enzymatically in vivo; therefore, they cannot be “activated” by the direct LOG-mediated cleavage, largely occurring with cytokinin ribonucleotides in plant cells. The contrasting effects exerted by deoxyribonucleosides of isoprenoid (true hormonal activity) and aromatic (almost no activity) cytokinins indicates a significant difference in the biosynthesis of these compounds.

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