Faculty Opinions recommendation of Loss-of-function mutations in the APX1 gene result in enhanced selenium tolerance in Arabidopsis thaliana.

2016 ◽  
Author(s):  
Wagner Araujo
Keyword(s):  
Arabidopsis Thaliana ◽  
Loss Of Function

scholarly journals The efficacy of CRISPR-mediated cytosine base editing with the RPS5a promoter in Arabidopsis thaliana

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Minkyung Choi ◽  
Jae-Young Yun ◽  
Jun-Hyuk Kim ◽  
Jin-Soo Kim ◽  
Sang-Tae Kim
Keyword(s):  
Arabidopsis Thaliana ◽  
Cytidine Deaminase ◽  
Biological Research ◽  
Loss Of Function ◽  
Nucleotide Substitutions ◽  
Viral Origin ◽  
Nonsense Mutations ◽  
Base Editing ◽  
Low Efficiency ◽  
Viable System

AbstractCRISPR/Cas9-mediated genome editing is an important and versatile technology in modern biological research. Recent advancements include base-editing CRISPR tools that enable targeted nucleotide substitutions using a fusion protein comprising a nickase variant of Cas9 and a base deaminase. Improvements in base editing efficiencies and inheritable of edited loci need to be made to make CRISPR a viable system in plants. Here, we report efficiency of cytosine base editors (CBEs) in Arabidopsis thaliana by applying the strong endogenous RPS5a promoter to drive the expression of nickase Cas9 and either rAPOBEC1 from rat (BE3) or the PmCDA1 activation-induced cytidine deaminase from sea lamprey (AIDv2). Compared with the strong heterologous CaMV35S promoter of viral origin, the RPS5a promoter improved CBE efficiency by 32% points with the number of T1 plants showing over 50% conversion ratio when the LFY gene was targeted. CBE induced nonsense mutations in LFY via C-to-T conversion, which resulted in loss-of-function lfy phenotypes; defects in LFY function were associated with the targeted base substitutions. Our data suggest that optimal promoter choice for CBE expression may affect base-editing efficiencies in plants. The results provide a strategy to optimize low-efficiency base editors and demonstrate their applicability for functional assays and trait development in crop research.

scholarly journals Control of auxin-regulated root development by the Arabidopsis thaliana SHY2/IAA3 gene

Development ◽  
1999 ◽  
Vol 126 (4) ◽  
pp. 711-721 ◽  
Author(s):  
Q. Tian ◽  
J.W. Reed
Keyword(s):  
Arabidopsis Thaliana ◽  
Lateral Root ◽  
Tissue Specificity ◽  
Plant Hormone ◽  
Root Formation ◽  
Auxin Transport ◽  
Feedback Regulation ◽  
Loss Of Function ◽  
Leaf Formation ◽  
Induced Genes

The plant hormone auxin controls many aspects of development and acts in part by inducing expression of various genes. Arabidopsis thaliana semidominant shy2 (short hypocotyl) mutations cause leaf formation in dark-grown plants, suggesting that SHY2 has an important role in regulating development. Here we show that the SHY2 gene encodes IAA3, a previously known member of the Aux/IAA family of auxin-induced genes. Dominant shy2 mutations cause amino acid changes in domain II, conserved among all members of this family. We isolated loss-of-function shy2 alleles including a putative null mutation. Gain-of-function and loss-of-function shy2 mutations affect auxin-dependent root growth, lateral root formation, and timing of gravitropism, indicating that SHY2/IAA3 regulates multiple auxin responses in roots. The phenotypes suggest that SHY2/IAA3 may activate some auxin responses and repress others. Models invoking tissue-specificity, feedback regulation, or control of auxin transport may explain these results.

scholarly journals Proxies of CRISPR/Cas9 Activity To Aid in the Identification of Mutagenized Arabidopsis Plants

2020 ◽  
Vol 10 (6) ◽  
pp. 2033-2042 ◽  
Author(s):  
Renyu Li ◽  
Charles Vavrik ◽  
Cristian H. Danna
Keyword(s):  
Arabidopsis Thaliana ◽  
Gene Function ◽  
Good Predictor ◽  
Gene Editing ◽  
Selection Strategy ◽  
Directed Mutagenesis ◽  
Loss Of Function ◽  
Editing Event ◽  
Endogenous Gene ◽  
Non Coding Rnas

CRISPR/Cas9 has become the preferred gene-editing technology to obtain loss-of-function mutants in plants, and hence a valuable tool to study gene function. This is mainly due to the easy reprogramming of Cas9 specificity using customizable small non-coding RNAs, and to the possibility of editing several independent genes simultaneously. Despite these advances, the identification of CRISPR-edited plants remains time and resource-intensive. Here, based on the premise that one editing event in one locus is a good predictor of editing event/s in other locus/loci, we developed a CRISPR co-editing selection strategy that greatly facilitates the identification of CRISPR-mutagenized Arabidopsis thaliana plants. This strategy is based on targeting the gene/s of interest simultaneously with a proxy of CRISPR-Cas9-directed mutagenesis. The proxy is an endogenous gene whose loss-of-function produces an easy-to-detect visible phenotype that is unrelated to the expected phenotype of the gene/s under study. We tested this strategy via assessing the frequency of co-editing of three functionally unrelated proxy genes. We found that each proxy predicted the occurrence of mutations in each surrogate gene with efficiencies ranging from 68 to 100%. The selection strategy laid out here provides a framework to facilitate the identification of multiplex edited plants, thus aiding in the study of gene function when functional redundancy hinders the effort to define gene-function-phenotype links.

scholarly journals Redox-Responsive Transcription Factor 1 (RRFT1) Is Involved in Extracellular ATP-Regulated Arabidopsis thaliana Seedling Growth

2020 ◽  
Vol 61 (4) ◽  
pp. 685-698 ◽  
Author(s):  
Ruojia Zhu ◽  
Xiaoxia Dong ◽  
Yingying Xue ◽  
Jiawei Xu ◽  
Aiqi Zhang ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Transcription Factor ◽  
Adenosine Triphosphate ◽  
Seedling Growth ◽  
Growth And Development ◽  
High Throughput Sequencing ◽  
Root Tip ◽  
Loss Of Function ◽  
Redox Responsive ◽  
Transcription Factor 1

Abstract Extracellular adenosine triphosphate (eATP) is an apoplastic signaling molecule that plays an essential role in the growth and development of plants. Arabidopsis seedlings have been reported to respond to eATP; however, the downstream signaling components are still not well understood. In this study, we report that an ethylene-responsive factor, Redox-Responsive Transcription Factor 1 (RRTF1), is involved in eATP-regulated Arabidopsis thaliana seedling growth. Exogenous adenosine triphosphate inhibited green seedling root growth and induced hypocotyl bending of etiolated seedlings. RRTF1 loss-of-function mutant (rrtf1) seedlings showed decreased responses to eATP, while its complementation or overexpression led to recovered or increased eATP responsiveness. RRTF1 was expressed rapidly after eATP stimulation and then migrated into the nuclei of root tip cells. eATP-induced auxin accumulation in root tip or hypocotyl cells was impaired in rrtf1. Chromatin immunoprecipitation and high-throughput sequencing results indicated that eATP induced some genes related to cell growth and development in wild type but not in rrtf1 cells. These results suggest that RRTF1 may be involved in eATP signaling by regulating functional gene expression and cell metabolism in Arabidopsis seedlings.

The COP9 signalosome influences the epigenetic landscape of Arabidopsis thaliana

2018 ◽  
Vol 35 (16) ◽  
pp. 2718-2723 ◽  
Author(s):  
Tamir Tuller ◽  
Alon Diament ◽  
Avital Yahalom ◽  
Assaf Zemach ◽  
Shimshi Atar ◽  
...  
Keyword(s):  
Gene Expression ◽  
Dna Methylation ◽  
Arabidopsis Thaliana ◽  
Gene Expression Regulation ◽  
Developmental Pathways ◽  
Cop9 Signalosome ◽  
Supplementary Information ◽  
Loss Of Function ◽  
Genome Wide ◽  
High Level

Abstract Motivation The COP9 signalosome is a highly conserved multi-protein complex consisting of eight subunits, which influences key developmental pathways through its regulation of protein stability and transcription. In Arabidopsis thaliana, mutations in the COP9 signalosome exhibit a number of diverse pleiotropic phenotypes. Total or partial loss of COP9 signalosome function in Arabidopsis leads to misregulation of a number of genes involved in DNA methylation, suggesting that part of the pleiotropic phenotype is due to global effects on DNA methylation. Results We determined and analyzed the methylomes and transcriptomes of both partial- and total-loss-of-function Arabidopsis mutants of the COP9 signalosome. Our results support the hypothesis that the COP9 signalosome has a global genome-wide effect on methylation and that this effect is at least partially encoded in the DNA. Our analyses suggest that COP9 signalosome-dependent methylation is related to gene expression regulation in various ways. Differentially methylated regions tend to be closer in the 3D conformation of the genome to differentially expressed genes. These results suggest that the COP9 signalosome has a more comprehensive effect on gene expression than thought before, and this is partially related to regulation of methylation. The high level of COP9 signalosome conservation among eukaryotes may also suggest that COP9 signalosome regulates methylation not only in plants but also in other eukaryotes, including humans. Supplementary information Supplementary data are available at Bioinformatics online.

Synthesis of complementary RNA by RNA-dependent RNA polymerases in plant extracts is independent of an RNA primer

2008 ◽  
Vol 35 (11) ◽  
pp. 1091 ◽  
Author(s):  
Lei Wang ◽  
Neil A. Smith ◽  
Lan Zhang ◽  
Elizabeth S. Dennis ◽  
Peter M. Waterhouse ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Rna Polymerase ◽  
Plant Extract ◽  
Plant Extracts ◽  
Callus Tissue ◽  
Polymerase Activity ◽  
Loss Of Function ◽  
Rna Dependent Rna Polymerase ◽  
Methyl Group ◽  
The Individual

RNA-dependent RNA polymerase (RDR) activities were readily detected in extracts from cauliflower and broccoli florets, Arabidopsis thaliana (L.) Heynh callus tissue and broccoli nuclei. The synthesis of complementary RNA (cRNA) was independent of a RNA primer, whether or not the primer contained a 3′ terminal 2′-O-methyl group or was phosphorylated at the 5′ terminus. cRNA synthesis in plant extracts was not affected by loss-of-function mutations in the DICER-LIKE (DCL) proteins DCL2, DCL3, and DCL4, indicating that RDRs function independently of these DCL proteins. A loss-of-function mutation in RDR1, RDR2 or RDR6 did not significantly reduce the amount of cRNA synthesis. This indicates that these RDRs did not account for the bulk RDR activities in plant extracts, and suggest that either the individual RDRs each contribute a fraction of polymerase activity or another RDR(s) is predominant in the plant extract.

scholarly journals A loss-of-function mutation in Calmodulin2 gene affects pollen germination in Arabidopsis thaliana

2010 ◽  
Vol 74 (3) ◽  
pp. 235-247 ◽  
Author(s):  
Michela Landoni ◽  
Alessandra De Francesco ◽  
Massimo Galbiati ◽  
Chiara Tonelli
Keyword(s):  
Arabidopsis Thaliana ◽  
Pollen Germination ◽  
Loss Of Function

scholarly journals Heat-induced transposition leads to rapid drought resistance of Arabidopsis thaliana

2021 ◽  
Author(s):  
Michael Thieme ◽  
Arthur Brêchet ◽  
Yann Bourgeois ◽  
Bettina Keller ◽  
Etienne Bucher ◽  
...  
Keyword(s):  
Arabidopsis Thaliana ◽  
Drought Resistance ◽  
Phenotypic Diversity ◽  
Plant Evolution ◽  
Loss Of Function ◽  
Copy Numbers ◽  
Drastic Increase ◽  
In The Wild ◽  
Rate Of Photosynthesis

Plant genomes comprise a vast diversity of transposable elements (TEs) (Tenaillon et al. 2010)⁠. While their uncontrolled proliferation can have fatal consequences for their host, there is strong evidence for their importance in fueling genetic diversity and plant evolution (Baduel et al. 2021)⁠. However, the number of studies addressing the role of TEs in this process is limited. Here we show that the heat-induced burst of a low-copy TE increases phenotypic diversity and leads to the rapid emergence of more drought-resistant individuals of Arabidopsis thaliana. We exposed TE-high-copy-(hc)lines (Thieme et al. 2017)⁠ with up to ~8 fold increased copy numbers of the heat-responsive ONSEN-TE (AtCOPIA78) (Ito et al. 2011; Cavrak et al. 2014; Tittel-Elmer et al. 2010)⁠ in the wild type background to desiccation as a straightforward and highly relevant selection pressure. We found evidence for a drastic increase of drought resistance in five out of the 23 tested hc-lines and further pinpoint one of the causative mutations to an exonic ONSEN-insertion in the ribose-5-phosphate-isomerase 2 gene. This loss-of-function mutation resulted in a decreased rate of photosynthesis and water consumption. This is one of the rare examples (Esnault et al. 2019)⁠ experimentally demonstrating the adaptive potential of mobilized stress-responsive TEs in eukaryotes. Our results further shed light on the complex relationship between mobile elements and their hosts and substantiate the importance of TE-mediated loss-of-function mutations in stress adaptation, particularly with respect to global warming.

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