Agrobacterium VirE2 Protein Modulates Plant Gene Expression and Mediates Transformation from Its Location in the Cytoplasm
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ABSTRACT:
PROVIDER: PRJNA722928 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR14272429.fastq.gz | Fastqsanger.gz | |||
| SRR14272430.fastq.gz | Fastqsanger.gz | |||
| SRR14272431.fastq.gz | Fastqsanger.gz | |||
| SRR14272432.fastq.gz | Fastqsanger.gz | |||
| SRR14272433.fastq.gz | Fastqsanger.gz |
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Project description:Agrobacterium effector protein VirE2 is important for plant transformation. VirE2 likely coats transferred DNA (T-DNA) in the plant cell and protects it from degradation. VirE2 localizes to the plant cytoplasm and interacts with several host proteins. Plant-expressed VirE2 can complement a virE2 mutant Agrobacterium strain to support transformation. We investigated whether VirE2 could facilitate transformation from a nuclear location by affixing to it a strong nuclear localization signal (NLS) sequence. Only cytoplasmic-, but not nuclear-localized, VirE2 could stimulate transformation. To investigate the ways VirE2 supports transformation, we generated transgenic Arabidopsis plants containing a virE2 gene under the control of an inducible promoter and performed RNA-seq and proteomic analyses before and after induction. Some differentially expressed plant genes were previously known to facilitate transformation. Knockout mutant lines of some other VirE2 differentially expressed genes showed altered transformation phenotypes. Levels of some proteins known to be important for transformation increased in response to VirE2 induction, but prior to or without induction of their corresponding mRNAs. Overexpression of some other genes whose proteins increased after VirE2 induction resulted in increased transformation susceptibility. We conclude that cytoplasmically-localized VirE2 modulates both plant RNA and protein levels to facilitate transformation.
2021-06-02 | GSE172314 | GEO
Project description:Agrobacterium tumefaciens is a special plant pathogen causing crown gall disease. This pathogen is well known for the technology Agrobacterium-mediated transformation. As a pathogen, Agrobacterium triggers plant immunity, and this affects transformation. But the signaling components and pathways in plant immunity to Agrobacterium remain elusive. We demonstrate two Arabidopsis MAPKKs MKK4/MKK5 and their downstream MAPKs MPK3/MPK6 play a major role in both Agrobacterium-triggered immunity and Agrobacterium-mediated transformation. Agrobacteria induce MPK3/MPK6 activity and plant defense responsive genes expression in a very early stage. This process is dependent on MKK4/MKK5 function. Loss of function of MKK4 and MKK5 or their downstream MPK3 and MPK6 abolishes plant immunity to agrobacteria, and increases the transformation frequency, while activation of MKK4 and MKK5 enhances the plant immunity and represses the transformation. Global transcriptome indicates agrobacteria induce various plant defense pathways, including ROS production, ethylene and SA-mediated defense responses, and MKK4/MKK5 is essential for these pathways induction. Activation of MKK4 and MKK5 promotes ROS production and cell death in agrobacteria infection process. Ethylene and SA act bypass of MKK4/MKK5 signaling to regulate transformation. Based on these results, we propose MKK4/5-MPK3/6 cascade is an essential signaling pathway to regulate Agrobacterium-mediated transformation by modulating Agrobacterium-triggered plant immunity.
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Project description:VIP2 is a transcription regulator and the C-terminal NOT2 domain of VIP2 interacts with VirE2. However, AtVIP2 overexpressor lines in Arabidopsis did not show an improvement in Agrobacterium-mediated stable root transformation. To investigate the role of VIP2 in Agrobacterium-plant interaction, we carried out transcriptome profiling by comparing a homozygous Arabidopsis AtVIP2 overexpressor line with wild-type Col-0 plants following Agrobacterium infection.
2019-09-26 | E-MTAB-8326 | biostudies-arrayexpress
Project description:Agrobacterium effector protein VirE2 is important for plant transformation. VirE2 likely coats transferred DNA (T-DNA) in the plant cell and protects it from degradation. VirE2 localizes to the plant cytoplasm and interacts with several host proteins. Plant-expressed VirE2 can complement a virE2 mutant Agrobacterium strain to support transformation. We investigated whether VirE2 could facilitate transformation from a nuclear location by affixing to it a strong nuclear localization signal (NLS) sequence. Only cytoplasmic-, but not nuclear-localized, VirE2 could stimulate transformation. To investigate the ways VirE2 supports transformation, we generated transgenic Arabidopsis plants containing a virE2 gene under the control of an inducible promoter and performed RNA-seq and proteomic analyses before and after induction. Some differentially expressed plant genes were previously known to facilitate transformation. Knockout mutant lines of some other VirE2 differentially expressed genes showed altered transformation phenotypes. Levels of some proteins known to be important for transformation increased in response to VirE2 induction, but prior to or without induction of their corresponding mRNAs. Overexpression of some other genes whose proteins increased after VirE2 induction resulted in increased transformation susceptibility. We conclude that cytoplasmically localized VirE2 modulates both plant RNA and protein levels to facilitate transformation.
| S-EPMC8213393 | biostudies-literature
Project description:Agrobacterium-mediated cassava transformation via friable embryogenic callus (FEC) has allowed the robust production of transgenic cassava. So far, the transformation has been performed mostly for 60444 (the model cassava variety for transformation) and African varieties due to the high ability of callus induction and good regeneration capacity from embryogenic tissues. It is important to develop the transformation methods for elite cassava varieties in Asian area which is one of main cassava production area in the world, however, the suitable transformation method for Asian elite variety via FEC has not been reported. Here, we developed the transformation method in Kasetsart 50 (KU50) which has the highest planting area in Thailand and Vietnam. In cassava transformation, the preparation of FEC is considered as a key step. The ability of FEC induction from KU50 was improved efficiently by the use of media with reduced nutrients and excess vitamin B1, even if the FEC-inducible efficiency was 5 times lower compared with cv. 60444. The optimal concentration of NAA for regeneration from FEC to cotyledon was 1.0 mg/L and the optimal concentration of BAP for shoot formation from cotyledon was 0.4 mg/L. The transformation efficiency was estimated as 45% for 60444 and 22% for KU50. Transcriptome analysis indicated that KU50 FEC is out of balance between cell wall production and assembly, because the expression of the genes related to loosing cell wall was upregulated in the FEC from KU50 compared with 60444. We hope that the developed technology will contribute to molecular breeding of useful cassava plants in Asia by the simultaneous use of genome-editing technology.
2022-03-26 | GSE169685 | GEO
Project description:Agrobacterium tumefaciens-mediated genetic transformation has been routinely used in rice for more than a decade. However, the transformation efficiency of the indica rice variety is still unsatisfactory and much lower than that of japonica cultivars. Further improvement on the transformation efficiency lies in the genetic manipulation of the plant itself, which requires a better understanding of the underlying process accounting for the susceptibility of plant cells to Agrobacterium infection as well as the identification of plant genes involved in the transformation process. In order to investigate the related genes affecting the transformation efficiency of embryogenic calli of different rice cultivars, we used Affymetrix GeneChip® Rice Genome Array to measure the global gene expression profiling just before transformation and at four different time points after transformation (1 h, 6 h, 12 h, 24 h) in both japonica rice cultivar Nipponbare and indica rice cultivar Zhenshan 97.
2012-01-01 | GSE32426 | GEO
Project description:As sessile organisms, plants require dynamic pathways in order to recognize pathogens and coordinate plant defenses by signalling. Agrobacterium tumefaciens C58 is able to avoid triggering plant defenses prior to entering the cell, and therefore is only detected once infection has begun making Agrobacterium a plant pathogen to numerous plant species. Understanding plant responses to Agrobacterium will be useful in improving plant defenses and potentially may also improve plant transformation efficiency. Microarrays were utilized for detailing the global gene expression pattern in A. thaliana Col-0 roots in response to A. tumefaciens C58 for the identification of differentially expressed genes.
2014-10-28 | GSE62750 | GEO
Project description:As sessile organisms, plants require dynamic pathways in order to recognize pathogens and coordinate plant defenses by signalling. Agrobacterium tumefaciens C58 is able to avoid triggering plant defenses prior to entering the cell, and therefore is only detected once infection has begun making Agrobacterium a plant pathogen to numerous plant species. Understanding plant responses to Agrobacterium will be useful in improving plant defenses and potentially may also improve plant transformation efficiency. Microarrays were utilized for detailing the global gene expression pattern in A. thaliana Col-0 leafs in response to A. tumefaciens C58 for the identification of differentially expressed genes.
2014-10-28 | GSE62749 | GEO
Project description:Plants transformed via agroinfiltration using a modified strain of Agrobacterium tumefaciens can produce antibody proteins for purification and application in antibody-based therapeutics. The rising demand for such drugs underscores the expansion of molecular farming and warrants the need to produce transformed plants at an industrial scale, requiring large quantities of A. tumefaciens culture, which is not feasible using traditional growth methods (e.g., shake flask). To overcome this limitation, we investigate the use of a bioreactor growth chamber system to meet production demands through growth optimization and efficient plant transformation. We observe differences in target antibody production associated with different growth conditions and aim to characterize the unique responses through profiling of protein-level changes specific to each condition. Our quantitative proteomic profiling defines growth-specific alterations in protein abundance and highlights the functional roles of these proteins. Moreover, we observe changes in the bacterial proteomes, which may influence bacterial processes important for effective agroinfiltration and transformation. Overall, our study provides insight into fundamental biological processes of A. tumefaciens influenced by growth conditions, which may influence plant transformation efficiency and outcome of antibody-based therapeutic production.
2020-12-10 | PXD018384 | Pride
Project description:Agrobacterium tumefaciens-mediated genetic transformation has been routinely used in rice for more than a decade. However, the transformation efficiency of the indica rice variety is still unsatisfactory and much lower than that of japonica cultivars. Further improvement on the transformation efficiency lies in the genetic manipulation of the plant itself, which requires a better understanding of the underlying process accounting for the susceptibility of plant cells to Agrobacterium infection as well as the identification of plant genes involved in the transformation process. In order to investigate the related genes affecting the transformation efficiency of embryogenic calli of different rice cultivars, we used Affymetrix GeneChipM-BM-. Rice Genome Array to measure the global gene expression profiling just before transformation and at four different time points after transformation (1 h, 6 h, 12 h, 24 h) in both japonica rice cultivar Nipponbare and indica rice cultivar Zhenshan 97. The mature embryo-derived embryogenic calli of Nipponbare (Nip) and Zhenshan 97 (ZS) were infected by Agrobacterium. Calli of Nip and ZS were sampled just before infection (0 h) and 1h, 6h, 12 h and 24h after infection, respectively. Three independent biological replications for each time point of the two varieties were used. To avoid the influence of polymorphisms between the probe sequence on the array and the genomes of the varieties used, we used a genomic DNA (gDNA)-based probe-selection strategy based on the hybridization efficiency of gDNA from Nip and ZS with the PM oligonucleotide probes on the rice array. The genomic DNA of Nip and ZS were extracted and hybridized to the Affymetrix Rice Genome Arrays. Three biological replications per cultivar were performed.
2012-01-01 | E-GEOD-32426 | biostudies-arrayexpress