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Insight in the transcriptome data of hairy root disease-causing bacterium-Agrobacterium rhizogenes.

ABSTRACT: Agrobacterium rhizogenes induce the production of the hairy root through the transformation of plant genomes. In this article, we executed the transcriptome of A. rhizogenes through RNA-sequencing. RNA-sequencing of A. rhizogenes generated a total of 2.6 Gb raw data with a 75?bp paired-end sequence. The raw data has been submitted to the SRA database of NCBI with accession number SRR5641651. Reads were generated 2946 unigenes and all unigenes were annotated in the database. The length of transcripts ranged from 90 to 6369?bp, with a median transcript length of 968. The transcripts were annotated through the number of databases to obtain information about SSRs, SNPs, Gene Ontology, Transcription factors, and pathways analysis .

SUBMITTER: Yadav A

PROVIDER: S-EPMC7334392 | biostudies-literature | 2020 Aug

REPOSITORIES: biostudies-literature

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Insight in the transcriptome data of hairy root disease-causing bacterium-Agrobacterium rhizogenes.

Yadav Akhilesh A Verma Hariom H Ansari Waquar Akhter WA Singh Asha Lata AL Singh Major M

Data in brief 20200621

Agrobacterium rhizogenes induce the production of the hairy root through the transformation of plant genomes. In this article, we executed the transcriptome of A. rhizogenes through RNA-sequencing. RNA-sequencing of A. rhizogenes generated a total of 2.6 Gb raw data with a 75 bp paired-end sequence. The raw data has been submitted to the SRA database of NCBI with accession number SRR5641651. Reads were generated 2946 unigenes and all unigenes were annotated in the database. ...[more]

PMID: 32642520

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Project description:BackgroundAgrobacterium-mediated genetic transformation is a widely used and efficient technique for gene functional research in crop breeding and plant biology. While in some plant species, including soybean, genetic transformation is still recalcitrant and time-consuming, hampering the high-throughput functional analysis of soybean genes. Thus we pursue to develop a rapid, simple, and highly efficient hairy root system induced by Agrobacterium rhizogenes (A. rhizogenes) to analyze soybean gene function.ResultsIn this report, a rapid, simple, and highly efficient hairy root transformation system for soybean was described. Only sixteen days were required for the whole workflow and the system was suitable for various soybean genotypes, with an average transformation frequency of 58-64%. Higher transformation frequency was observed when wounded cotyledons from 1-day-germination seeds were inoculated and co-cultivated with A. rhizogenes in 1/2 B5 (Gamborg' B-5) medium. The addition of herbicide selection to root production medium increased the transformation frequency to 69%. To test the applicability of the hairy root system for gene functional analysis, we evaluated the protein expression and subcellular localization in transformed hairy roots. Transgenic hairy roots exhibited significantly increased GFP fluorescence and appropriate protein subcellular localization. Protein-protein interactions by BiFC (Bimolecular Fluorescent Complimentary) were also explored using the hairy root system. Fluorescence observations showed that protein interactions could be observed in the root cells. Additionally, hairy root transformation allowed soybean target sgRNA screening for CRISPR/Cas9 gene editing. Therefore, the protocol here enables high-throughput functional characterization of candidate genes in soybean.ConclusionA rapid, simple, and highly efficient A. rhizogenes-mediated hairy root transformation system was established for soybean gene functional analysis, including protein expression, subcellular localization, protein-protein interactions and gene editing system evaluation.

Dataset Information

Insight in the transcriptome data of hairy root disease-causing bacterium-Agrobacterium rhizogenes.

Publications

Insight in the transcriptome data of hairy root disease-causing bacterium-<i>Agrobacterium rhizogenes</i>.

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