ArticlePlant cell reports2012
An efficient and high-throughput protocol for Agrobacterium-mediated transformation based on phosphomannose isomerase positive selection in Japonica rice (Oryza sativa L.).
Article in Plant cell reports, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
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25 citing papers in PubMed, 71 citations in OpenAlex.
- Agrobacterium-Mediated Transformation for the Development of Transgenic Crops; Present and Future Prospects.Molecular biotechnology · 2024Review
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- Glucomannan inGenes · 2022Review
- Plant Growth-Promoting Activity of Bacteria Isolated from Asian Rice (Oryza sativa L.) Depends on Rice Genotype.Microbiology spectrum · 2022Article
- Methylglyoxal-glyoxalase system as a possible selection module for raising marker-safe plants in rice.Physiology and molecular biology of plants : an international journal of functional plant biology · 2021Article
- The trihelix transcription factor OsGTγ-2 is involved adaption to salt stress in rice.Plant molecular biology · 2020Article
- Neo-functionalization of a Teosinte branched 1 homologue mediates adaptations of upland rice.Nature communications · 2020Article
- Efficient Biolistic Transformation of Immature Citrus Rootstocks Using Phosphomannose-isomerase Selection.Plants (Basel, Switzerland) · 2019Article
- Isolation of five rice nonendosperm tissue-expressed promoters and evaluation of their activities in transgenic rice.Plant biotechnology journal · 2018Article
- Article
- Characterization of Transcription Factor GeneFrontiers in plant science · 2018Article
- Highly phosphorylated functionalized rice starch produced by transgenic rice expressing the potato GWD1 gene.Scientific reports · 2017Article
- Generation of targeted mutant rice using a CRISPR-Cpf1 system.Plant biotechnology journal · 2017Article
- Plant phosphomannose isomerase as a selectable marker for rice transformation.Scientific reports · 2016Article
- Identification of a regulatory element responsible for salt induction of rice OsRAV2 through ex situ and in situ promoter analysis.Plant molecular biology · 2016Article
- Functional Studies of Heading Date-Related Gene TaPRR73, a Paralog of Ppd1 in Common Wheat.Frontiers in plant science · 2016Article
- High-throughput transformation pipeline for a Brazilian japonica rice with bar gene selection.Protoplasma · 2015Article
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Authors and funding
11 authors at 4 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
unlabelledA number of Agrobacterium-mediated rice transformation systems have been developed and widely used in numerous laboratories and research institutes. However, those systems generally employ antibiotics like kanamycin and hygromycin, or herbicide as selectable agents, and are used for the small-scale experiments. To address high-throughput production of transgenic rice plants via Agrobacterium-mediated transformation, and to eliminate public concern on antibiotic markers, we developed a comprehensive efficient protocol, covering from explant preparation to the acquisition of low copy events by real-time PCR analysis before transplant to field, for high-throughput production of transgenic plants of Japonica rice varieties Wanjing97 and Nipponbare using Escherichia coli phosphomannose isomerase gene (pmi) as a selectable marker. The transformation frequencies (TF) of Wanjing97 and Nipponbare were achieved as high as 54.8 and 47.5%, respectively, in one round of selection of 7.5 or 12.5 g/L mannose appended with 5 g/L sucrose. High-throughput transformation from inoculation to transplant of low copy events was accomplished within 55-60 days. Moreover, the Taqman assay data from a large number of transformants showed 45.2% in Wanjing97 and 31.5% in Nipponbare as a low copy rate, and the transformants are fertile and follow the Mendelian segregation ratio. This protocol facilitates us to perform genome-wide functional annotation of the open reading frames and utilization of the agronomically important genes in rice under a reduced public concern on selectable markers. KEY MESSAGE: We describe a comprehensive protocol for large scale production of transgenic Japonica rice plants using non-antibiotic selectable agent, at simplified, cost- and labor-saving manners.
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