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Related Concept Videos

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Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
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Transposons make up a significant part of genomes of various organisms. Therefore, it is believed that transposition played a major evolutionary role in speciation by changing genome sizes and modifying gene expression patterns. For example, in bacteria, transposition can lead to conferring antibiotic resistance. Movement of transposable elements within the genetic pool of pathogenic bacteria can aid in transfer of antibiotic-resistant genetic elements. In eukaryotes, transposons can carry out...
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Related Experiment Video

Updated: Nov 6, 2025

Agrobacterium-Mediated Genetic Transformation, Transgenic Production, and Its Application for the Study of Male Reproductive Development in Rice
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Generation of Marker-Free Transgenic Rice Resistant to Rice Blast Disease Using Ac/Ds Transposon-Mediated Transgene

Xin Li1, Longyu Pan1,2, Dongling Bi1

  • 1State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou, China.

Frontiers in Plant Science
|May 7, 2021
PubMed
Summary

This study developed marker-free transgenic rice resistant to rice blast disease using maize Activator/Dissociation transposon technology. The resulting rice lines showed enhanced disease resistance and reduced Pi21 gene expression.

Keywords:
Ac/Ds transposable elementdisease resistance genemarker-free transgenic plantricerice blast (Magnaporthe oryzae)selection marker

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Generating Transgenic Plants with Single-copy Insertions Using BIBAC-GW Binary Vector
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Area of Science:

  • Plant Biotechnology
  • Genetics
  • Agricultural Science

Background:

  • Rice blast poses a significant threat to global rice production.
  • Developing disease-resistant rice varieties is crucial for food security.
  • Transgenic approaches offer a faster alternative to traditional breeding for disease resistance.

Purpose of the Study:

  • To generate marker-free transgenic rice with enhanced resistance to rice blast.
  • To utilize maize Activator/Dissociation (Ac/Ds) transposon system for efficient transgene integration and removal.
  • To validate the efficacy of marker-free transgenic rice in conferring blast resistance.

Main Methods:

  • Employed maize (Zea mays) Ac/Ds transposon vectors with dual fluorescent protein markers (GFP and mCherry) for generating marker-free transgenic rice.
  • Used double fluorescent protein-aided counterselection and PCR-based screening to identify marker-free progeny.
  • Cloned a Pi21 gene RNAi expression cassette into the Ds element as the gene of interest (GOI).
  • Verified marker-free status and transgene integration using PCR, Southern hybridization, and TAIL-PCR.

Main Results:

  • Successfully generated marker-free T1 transgenic rice plants from 13 independent lines.
  • Confirmed the absence of transgenic markers and T-DNA sequences in selected lines via molecular analyses.
  • Demonstrated reduced Pi21 gene expression and increased resistance to rice blast in marker-free transgenic lines.
  • Identified transgene reintegration into intergenic regions of the rice genome.

Conclusions:

  • The Ac/Ds transposon system with dual fluorescent markers provides a reliable method for screening marker-free transgenic rice.
  • Marker-free transgenic rice lines exhibit improved resistance to rice blast.
  • This technology can be applied to develop rice with enhanced disease resistance and other desirable agronomic traits.