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RNA interference (RNAi) is a cellular mechanism that inhibits gene expression by suppressing its transcription or activating the RNA degradation process. The mechanism was discovered by Andrew Fire and Craig Mello in 1998 in plants. Today, it is observed in almost all eukaryotes, including protozoa, flies, nematodes, insects, parasites, and mammals. This precise cellular mechanism of gene silencing has been developed into a technique that provides an efficient way to identify and determine the...
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Direct Agroinoculation of Maize Seedlings by Injection with Recombinant Foxtail Mosaic Virus and Sugarcane Mosaic Virus Infectious Clones
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Efficient and high-throughput pseudorecombinant-chimeric Cucumber mosaic virus-based VIGS in maize.

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A new Cucumber mosaic virus (CMV)-based virus-induced gene silencing (VIGS) system efficiently silences maize genes for up to 105 days. This improved VIGS tool facilitates large-scale functional genomics in maize.

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Area of Science:

  • Plant molecular biology
  • Genomics
  • Virology

Background:

  • Virus-induced gene silencing (VIGS) is crucial for plant gene function studies.
  • Existing VIGS methods for maize have limitations including low efficiency and instability.

Purpose of the Study:

  • To develop an efficient and stable VIGS system for maize (Zea mays).
  • To overcome limitations of current VIGS approaches in maize.

Main Methods:

  • Developed a pseudorecombinant-chimeric (Pr) CMV-based VIGS system (Pr CMV VIGS).
  • Utilized mechanical inoculation of Nicotiana benthamiana sap onto young maize leaves.
  • Employed a ligation-independent cloning (LIC) strategy for vector modification.

Main Results:

  • Pr CMV VIGS demonstrated high infection efficacy and mild symptoms in maize.
  • Efficient systemic gene silencing (e.g., ZmIspH) observed as early as 5 days post-inoculation.
  • Silencing was maintained effectively throughout the entire maize growth period (up to 105 days).

Conclusions:

  • The Pr CMV VIGS system offers a significant improvement for maize gene silencing.
  • The developed system facilitates efficient, long-duration silencing for high-throughput functional genomics.
  • The pseudorecombination-chimera strategy can be applied to create efficient VIGS systems in other plants.