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This chapter focuses on initiating and establishing in vitro shoot cultures, emphasizing optimal growth conditions and sub-culturing techniques. The transformation of shoot cultures using Agrobacterium tumefaciens, aiming to induce shooty teratomas and evaluate transformation efficiency is discussed. The spectrophotometric estimation of nicotine from these cultures is provided, while gravimetric method for nicotine isolation and quantification is also discussed. The results from these protocols aim to optimize conditions for shoot culture growth, transformation, and nicotine production.

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Practical Techniques for Shoot Cultures

  • 章节

  • 发表日期：

    2025年

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  • M. Anuradha,
  • P. Shilpa,
  • Umalatha,
  • S. Balasubramanya

摘要

This chapter focuses on initiating and establishing in vitro shoot cultures, emphasizing optimal growth conditions and sub-culturing techniques. The transformation of shoot cultures using Agrobacterium tumefaciens, aiming to induce shooty teratomas and evaluate transformation efficiency is discussed. The spectrophotometric estimation of nicotine from these cultures is provided, while gravimetric method for nicotine isolation and quantification is also discussed. The results from these protocols aim to optimize conditions for shoot culture growth, transformation, and nicotine production.