Scholarlink

<p>Rice feeds nearly half of the world’s population and underpins global food security. Climate change now poses a major threat to rice productivity worldwide. Genome editing has reshaped crop improvement strategies. Among these tools, the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein (CRISPR/Cas) system stands out for its precision, efficiency, and scalability. However, <i>Agrobacterium</i>-mediated transformation efficiency is often low, particularly in indica rice varieties. Here, we optimized an <i>Agrobacterium</i>-mediated transformation protocol for indica rice cultivars. The method was established in Lalat and MTU-1010. Seed-derived embryogenic calli were used to introduce the thermosensitive genic male sterile <i>(OsTMS5)</i> gene. A CRISPR/Cas9 vector carrying a gRNA and the selectable marker <i>hptII</i> was used for transformation. Callus induction reached 96.87% in MTU-1010 and 93.30% in Lalat MS medium supplemented with 3&#xa0;mg/L 2,4-D and 0.5&#xa0;mg/L BAP. In contrast, regeneration efficiency was higher in Lalat (90.28%) than in MTU-1010 (87.51%) on MS medium supplemented with 0.25&#xa0;mg/L NAA, 0.5&#xa0;mg/L kinetin, and 2&#xa0;mg/L BAP. In addition, PCR analysis further verifies the integration of the transgene. Subsequently, the transformation efficiency was 37.20% in Lalat and 29.62% in MTU-1010. Therefore, this protocol provides a robust platform for gene function analysis and trait editing in rice. Its application may accelerate yield improvement and enhance stress tolerance under changing climatic conditions.</p> Graphical Abstract <p></p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

An improved Agrobacterium-mediated transformation method for genome editing using CRISPR/Cas9 in elite indica rice (Oryza sativa L.)

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年4月7日

查看全文
  • Laxmipreeya Behera,
  • Kailash Ch. Samal,
  • Parameswaran C,
  • Pawan Kumar Agrawal,
  • V. Mohan Murali Achary,
  • Manasi Dash,
  • Ashok Mishra,
  • Manjusha Rani,
  • Fred Bwayo Masika,
  • Gurunatham Sai Deekshith Goud,
  • Mahipal Singh Kesawat,
  • Sanghamitra Samantaray

摘要

Rice feeds nearly half of the world’s population and underpins global food security. Climate change now poses a major threat to rice productivity worldwide. Genome editing has reshaped crop improvement strategies. Among these tools, the Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein (CRISPR/Cas) system stands out for its precision, efficiency, and scalability. However, Agrobacterium-mediated transformation efficiency is often low, particularly in indica rice varieties. Here, we optimized an Agrobacterium-mediated transformation protocol for indica rice cultivars. The method was established in Lalat and MTU-1010. Seed-derived embryogenic calli were used to introduce the thermosensitive genic male sterile (OsTMS5) gene. A CRISPR/Cas9 vector carrying a gRNA and the selectable marker hptII was used for transformation. Callus induction reached 96.87% in MTU-1010 and 93.30% in Lalat MS medium supplemented with 3 mg/L 2,4-D and 0.5 mg/L BAP. In contrast, regeneration efficiency was higher in Lalat (90.28%) than in MTU-1010 (87.51%) on MS medium supplemented with 0.25 mg/L NAA, 0.5 mg/L kinetin, and 2 mg/L BAP. In addition, PCR analysis further verifies the integration of the transgene. Subsequently, the transformation efficiency was 37.20% in Lalat and 29.62% in MTU-1010. Therefore, this protocol provides a robust platform for gene function analysis and trait editing in rice. Its application may accelerate yield improvement and enhance stress tolerance under changing climatic conditions.

Graphical Abstract