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

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dc.contributor.authorBehera, Laxmipreeya
dc.contributor.authorSamal, Kailash Ch.
dc.contributor.authorParameswaran C,
dc.contributor.authorAgrawal, Pawan Kumar
dc.contributor.authorAchary, V. Mohan Murali
dc.contributor.authorDash, Manasi
dc.contributor.authorMishra, Ashok
dc.contributor.authorRani, Manjusha
dc.contributor.authorMasika, Fred Bwayo
dc.contributor.authorGoud, Gurunatham Sai Deekshith
dc.contributor.authorKesawat, Mahipal Singh
dc.contributor.authorSamantaray, Sanghamitra
dc.date.accessioned2026-04-19T08:05:07Z
dc.date.available2026-04-19T08:05:07Z
dc.date.issued2026-04-07
dc.descriptionThis study advances genomic research to improve oil palm productivity and resilience, supporting the development of high-yield, locally adapted varieties. The findings contribute to SDG 2 (Zero Hunger), SDG 9 (Industry, Innovation and Infrastructure), and SDG 12 (Responsible Consumption and Production) by promoting agricultural innovation and sustainable resource use. The results also align with Uganda’s National Development Plan IV, focusing on agro-industrialisation, agricultural research, and innovation to strengthen food security and economic growth.
dc.description.abstractRice 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.
dc.description.sponsorshipDepartment of Science and Technology, Government of India, New Delhi
dc.identifier.citationBehera, L., Samal, K. C., Agrawal, P. K., Achary, V. M. M., Dash, M., Mishra, A., ... & Samantaray, S. (2026). An improved Agrobacterium-mediated transformation method for genome editing using CRISPR/Cas9 in elite indica rice (Oryza sativa L.). BMC genomics.
dc.identifier.issn1471-2164
dc.identifier.urihttps://dir.muni.ac.ug/handle/20.500.12260/962
dc.language.isoen
dc.publisherSpringer Nature
dc.subjectRice
dc.subjectAgrobacterium-mediated transformation
dc.subjectCallus
dc.subjectMS medium
dc.subjectgRNA
dc.subjectPCR
dc.subjectCRISPR/Cas.
dc.titleAn improved Agrobacterium-mediated transformation method for genome editing using CRISPR/Cas9 in elite indica rice (Oryza sativa L.)
dc.typeArticle

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