Please use this identifier to cite or link to this item: http://223.31.159.10:8080/jspui/handle/123456789/1564
Title: A simplified and improved protocol of rice transformation to cater wide range of rice cultivars
Authors: Rengasamy, Balakrishnan
Manna, Mrinalini
Jonwal, Sarvesh
Sathiyabama, Muthukrishnan
Thajuddin, Nargis Begum
Sinha, Alok Krishna
Keywords: Rice transformation
Agrobacterium tumefaciens
Indica rice
Japonica rice
CRISPR-Cas9
sgRNA
Regeneration efciency
Transformation efciency
Editing efciency
Issue Date: 2024
Publisher: Springer Nature Publishing AG
Citation: Protoplasma, 261(4): 641-654
Abstract: The latest CRISPR-Cas9-mediated genome editing technology is expected to bring about revolution in rice yield and quality improvement, and thus validation of rice transformation protocols using CRISPR-Cas9-gRNA constructs is the need of the hour. Moreover, regeneration of more number of transgenic rice plants is prerequisite for developing genome-edited rice lines, as recalcitrant rice varieties were shown to have lower editing efciencies which necessities screening of large number of transgenic plants to fnd the suitable edits. In the present study, we have simplifed the Agrobacterium-mediated rice transformation protocol for both Indica and Japonica rice cultivars using CRISPR/Cas9 empty vector construct, and the protocols have been suitably optimized for getting large numbers of the regenerated plantlets within the shortest possible time. The Japonica transgenic lines were obtained within 65 days and for the Indica cultivars, it took about 76–78 days. We also obtained about 90% regeneration efciency for both Japonica and Indica cultivars. The transformation efciency was about 97% in the case of Japonica and 69–83% in the case of Indica rice cultivars. Furthermore, we screened the OsWRKY24 gene editing efciency by transforming rice cultivars with CRISPR/Cas9 construct harbouring sgRNA against OsWRKY24 gene and found about 90% editing efciency in Japonica rice cultivars, while 30% of the transformed Indica cultivars were found to be edited. This implicated the presence of a robust repair mechanism in the Indica rice cultivars.
Description: Accepted date: 24 December 2023
URI: https://link.springer.com/article/10.1007/s00709-023-01925-8
http://223.31.159.10:8080/jspui/handle/123456789/1564
ISSN: 1615-6102
0033-183X
Appears in Collections:Institutional Publications

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