ICAR-Indian Institute of Millets Research (IIMR)

Collaborating Centre Principal Investigator (CC-PI)

Dr. D Balakrishna

Principal Scientist

Principal Investigator involved in the project

Dr. D Balakrishna

Principal Scientist

Co-Principal Investigators (Co-PIs)

Dr. T Nepolean

Principal Scientist

Dr. B Gangaiah

Principal Scientist

Dr. VM Malathi

Scientist

Dr. Avinash Singode

Senior Scientist

Dr. Jinu Jacob

Senior Scientist

Target Traits in Sorghum & Pearl millet

Research Methods & Expected Outcomes

Genome editing in sorghum and pearl millet was initiated using CRISPR/Cas9 by cloning multiple sgRNAs targeting traits related to herbicide tolerance and grain quality improvement. A total of twelve guide RNAs (three per target) were designed and individually cloned into the binary vector pBUN421, driven by the maize TaU3 promoter. This construct also carried the Cas9 coding region under the control of the maize codon-optimized Ubiquitin (Ubi) promoter, along with a plant-selectable marker (Bar gene) driven by the 35S promoter. These twelve binary vectors were further mobilized individually into Agrobacterium tumefaciens strain LBA4404 for plant transformation using shoot meristem/shoot apices explants.

In pearl millet, a total of 300 phosphinothricin (3 mg/l) resistant putative transformants were developed out of 1800 shoot tip explants (200 explants/vector) of the Dhanshakti cultivar infected through Agrobacterium. These plants were successfully transferred to the glasshouse for further growth and molecular analysis. In sorghum, 208 phosphinothricin (3 mg/l) resistant plants were regenerated out of 600 shoot tip explants (200 explants/vector) of the C43 cultivar following Agrobacterium-mediated transformation. The regenerated plants were subsequently transferred to rooting media for proper root development.