Optimization of biolistic transformation of rice
Student name: Ms Neha Rajkhowa
Guide: Dr Chaithanya Madhurantakam
Year of completion: 2017
Host Organisation: ICGEB, New Delhi
Supervisor (Host Organisation): Dr Tanushri Kaul
Abstract: An efficient protocol is required to generate transgenic rice plant from transformed calli. In this study, embryogenic, compact calli were harvested from mature seeds of indica rice variety Samba Mahsuri and transformed by particle bombardment and placed on regeneration medium. MS B5 medium supplemented with 30g of maltose, 10g of glucose, 1g of proline and plant growth regulators l-asparagine, silver nitrate, cysteine HCl, glutamine, dicamba and 2,4 D gelled with 4g/L Phytagel gave best results for callus induction. Selected embryogenic calli were transformed using the biolistic method using Cas9 construct. The bombarded calli were then placed on regeneration medium. Small green shoots have been observed in putative transformants. CRISPR/Cas9 based targeted mutagenesis is an excellent substitute to traditional plant breeding and transgene integration for crop improvement. CRISPR/Cas9 editing mechanism can be traced back to type II CRISPR/Cas9 immune system found in prokaryotic bacteria that enables precise genome editing by a single guide RNA (sgRNA), resulting in mutations by NHEJ or HDR mechanisms.
Keywords: Optimization, Biolistic Transformation, CRISPR, Cas9, Samba Mahsuri, indica.
