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Volume 12
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10.3390/agronomy12010179
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Article

Efficient Genetic Transformation of Rice for CRISPR/Cas9 Mediated Genome-Editing and Stable Overexpression Studies: A Case Study on Rice Lipase 1 and Galactinol Synthase Encoding Genes

by
Tanika Thakur
1,
Kshitija Sinha
2,3,
Tushpinder Kaur
1,
Ritu Kapoor
1,
Gulshan Kumar
4,*,
Rupam Kumar Bhunia
2,* and
Prafull Salvi
1,*
1
Agri-Biotechnology, National Agri-Food Biotechnology Institute, Mohali 140306, India
2
Plant Tissue Culture and Genetic Engineering, National Agri-Food Biotechnology Institute, Mohali 140306, India
3
Department of Biotechnology, Panjab University, Sector-25, Chandigarh 160014, India
4
Food and Nutritional Biotechnology, National Agri-Food Biotechnology Institute, Mohali 140306, India
*
Authors to whom correspondence should be addressed.
Agronomy 2022, 12(1), 179; https://doi.org/10.3390/agronomy12010179
Submission received: 30 October 2021 / Revised: 24 November 2021 / Accepted: 31 December 2021 / Published: 12 January 2022
(This article belongs to the Special Issue Applications of Transgenic and Targeted Genome Editing in Rice Improvement)
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Abstract

Rice is a staple food crop for almost half of the world’s population, especially in the developing countries of Asia and Africa. It is widely grown in different climatic conditions, depending on the quality of the water, soil, and genetic makeup of the rice cultivar. Many (a)biotic stresses severely curtail rice growth and development, with an eventual reduction in crop yield. However, for molecular functional analysis, the availability of an efficient genetic transformation protocol is essential. To ensure food security and safety for the continuously increasing global population, the development of climate-resilient crops is crucial. Here, in this study, the rice transformation protocol has been effectively optimized for the efficient and rapid generation of rice transgenic plants. We also highlighted the critical steps and precautionary measures to be taken while performing the rice transformation. We further assess the efficacy of this protocol by transforming rice with two different transformation constructs for generating galactinol synthase (GolS) overexpression lines and CRISPR/Cas9-mediated edited lines of lipase (Lip) encoding the OsLip1 gene. The putative transformants were subjected to molecular analysis to confirm gene integration/editing, respectively. Collectively, the easy, efficient, and rapid rice transformation protocol used in this present study can be applied as a potential tool for gene(s) function studies in rice and eventually to the rice crop improvement.
Keywords: rice (Oryza sativa); galactinol synthase; lipase; Agrobacterium tumefaciens; transformation rice (Oryza sativa); galactinol synthase; lipase; Agrobacterium tumefaciens; transformation

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MDPI and ACS Style

Thakur, T.; Sinha, K.; Kaur, T.; Kapoor, R.; Kumar, G.; Bhunia, R.K.; Salvi, P. Efficient Genetic Transformation of Rice for CRISPR/Cas9 Mediated Genome-Editing and Stable Overexpression Studies: A Case Study on Rice Lipase 1 and Galactinol Synthase Encoding Genes. Agronomy 2022, 12, 179. https://doi.org/10.3390/agronomy12010179

AMA Style

Thakur T, Sinha K, Kaur T, Kapoor R, Kumar G, Bhunia RK, Salvi P. Efficient Genetic Transformation of Rice for CRISPR/Cas9 Mediated Genome-Editing and Stable Overexpression Studies: A Case Study on Rice Lipase 1 and Galactinol Synthase Encoding Genes. Agronomy. 2022; 12(1):179. https://doi.org/10.3390/agronomy12010179

Chicago/Turabian Style

Thakur, Tanika, Kshitija Sinha, Tushpinder Kaur, Ritu Kapoor, Gulshan Kumar, Rupam Kumar Bhunia, and Prafull Salvi. 2022. "Efficient Genetic Transformation of Rice for CRISPR/Cas9 Mediated Genome-Editing and Stable Overexpression Studies: A Case Study on Rice Lipase 1 and Galactinol Synthase Encoding Genes" Agronomy 12, no. 1: 179. https://doi.org/10.3390/agronomy12010179

APA Style

Thakur, T., Sinha, K., Kaur, T., Kapoor, R., Kumar, G., Bhunia, R. K., & Salvi, P. (2022). Efficient Genetic Transformation of Rice for CRISPR/Cas9 Mediated Genome-Editing and Stable Overexpression Studies: A Case Study on Rice Lipase 1 and Galactinol Synthase Encoding Genes. Agronomy, 12(1), 179. https://doi.org/10.3390/agronomy12010179

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