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

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

Effect of Pre-milling Treatments on Green Gram Dhal Recovery

Effect of oil and thermal treatments on milling quality of green gram was studied at 8 to 12% moisture content (w.b) with 2.40 to 4.00 g.kg-1 oil for treatment and holding time of 5 to 10 h, thermal treatment at temperature of 80 to 120°C and roasting duration of 1 to 5 min. These pretreatments showed dhal recovery of 70.01 to 74.93% in case of oil treatment, and 71.18 to 78.12% in case of thermal treatment as against 61.19 to 65.64% without any pretreatment. Optimum conditions for oil treatment for maximum dhal recovery were 10.26% (w.b), oil quantity of 3.17 g.kg-1 and holding for 7.72 h, roasting for 2.86 min at temperature of 99.24°C and 10.25% moisture content (w.b) for thermal treatment of green gram