TY - JOUR AU - Nhu Thao, Le Thi AU - Hong Nhung, Nguyen AU - Quang Huy, Le AU - Phuong Thao, Bui AU - Thu Ngoc, Le AU - Bich Ngoc, Pham AU - Hoang Ha, Chu AU - Tien Phat, Do PY - 2021/10/13 Y2 - 2024/03/29 TI - Development of an in vitro hairy root induction system in different soybean cultivars for gene expression and genome editing studies JF - Vietnam Journal of Biotechnology JA - Vietnam J. Biotechnol. VL - 19 IS - 3 SE - Articles DO - 10.15625/1811-4989/15421 UR - https://vjs.ac.vn/index.php/vjbt/article/view/15421 SP - 459-470 AB - <p>Hairy root induction system has been widely applied for studies of gene function, gene expression, and genome editing in numerous plant species. In this study, we developed and evaluated the performance of an <em>in vitro</em> hairy root induction system <em>via</em> <em>Agrobacterium rhizogenes </em>on several Vietnamese and international soybean cultivars. The efficacy <em>of in vitro</em> hairy root induction and of transformation using this system was varied and depended on soybean cultivars as well as transgenic constructs. The hairy root induction frequency of different soybean cultivars ranged from 61.67% to 100% after 5 days on culture medium. From 43.8% to 79.8% of hairy roots transformed with GFP-expressing construct showed transgene expression, while that for the construct with the <em>gus</em> gene was from 38.07% to 72.33%. Among tested soybean cultivars, DT26 demonstrated the highest transformation and gene expression efficacy with both investigated vectors. This hairy root induction system was further utilized for targeted knockout mutagenesis <em>via </em>CRISPR/Cas9 of two genes which are<em> G03</em> and <em>G09</em> in the DT26 soybean cultivar. Successful mutagenesis in the regions of targeted genes was confirmed by shifted and multiple bands compared to which of non-transgenic hairy root in PAGE analysis. Sequencing results of targeted regions presented various nucleotide deletions ranging between -3 bp and -25 bp in size in both two genes of interest. This study laid an important basis for future gene function studies and targeted gene editing investigations on soybean plants in Vietnam.</p> ER -