Rapid and Efficient Optimization Method for a Genetic Transformation System of Medicinal Plants Erigeron breviscapus

• Published in: International journal of molecular sciences Vol. 24; no. 6; p. 5611
• Main Authors: Zhao, Yujun, Yu, Yifan, Guo, Juan, Zhang, Yifeng, Huang, Luqi
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 15.03.2023; MDPI
• Subjects: Alzheimer's disease; biolistic; Bombardment; Callus; Cerebrovascular diseases; Cerebrovascular Disorders; Efficiency; Erigeron - genetics; Erigeron breviscapus; Genes; Genetic engineering; Genetic transformation; Hemorrhage; Herbal medicine; Hygromycin; Hygromycin B; Magnesium chloride; medicinal plant; optimization strategies; Particle bombardment; Plant Breeding; Plants, Medicinal - genetics; Plasmids; regeneration; transformation; Transformation, Genetic; Transformations; Traumatic brain injury; Variance analysis; biolistic; medicinal plant; optimization strategies; transformation; regeneration; Erigeron breviscapus
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms24065611

is an important medicinal plant with high medicinal and economic value. It is currently the best natural biological drug for the treatment of obliterative cerebrovascular disease and the sequela of cerebral hemorrhage. Therefore, to solve the contradiction between supply and demand, the study of genetic transformation of is essential for targeted breeding. However, establishing an efficient genetic transformation system is a lengthy process. In this study, we established a rapid and efficient optimized protocol for genetic transformation of using the hybrid orthogonal method. The effect of different concentrations of selection pressure (Hygromycin B) on callus induction and the optimal pre-culture time of 7 days were demonstrated. The optimal transformation conditions were as follows: precipitant agents MgCl + PEG, target tissue distance 9 cm, helium pressure 650 psi, bombardment once, plasmid DNA concentration 1.0 μg·μL , and chamber vacuum pressure 27 mmHg. Integration of the desired genes was verified by amplifying 1.02 kb of gene from the T0 transgenic line. Genetic transformation of was carried out by particle bombardment under the optimized conditions, and a stable transformation efficiency of 36.7% was achieved. This method will also contribute to improving the genetic transformation rate of other medicinal plants.