Su Yuekai, Lin Caicai, Zhang Jin, Hu Bei, Wang Jie, Li Jingyu, Wang Shiqi, Liu Ruihao, Li Xia, Song Zhenqiao, Wang Jianhua
State Key Laboratory of Crop Biology, Shandong Key Laboratory of Crop Biology, College of Agronomy, Shandong Agricultural University, Tai'an, China.
Taishan Academy of Forestry Sciences, Tai'an, China.
Front Plant Sci. 2022 May 20;13:913985. doi: 10.3389/fpls.2022.913985. eCollection 2022.
is a traditional Chinese medicinal plant of Labiatae, which has been widely utilized to treat a variety of cardiovascular and cerebrovascular diseases. However, due to the long growth cycle, low content of active ingredients, and serious quality deterioration of , the use of biotechnology to improve to meet the growing demand for clinical applications has become a research hotspot. In this study, a novel one-step hairy root regeneration method was developed, which could rapidly obtain hairy roots and regenerated plants with high tanshinone content. By optimizing the parameters of transformation in , it was finally established that the explants were infected in (OD = 0.6) for 10 min, co-cultured for 3 days, and then screened on the screening medium containing 7.5 mg/l hygromycin, the maximum transformation frequency can reach 73.85%. GFP and PCR detection yielded a total of 9 positive transgenic hairy root lines and 11 positive transgenic regenerated plants. was successfully overexpressed in positive transgenic regenerated plants, according to the results of qRT-PCR. The content of tanshinone IIA and cryptotanshinone were dramatically enhanced in transgenic regenerated plants and hairy roots by Ultra Performance Liquid Chromatography analysis. Based on the Agrobacterium-mediated transformation of , this study developed a new method for regenerating plants with transgenic hairy roots. This method provides a foundation for the breeding of and the sustainable development of medicinal plant resources, as well as provides a useful reference for the application of other species.
是唇形科的一种传统中药材，已被广泛用于治疗多种心脑血管疾病。然而，由于其生长周期长、活性成分含量低以及严重的品质劣化问题，利用生物技术改良以满足临床应用不断增长的需求已成为研究热点。在本研究中，开发了一种新颖的一步法毛状根再生方法，该方法能够快速获得丹参酮含量高的毛状根和再生植株。通过优化丹参转化参数，最终确定外植体在OD=0.6的农杆菌菌液中侵染10分钟，共培养3天，然后在含有7.5mg/l潮霉素的筛选培养基上筛选，最大转化频率可达73.85%。GFP和PCR检测共获得9个阳性转基因毛状根系和11个阳性转基因再生植株。根据qRT-PCR结果，在阳性转基因再生植株中成功过表达。通过超高效液相色谱分析，转基因再生植株和毛状根中丹参酮IIA和隐丹参酮的含量显著提高。基于农杆菌介导的丹参转化，本研究开发了一种转基因毛状根再生植株的新方法。该方法为丹参育种和药用植物资源的可持续发展提供了基础，也为其他物种的应用提供了有益参考。
