Djerdjouri Amina, Abbad Mohamed, Boumrah Yacine, Malik Sonia, Makhzoum Abdullah, Lakhdar Khelifi
École Nationale Supérieure Agronomique (ES1603), Laboratoire des Ressources Génétiques et Biotechnologie, 16200 Alger, Algérie.
Université de Blida1, Faculté des Sciences de la Nature et de la Vie, Département des Biotechnologies, Laboratoire de biotechnologie des productions végétales, Blida, BP 270, Route de Soumaâ, 09000 Algérie.
3 Biotech. 2024 Sep;14(9):199. doi: 10.1007/s13205-024-04035-1. Epub 2024 Aug 12.
The present work deals with the establishment of hairy root cultures from different explants of using strain A4. A high transformation frequency (95%) was obtained from leaves followed by cotyledons (81.6%) and hypocotyls (38.3%). Genetic transformation of hairy roots was confirmed through PCR by amplifying a 400 bp fragment of the gene. Hairy roots were highly branched, possessed plagiotropic and rapid growth on hormone-free ½ B5 medium. Ten cardiac glycosides, including calotropagenin, calotoxin, frugoside, coroglaucigenin, calotropin, calactin, uzarigenin, asclepin, uscharidin, and uscharin, based on their specific masses and fragmentation properties were identified in ethanolic extracts of hairy roots by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry UHPLC/QTOF-MS. This protocol could be used as a powerful tool for large-scale in vitro production of highly valued cardiac glycosides and for further transcriptomics or metabolomics studies.
本研究利用发根农杆菌A4菌株，从[植物名称]的不同外植体建立毛状根培养物。叶片的转化频率最高（95%），其次是子叶（81.6%）和下胚轴（38.3%）。通过PCR扩增[基因名称]的400 bp片段，证实了毛状根的遗传转化。毛状根高度分支，在无激素的1/2 B5培养基上呈斜生且生长迅速。通过超高效液相色谱-四极杆飞行时间质谱联用技术（UHPLC/QTOF-MS），基于其特定质量和碎片性质，在毛状根的乙醇提取物中鉴定出10种强心苷，包括牛角瓜苷元、牛角瓜毒素、呋甾皂苷、紫花洋地黄苷元、牛角瓜苷、卡拉亭、乌沙苷元、阿斯科勒平、乌沙立定和乌沙林。该方案可作为大规模体外生产高价值强心苷以及进一步进行转录组学或代谢组学研究的有力工具。
