Ismail Hammad, Kayani Sania Sabahat, Kayani Sadaf Ilyas, Mirza Bushra, Waheed Mohammad Tahir
1Department of Biochemistry and Biotechnology, University of Gujrat, Gujrat, 50700 Pakistan.
2Department of Biochemistry, Quaid-I-Azam University, Islamabad, 45320 Pakistan.
3 Biotech. 2019 Sep;9(9):339. doi: 10.1007/s13205-019-1870-x. Epub 2019 Aug 22.
In vitro suspension culture techniques are cost effective for large-scale production of secondary metabolites. In the present study, firstly, suspension cultures of untransformed were prepared using different hormonal combinations and were subjected to different pH, temperature and salt concentrations. Maximum biomass was obtained for suspensions supplemented with 1.5 mg/L BAP and 0.1 mg/L NAA, at pH 5.8, temperature 28 °C and 0 mM NaCl concentration. Using these parameters, suspensions were produced for and -transformed lines of . All the transgenic lines showed prominent increase in fresh weight (FW) and dry weight (DW) with maximum values for 2 line producing 169.8 mg/mL FW and 25.3 mg/mL DW. The exudates of transformed and untransformed plants were tested for the antioxidant activity and in vivo assays on rats. Maximum phenolic content (261 μg/mL) and flavonoid content (637.6 μg/mL) were obtained for 1 transgenic line. Total antioxidant capacity was found maximum (1451.7 μg/mL) for untransformed lettuce, whereas 1 showed maximum total reducing power activity (637.6 μg/mL). In DPPH assay, maximum activity (104.7 μg/mL) was shown by 3 line. In rats analgesic assay, maximum activity (74.9%) was shown by 2. Line 1 showed maximum anti-inflammatory activity (69.2%) and maximum antidepressant activity (minimum immobility time of 55 s). Maximum anticoagulant activity was observed for 2 with maximum clotting time of 130 s. The present study could help in using lettuce suspension culture as platform for the enhanced production of important metabolites.
体外悬浮培养技术对于大规模生产次生代谢产物具有成本效益。在本研究中,首先,使用不同的激素组合制备未转化植株的悬浮培养物,并使其处于不同的pH值、温度和盐浓度条件下。在pH 5.8、温度28°C和0 mM NaCl浓度下,添加1.5 mg/L BAP和0.1 mg/L NAA的悬浮培养物获得了最大生物量。利用这些参数,为[植物名称]的未转化植株以及转化植株系制备了悬浮培养物。所有转基因株系的鲜重(FW)和干重(DW)均显著增加,其中2号株系的FW最大值为169.8 mg/mL,DW最大值为25.3 mg/mL。对转化和未转化植株的渗出物进行了抗氧化活性测试以及对大鼠的体内试验。1个转基因株系的酚类含量最高(261 μg/mL),黄酮类含量最高(637.6 μg/mL)。未转化的生菜总抗氧化能力最高(1451.7 μg/mL),而1号株系的总还原能力活性最高(637.6 μg/mL)。在DPPH测定中,3号株系的活性最高(104.7 μg/mL)。在大鼠镇痛试验中,2号株系的活性最高(74.9%)。1号株系的抗炎活性最高(69.2%),抗抑郁活性最高(最短不动时间为55秒)。2号株系的抗凝活性最高,凝血时间最长为130秒。本研究有助于将生菜悬浮培养作为增强重要代谢产物生产的平台。