Cingoz Gunce Sahin, Verma Sandeep Kumar, Gurel Ekrem
Abant Izzet Baysal University, Department of Biology, Bolu 14280, Turkey.
Abant Izzet Baysal University, Department of Biology, Bolu 14280, Turkey.
Plant Physiol Biochem. 2014 Sep;82:89-94. doi: 10.1016/j.plaphy.2014.05.008. Epub 2014 May 28.
The effect of hydrogen peroxide (H2O2) on callus cultures of four Digitalis species (Digitalis lamarckii, Digitalis trojana, Digitalis davisiana and Digitalis cariensis) increased catalase (CAT), superoxide dismutase (SOD), total phenolic, proline activity and cardiotonic glycoside production. Callus derived from hypocotyl explants was cultured on Murashige and Skoog medium supplemented with 0.25 mg L(-1) indole-3-acetic acid (IAA) and 0.5 mg L(-1) thidiazuron (TDZ). After a month of culture, callus was transferred to MS medium containing 10 mM H2O2 and then incubated for 6 h. The amount of five cardenolides (Lanatoside C, Digitoxin, Digoxigenin, Gitoxigenin and Digoxin) as well as CAT, SOD, total phenolic, proline activity from Digitalis species were compared. No digoxin was detected in all treatments and control groups. The total cardenolides estimated were in the order of D. lamarckii (586.65 μg g(-1) dw), D. davisiana (506.79 μg g(-1) dw), D. cariensis (376.60 μg g(-1) dw) and D. trojana (282.39 μg g(-1) dw). It was clear that H2O2 pre-treatment resulted in an increase in enzymatic and nonenzymatic antioxidants. However, a significant negative relationship between cardenolides production and overall activities of CAT, SOD, total phenolic and proline was evident. The described protocol here will be useful for the development of new strategies for a large-scale production of cardenolides.
过氧化氢（H₂O₂）对四种毛地黄属植物（拉马克毛地黄、特罗亚纳毛地黄、戴维斯毛地黄和卡里恩斯毛地黄）愈伤组织培养物的影响提高了过氧化氢酶（CAT）、超氧化物歧化酶（SOD）、总酚、脯氨酸活性以及强心苷的产量。将来自下胚轴外植体的愈伤组织接种在添加了0.25 mg L⁻¹吲哚-3-乙酸（IAA）和0.5 mg L⁻¹噻二唑隆（TDZ）的Murashige和Skoog培养基上。培养一个月后，将愈伤组织转移至含有10 mM H₂O₂的MS培养基中，然后孵育6小时。比较了四种毛地黄属植物中五种强心甾类化合物（毛花苷C、洋地黄毒苷、地高辛配基、吉他洛苷配基和地高辛）以及CAT、SOD、总酚、脯氨酸活性的含量。在所有处理组和对照组中均未检测到地高辛。估计的总强心甾类化合物含量顺序为：拉马克毛地黄（586.65 μg g⁻¹干重）、戴维斯毛地黄（506.79 μg g⁻¹干重）、卡里恩斯毛地黄（376.60 μg g⁻¹干重）和特罗亚纳毛地黄（282.39 μg g⁻¹干重）。显然，H₂O₂预处理导致酶促和非酶促抗氧化剂增加。然而，强心甾类化合物产量与CAT、SOD、总酚和脯氨酸的总体活性之间存在明显的负相关关系。本文所述的方案将有助于开发大规模生产强心甾类化合物的新策略。
