This is the first report on identification and quantification of important hepatoprotective and anticancer polyphenolic lignans such as phyllanthin (PH), hypophyllanthin (HPH), niranthin (NH) and phyltetralin (PT) in natural plant and in vitro cultures of Phyllanthus tenellus Roxb. The identification of lignans was carried out by Liquid Chromatography–High Resolution Mass Spectrometry (LC–HRMS) and quantified using High-Performance Liquid Chromatography (HPLC). In addition, an efficient protocol has been developed for multiple shoot induction in nodal explants of in vitro derived shoots of P. tenellus. Maximum number of shoot regeneration (7.83 ± 0.15) was achieved on medium incorporated with 1.0 mg/l 6-Benzylaminopurine (BAP). The medium containing Indole-3-acetic acid (IAA) 2 mg/l was superior for induction of rooting in in vitro raised shoots. The plantlets were acclimatized to the field condition with 100% survival. The quantitative HPLC analysis showed that the lignan content was variable with the auxins and cytokinins incorporated in the medium. The lignan content was higher in callus grown on Murashige and Skoog (MS) medium + 2.0 mg/l Naphthaleneacetic acid (NAA). The reported protocol can be used for mass propagation and application of biotechnological approaches for improvement of P. tenellus. The results indicate intriguing possibilities for the utilization of P. tenellus plant parts as an alternative source and of callus culture to scale up bioactive lignan production for pharmaceutical applications.
Chloride and sodium constitute as the major ions in most saline soils, contributing to salt-induced damage in plants. Research on salt tolerance has mostly concentrated on the sodium toxicity; however, chloride toxicity also needs to be considered to understand the physiological, biochemical, and metabolite changes under individual and additive salts. In this study, we investigated the effect of individual Na + and/or Cl − ions (equimolar 100 mM NaCl, Na + and Cl − salts) using in vitro cultures of four soybean genotypes with contrasting salt tolerance. In general, all the treatments significantly induced antioxidant enzymes activities such as catalase, ascorbate peroxidase, glutathione reductase, guaiacol peroxidase, and superoxide dismutase and osmolytes including proline, glycine betaine, and total soluble sugar (TSS). Both individual (Na + , Cl −) and additive (NaCl) stresses induced more pronounced activation of antioxidant enzyme machinery and osmolytes accumulation in the tolerant genotypes (MAUS-47 and Bragg). The sensitive genotypes (Gujosoya-2 and SL-295) showed higher accumulation of Na + and Cl − , while the tolerant genotypes were found to maintain a low Na + /K + and high Ca 2+ level in combination with enhanced antioxidant defense and osmotic adjustment. Gas chromatography-mass spectrometry (GC-MS)-based metabolomic profiling depicted the association of certain metabolites under individualistic and additive salt effects. The genotypespecific metabolic changes indicated probable involvement of azetidine, 2-furanmethanol, 1,4-dioxin, 3-fluorothiophene, decanoic acid and 2-propenoic acid methyl ester in salt-tolerance mechanism of soybean.
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