ENDOPHYTIC FUNGI FROM PHYLLANTHUS AMARUS SCHUM. & THONN. CAPABLE OF PRODUCING PHYLLANTHIN, HYPOPHYLLANTHIN AND/OR RELATED COMPOUNDS
Keywords:Phyllanthus amarus, Endophytic Fungi, Phyllanthin, Hypopllanthin, HPLC, HPTLC
Objective: To isolate endophytic fungi from various parts of Phyllanthus amarus (family Phyllanthaceae) and to probe for production of the bioactive compounds Phyllanthin and Hypophyllanthin.
Methods: The fungi were isolated using Potato Dexotrose Agar (PDA) and a novel P. amarus powder amended (2 g/l) PDA (PPDA). The isolates were cultured in liquid medium and after liquid-liquid extraction they were screened for production of Phyllanthin, Hypophyllanthin and/or other related compounds using Shimadzu HPLC system equipped with PDA and UV detector at 230 nm using phosphate buffer (pH 2.8) and acetonitrile in a gradient elution at a flow rate of 1.5 ml/min. Isolates which showed close overlaps were subjected to HPTLC in Pre-coated silica gel F60254 on aluminium plates using Linomat V, Camag TLC applicator and Toluene: Ethyl acetate (2: 1) as solvent system. Developed plates were sprayed with 10% v/v sulphuric acid in methanol. Isolates which showed promising results were identified using morphological characters and ITS sequences. The sequences were submitted to GenBank.
Results: Twenty three endophytic fungal isolates embracing eighteen from PDA and five from PPDA were obtained with roots serving as major home. HPLC analyses resulted in shortlisting of four isolates SATR2-A, SATR1-A, KOPS4-A and SKTS2-B and HPTLC analyses exposed further agreement. Bands apprise putative role of these endophytes in bi Â°Chemical synthesis of phyllanthin and hypophyllanthin. Morphological characters and ITS sequences identified the isolates as Fusarium oxysporum (SATR2-A), Gibberella moniliformis (Synonym: Fusarium verticillioides, SATR1-A), Alternaria sp. (KOPS4-A) and Edenia gomezpompae (SKTS2-B).
Conclusion: Phyllanthin and Hypophyllanthin, the twin magic bullets from P. amarus has widespread clinical importance and especially as hepatoprotectives. The study enlightens the role of endophytic fungi from P. amarus which can synthesize these bioactive compounds. This is the first such report from this plant and further research will result in confirming the potential.
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