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Diospyros kaki L. is considered one of the most important economical medicinal plant in Chinese herbal medicine belonging to family Ebenaceae as it contains varied secondary metabolites and used in treatment of many diseases. For there is an efficient and promising protocol for in vitro propagation of Diospyros kaki L. cv. Costata was established Diospyros kaki L. cv. Costata although it is difficult to initiate it in sterilized artificial media due to the browning of tissue caused by oxidation of phenolics. An efficient sterilizing method for leaf and internode explants was obtained by using 0.2% mercuric chloride (Hg2Cl2) for 5 min. Woody plant medium (WPM) supplemented with 2 mg/l zeatin (Zt) + 5 mg/l isopentenyl adenine (2 iP). Calli were induced on Murashige and Skoog medium (MS) augmented with 10 mg/l Zt + 10 mg/l indole-3-acetic acid (IAA) + 500 mg/l polyvinylpyrolidone (PVP) + 0.1 mg/l thiamine HCL from internode explants. However, the regeneration efficiency was obtained with ½ MS-media fortified with 1 mg/l Zt + 2 mg/l IAA + 4 mg/l benzylaminopurine (BAp) + 0.5 g/l PVP from internode calli explants. The highest amounts of scopoletin 57.08, 26.42, and 25.30 (μg/g DW) were detected using reversed phase of high-performance liquid chromatography (RP-HPLC) in leaves extract of intact plant followed by regenerated and calli cultures of internod explants, respectively. This study is the first record for in vitro propagation and production of secondary metabolites from Diospyros kaki L. using biotechnology techniques. Chemical anaysis were carried out using HPLC technique.

Biotechnological and chemical analysis of Egyptian Diospyros kaki L. cv. Costata grown in Egypt