Ruta graveolens is a widespread plant in moderate and semiarid regions with several medicinal properties that are overturning and is threatened due to destruction and overuse for the pharmaceutical and food industry. Moreover, conventional propagation through seeds is not sufficient enough to produce due to seed dormancy and poor seed viability for which clonal propagation is the need of the hour. Therefore, the current study outlines a simple approach for clonal propagation and germplasm conservation of R. graveolens. Seeds were firstly surface decontaminated and germinated on agar water media, then micro shoots were cultivated on Murashige and Skoog (MS) medium contains 3% sucrose. The addition of 1.5 mg/L 6-Benzylaminopurine (BAP) produced a maximum multiplication rate (4.2 shoots per explant). Root formation was developed at the base of micro shoot on MS containing Indole-3- Butyric acid (IBA), Indole Acetic Acid (IAA), or Naphthalene Acetic Acid (NAA). Maximum root number and root length produced at 0.9 mg/L NAA (2.25 roots per explants, and 2.69 mm length). The survival rate after acclimatization reached 90% under the greenhouse conditions when rooted plantlets were adopted to ex vitro employing in 1 soil: 1 perlite: 1 peat mixture. Micro shoots were kept for up to 16 weeks without serious losses at 6 ± 2 °C utilizing MS medium supplemented with sucrose, glucose, fructose, or sorbitol at different concentrations. Media supplemented with 6% sucrose resulted in the maximum number of shoots (2.69 shoots per explants) in the two methods of conservation. Further studies are still needed on medium-term conservation to enhance the survival percentages of different plant material types.
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