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Caucasian whortleberry (Vaccinium arctostaphylos L.) plant traditionally has been included among important medicines as remedy for diabetic patients and also in modern medicine found to be effective against cardiovascular diseases. V. arctostaphylos leaves are known to be rich in phytochemicals, however, no comprehensive and reliable protocol has been developed to date in order to produce plant materials for conservation purposes and/or to generate secondary metabolites under in vitro conditions. Therefore, present study was designed to develop a micropropagation system having high genetic fidelity and considerable potential to produce secondary chemicals from V. arctostaphylos. No morphological dissimilarities have been observed in tissue culture-produced shoots after their successfully placement in a greenhouse with high survival rate (≥ 90%). At the end of 2nd subculture, woody plant medium (WPM) containing moderate level of plant growth regulators (PGRs) such as Zeatin (2 ppm) and Auxin (Indole-3-butyric acid - IBA; 0.1 ppm) has significantly increased shoot length and shoot numbers per explants, whereas, other vegetative growth characteristics have shown significant increase in mediums containing the highest Zeatin and IBA concentrations. Half-strength WPM medium containing IBA (1 ppm) and activated charcoal (AC; 0.2%) showed maximum root length, number of roots and rooted explants and that the increasing concentrations of IBA and AC have positively improved root induction. Genetic homogeneity of the micropropagated plants has been analyzed using four pairs of simple sequence repeats (SSR) markers. Proliferated plants showed monomorphic bands identical to the donor plant and manifested the genetic stability of the newly produced plants. Different assays were used to measure the antioxidant potential and phenolic compound from in vitro, acclimatized and mother plant leaves. In all experiments, PGRs esp., Zeatin has shown significant effects on antioxidant scavenging, phenolic and flavonoid content of micropropagated plants. Metabolite screening using Infrared Spectroscopy (IR) indicated the presence of various functional groups such as amines, amides, carboxylic acids, esters, alkanes and aliphatic amine similar to mother plants. Results from present study have clearly indicated that the acclimatized plants of V. arctostaphylos could have been a reliable source for in vitro or ex-situ conservation and breeding program and, therefore, its conservation may highly be important due to availability of medicinally valuable plant secondary metabolites. |
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