Abstract
Bacopa monnieri (L.) Wettst. (Scrophulariaceae) is a highly sought after medicinal plant with therapeutic properties as cognition enhancer as well as for other brain and body functions. Research was conducted to optimize a thin cell layer explant based micropropagation system to assist mass propagation. Thin cell layers (TCL) derived from leaf and internode segments were used as explants. Murashige and Skoog medium was used to formulate shoot induction, elongation, and rooting media. Shoot induction media were prepared by supplementing three concentrations (0.1, 1.0, and 10.0 μM) of four cytokinins 6-benzylaminopurine, 2-isopentenyl-adenine, 6-3-Hydroxybenzylaminopurine, and thidiazuron to study adventitious shoot bud induction response. An optimum shoot bud induction response was observed on MS medium supplemented with 10.0 μM 6-benzylaminopurine for both leaf and stem transverse thin cell layer (tTCL) explants. The average number of shoot buds from leaf tTCL explants was 59, whereas, on an average, 33 shoot buds were regenerated from internode tTCL explants. Elongation of adventitious shoot buds was achieved best in a liquid medium using Liquid Lab Rocker® system. Elongated shoots recorded 100% rooting in MS medium supplemented with 5 μM indole butyric acid. Bacopa micropropagation employing tTCL explants for initial shoot bud induction and using LLR® boxes in subsequent elongation step can achieve cost effective way to regenerate high volume of plantlets and biomass required for herbal industry. Leaf and stem tTCL explants both were suitable for Agrobacterium tumefaciens (EHA105) mediated genetic transformation. Successful transformation was scored within three days of co-cultivation with Agrobacterium suspension on the basis of Enhanced Green Fluorescent Protein (EGFP) expression as an early and non-destructible screening device. Transformation frequencies of 83% and 76% were accomplished for leaf and stem tTCL explants, respectively. Greenhouse grown Bacopa plants were analyzed as fresh and dry methanolic extracts for total polyphenol content (811.93 ± 7.98 and 814 ± 17.64 GAE mg g-1) and the Trolox Equivalent Antioxidant Capacity values were 1918.25 ± 173.12 and 3163.14 ± 403.25 μmol/g, respectively.
Highlights
Bacopa monnieri (L.) Wettst. (Bacopa monniera, family Scrophulariaceae) is a well-documented herb in the traditional Ayurveda medical system attributed with medicinal properties improving cognition, such as memory retention, concentration, and learning [1]
Organogenesis Using transverse thin cell layer (tTCL) of B. monnieri Adventitious Shoot Organogenesis and Elongation A preliminary investigation was carried out to test the suitability of Murashige and Skoog (MS) and Gamborg’s B5 medium [39] for tTCL based micropropagation
Morphogenic response of leaf and stem tTCL explants has been shown in the Figure 1
Summary
Bacopa monnieri (L.) Wettst. (Bacopa monniera, family Scrophulariaceae) is a well-documented herb in the traditional Ayurveda medical system attributed with medicinal properties improving cognition, such as memory retention, concentration, and learning [1]. (Bacopa monniera, family Scrophulariaceae) is a well-documented herb in the traditional Ayurveda medical system attributed with medicinal properties improving cognition, such as memory retention, concentration, and learning [1]. Though Bacopa (locally known as Brahmi) is indigenous to India, it has been found in wild in countries as far away as Argentina [2]. Bacopa species are commercially sold as an aquatic plant for the aquarium. The bioactivity and medicinal properties have been assigned to the presence of alkaloids (nicotine, brahmine, herpestine), saponins (hersaponin, betulic acid, bacosides A, B, C and D) and other chemicals like stigmastanol, bsitosterol and stigmasterol [4]-[6]. Clinical studies have further evaluated the nootropic potential of Bacopa monnieri and confirmed its cognition-enhancing effects, attributing the effects to bioactive compounds collectively known as “bacosides” [7]. Bacosides are a type of saponin triterpene, and the bioactive compounds Bacosides A and B aid in the repair of damaged neurons by enhancing kinase activity, neuronal synthesis, restoration of synaptic activity, and nerve impulse transmission [8]
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