Abstract
An effective and consistent in vitro propagation technique for Boerhaavia diffusa L. was developed using different types of explants and media compositions. Shoot apex and nodal explants of field grown plants were aseptically cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of auxins (IAA and NAA) and cytokinins (BAP and Kn). The maximum number of multiple shoot buds (6.65 ± 0.09) obtained from nodal explants on MS medium containing 2.0 mg/l BAP + 0.5 mg/l IAA. Multiple shoot buds underwent rapid elongation (4.24 ± 0.06 cm) on MS media fortified with 3.0 mg/l BAP + 0.5 mg/l NAA. Half strength MS media supplemented with 1.0 mg/l IBA + 0.5 mg/l IAA was better for induction and proliferation (6.42 ± 0.07) of roots and 75% of plantlets were successfully acclimatized to ex vitro condition, exhibiting a normal development. Somatic chromosome number of in vivo and in vitro grown plants were confirmed to be 2n = 26. Chromosome length ranged from 1.40 to 2.43 μm in the mother plants and 1.34 to 2.31 μm for in vitro grown plants. The total form percent (TF%) of mother and in vitro grown plants was 42.91% and 41.16%, respectively. The karyotype formula of in vivo grown plants was 2n = 26 = 4Lsm + 6Msm + 2Mm + 14Sm, whereas that of the in vitro grown plants was 2n = 26 = 8Lsm + 4Msm + 2Mm + 12Sm. The frequency of the chromosome having arm more than 2:1 was 0.08 for in vivo grown plants and 0.15 for in vitro raised plants. Thus, according to Stebbins classification (1971) the karyotype of both plants falls into 2A symmetrical type.
Highlights
Boerhaavia diffusa L., generally known as Punarnava, is a perennial creeping, diffusely branched herb belonging to the Nyctaginaceae family
In order to establish a suitable protocol for in vitro mass scale propagation of Boerhaavia diffusa L., shoot apices and nodal segments of field grown plants were collected and aseptically cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of cytokinin (BAP and Kn) and auxin (NAA, IAA and IBA) in different combinations
Different concentrations of cytokinin BAP (0.5 - 3.0 mg/l) or Kn (0.5 - 3.0 mg/l) alone and in combination with different concentrations of NAA or IAA were used in MS medium for induction and development of MSBs in nodal segments and shoot apices of B. diffusa
Summary
Boerhaavia diffusa L., generally known as Punarnava, is a perennial creeping, diffusely branched herb belonging to the Nyctaginaceae family. Punarnava leaves are usually 1.25 - 5 cm long, thick, broadly ovate or suborbicular, rounded at the apex, usually white beneath. The Boerhaavia diffusa plant contains many bioactive compounds like tannins, flavonoids, alkaloids (punarnavine), glycosides, steroids, terpenoids, phenolic compounds, rotenoids (boeravinones A-O) etc. B. diffusa has various types of pharmacological properties like antilymphoproliferative, antiurethritis, antiasthmatic, antibacterial, anti-inflammatory, antileprosy, antidiabetic, immune-modulation and anti-metastatic [4]. In Bangladesh, there is virtually no cultivation of medicinal plants on any significant scale [5] for commercial purpose. It is essential to take urgent steps for large scale propagation of this medicinal plant species, to conserve the plant and meet the demand of ayurvedic industries
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