title: In vitro clonal propagation of Achyranthes aspera L. and Achyranthes bidentata Blume using nodal explants.
abstract: OBJECTIVE
  To develop the reproducible in vitro propagation protocols for the medicinally important plants viz., Achyranthes aspera (A. aspera) L. and Achyranthes bidentata (A. bidentata) Blume using nodal segments as explants.
  
  
  METHODS
  Young shoots of A. aspera and A. bidentata were harvested and washed with running tap water and treated with 0.1% bavistin and rinsed twice with distilled water. Then the explants were surface sterilized with 0.1% (w/v) HgCl2 solutions for 1 min. After rinsing with sterile distilled water for 3-4 times, nodal segments were cut into smaller segments (1 cm) and used as the explants. The explants were placed horizontally as well as vertically on solid basal Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% (w/v) agar (Hi-Media, Mumbai) and different concentration and combination of 6-benzyl amino purine (BAP), kinetin (Kin), naphthalene acetic acid (NAA) and indole acetic acid (IAA) for direct regeneration.
  
  
  RESULTS
  Adventitious proliferation was obtained from A. aspera and A. bidentata nodal segments inoculated on MS basal medium with 3% sucrose and augmented with BAP and Kin with varied frequency. MS medium augmented with 3.0 mg/L of BAP showed the highest percentage (93.60±0.71) of shootlets formation for A. aspera and (94.70±0.53) percentages for A. bidentata. Maximum number of shoots/explants (10.60±0.36) for A. aspera and (9.50±0.56) for A. bidentata was observed in MS medium fortified with 5.0 mg/L of BAP. For A. aspera, maximum mean length (5.50±0.34) of shootlets was obtained in MS medium augmented with 3.0 mg/L of Kin and for A. bidentata (5.40±0.61) was observed in the very same concentration. The highest percentage, maximum number of rootlets/shootlet and mean length of rootlets were observed in 1/2 MS medium supplemented with 1.0 mg/L of IBA. Seventy percentages of plants were successfully established in polycups. Sixty eight percentages of plants were well established in the green house condition. Sixty five percentages of plants were established in the field.
  
  
  CONCLUSIONS
  The results have shown that use of nodal buds is an alternative reproducible and dependable method for clonal propagation of A. aspera and A. bidentata. The high rate of direct shoot-root multiplication and their high rate of post-hardening survival indicate that this protocol can be easily adopted for commercial large scale cultivation.
authors.affiliation: Department of Botany, Arignar Anna Government Arts College, Namakkal, Tamil Nadu, India.
authors.forename: Wesely Edward
authors.initials: WE
authors.lastname: Gnanaraj
authors.forename: Johnson Marimuthu
authors.initials: JM
authors.lastname: Antonisamy
authors.forename: Mohanamathi
authors.initials: M
authors.lastname: R B
authors.forename: Kavitha Marappampalyam
authors.initials: KM
authors.lastname: Subramanian
mesh_terms: D029044:Achyranthes / Q000254:growth & development*; D003470:Culture Media / Q000737:chemistry; D018517:Plant Roots / Q000254:growth & development; D018520:Plant Shoots / Q000254:growth & development; D010946:Plants, Medicinal / Q000254:growth & development*; D016019:Survival Analysis
chemical_list: D003470:Culture Media
keywords: Achyranthes aspera; Achyranthes bidentata; BAP; Clonal propagation; IAA; In vitro; Kinetin; MS medium; NAA; Nodal culture; Nodal explant; Nodal segment; Plant regeneration; Reproducible; Rootlet; Shootlet; Shoots; Tissue culture