G. lotoides L. is a threatened plant that is frequently harvested for medicinal purpose. However, its distribution in the world is limited because of short period of seed viability and poor seed germination. The objective of this study was to develop in vitro propagation protocol for G. lotoides through callus induction. For callus induction, different concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), NAA (α-naphthalene acetic acid) and BAP (6-benzyl amino purine) were used. Seeds were sown on growth regulator-free MS medium and shoots from the in vitro germinated seedlings were excised and cultured on MS medium containing 0.5 mg/l BAP. Young leaves from these shoots were used as explant for callus induction and shoot regeneration. Maximum callus induction (100%) was observed on medium containing 2,4-D (0.5, 2.0, 3.5 mg/l) or NAA (2.0, 2.5 mg/l) in combination with 0.5 mg/l BAP. However, 2,4-D was the best in overall callus induction. The highest regeneration (20%) frequency was achieved on the medium containing 0.5 mg/l BAP. Highest number of shoot (2.83 ± 1.22) and shoot length (2.16 ± 0.87 cm) per explant were obtained in the presence of 0.25 mg/l BAP + 0.5 mg/l KIN (Kinetin). In shoot multiplication media, maximum mean (6.43 ± 0.87) of shoot was observed on MS medium containing 0.5 mg/l BAP. The best shoot length (1.70 ± 0.14 cm) was recorded on control medium. The highest (95%), maximum root number (14.10 ± 1.47) and root length (1.01 ± 0.10 cm) were obtained on a medium supplemented with 1.5 mg/l Indole-3-butyric acid (IBA). All the plants (100%) were survived after acclimatization in greenhouse. The present study can be useful for callus induction and indirect shoot regeneration form G. lotoides.

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Glinus lotoides L. is a valuable medicinal herb for the treatment of human and livestock ailments. However, it was listed under endangered plant species due to over utilization and lower seed viability. The purpose of this study has been to develop a micropropagation method from nodal segments of G. lotoides. MS containing various concentrations of Kn was used for culture induction. Best culture establishment (95%) and highest mean shoot number (2.45 ± 0.37) were achieved on a medium containing 1.5 mg/l Kn. Even though different concentrations of 6-benzyl aminopurine (BAP), Kn and α-naphthalene acetic acid (NAA) were utilized for shoot proliferation, the highest number of shoots per nodal segment (3.66 ± 0.61) was attained in the medium containing 1.0 mg/l BAP in combination with 0.2 mg/l NAA. Shoots were rooted on half strength MS containing IBA, IAA and NAA. In a medium containing 1.5 mg/l IBA, 5.80 ± 1.10 roots per shoot, 1.05 ± 0.10 cm root length and 100% root generation frequency were obtained. Plantlets were successfully established under greenhouse conditions with 94% survival rate and no aberrant plants were detected.