Development of efficient micropropagation protocol for Andrographis paniculata through epicotyledonary node

ABOUT AUTHOR
Bhanwar Lal Jat*¹, Brij Mohan Singh², C R Choudhary³, Raaz K Maheshwari^4 
1Department of Botany, SBRM Govt PG College, Nagaur, Rajasthan
²Department of Botany, SRKP, Govt PG College Kishangarh-Ajmer, Rajasthan
³Pro-President of Mewar University, Gangrar, Chittorgarh, Rajasthan
⁴Department of Chemistry, SBRM Govt PG College, Nagaur, Rajasthan

Abstract
Andrographis paniculata seeds were cultured on a basal medium of half strength MS medium with activated charcoal and 3% sucrose, incubated at 25 + 2˚C and under a light intensity of about 3000 Lux. The seeds were germinated and elongated after 4-5 weeks. The seedlings (epicotyledonary node) were used as explants. The explants (epicotyledonary node) showed swelling at nodal region which was followed by the emergence of shoot buds. Multiple shoot formation was achieved from per-existing meristems of nodal region of explant. The time taken for shoot initiation from explant was independent of growth hormones and nutrient media but the no. of shoots per explants was dependant on the concentration of growth hormones. The epicotyledonary node explants of Andrographis paniculata were inoculated on MS medium containing BAP with IAA and kinetin NAA. The no. of shoots produced per explants varied in different concentration of plant growth regulator but the time period required for shoot induction in all the treatment was similar. As the BAP concentration increased from 2.21µM/lit to 13.31µM/lit the no. of shoot buds were also increased. The maximum number of shoots (20 shoots with 33.5mm length) was observed on BAP (13.31µM/lit) with combination of IAA (5.70µM/lit). As far as the effect of various concentration of BAP, and kinetin with IAA and NAA on shoots regeneration of explants (epicotyledonary node) of Andrographis paniculata, it was concluded that BAP with combination of IAA were suitable for induction of maximum multiple shoots but the concentration of BAP higher than 13.31µM/lit with IAA 5.70 µM/lit, the shoots no. was reduced. On the medium containing kinetin 13.93µM/lit and NAA 4.83µM/lit the growth of shoot bud enhanced. The lower concentration of NAA was ineffective for growth of shoots where as the concentration of NAA higher than 1.07µM/lit inhibited the shoot growth and have induced callusing at the base of shoots. Excised shoots regenerated on MS medium containing BAP 13.31µM/lit and IAA 2.85µM/lit were transferred on to the rooting medium individually. The IBA, NAA and IAA were used individually and in combination to induce root in the regenerated shoots. Although root induction was achieved on auxin free MS medium but these roots were of poor growth. The effect of IBA (4.92µM/lit to 24.60µM/lit) was also studied which revealed that on 19.68µM/lit IBA root induction was 95% within 4-5 weeks on the lower concentration of IBA rooting reduced to 41% and in the concentration of IBA higher than 19.68µM/lit only 90% shoots were rooted with callus. When NAA with combination of IAA in various concentration was used. NAA (10.74µM/lit) and IAA (11.41µM/lit) in MS medium were found to be optimum for achieving maximum (74.8%) rooting in micro shoots of Andrographis paniculata. The rooted plantlets were hardened by keeping them in hardening chamber for 3-4 weeks. After 3-4 weeks of hardening 100 plantlets were transferred to the field conditions where 75 plants could survive.
Keywords: Andrographis paniculata media, in vitro, seedling segment