Bhanwar Lal Jat*1, Ramesh Joshi2, C R Choudhary3, Raaz K Maheshwari4
1Department of Botany, SBRM Govt. PG College, Nagaur, Rajasthan, India
2Department of Botany, Govt. PG College Ajmer, Rajasthan, India
3Pro-President of Mewar University, Gangrar, Chittorgarh, Rajasthan, India
4Department of Chemistry, SBRM Govt. PG College, Nagaur, Rajasthan, India
The Murraya koenigii have very poor rate of multiplication through conventional method under natural conditions. Considering that the proposed plants species have proven valuable medicinal importance for their chemical properties. Micropropagation of this plant through standardized tissue culture technologies will compensate their poor propagation under natural conditions. Present investigation is mainly focused on feasible research on development of micro propagation protocol through axillary buds. Juvenile and mature stem explants of Murraya koenigii were inoculated aseptically on MS semi solid as well as on liquid media containing cytokinis and auxins in the different concentrations and combinations. The shoot initiation was observed on both liquid and semi solid media but in semi solid media browning of shoots and explants with medium and leaf fall and chlorosis was observed within one week of culture. Due to the excessive browning liquid media (without agar-agar) were used in all the experiments, conducted for shoot induction. The adenine sulphate was added in the concentration ranges between 13.57µM to 81.44µM. On the optimum physico-chemical conditions for induction and growth of shoots from axillary bud explant were defined, the in vitro shoot cultures were excised as such in the form of a cluster and were cut into two or more bunches each consisted of 4-5 shoots and each bunch was called a propagule. In this way if one mother propagule produced ten shoots after a period of 30-35 days incubation then the same propagule was cut into two daughter propagules the number of daughter propagules (4-5 shoots) achieved and from each mother propagule was presented as rate of multiplication to multiplication (M-M). At the time of sub-culture of mother propagule the elongated shoots (3-4cm) were excised individually and were transferred for rooting. The number of shoots harvested for rooting from each mother propagule was presented as rate of multiplication to rooting (M-R) in the results. The experiments for shoot multiplication were conducted in liquid as well as in semi-solid (with agar-agar) MS media but chlorosis and browning of shoots were not observed in semi solid media in multiplication experiments, therefore, only semisolid MS media were used for subsequent experimentations of multiplication. The ADS was added in concentration range of 13.57µM to 81.44µM with pre-optimized BAP concentration 13.31µM and kinetin 13.93µM. At lower concentration of ADS was not effectively enhanced the M-M and M-R. From the results of all the experiments conducted for multiplication of cultures from axillary bud explants of Murraya koenigii, it was found that BAP 13.31µM and kinetin 13.93µM and ADS 81.44µM in MS semisolid medium was found to be most suitable for high frequency multiplication of shoots for M-M and M-R. During the experiments for shoot regeneration from axillary bud explant it was observed that on the media in which ADS was added in some of this shoot buds were developed from the inter nodal region of stem explants.
Keywords: Murraya koenigii, media, in vitro, axillary bud, Adenine sulphate, Aromatic plant.