In vitro mass production of Solanum surattense Burm. f. - A medicinal plant through seed culture Archana Pamulaparthi, Archana Kurra, Himabindu Kurra, Rakesh Basani and Rama Swamy Nanna*. Department of Biotechnology, Kakatiya University Warangal, Andhra Pradesh, India. swamynr.dr@gmail.com Abstract Aim: An efficient, reproducible protocol for multiple shoot induction and plantlet establishment was achieved through seed culture in Solanum surattense Burm. f. (Indian Solanum) an important medicinal herb. Methods: Seeds of S. surattense were germinated on MS (Murashige and Skoog’s) medium supplemented with different concentrations of BAP (6-Benzylaminopurine 0.5 - 3mg/L). Results: Maximum number of multiple shoot formation (31.83±0.53) was observed on MS medium containing 2.5 mg/L BAP. The highest percentage of germination was also recorded at the same concentration of BAP with an average number of 5 days for seed germination. When 2-4 shoots were subcultured on fresh medium containing 2.5 mg/L BAP, continual enhancement in shoot production was observed for every subculture. The microshoots were rooted on MS medium supplemented with 1.0mg/L IAA (Indole-3- Acetic Acid). The regenerants were acclimatized and the survival percentage was found to be 70% with normal flowering and fruiting. Conclusion: The present protocol can be used for in planta transformation using Agrobacterium tumefaciens to introduce gene(s) of interest. Key Words: Indian solanum, In vitro rooting, Kantakari, Multiple shoots, Solanum surattense, Seed germination. Abbreviations: BAP: 6-Benzylaminopurine; IAA: Indole-3- Acetic Acid; MS: Murashige and Skoog. Introduction Solanum surattense Burm. f. (syn. S.xanthocarpum Schrad and Wendl), commonly known as yellow berried nightshade, Indian solanum or kantakari belongs to family Solanaceae is a potent medicinal plant. The fruits of plant are used as vegetable apart from their use in the treatment of cough, asthma, rheumatism and chest pain [1]. A number of sterols [2], alkaloids [3,4] were reported in the plant. The plant is also known to possess antibacterial, antifungal [5], antioxidant[6], hypoglycemic[7] and larvicidal[8] activities. In view of its medicinal importance the species is being over exploited and there is a need to conserve the species using different in vitro techniques available. A number of regeneration studies from leaf [9], root [10] , anther [11], nodal and shoot tip explants[12,13] were reported. A protocol on plantlet regeneration through somatic embryogenesis from cotyledon and leaf explants and streptomycin resistant plantlets using in vitro mutagenesis was also developed in S.surattense [14,15]. Agrobacterium mediated genetic transformation using leaf explants of S.surattense was also reported by Rama Swamy[16]. However, there is no report on in vitro micropropagation using the seeds as an explant. We report in this communication on multiple shoots formation and plantlet establishment in S.surattense for the first time through seed culture. Materials and Methods Plant Material The seeds of S.surattense were collected from the plants growing in the research field, Department of Biotechnology, Kakatiya University, Warangal, India. The seeds were washed under running tap water for 30 minutes followed by soaking for 15-20 minutes in 2% (w/v) Bavistin and subsequently washed 5-6 times with sterile distilled water. These seeds were sterilized with 0.1% (w/v) HgCl 2 for 2-3 minutes followed by 3 rinses in sterile distilled water under aseptic conditions, blotted dry on sterile tissue paper and were inoculated on MS [17] medium. MS medium containing 3% (w/v) sucrose supplemented with various concentrations of N 6 - benzylaminopurine (BAP) and 0.8% (w/v) agar (Hi-Media, India) was used for seed germination and multiple shoot induction. A control was also maintained without plant growth regulators (MS basal). The P H of the media was adjusted to 5.7±0.1 either with 0.1N HCl or 0.1N NaOH before autoclaving at 121 o C for 15-20 minutes. Multiple shoot induction The explants with multiple shoots were transferred onto the fresh MS medium supplemented with 2.5 mg/L BAP. After 4 weeks of the transfer of the explant with proliferating shoots was divided into pieces Archana Pamulaparthi et al. / International Journal of Pharmaceutical and Biological Research (IJPBR) ISSN : 0976- 285X Vol 3 Issue 4 Aug-Sep 2012 176