Indian Journal of Biotechnology Vol 6, April 2007, pp 274-276 In vitro propagation of tikhur (Curcuma angustifolia Roxb.): A starch yielding plant S K Shukla*, Susmita Shukla, Vijaya Koche and S K Mishra PTC Laboratory, School of Life Sciences Pt Ravishankar Shukla University, Raipur 492 010, India Received 23 September 2005; revised 4 May 2006; accepted 25 July 2006 In vitro regeneration of Curcuma angustifolia Roxb. was achieved through shoot meristem culture. The shoot buds (2-3 cm long) from rhizome were inoculated on MS medium supplemented with 3.0 mg/L BAP for initiation and elongation of shoots. As a result, 1.87±0.28 shoots per explant were produced These shoots were transferred on MS medium supplemented with 3.0 mg/L BAP and 25 mg/L adenine sulfate for further shoot multiplication. About 6.9±0.69 micro-shoots per explant were produced with in 6 wk. The roots appeared from shoots on shoot establishment as well as multiplication media. The rooted plants were transferred to pots and acclimatized, which showed 83% survival with normal growth. Keywords: Curcuma angustifolia, meristem culture, micropropagation, shoot bud IPC Code: Int. Cl. 8 A01H4/00 Curcuma angustifolia Roxb. (Family: Zingiberaceae) produces edible rhizome rich in starch content 1 . The rhizome is processed to obtain tikhur, which is sold in the market. Tikhur flour is cooked and consumed in many parts of India 2 . The plant grows wild in its natural habitat and usually perpetuates through vegetative reproduction. Overexploitation has made tikhur scarce in natural habitat and costly in the market. Cultivation of superior clones would greatly enhance its production and quality. In nature, propagation of tikhur occurs through rhizome, which is a slow process. Tissue culture techniques offer opportunity for fast multiplication of superior clones in relatively small space and time. Tissue culture of some Curcuma spp. has been reported earlier. In vitro plantlet regeneration has been achieved in C. amada 3-5 , C. aromatica 6 , C. longa 7-11 , C. zedoaria 12 and Curcuma sp. 13,14 . However, the reports are lacking on in vitro propagation of C. angustifolia. Here, we report a protocol of this important species. The rhizomes of C. angustifolia were collected from natural forests of Pithora, about 100 km from Raipur, India. After washing, the rhizomes were planted on sand beds of a greenhouse in the month of July. The shoot buds appeared within 15-20 d. The sprouted rhizomes collected from sand beds were washed thoroughly in running tap water to remove sand particles. The shoot buds, 2-3 cm long, were excised from rhizome under aseptic condition, sterilized in 0.2% mercuric chloride for 15 min and rinsed 3-4 times in sterilized water. The outer leaves were removed from shoot buds and shoot meristems with remaining inner leaves were used as explants. These shoot bud explants were inoculated on establishment medium of Murashige and Skoog 15 (MS) medium supplemented with 0.0-5.0 mg/L BAP for initiation and elongation of shoots. After 6 wk, the elongated shoots along with original explants were placed on shoot multiplication medium of MS+3.0 mg/L BAP+0-100 mg/L adenine sulfate. All the media adjusted at pH 5.7 were sterilized at pressure 1.05 kg/c m 2 for 25 min. After inoculation, the cultures were placed in tissue culture room at 25±2 o C under 16/8 h (light/dark) photoperiod with 1000 lux light intensity. Regenerated plants were acclimatized in the greenhouse at 12 h light/dark regime, 70% relative humidity and 25-30 o C temperature. The optimum shoot bud initiation (80%), number of shoots (1.87±0.28) and shoot elongation (48±5.5) occurred in explants placed on MS medium supplemented with 3.0 mg/L BAP (Table 1). On this establishment medium, shoots also produced optimum number of leaves (2.7±0.35/shoot) having maximum width (18±1.7 mm/leaf) and roots (5.5±0.41/shoot). However, further increase in BAP concentration of medium had adverse effects. Poor growth at higher concentration of BAP and Kn was also reported in C. longa 7 . The shoots that grew from explants on MS+ 3.0 mg/L BAP were further used for shoot multiplication. The shoots placed on MS medium supplemented with 3.0 mg/L BAP and 25 mg/L adenine sulfate exhibited maximum shoot proliferation as well as root __________ *Author for correspondence: Tel.: 91-771-2262631 E-mail: shuklashivkant@yahoo.co.in