In Vitro Regeneration Potentiality of Potato under Different Hormonal Combination

The present investigation was conducted to develop a protocol for rapid plant regeneration in potato. The newly emerge sprout of six potato cultivars were used as explant. Varietal response on in vitro regeneration under different hormonal concentration and combination were studied. MS medium supplemented with 0.5, 1.0, 1.5, 2.0, 4.0 mg L of KIN and IAA in combination of both were employed for in vitro regeneration. Among the 1 different treatments the combination 2.0 mg L of KIN and IAA showed the best response to multiple shoot 1 and root regeneration. The same concentration also takes minimum time for regeneration. The interaction between hormonal concentration and varietal potentiality for all the parameter showed significant difference at 5% level of probability. The in vitro regeneration and multiplication potentiality was the highest in the variety Granulla followed by Cardinal and Diamont. Key word: Potato Indole acetic acid (IAA) Kinetin (KIN) Regeneration INTRODUCTION imported basis seeds. It also reduce 50% seed cost as Potato (Solanum tuberosum L.) is one of the most regeneration protocol was establish in many laboratories important vegetable crop in Bangladesh and as well in the world but still there are very less information as world contest. It has multipurpose use in daily regarding the comparative studies among the different consumption and also industrial purpose. Potato varieties of potato. Which genotypes is more efficient for propagated vegetatively by means of tuber. Potato rapid multiplication is need to identified. Need to asses production with seed tuber is constrained by the the regeneration potentiality of available release varieties accumulation of pathogen, physiological decline and of potato for better utilization in commercial purpose. low multiplication rates. Seed tuber is most expensive Growth regulators play an important role in potato input in potato production. Atlest 35-40% total cost of regeneration. Individual hormone has its on effect on potato production is covered by seed tuber. The above regeneration. Combine effect of rooting and shooting major constrains can be over come by using tissue culture growth regulator is need to study for better regeneration technique for rapid multiplication of plantlet production. of potato. Therefore, the present investigation was carried Successful in vitro plant regeneration has been achieved out to identify the best hormonal combination and from different explant like leaf and stem [1]. In vitro concentration for potato regeneration. As well as to produced disease free potato clones combined with detect the best response to regeneration among the conventional multiplication methods has become an different varieties of potato. integral part of seed production in many countries [2]. Meristem culture derived micro propagated potato MATERIALS AND METHODS plantlets are free from the risk of speading bacteria, fungi and insect transmitted virus. In vitro regeneration The present study was carried out at the Department technique is adopted commercially in many developing of Biotechnology, Sher-e-Bangla Agricultural University, countries. The seed potato produced by tissue culture Sher-e-Bangla Nagar, Dhaka, Bangladesh, during the method proved better performance as compare to period September, 2008 to June, 2009. The experimental composed to imported potato [3]. Although potato World J. Agric. Sci., 6 (6): 660-663, 2010 661 materials (six potato varieties) were collected from Tuber Crops Research Center (TCRC), Bangladesh Agricultural Research Institute (BARI), Gazipur-1701, Bangladesh. The cultivars are as followsDiamant, Cardinal, Granulla, Ultra, Dheera and Provinto. Disease-free healthy potato tuber were treated with 50 ppm GA for sprouting. The sprouted 3 buds were used as explant. The experiment was design on two factor factorial condition. The different hormonal concentration was treated as factor 1 and the different varieties of potato were used as factor 2. The experiment was laid out in Completely Randomized Design (CRD) with 3 replications. Ten explants were considered in each treatment and in each varieties. The sprouts were surface sterilized by immersing in 0.2% HgCl solution for 2-3 min 2 and then washed several times with sterile distilled water. The explant (sprouts) were cultured in MS semi-solid medium (pH 5.8) on different culture vasel. The medium were autoclaved at 121°C with 1.16 kg cm pressure for -2 25 minute. The treatment concentration of Kinetin (KIN) and Indole Acetic Acid (IAA) were viz. 0.5, 1.0, 1.5, 2.0 and 4.0 mg L . The concentration were applied in 1 combination of both the hormone. The cultures were incubated at 25±2°C under white light intensity (2500-3000 Lux). Data were recorded on the following parameters, days to shoot regeneration, number of shoot per explant, days to root induction, number of root per explant. Two factor factorial analysis of variance were performed and mean were compared by the Duncan’s Multiple Range Test (DMRT). RESULTS AND DISCUSSION The six potato varieties viz. Diamant, Cardinal, Granulla, Ultra, Dheera and Provinto were cultured on MS medium supplemented with different concentration of KIN in combination with IAA. Data were analyzed on different parameters. The results of each treatment and varietal interaction were presented below under the sub-handing. Effects on Shoot Regeneration: It was observed that the treatment 2.0 mg L of both IAA + KIN required 1 minimum time (4.28 days) for shoot regeneration which was statistically significant from rest of the treatment. The maximum time (8.07 days) required in simple MS media. The variety Ultra take less time (2.66 days) for regeneration in the same concentration (Table 1). Average varietal response on days to shoot regeneration was the lowest in case of Ultra (5.41 days) followed by Granulla, Cardinal and Dheera. The variety Provinto take highest time (6.26 days) which is statistically similar with Diamant, Cardinal and Dheera. Fig. 1: Multiple shooting of regenerated potato Fig. 2: Multiple rooting of regenerated potato Effect on Multiple Shooting: Huge number of shoot was regenerated due to application of both shooting and rooting hormone. It means that, the rooting hormone enhance the shooting ability in potato genotypes. The variety Granulla produced the maximum number (14.07) of shooting in the concentration of 2.0 mgL hormone for 1 both IAA and KIN. Average number of shoot also higher in the same treatment, which is statistically different from rest of the treatment. It was noticed that, in higher concentration of both the hormone the number of shoot World J. Agric. Sci., 6 (6): 660-663, 2010 662 Table 1: Combined effect of IAA + KIN on days to shoot regeneration in different potato varieties Varieties -------------------------------------------------------------------------------------------------Treatments IAA + KIN (mg L ) Diamant Cardinal Granulla Ultra Dheera Provinto Treatment mean and DMRT ranking 1 T : Simple MS 8.73 8.40 7.83 7.93 8.00 7.53 8.07a 1 T : 0.5 + 0.5 7.73 7.93 6.83 6.86 7.73 7.86 7.49b 2 T : 1.0 + 1.0 6.73 7.23 5.86 5.90 6.93 6.93 6.60c 3 T : 1.5 + 1.5 5.80 5.40 5.33 5.40 5.80 5.50 5.53d 4 T : 2.0 + 2.0 2.83 3.40 2.83 2.66 3.46 4.50 4.28f 5 T : 4.0 + 4.0 5.16 4.20 3.93 3.73 4.66 5.26 4.49e 6 Variety mean and DMRT ranking 6.16a 6.09a 5.43b 5.41b 6.10a 6.26a * Significant at 5% level Table 2: Combined effect of IAA + KIN on multiple shooting in potato varieties Varieties -------------------------------------------------------------------------------------------------Treatments IAA + KIN (mg L ) Diamant Cardinal Granulla Ultra Dheera Provinto Treatment mean and DMRT ranking 1 T : Simple MS 1.73 1.66 2.86 0.96 1.26 1.33 1.63e 1 T : 0.5 + 0.5 4.06 5.86 6.93 4.40 6.00 4.86 5.35d 2 T : 1.0 + 1.0 4.93 5.80 9.46 5.86 7.73 5.00 6.46c 3 T : 1.5 + 1.5 5.86 11.07 11.53 7.40 9.86 7.53 8.87b 4 T : 2.0 + 2.0 11.40 11.53 14.07 9.86 10.40 9.73 11.17a 5 T : 4.0 + 4.0 10.83 9.60 10.70 6.73 9.20 7.53 9.10b 6 Variety mean and DMRT ranking 6.47c 7.58b 9.26a 5.87d 7.41b 6.00d * Significant at 5% level Table 3: Combined effect IAA + KIN on days to root induction in potato varieties Varieties -------------------------------------------------------------------------------------------------Treatments IAA + KIN (mg L ) Diamant Cardinal Granulla Ultra Dheera Provinto Treatment mean and DMRT ranking 1 T : Simple MS 16.07 14.00 11.40 15.33 14.87 15.40 14.51a 1 T : 0.5 + 0.5 12.00 11.73 9.80 11.67 12.07 12.47 11.62b 2 T : 1.0 + 1.0 8.60 9.86 8.53 10.13 10.75 10.20 9.67c 3 T : 1.5 + 1.5 7.73 7.80 7.40 8.73 8.86 8.40 8.15e 4 T : 2.0 + 2.0 7.00 7.06 7.00 7.80 7.40 7.46 7.28f 5 T : 4.0 + 4.0 8.06 7.86 8.06 8.00 9.00 9.73 8.48d 6 Variety mean and DMRT ranking 9.91c 9.72c 8.70d 10.28b 10.49ab 10.64a * Significant at 5% level Table 4: Combined effect of IAA + KIN on multiple rooting in potato varieties Varieties -------------------------------------------------------------------------------------------------Treatments IAA + KIN (mg L ) Diamant Cardinal Granulla Ultra Dheera Provinto Treatment mean and DMRT ranking 1 T : Simple MS 1.73 1.20 2.00 0.86 0.73 1.13 1.27f 1 T : 0.5 + 0.5 9.73 8.06 10.73 7.06 8.20 6.46 8.37e 2 T : 1.0 + 1.0 9.86 9.46 10.87 8.60 9.73 8.06 9.43d 3 T : 1.5 + 1.5 11.00 11.07 11.80 10.07 10.80 10.03 10.79c 4 T : 2.0 + 2.0 15.07 14.20 16.20 11.47 11.37 11.13 13.30a 5 T : 4.0 + 4.0 12.03 14.40 12.73 10.00 9.66 9.80 11.44b 6 Variety mean and DMRT ranking 9.90b 9.73b 10.72a 8.01d 8.47d 7.77d * Significant at 5% level World J. Agric. Sci., 6 (6): 660-663, 2010 663 regeneration was decreased. Its mean that, higher higher in combination of auxin group (IAA). Rapid shoot concentration of hormone has negative effect of rootregeneration and root induction was achieved when the multiplication. Among the variety Granulla response explant supplemented with 2.0 mg L of KIN and IAA. well in all the treatment. It was followed by Cardinal and Dheera (Table 2). Hardening Efficiency: The fully developed plantlet were Effect of Root Induction: It reveals that, 2.0 mg L of KIN sterilized soil in the pot. Fifty plantlet were transferred1 and IAA takes less time (7.28 days) for root induction for all the variety. Among the different genotypeson average treatment combination. The highest time highest percent (86.01%) of plantlet in case of Granulla(14.51 days) was recorded in case of control condition was survive in the soil and establish as healthy seeding.(Table 3). The minimum days required (8.70 days) for Lowest percentage (39.48%) of hardening efficiency wasroot induction in the variety Granulla and followed by recorded in case of Provinto followed by Dheera (48.03%)Cardinal and Diamant and the entry Provinto takes and Ultra (52.19%).maximum days (10.64) for root induction in average ofall the treatments. It is interesting to note that, all theREFERENCEScultivar showed minimum time for root induction onthe concentration of 2.0 mg L of hormone application. 1. Cearley, T.A. and M.G. Bolyard, 1997. Regeneration1 Higher concentration of hormone showed negativeof Solanum tuberosum cv. katahdin from leafeffect on root induction.explants in vitro. An Potato J., 74: 125-129. Effect on Multiple Rooting: The maximum numberand conservation of genetic resources in potato. In:(13.30) root was initiated in the concentration of 2.0K.L. Chadha, P.N. Ravindran and S. Leela (eds)mg L in both IAA + KIN. 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