EFFECT OF DIFFERENT PLANT HORMONES ON NODE CULTURE IN POINTED GOURD

Journal of Cell and Tissue Research Vol. 14(2) 4323-4331 (2014) (Available online at www.tcrjournals.com) ISSN: 0973-0028; E-ISSN: 0974-0910 Original Article EFFECT OF DIFFERENT PLANT HORMONES ON NODE CULTURE IN POINTED GOURD AHMAD, T., KUMAR, V., PATEL, N. L., KAPADIA C. V., SUTHAR, K. P. AND SINGH, D.? Department of Plant Molecular Biology and Biotechnology, ASPEE College of Horticulture and Forestry, Navsari Agricultural University, Navsari 396450, Gujarat. E mail: chintan_bt@yahoo.co.in Received: April 30, 2014; Revised: May 15, 2014; Accepted: May 21, 2014 Abstract: In India pointed gourd (Trichosanthes dioca Roxb.) is extensively cultivated in eastern Uttar Pradesh, Bihar and West Bengal and to some extent in Assam, Orissa, Madhya Pradesh, Maharashtra and Gujarat. Propagation of pointed gourd from seed is not desirable due to poor germination as well as dioecism which results in 50% unproductive male plants. Traditionally pointed gourd is multiplied through stem cuttings and root suckers. For planting one hectare land, 6,000-7,500 cuttings are required, however cuttings are not easily available and therefore tissue culture offers a promise for the bulk supply of planting material which can be transplanted in a proper sex ratio in the field. Effect of various treatments of plant hormones supplemented in MS medium was examined for node culture in four genotypes of pointed gourd viz. Calcutti Female, Calcutti Male, Hybrid Female, and Hybrid Male. Four different parameters viz. (i) per cent establishment, (ii) number of shoots per culture, (iii) shoot length and (iv) number of nodes per culture was studied. The medium supplemented with 1.0 mg/l IAA + 0.2 mg/l BA showed longest shoot with better and quicker establishment and little or no root formation in all the genotypes of pointed gourd. This was found to be best culture establishment medium. Among different genotypes, in vitro response of Calcutti Female was best followed by Hybrid Female, Calcutti Male and Hybrid Male. Of the various treatment of plant hormones examined in MS medium, the same treatment 1.0 mg/l IAA + 0.2 mg/l BA was found to be best shoot proliferation medium for continuous sub culturing. Rooting of in vitro shoots in pointed gourd was not a problem as rooting occurred in many treatments during shoot establishment itself. Half strength MS medium supplemented with 0.1mg/l IAA + 0.2 mg/l IBA was found most effective for rooting of in vitro raised shoots of pointed gourd as it gave highest number of longest root with maximum frequency. Key words: Node culture, pointed gourd, Trichosanthes dioca Roxb. INTRODUCTION Among cucurbits, pointed gourd (Trichosanthes dioca Roxb.) known as parwal, patal, palwal or parmal is an important vegetable crop and becoming more popular now a days. Few years back it was a neglected vegetable, however, due to its high medicinal value it is becoming popular day by day not only in the Gujarat. but also in country. Its cultivation is highly profitable [1]. It is an under exploited important summer cucurbit vegetable. The Bengal and Assam region of India is the primary centre of its origin. It is cultivated in subtropical and tropical regions around the globe. It is extensively cultivated in eastern Uttar Pradesh, Bihar and West Bengal and to some extent in Assam, Orissa, Madhya Pradesh, Maharashtra and Gujarat. In the diara lands (lands prone to submergence by flood during the wet 4323

J. Cell Tissue Research season) of Bihar and parts of eastern Uttar Pradesh, the crop is rainfed. Propagation of pointed gourd from seed is not desirable due to poor germination as well as dioecism which results in around 50% unproductive male plants [2]. Traditionally pointed gourd is multiplied through stem cuttings and root suckers. For planting one hectare land, 6,000-7,500 cuttings are required which are not easily available. Tissue culture offers a promise for the supply of planting material which can be raised in a proper sex ratio in the field. MATERIALS AND METHODS The most widely accepted MS medium [3] was used as basal medium supplemented with 30g/l sucrose and solidified with 8 g/l agar. Different concentrations and combinations of plant hormones viz. BA, Kinetin NAA, IAA, IBA and Adenine sulphate were incorporated in the basal medium. Fourteen different treatments were tried to standardize establishment and shoot proliferation medium for node culture of four pointed gourd genotypes viz. Calcutti female (CF), Calcutti male (CM), Hybrid Female (HM) and Hybrid male (HM) as shown in Table 1.The ph of the medium was adjusted at 5.8. After ph of the media had been adjusted, the known volumes of media were distributed in culture vessels (20 to 25 ml). The mouth of Mc Cartney bottles were tightly closed with aluminium caps, and that of rimless test tubes were closed with polypropylene caps and plugs made of non-absorbent cotton wrapped in gauze cloth. The cotton plugs on the mouth of test tubes were further wrapped with double layered ordinary paper to protect them from condensing water vapour. The culture vessels were then subjected to wet sterilization using autoclave. Autoclaving was carried out at a pressure of 15 lb/inch 2 at 121 0 C temperature for 20 minutes after which the culture vessels containing autoclaved media were transferred to an air-conditioned room. They were stored for a minimum period of 4 days before use. Explants were collected from male and female lines of two local cultivars of pointed gourd viz. Calcutti and Hybrid grown in experimental farm of Agriculture Experiment Station, Paria. These were considered as four genotypes/lines namely (a) Calcutti Female (b) Calcutti Male (c) Hybrid Female and (d) Hybrid Male. To control endophytic contamination of nodal segment, 200 mg/l carbendazim + 200 mg/l streptocyclin treatment was given for 1 hour. Endophytic contamination control was followed by surface sterilization. As far as surface sterilization is concerned, 0.1% mercuric chloride treatment for 6 minutes was given to nodal segments. All the cultures were incubated in a culture room at a temperature of 25 ±2 o C with relative humidity at 55 ± 5 per cent and were exposed to 16 hr photoperiod of approximately 2000 lux/m 2 light by 40W cool-white fluorescent tubes kept 15 inches above bench surface. RESULTS Effect of fourteen different concentrations and combinations of plant hormones supplemented in MS medium was examined on four genotypes of pointed gourd. Four different parameters viz. (i) per cent establishment, (ii) number of shoots per culture, (iii) shoot length and (iv) number of nodes per culture were studied (Fig.1 A-N). i) Effect of different treatments on per cent establishment: The data in Table 1 showed that among different treatments significantly maximum mean establishment (93.33 %) was recorded on treatment F 12 (1.0 mg/l IAA + 0.2 mg/l BA), which Explanation of figures: plates 1,2,3 Fig.1: Response of nodal segments of pointed gourd genotypes (a:-calcutti Female, b:- Calcutti Male, c:- Hybrid Female d:- Hybrid Male) on MS medium supplemented with different concentrations and combinations of plant hormones on shoot establishment. A) Treatment F 1 [NAA (0.2mg/l) + BA (1.0 mg/l)], B) Treatment F 2 [NAA (0.2 mg/l) + BA (2.0 mg/l)], C) Treatment F 3 [NAA (1.0 mg/l) + BA (1.0 mg/)], D) Treatment F 4 [NAA (1.0 mg/l) + BA (0.2 mg/l)], E) Treatment F 5 [NAA (0.2 mg/l) + BA (2.0) + AS (10.0 mg/l)], F), Treatment F 6 [NAA (0.2 mg/l) + BA (2.0 mg/l) + AS 40.0 mg/l)], G) Treatment F 7 [NAA (0.2 mg/l) + K (2.0 mg/l)], H) Treatment F 8 [BA (2.0 mg/l)], I) Treatment F 9 [BA (3.0 mg/l)], J) Treatment F 10 [BA (4.0 mg/l)], K) Treatment F 11 [IAA (0.2 mg/l) + BA (2.0 mg/l)], L) Treatment F 12 [IAA (1.0 mg/l) + BA (0.2 mg/l)], M) Treatment F 13 [IAA (0.2 mg/l) + IBA (0.04 mg/l)], N) Treatment F 14 [IAA (0.3 mg/l) + IBA (0.06 mg/l)] 4324

Ahmad et al. A B C D E F Plate 1 4325

J. Cell Tissue Research Table 1: Effect of different plant hormones concentrations and combinations on per cent culture establishment in node culture of pointed gourd genotypes. Figures in parentheses are original values No. Treatment (mg/l) Genotypes Calcutti Female Calcutti Male Hybrid Female Hybrid Male Mean F 1 NAA 0.2 + BA 1.0 66.81 (83.33) 58.98 (73.33) 61.12 (76.67) 55.95 (68.33) 60.71 (75.41) F 2 NAA 0.2 + BA 2.0 60.05 (75.00) 53.74 (65.00) 58.91 (73.33) 53.74 (65.00) 56.61 (69.58) F 3 NAA 1.0 + BA 1.0 71.92 (90.00) 64.97 (81.67) 67.38 (85.00) 61.12 (76.67) 66.35 (83.33) F 4 NAA 1.0 + BA 0.2 69.21 (86.67) 61.12 (76.67) 64.97 (81.67) 57.84 (71.67) 63.29 (79.17) F 5 NAA 0.2 + BA 2.0+ AS 10 56.82 (70.00) 49.87 (58.33) 54.81 (66.67) 47.86 (55.00) 52.34 (62.50) F 6 NAA 0.2 + BA 2.0+ AS 40 55.95 (68.33) 48.85 (56.67) 57.84 (71.67) 50.77 (60.00) 53.35 (64.17) F 7 NAA 0.2 + K 2.0 46.93 (53.33) 39.19 (40.00) 0.64 (00.00) 0.64 (00.00) 21.85 (46.66) F 8 BA 2.0 62.76 (78.33) 55.95 (68.33) 63.52 (80.00) 56.82 (70.00) 59.76 (74.16) F 9 BA 3.0 50.77 (60.00) 44.02 (48.33) 49.87 (58.33) 43.06 (46.67) 46.93 (53.33) F 10 BA 4.0 48.85 (56.67) 41.11 (43.33) 51.79 (61.67) 44.02 (48.33) 46.44 (52.50) F 11 IAA 0.2 + BA 2.0 52.86 (63.33) 45.95 (51.67) 53.74 (65.00) 46.93 (53.33) 49.87 (58.33) F 12 IAA 1.0 + BA 0.2 85.23 (98.33) 71.92 (90.00) 81.14 (96.67) 70.09 (88.33) 77.90 (93.33) F 13 IAA0.2+IBA0.04 81.14 (96.67) 70.09 (88.33) 79.30 (95.00) 69.21 (86.67) 75.74 (91.66) F 14 IAA0.3+IBA0.06 77.46 (93.33) 67.38 (85.00) 70.09 (88.33) 63.52 (80.00) 69.61 (86.66) Mean 63.34 (76.66) 55.45 (66.19) 58.22 (71.43) 51.77 (62.14) Source S.Em. C.D.at 5% Treatment 1.6525 4.6337 Genotype 0.8833 2.4768 C.V.% =10.01 Treatment X Genotype 3.3050 9.2674 Table 2: Effect of different plant hormones concentrations and combinations on number of shoots per culture in node culture of pointed gourd genotypes. Figures in parentheses are original values No. Treatment (mg/l) Genotypes Calcutti Female Calcutti Male Hybrid Female Hybrid Male Mean F 1 NAA 0.2 + BA 1.0 1.49 (1.73) 1.31 (1.23) 1.46 (1.63) 1.28 (1.13) 1.38 (1.43) F 2 NAA 0.2 + BA 2.0 1.69 (2.37) 1.36 (1.37) 1.62 (2.13) 1.31 (1.23) 1.50 (1.77) F 3 NAA 1.0 + BA 1.0 1.22 (1.00) 1.22 (1.00) 1.22 (1.00) 1.22 (1.00) 1.22 (1.00) F 4 NAA 1.0 + BA 0.2 1.22 (1.00) 1.22 (1.00) 1.22 (1.00) 1.22 (1.00) 1.22 (1.00) F 5 NAA 0.2 + BA 2.0+ AS 10 1.91 (3.17) 1.71 (2.43) 1.83 (2.87) 1.76 (2.63) 1.81 (2.77) F 6 NAA 0.2 + BA 2.0+ AS 40 2.13 (4.07) 1.81 (2.77) 1.83 (2.87) 1.50 (1.77) 1.82 (2.87) F 7 NAA 0.2 + K 2.0 1.22 (1.00) 1.22 (1.00) 0.71 (0.00) 0.70 (0.00) 0.96 (0.50) F 8 BA 2.0 2.08 (3.83) 1.72 (2.47) 2.05 (3.70) 1.59 (2.03) 1.86 (3.01) F 9 BA 3.0 1.74 (2.53) 1.57 (1.97) 1.62 (2.13) 1.52 (1.83) 1.61 (2.11) F 10 BA 4.0 2.22 (4.43) 1.77 (2.63) 1.82 (2.83) 1.68 (2.33) 1.87 (3.05) F 11 IAA 0.2 + BA 2.0 2.13 (4.03) 2.04 (3.67) 2.08 (3.83) 1.77 (2.63) 2.00 (3.54) F 12 IAA 1.0 + BA 0.2 1.39 (1.43) 1.22 (1.00) 1.34 (1.30) 1.28 (1.13) 1.31 (1.21) F 13 IAA0.2+IBA0.04 1.24 (1.03) 1.24 (1.03) 1.22 (1.00) 1.25 (1.07) 1.24 (1.03) F 14 IAA0.3+IBA0.06 1.25 (1.07) 1.26 (1.10) 1.24 (1.03) 1.25 (1.07) 1.25 (1.07) Mean 1.64 (2.33) 1.48 (1.76) 1.52 (1.95) 1.38 (1.49) Source S.Em. C.D. at 5% Treatment 0.0208 0.0582 Genotype 0.0111 0.0311 C.V.% = 4.78 Plant hormones X Genotype 0.415 0.1164 was at par with treatment F 13 (0.2 mg/l IAA + 0.04 mg/l IBA) which recorded 91.66 % mean establishment. Among genotypes, significantly maximum mean establishment was found in Calcutti Female (76.66 %) followed by Hybrid Female (71.43 %), Calcutti Male (66.19%) and Hybrid Male (62.14 %). In Calcutti Female genotype, significantly maximum establishment (98.33%) was observed on treatment F 12 followed by 96.67% on treatment F 13 and 93.33 % on treatment F 14 (0.3 mg/l IAA+0.06 mg/l IBA). In Calcutti Male genotype, significantly maximum establishment (90.00 %) was recorded on treatment F 12 followed by 88.33 % on treatment F 13 and 85.00 % on treatment F 14. In Hybrid Female genotype, significantly maximum establishment (96.67 %) was achieved on treatment F 12 followed by 95.00 % on treatment F 13. In Hybrid Male genotype significantly maximum establishment (88.33 %) was achieved on treatment F 12 which was at par (86.67 %) with treatment F 13. Over all best performance in all the genotypes was obtained on MS medium supplemented with1.0 mg/l IAA + 0.2 mg/l BA (F 12 ) as far as establishment of shoot was concerned. ii) Effect of different treatments on number of shoots: Out of fourteen plant hormones combinations tried to see their effect on number of shoots during node culture in four pointed gourd genotypes, the treatment F 11 (0.2 mg/l IAA + 2.0 mg/l BA) showed maximum mean number of shoots (3.54) (Table 2) 4326

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J. Cell Tissue Research Table 3: Effect of different plant hormones concentrations and combinations on shoot length in node culture of pointed gourd genotypes. Figures in parentheses are original Sr. No. Treatment (mg/l) Calcutti Female Genotypes (cm) Calcutti Male Hybrid Female Hybrid Male F 1 NAA 0.2 + BA 1.0 2.46 (5.55) 2.22 (4.43) 2.35 (5.05) 2.09 (3.86) 2.28 (4.72) F 2 NAA 0.2 + BA 2.0 1.77 (2.63) 1.71 (2.43) 1.73 (2.51) 1.69 (2.36) 1.73 (2.48) F 3 NAA 1.0 + BA 1.0 3.16 (9.51) 2.99 (8.46) 3.13 (9.32) 2.63 (6.43) 2.98 (8.43) F 4 NAA 1.0 + BA 0.2 2.76 (7.13) 2.43 (5.41) 2.57 (6.13) 2.19 (4.32) 2.49 (5.75) F 5 NAA 0.2 + BA 2.0+ AS 10 1.74 (2.53) 1.62 (2.14) 1.75 (2.58) 1.59 (2.03) 1.68 (2.32) F 6 NAA 0.2 + BA 2.0+ AS 40 1.48 (1.71) 1.42 (1.54) 1.46 (1.64) 1.31 (1.23) 1.42 (1.53) F 7 NAA 0.2 + K 2.0 1.28 (1.13) 1.24 (1.04) 0.71 (0.00) 0.71 (0.00) 0.98 (0.54) F 8 BA 2.0 2.03 (3.63) 1.93 (3.24) 1.52 (1.82) 1.49 (1.72) 1.74 (2.60) F 9 BA 3.0 1.24 (1.05) 1.23 (1.02) 1.24 (1.03) 1.20 (0.95) 1.23 (1.01) F 10 BA 4.0 1.16 (0.85) 1.10 (0.73) 1.14 (0.81) 1.06 (0.62) 1.12 (0.75) F 11 IAA 0.2 + BA 2.0 1.45 (1.61) 1.35 (1.32) 1.39 (1.43) 1.27 (1.11) 1.37 (1.37) F 12 IAA 1.0 + BA 0.2 3.19 (9.67) 2.85 (7.62) 3.13 (9.32) 2.84 (7.56) 3.00 (8.54) F 13 IAA0.2+IBA0.04 3.20 (9.75) 2.66 (6.58) 3.02 (8.67) 2.58 (6.18) 2.87 (7.79) F 14 IAA0.3+IBA0.06 3.18 (9.64) 2.83 (7.51) 2.98 (8.41) 2.26 (4.61) 2.83 (7.54) Mean 2.29 (4.74) 2.07 (3.82) 2.17 (4.20) 1.88 (3.07) Source S.Em. C.D. at 5% Treatment 0.0166 0.0464 Genotype 0.0089 0.0249 C.V.% = 2.90 Treatment X Genotype 0..0332 0.0929 Table 4: Effect of different plant hormones concentrations and combinations on number of nodes in node culture of pointed gourd genotypes. Figures in parentheses are original No. Treatment (mg/l) Genotypes Calcutti Female Calcutti Male Hybrid Female Hybrid Male Mean F 1 NAA 0.2 + BA 1.0 3.27 (10.23) 2.78 (7.27) 2.66 ( 6.57) 2.42 (5.37) 2.78 ( 7.36) F 2 NAA 0.2 + BA 2.0 3.12 ( 9.23) 2.74 (7.07) 2.62 ( 6.37) 2.56 (6.07) 2.76 ( 7.18) F 3 NAA 1.0 + BA 1.0 3.41 (11.13) 2.55 (6.00) 2.98 ( 8.40) 2.52 (5.87) 2.86 ( 7.85) F 4 NAA 1.0 + BA 0.2 2.66 ( 6.57) 2.24 (4.53) 2.57 ( 6.17) 2.03 (3.63) 2.38 ( 5.22) F 5 NAA 0.2 + BA 2.0+ AS 10 2.91 ( 7.97) 2.56 (6.07) 2.77 ( 7.17) 2.66 (6.57) 2.72 ( 6.94) F 6 NAA 0.2 + BA 2.0+ AS 40 3.26 (10.17) 2.99 (8.43) 3.05 ( 8.83) 2.04 (3.67) 2.83 ( 7.77) F 7 NAA 0.2 + K 2.0 1.59 ( 2.03) 1.46 (1.63) 0.71 ( 0.00) 0.71 (0.00) 1.11 ( 0.91) F 8 BA 2.0 3.97 (15.33) 2.59 (6.23) 3.40 (11.13) 2.36 (5.07) 3.08 ( 9.44) F 9 BA 3.0 2.36 ( 5.07) 2.15 (4.13) 2.20 ( 4.33) 1.68 (2.33) 2.10 ( 3.96) F 10 BA 4.0 2.23 ( 4.47) 1.72 (2.47) 1.79 ( 2.73) 1.62 (2.13) 1.54 ( 2.95) F 11 IAA 0.2 + BA 2.0 3.54 (12.07) 3.39 (11.07) 3.46 (11.47) 2.88 (7.80) 3.32 (10.60) F 12 IAA 1.0 + BA 0.2 3.58 (12.33) 3.10 (9.13) 3.44 (11.37) 3.00 (8.53) 3.28 (10.34) F 13 IAA0.2+IBA0.04 3.18 ( 9.63) 2.50 (5.77) 2.76 ( 7.13) 2.48 (5.63) 2.73 ( 7.04) F 14 IAA0.3+IBA0.06 2.60 ( 6.27) 2.31 (4.83) 2.50 ( 5.77) 2.35 (5.03) 2.44 ( 5.47) Mean 2.98 ( 8.75) 2.51 (6.04) 2.64 ( 6.96) 2.24 (4.83) Source S.Em. C.D.at 5% Treatment 0.0368 0.1031 Genotype 0.0197 0.0551 C.V.% = 4.92 Treatment X Genotype 0.0735 0.2062 Mean (Fig.1K). This treatment recorded significantly highest number of shoots in three pointed gourd genotypes viz. Hybrid Female (3.83), Calcutti Male (3.67) and Hybrid Male (2.63). On the other hand, in case of Calcutti Female genotype, maximum number of shoots (4.43) was seen on the treatment F 10 (4.0 mg/l BA) (Fig.1J). Among genotypes, significantly maximum mean number of shoots was found in Calcutti Female (2.33) followed by Hybrid Female (1.95), Calcutti Male (1.76) and Hybrid Male (1.49). iii) Effect of different treatments on shoot length: Among various treatments tested, significantly maximum mean shoot length (8.54 cm) was observed on the treatment F 12 (1.0 mg/l IAA + 0.2 mg/l BA) (Table 3) (Fig.1 L). Among genotypes, significantly maximum mean length of shoots was found in Calcutti Female (4.74 cm) followed by Hybrid Female (4.20) cm), Calcutti Male (3.82 cm) and Hybrid Male (3.07 cm). In the genotype Calcutti Female longest shoot (9.75 cm) was seen on the treatment F 13 (0.2 mg/l IAA + 0.04 mg/l IBA) (Fig.1 M) which was at par with treatment F 12 (1.0 mg/l IAA + 0.2 mg/l BA) and F 14 (0.3 mg/l IAA + 0.06 mg/l IBA) which recorded 9.67 and 9.64 cm long shoot, respectively. The treatment F 12 gave longest 4328

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J. Cell Tissue Research shoot in other three genotypes viz. Hybrid Female (9.32 cm), Calcutti Male (7.62 cm) and Hybrid Male (7.56 cm). iv) Effect of different treatments on number of nodes per culture: Significantly maximum mean number of nodes per culture (10.60) was seen on the treatment F 11 (0.2 mg/l IAA + 2.0 mg/l BA) followed by the treatment F 12 (1.0 mg/l IAA + 0.2 mg/l BA) which showed 10.34 mean number of nodes (Table 4). The treatment F 11 gave maximum number of nodes per culture in pointed gourd genotypes viz. Hybrid Female (11.47) followed by Calcutti Male (11.07) and Hybrid Male (7.80), (Fig.1 L) whereas in case of genotype Calcutti Female maximum number of nodes (15.33) was observed in the treatment F 8 (2.0 mg/l BA) (Fig.1 H). Among genotypes maximum mean number of nodes per culture was achieved in Calcutti Female (8.75) followed by Hybrid Female (6.96), Calcutti Male (6.04) and Hybrid Male (4.83). Considering the above four parameters it can be concluded that the medium supplemented with 1.0 mg/l IAA + 0.2 mg/l BA showed longest shoot with better and quicker establishment and little or no root formation in all the genotypes of pointed gourd. This was found to be best culture establishment medium. Among different genotypes, in vitro response of Calcutti Female was best followed by Hybrid Female, Calcutti Male and Hybrid Male. Of the various treatment of plant hormones examined in MS medium, the same treatment 1.0 mg/l IAA + 0.2 mg/l BA was found to be best shoot proliferation medium for continuous sub culturing. Rooting of in vitro shoots in pointed gourd was not a problem as rooting occurred in many treatments during shoot establishment itself. Half strength MS medium supplemented with 0.1mg/l IAA + 0.2 mg/l IBA was found most effective for rooting of in vitro raised shoots of pointed gourd as it gave highest number of longest root with maximum frequency. DISCUSSION Propagation of pointed gourd is not feasible using seeds due to poor germination and dioecism which results in 50% unproductive male plants. Traditionally it is propagated through stem cuttings and root suckers which are a labour intensive technique and require bulk amount of vines and roots, restricting their multiplication at commercial level [4-6] and [7]. Therefore, in vitro propagated pointed gourd provides excellent advantage over traditional propag-ation with a very high multiplication rate followed by year round availability of disease free planting material.nodal segments of four pointed gourd genot-ypes showed varied response to shoot establishment and growth on MS medium supplemented with different concentrations and combinations of BA, K, NAA, IAA, IBA or adenine sulphate. In the present investigation, medium supplemented with 1.0 mg/l IAA + 0.2 mg/l BA recorded longest shoot in all the genotypes of pointed gourd (Fig.1 L).The medium supplemented with 0.2 mg/l IAA+0.04 mg/l IBA gave long shoots in cv. Calcutti Female. There is report on regeneration of longer shoots from nodal segments of pointed gourd on MS medium supplemented with IAA (0.175 mg/l) and IBA (0.04 mg/l) [5]. In pointed gourd, IAA appeared to play an important role in longer shoot regeneration from nodal segments. However, IAA along with BA gave lesser root formation, but more roots were regenerated when IAA was supplemented along with IBA. In the present study, maximum number of shoots and nodes per culture were recovered on medium supplemented with 0.2 mg/l IAA + 2.0 mg/l BA, however shoots were smaller in this plant hormones combination. The medium containing 2.0 mg/l BA recorded highest number of shoots. In cucurbitaceous taxa, different levels of BA was found effective to induce optimum shoot regeneration from nodal segments viz. 2.0 mg/l in pointed gourd [8], 8.0 mg/ l in musk melon [9], 0.225 mg/l in cucumber [10] and 0.5 mg/l in Trichosanthes cucumerina [11]. The response of medium fortified with different concentrations of NAA + BA appeared poor for the establishment and growth of shoots from nodal explants in the present investigation. In oriental melon and muskmelon (Cucumis melo) addition of NAA in the medium containing 0.2-1.0 mg/l BA retarded shoot growth from nodal segments [12]. Surprisingly shoot establishment was not obtained in the medium fortified with 0.2 mg/l NAA+2.0 mg/l Kinetin in Hybrid Female and Hybrid Male genotypes as compared to Calcutti genotypes which showed smaller shoots. This plant hormone combination i.e. NAA+ kinetin appeared to be highly sensitive to pointed gourd genotypes for shoot establishment., whereas NAA+BA combination did not show such a behavior. Further, kinetin did not appear specific for male or female genotype. BA 4330

Ahmad et al. was a more effective cytokinin than kinetin for shoot formation from nodal segments of cucumber [10]. In view of above discussion, MS medium supplemented with 1.0 mg/l IAA + 0.2 mg/l BA appeared best for shoot establishment of nodal segments of Calcutti as well as Hybrid genotypes of pointed gourd. CONCLUSIONS In node culture, effect of various treatments of plant hormones supplemented in MS medium on four genotypes of pointed gourd viz. Calcutti Female, Calcutti Male, Hybrid Female, and Hybrid Male indicated that the medium supplemented with 1.0 mg/ l IAA + 0.2 mg/l BA showed longest shoot with better and quicker establishment and little or no root formation in all the genotypes of pointed gourd. This combination was found to be best culture establishment medium. Among different genotypes, in vitro response of Calcutti Female was best followed by Hybrid Female, Calcutti Male and Hybrid Male. Of the various treatment of plant hormones examined in MS medium, the treatment 1.0 mg/l IAA + 0.2 mg/l BA was found to be best shoot proliferation medium for continuous sub culturing. Rooting of in vitro shoots occurred during shoot establishment and proliferation. In case of poor rooted shoots, ½ MS medium supplemented with 0.1mg/l IAA + 0.2 mg/l IBA was found to be effective in inducing good rooting. During hardening, maximum survival was recorded from 20 days old plantlet in potting mix (Soil: FYM: Coco peat) (1:1:1v/v/v). These media combinations can be used for large scale micropropagation of pointed gourd. New Crops and New Uses (Janick, J. eds.): ASHS Press. Alexandria, VA, pp 397-399 (1999). [7] Hoque, A.; Islam, R. and Arima, S.: Phytomorphology, 50: 267-272 (2000). [8] Kumar, S., Singh, M., Singh, A.K., Srivastava, K. and Banerjee, M.K.: Cucurbit Genetics Cooperative, 26: 74-75 (2003). [9] Chan, L.K. and Lok, K.H.: Biotechnology, 4(4): 354-357 (2005). [10] Ahmad, N. and Anis, M.: Turk J. Bot., 29: 237-240 (2005). [11] Sankar, M. A., Sajitha, K.K., Alice Kurian, Salmath, K.K., Joseph, L. and Nybe, E.V.: In vitro multiplication of Trichosanthes cucumerina. In: Recent trends in horticultural biotechnology, Vol. I and II. ICAE National Symposium on Biotechnological Interventions for Improvement of Horticultural Crops: Iissues and Strategies, (Keshavachandran, R.; Nazeem, P.; Girija, D.; John, P. S. and Peter, K. V. eds.) 10-12 January, Vellanikkara, Kerala, India, pp. 259-263 (2007). [12] Lee, W..S., Lee H.Y., Kwon, H. J., Hwang H.S., Mo H.A. and Hong, S.Y.: J. Korean Society Horticul. Sci., 36 (2):199-210 (1995). REFERENCES [1] Khunt, K.A. and Desai, D.B. Financing Agriculture, 28: 19-24 (1996). [2] Singh, K.: Pointed gourd (Trichosanthes dioica Roxb.). Indian Hort.33 & 34: 34 37 (1989). [3] Murashige, T. and Skoog, F.: Physiol. Plant., 15: 473-97 (1962). [4] Peter, K.V., Sadhu, M.K., Raj, M. and Prasanna, K.P.: Improvement and cultivation: Bitter gourd, snake gourd, pointed gourd, and ivy gourd. In: Cucurbits (Nayar, N.M. and More, T.A. eds.), Oxford / IBH, New Delhi, pp 187 198 (1998). [5] Mythili, J.B. and Thomas, P.: Scientia Horticul., 79 (1/ 2): 87-90 (1999). [6] Singh, B.P. and Whitehead, W.F.: Pointed gourd: Potential for temperate climates. In: Perspectives on 4331