Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402

International Journal of Current Microbiology and Applied Sciences ISSN: 2319-7706 Volume 6 Number 4 (2017) pp. 2392-2402 Journal homepage: http://www.ijcmas.com

Original Research Article https://doi.org/10.20546/ijcmas.2017.604.279

In vitro Protocol for Direct Organogenesis in cymbalaria. Fenzl

T. Devi, V. Rajasree*, V. Premalakshmi, K. Hemaprabha and S. Praneetha

Department of Vegetable Crops, HC & RI, Agricultural University, Coimbatore - 641003, Tamil Nadu, *Corresponding author

ABSTRACT

Momordica cymbalaria Fenzl. is a perennial herbaceous climber belongs to

family and the genus of bitter gourd (). Fruits are used as vegetables, it possess hypoglycaemic, hypolipidemic, cardio protective, hepatoprotective, nephroprotective and antioxidant properties. It is rich in crude fiber (6.42 g/100 g), K e yw or ds calcium (72 g/100g), potassium, sodium and vitamin C (290 g/100g).The major constraints in propagation of Momordica cymbalaria are non-availability of standardized propagation M. cymbalaria, methods and low yield. In vitro propagation technique could be a valuable alternative to Micropropagation, conventional methods for mass multiplication. To achieve this goal, investigation on Direct organonesis, micropropagation of Momordica cymbalaria was carried out at the Tissue Culture Shoot regeneration, Laboratory of Horticultural College and Research Institute, TNAU Coimbatore. For direct Rooting, Hardening -1 -1 organogenesis, MS medium supplemented with 2.0 mgl of BAP and 3.0 mgl of kinetin Article Info was found to be the most suitable concentration for maximum shoot regeneration

percentage and shoot length for both axillary bud and shoot tip. Efficient rooting was Accepted: -1 -1 20 March 2017 observed in half MS medium fortified with 1.0 mgl IBA+ 0.1 mgl of NAA. Rooted plantlets were hardened with various media among which the sterilized pot mixture, coco Available Online: 10 April 2017 peat was found to be the most ideal hardening medium. The present study revealed that

micro propagation through direct organogenesis using axillary bud could serve as a viable -1 protocol, when MS medium is augmented with 2.0 mgl BAP is used for shoot regeneration. These shoots multiplied to 7.48 numbers in MS medium supplemented with -1 3.0 mgl BAP, followed by maximum rooting percentage in half MS medium fortified with 1.0 mgl-1IBA + 0.1mgl-1 of NAA and finally acclimatized in pot mixture, coco peat

which afford mass multiplication of Momordica cymbalarias.

Introduction

Momordica cymbalaria Fenzl. is a perennial hypoglycaemic, hypolipidemic, cardio herbaceous climber, which belongs to protective, hepatoprotective, nephroprotective cucurbitaceae family and the genus of bitter and antioxidant properties (Jeyadevi et al., gourd (Momordica charantia). It is widely 2012). The roots of this have been used distributed in the tropical regions of Africa by the natives of north Karnataka and Andhra and India. It is found in the south Indian states Pradesh to treat some gynecological ailments of Andhra Pradesh, Karnataka, Madhya and also to induce abortions. The tuber is used Pradesh, and Tamil Nadu. Fruits as an abortifacient (Parvathi and Kumar are used as vegetables it possess 2002).

2392

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402

The minimum requirement of vegetable dioica, Ugandhar et al., 2011 in Cucumis consumption is 280g/day/adult. However in sativus, Jeyakumar et al.,2013 in Cucumis many developing countries vegetable angurina, Debnath et al., (2013) and Rathod consumption is far below the normal (2015) in Momordica dioica. requirement, which may be attributed to the low availability or unaffordable cost. The To date there is not much reports on tissue feasible alternative is to explore additional culture system of Momordica cymbalaria. source of food and to minimize the load on Hence the present study was under taken to production of conventional food . The standardize the suitable explants for micro fruits consumed as vegetable are rich in crude propagation and suitable culture media for in fiber (6.42 g/100 g), calcium (72 g/100g), vitro shoot regeneration and rooting of shoots. potassium, sodium and vitamin C (290g/100g) as compared to bitter gourd and Materials and Methods its fruits have medicinal value. The major constraints in propagation of Momordica Tuber materials were collected from the cymbalaria are non-availability of farmer’s field in Ramanathapuram, planted in standardized propagation methods and low college orchard, Tamil Nadu agricultural yield. A limited number of perennial tubers university, Coimbatore for explant collection. survive in soil and produce single plant in the The explants used were nodal segments and next season. The population is reduced by shoot tip. The explants were washed with consumption of tubers by animals and water teepol to remove the soil debris and pretreated logging conditions, respectively. The fruits with 0.2% bavistin and 0.1% streptomycin for had few seeds that remain dormant and low 30 min on rotatory shaker and thoroughly germination rate further curtails the washed with distilled water. The explants propagation through seeds. It can be were then surface sterilized by using 70% overcome by biotechnological approach ethanol for 30 sec. followed by 0.2% HgCl2 through micropropagation. for 5min. The explants were then washed with sterile distilled water to remove trace of To achieve this goal, the first important step is mercuric chloride. to establish a reliable tissue culture system for plant regeneration. The controlled conditions The axillary bud and shoot tip were placed on provide the culture an environment conducive MS medium basal medium (Murashige and for growth and multiplication. These Skoog, 1962) with 3% sucrose was used for conditions include proper supply of nutrients, breaking the dormancy of shoot buds as well pH medium, adequate temperature, proper as for proliferation. The pH of the medium light environment. Plant tissue culture was adjusted to 5.7 using 0.1 N KOH or 0.1 N technology is being widely used for large HCl as and when required and 0.7% (w/v) scale plant multiplication. Endangered, agar (Merck) before autoclaving. threatened and rare species have successfully Approximately 20 ml of molten medium was been grown and conserved by micro dispensed into each culture tube (25×150 propagation because of high coefficient of mm) and plugged with non-absorbent cotton. multiplication and small demands on number The tubes steam sterilized at 15 lb inch 20 of initial plants and space. There has been min. All the cultures were grown at 24±2 °C progress in tissue culture studies in many temperature and 55±5% relative humidity cucurbitaceae members such as under 16 h photoperiod supplied by Philips Venkateshwaralu (2009) in Coccinia indica, fluorescent tubes. Karim and Ahmed (2010) in Momordica 2393

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402

In vitro shoot regeneration and Results and Discussion multiplication Effect of different concentrations of (BAP) For in vitro shoot multiplication, the explants cytokinin on shoot regeneration were cultured on MS medium alone as control and MS medium with different concentrations Table 1 represents the shoot regeneration of of cytokinins (BAP, Kn). The effects of nodal segment, shoot tip explant to various different plant growth regulators were tested concentrations of BAP and Kn. It was on multiple shoot induction, shoot elongation, observed that maximum response was and were recorded periodically. observed in nodal segment explants compared to shoot tip in both the cytokinins Minimum In vitro root induction days (4.60) was taken for bud sprout in MS medium supplemented with 3.0 mg/l of BAP For rooting, in vitro grown shoots with more followed by 2.5 mg/l of BAP when nodal than four leaves were excised and implanted segment used as explant and maximum days on half MS basal media and with different (19.4 days) taken for bud sprout was recorded concentration of IBA and NAA. The cultures in control. Maximum shoot length was were examined every week and were recorded observed in MS augmented with 2.5 mg/l of on the basis of visual observations. BAP (5.62 cm) followed by MS supplemented with 2.0mg/l of BAP (5.46 cm). Hardening of plants The maximum shoot regeneration percentage (97.7%) was recorded in MS medium For hardening, at first the tissue culture supplemented with 2.0mg/l of BAP followed derived healthy rooted plantlets were removed by MS medium fortified with 2.5 mg/l of from the agar medium, washed thoroughly BAP (95.5%) and minimum shoot under running tap water and transferred to regeneration percentage was observed in MS earthen pots containing sterilized media basal medium (12.5%). composition. To preserve moisture, the potted plantlets were covered with transparent Effect of different concentration of Kn on polythene bag and placed in the green house. shoot regeneration After one month, plantlets were transferred to the main field. It was observed that minimum days (4.46) taken for bud sprouting was observed in MS Statistical analysis medium enriched with 3.0 mg/l of Kn and the maximum days (8.23) taken for bud sprout The experiment design was Completely was recorded in control. Maximum shoot Randomized block Design and Factorial length (6.76) and maximum shoot induction (FCRD) as per the standard procedures of percentage (75.3%) was observed in MS Gomez and Gomez (1984). Observations medium supplemented with 3.0 mg/l of Kn. recorded as percentage were subjected to The MS basal medium showed less response angular transformation (Snedecor and (Table 3). Cochran, 1980). Analysis was carried out with AGRES software package and MS Excel In the present study, MS medium spreadsheet. The means of different supplemented with 3.0 mgl-1 of BAP recorded experiments were compared using DMRT the minimum number of days taken for bud (Duncan’s Multiple Range Test) at the 5% emergence for both shoot tip and axillary bud level. 2394

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402 explant followed by the MS medium concentration for the better performance of supplemented with 2.5 mgl-1 of BAP. It may this particular hormone. Any high or low due to the effect of cytokinin to overcome from this normal concentration of the growth apical dominance, release lateral buds from regulator exacerbated the response. The fact dormancy, and promote shoot formation. The that higher doses failed to manifest their results are in conformity with findings of effect could be attributed to an obnoxious Mahzabin et al., 2008 in Cucurbita maxima, effect at higher concentration, whereas, the Ugandhar et al., (2011) in Cucumis sativus, ineffectiveness of the lower dose indicated Rani et al., 2012 in Murraya koenigii, Munsur inadequate dose of hormone as a consequence et al., (2007) in Momordica charantia. The indicating poor performance. These findings shoot length was higher in MS medium can be affirmatively compared with the supplemented with 2.5 mgl-1 of BAP followed observations of Ugandhar et al., (2011) in by MS medium supplemented with 2.0 mgl-1 cucumber and Venkeshwaralu (2012) in of BAP. But interestingly shoot induction Cucumis melo. Many literatures suggest that percentage was higher in MS+2.0 mgl-1 of BAP is most active at the concentration of 1.0 BAP. This may due to increasing to 2.0 mgl-1 in many plant species concentration of BAP cause antagonistic (Senthilkumar et al., 2007; Karupusamy et effect. It could be inferred from above results al., 2006; Arulanandam et al., 2011). that 2.0 mgl-1 of BAP is the optimum

Table.1 Effect of different concentrations of BAP on shooting in Momordica cymbalaria

Days taken for bud Length of shoot (cm) Percentage of shoot sprout induction (%) Treatment details Axillary Shoot tip Axillary Shoot Axillary Shoot tip Treatme bud (E1) (E2) bud (E1) tip (E2) bud (E1) (E2) nt code 16.5g 12.5g f g f g T0 MS media 19.4 15.4 2.83 2.40 (23.9) (20.7) MS + BAP 1.0 89.9c 34.4f T1 mgl-1 8.23e 7.06d 5.02b 4.18e (71.7) (35.9) bc e T2 MS + BAP 1.5 92.2 38.8 mgl-1 7.86de 6.70d 5.16b 4.29e (74.2) (42.4) a d T3 MS + BAP 2.0 97.7 52.2 mgl-1 6.00bc 6.13c 5.46a 4.62d (82.7) (49.4) ab d T4 MS + BAP 2.5 95.5 49.9 mgl-1 5.56b 5.43b 5.62a 4.73cd (77.9) (47.5) e f T5 MS + BAP 3.0 45.5 27.7 mgl-1 4.60a 4.73a 4.66bc 4.97bc (42.4) (33.8) (Note values in parenthesis are the angular transformed values of the original percentile data

2395

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402

Table.2 Effect of different concentrations of kinetin on shooting in Momordica cymbalaria

Treatment Days taken for bud Length of shoot (cm) Shoot induction percent Treatment details sprout (%) code Axillary Axillary bud Axillary Axillary bud Axillary Axillary bud bud (E1) (E1) bud (E1) (E1) bud (E1) (E1) i h h h T0 MS medium 19.4 15.4 2.80 2.43 23.9 20.7 MS + KN 1.0 T1 mgl-1 8.00g 6.10e 3.94 3.90 60.4c 31.7g T2 MS + KN 1.5mgl-1 7.20f 5.83de 5.31 4.71 64.1bc 33.8fg T3 MS + KN 2.0mgl-1 5.80de 5.40cd 5.86 5.17 65.7b 37.9ef T4 MS + KN 2.5mgl-1 5.33cd 5.00bc 6.30 5.53 72.7a 41.1de T5 MS + KN 3.0mgl-1 4.46a 4.60ab 6.76 5.92 75.3a 43.0d

Table.3 Effect of different concentrations of BAP on shooting in Momordica cymbalaria

Treatme Days taken for Length of shoot No. of shoots per Percentage of Treatme nt details multiple shoot (cm) explant multiple shoot nt code induction induction Axillary Axillary Axillary Axillary Axillary Axillary Axillary Axillary bud (E1) bud (E1) bud (E1) bud (E1) bud (E1) bud (E1) bud (E1) bud (E1) MS 30.5i 27.1i g h i j T0 medium 61.4 66.5 1.51 1.44 1.26 0.80 (33.5) (31.4) MS + BAP 2.0 77.7c 53.3g T1 mgl-1 52.0f 54.0e 4.00 3.53 5.51c 2.30g (61.8) (46.9) MS + b fg T2 BAP 2.5 86.6 58.4 mgl-1 47.4d 48.2d 4.76 4.50 6.49b 2.73f (68.6) (49.8) T3 MS + BAP 3.0 93.3a 61.1ef mgl-1 34.5c 36.0c 6.13 6.05 7.48a 4.20d (75.3) (51.4) T4 MS + BAP 3.5 72.2d 45.5h mgl-1 31.8b 32.5b 2.53 2.00 3.53e 2.00g (58.2) (42.4) T5 MS + BAP 4.0 64.4e 42.5h mgl-1 30.2a 30.9ab 1.83 1.63 2.26g 1.86h (53.4) (40.7) (Note: Values in parenthesis are the angular transformed values of the original percentile data)

2396

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402

Table.4 Effect of different concentrations of IBA and NAA on rooting

Treatment details No of Days No of Root Root Treatment taken for root roots per length induction code induction plant percentage 0.50g 34.0i f h T0 Half MS media 24.9 2.33 (34.7) Half MS+ IBA 0.5 mg 0.67g 63.4h T1 l-1 10.5e 6.37g (52.8) T2 Half MS+ IBA 0.5+ 1.33f 66.8g NAA 0.1mg l-1 8.27d 7.30fg (54.8) e T3 1.80 Half MS+ IBA 0.5+ 73.4f NAA 0.2mg l-1 8.13cd 7.80fg (58.9) d T4 2.20 Half MS+ IBA 0.5+ 77.7e NAA 0.3mg l-1 7.90cd 8.20f (61.8) b T5 3.20 Half MS+ IBA 1.0+ 83.5c NAA 0.1mg l-1 7.57bc 33.6a (66.0) 3.57a Half MS+ IBA 1.0+ 93.5a -1 b a T6 NAA 0.2mg l 7.27 33.3 (75.2) 3.67a Half MS+ IBA 1.0+ 85.9b -1 b b T7 NAA 0.3mg l 7.17 29.6 (67.9) e T8 1.90 Half MS+ IBA 1.5+ 80.5d NAA 0.1mg l-1 5.30a 25.0c (63.8) d e T9 Half MS+ IBA 1.5+ 2.23 75.9 NAA 0.2mg l-1 5.40a 22.0d (60.6) d f T10 Half MS+ IBA 1.5+ 2.14 72.5 NAA 0.3mg l-1 5.20a 19.0e (58.3) (Note: values in parenthesis are the angular transformed values of the original percentile data)

2397

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402

Table.5 Effect of hardening media on ex vitro plantlet of Momordica cymbalaria

Treatment Treatment detail Ex vitro plantlet Days for emergence of Number of new code survival (%) 1st new leaf leaves H1 Pot mix 91.2b (1:1:1 of soil: sand: (72.7) FYM) 18.0b 5.62b d H2 Sand 44.7 (41.9) 21.6c 2.61c c H3 Cocopeat 53.7 (47.1) 25.0d 1.30d a H4 Cocopeat +pot 95.0 mix(1+1) (77.0) 16.0a 6.30a (Note: Values in parenthesis are the angular transformed values of the original percentile data) ** - High Significant

Figure.1 Effect of different concentrations of BAP on shoot multiplication

2398

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402

Figure.2 Effect of different concentrations of IBA and NAA on rooting

Figure.3 Hardening of in vitro grown M. cymbalaria

2399

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402

Effect of different concentration of BAP on Identification of viable explant shoot multiplication Within the explants compared in the present Different concentration of BAP showed study the axillary bud was found to be better significant difference on shoot multiplication and more convenient than shoot tip with present in table 2. respect to overall response for in vitro regeneration and shoot proliferation. This The minimum days (30.2) taken for multiple may due to the apical dominance of shoot tip shoot induction was recorded in MS medium which produce lesser shoot multiplication rate supplemented with 4.0 mg/l of BAP. than axillary bud. The results of the present Maximum multiple shoot induction study are in corroboration with that of Verma percentage (93.3%) and maximum number et al., (2014). (7.48) of shoot per explant was recorded in MS medium fortified with 3.0 mg/l of BAP Effect of different concentration of NAA and minimum shoot induction percentage and IBA on rooting (30.5%) and shoots (1.26) per explant was recorded in MS basal medium. The in vitro rooting of regenerated shoot was presented in table 3. The minimum of 5.20 Among the two cytokinins, BAP proved to be days taken for root initiation was recorded in better than kinetin in overall response of in half MS medium supplemented with 1.5 mg/l vitro shoot regeneration of axillary bud and of IBA+0.3 mg/l of NAA and maximum of shoot tips of Momordica cymbalaria. 24.9 days taken for root initiation was observed in half MS basal medium. The above results are also in line with the Maximum number (33.6) of roots per shoot reports Ugardhar et al., 2011 in Cucumis was recorded in MS medium enriched with sativus. In the present study, the minimum Half MS+ IBA 1.0+ NAA 0.1mg/l. Maximum number of days taken for multiple shoot root length of 3.67 cm was observed in MS induction was observed in MS supplemented medium enriched with Half MS+ IBA 1.0+ with 4.0 mgl-1of BAP. Where Debnath et al., NAA 0.3mg/l which was on par with Half (2013) in Momordica dioica reported the MS+ IBA 1.0+ NAA 0.2mg/l (3.57cm). same. But the main factors which directly Maximum root induction percentage (93.5%) influence the maximum shoot multiplication was recorded in Half MS+ IBA 1.0+ NAA rate were shoot length, number of multiple 0.2mg/l followed by Half MS+ IBA 1.0+ shoot and multiple shoot induction percentage NAA 0.3mg/l showed 85.9 % of root was maximum in MS + 3.0 mgl-1 BAP. induction. Minimum root length (0.50 cm), Above this concentration number of multiple least number of root (2.33) and minimum root shoots, multiple shoot induction percentage induction percentage (34.0%) was recorded in gets reduced drastically. This may due to Half MS basal medium. The most commonly increasing the concentration of BAP in MS used auxin for root induction is IBA. In our showed abnormality development of the buds. present study, among the ten treatments Such observations are in corroboration with minimum days taken for root induction were those of Munsur et al., (2007) in Momordica observed in ½ MS medium + 1.5 mgl-1 IBA+ charantia, Mahzabin et al., (2008) in 0.1 mgl-1 NAA which were on par with ½ MS Cucurbita maxima and Sridhar and Naidu medium + 1.5 mgl-1 IBA+ 0.2 mgl-1 NAA and (2011) in Solanum nigrum. ½ MS medium + 1.5 mgl-1 IBA+ 0.3 mgl-1 NAA. The number of root per shoot was

2400

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402 higher in ½ MS medium + 1.0 mgl-1 IBA+ 0.1 pot mixture alone or in combination with mgl-1 NAA which were on par with ½ MS cocopeat was found to be the most ideal medium + 1.0 mgl-1 IBA+ 0.2 mgl-1 NAA. hardening medium. The results of the present study are in line with those of The highest root length was observed in ½ Thiripurasundhari and Rao (2011) in Coccinia MS medium + 1.0 mgl-1 IBA+ 0.2 mgl-1 NAA indica observed that in pot mixture alone or in which were on par with ½ MS medium + 1.0 combination with cocopeat gave the highest mgl-1 IBA+ 0.3 mgl-1 NAA. The percentage of plant survival rates. root induction percentage was higher in ½ MS medium + 1.0 mgl-1 IBA+ 0.2 mgl-1NAA In conclusion the present study revealed that followed by ½ MS medium + 1.0 mgl-1 IBA+ micropropagation through direct 0.3 mgl-1 NAA. Anisley and Lee (2010) found organogenesis using axillary bud could serve IBA is the best rooting hormone. as a viable explant when MS medium is augmented with 2.0 mgl-1 BAP for shoot Contrarily, Reddy et al., (2012) in Asclepias regeneration. These shoots multiplied to 7.48 curassavica, observed the highest number of numbers in MS medium supplemented with root in ½ MS medium supplemented with 1.0 3.0 mgl-1 BAP, followed by maximum rooting mgl-1+ 0.2 mgl-1 kinetin. Anbazhagan et al., percentage in half MS medium fortified with (2014), for rooting in banana ½ MS medium 1.0 mgl-1 IBA + 0.1 mgl-1 of NAA and finally supplemented with 1.0 mgl-1 IBA. Pal et al., acclimatized in pot mixture, coco peat afford (2007) in summer squash observed 70% mass multiplication of Momordica rooting in ½ MS medium supplemented with cymbalaria 1.0 mgl-1 IBA. The role of IBA in favoring the conjugation between endogenous IAA and References amino acids that leads to the synthesis of the specific protein necessary for the formation of Debnath, B., Sinha,S. and Sinha, R.K. 2013. root initiation. Rapid in vitro differentiation and regeneration of Momordica dioica Effect of hardening media on ex vitro Roxb. Indian J. Plant Sci., 2(3): 43-47. plantlet of Momordica cymbalaria Gomez, K.A .and Gomez, A.A. 1984.Statistically procedure for Effect of different media concentrations on agricultural research. An IRRI book, hardening of in vitro grown Momordica Wyley inter science application cymbalaria was furnished in table 4. Among pulication, John Wyley and Sons, the media concentration used, cocopeat + pot Newyork. USA. pp.680 mix (1+1) showed minimum days for Jeyadevi, R., Sivasudha, T., Rameshkumar, emergence of 1st new leaf and the maximum A., Sangeetha, B., Ananth, D.A. and number of new leaves and survival percentage Aseervatham, G.S.B. 2012. was also recorded in cocopeat + pot mix Nutritional constituents and medicinal (1+1) followed by pot mix alone. values of Momordica cymbalaria. Asian Pacific J. Tropical Hardening / acclimatization Biomedicine.S456-S461. Jeyakumar, J.J.,Kamaraj, M. and The experiment was taken up to study the Thiruvengadam. 2013. Biochemical effect of hardening media for acclimatization analysis of in vitro induced callus of of the in vitro regenerated plantlets of Cucumis anguria L. Int. J. Pharm Bio Momordica cymbalaria. In the present study, 2401

Int.J.Curr.Microbiol.App.Sci (2017) 6(4): 2392-2402

Sci., 4(3): 484 – 489. Sridhar, T.M. and Naidu, C.V. 2011. In Vitro Karim, M.A and Ahmed, S.U. 2010. Somatic Direct Shoot Organogenesis from Leaf embryogenesis and micro propagation Explants of Solanum nigrum (L.) An in teasle Gourd. Int. J. Environ. Sci. Important Antiulcer Medicinal Plant. Dev., 1(1): 10-14. J. Phytol., 3(5): 29-35. Mahzabin, F., Parvez, S. and Alam, M.F. Ugandhar, T., Venkateshwarrlu, M., Begum, 2008. Micropropagation of Cucurbita G., Srilatha, T. and Jaganmohanreddy, maxima Duch. through shoot tip K. 2011. In Vitro plant regeneration of culture. J. Bio-Sci.,16: 59-65. Cucumber (Cucumis sativum (L.) from Munsur, M. A. Z., Haque, M.S., Nasiruddin, cotyledon and hypocotyl explants. Sci. K.M. and Hasan, M.J. 2007. Res. Rep., 1(3): 164-169. Regeneration of bitter gourd Venkateshwarlu, M. 2009. Direct multiple (Momordica charantia L.) fromleaf shoots proliferation of muskmelon segments and root tips. Prog. Agric., (Cucumis melo L.) from shoot tip 18(2): 1-9. explants. Int. J. Pharma and Biosci., Paravathi, S. and Kumar. V.J.F. 2002. Studies 2(3): 645-652. on chemical composition and Verma, A.K., Kumar, M., Tarafdar, S., Singh, utilization of wild edible vegetable R. and Thakur, S. 2014. Development athalakkai (Momordica tuberosa). Pl. of protocol for micro propagation of Foods Hum Nutr., 57: 215-222. gynoecious bitter gourd (Momordica Rathod, V. 2015. Plant regeneration in charantia L). Int. J. Plant, Animal and Momordica Dioica (Roxb) by root Environ. Science. explant.J. Pharmacy and Bio Sci., 10(2): 80-83.

How to cite this article:

Devi, T., V. Rajasree, V. Premalakshmi, K. Hemaprabha and Praneetha, S. 2017. In vitro Protocol for Direct Organogenesis in Momordica cymbalaria. Fenzl. Int.J.Curr.Microbiol.App.Sci. 6(4): 2392-2402. doi: https://doi.org/10.20546/ijcmas.2017.604.279

2402