International Journal of Agricultural Science and Research (IJASR) ISSN 2250-0057 Vol.

3, Issue 1, Mar 2013, 189-196 TJPRC Pvt Ltd.

IN VITRO MULTIPLICATION POTENTIAL OF COTYLEDONARY NODE EXPLANTS IN PEANUT (ARACHIS HYPOGAEA L.)
RAJINIKANTH MARKA, SRINIVAS PENCHALA, SAMATHA TALARI, SRIKANTH KAGITHOJU & RAMA SWAMY NANNA Plant Biotechnology Research Laboratory, Department of Biotechnology, Kakatiya University, Warangal, India

ABSTRACT
Among the leguminous crops Arachis hypopgaea is considered as The king of oil seeds, as it is globally valued for its protein content and oil quality. Which plays an important role in the economies of semi-arid tropics. We report the reproducible regeneration protocol from cotyledonary nodal explants of peanut cvs ICG 7827 and 13942. The cotyledonary nodal explants excised from 10-12 day old seedlings were inoculated on shoot induction medium (SIM) containing MS salts with B5 vitamins+3% sucrose medium supplemented with different concentrations and combinations of plant growth regulators (BAP/KIN/BAP+KIN/NAA+BAP/KIN/BAP+KIN). Maximum number of multiple shoots formation was found at 20 mg/L BAP (47.30.57 shoots/explant) in cv ICG 13942 and at 15mg/L BAP (45.50.35 shoots/explant) in cv ICG 7827 followed by 25mg/L KIN (42.30.91 shoots/explant). In comparison to all other concentrations and combinations of plant growth regulators (PGRs) used. The maximum shoot length (2.60.06cm) was observed at 0.5mg/L NAA+15mg/L BAP+15mg/L KIN in cv ICG 7827. Shoot buds were elongated on SEM (Shoot elongation medium) fortified with 0.5mg/L BAP. In vitro rooting was observed on RIM (Root induction medium) supplemented with NAA/IBA/NAA+IBA. The in vitro rooted plantlets were acclimatized in the plant growth chamber. These were transferred to plastic cups containing a mixture of soil: sand: vermicompost (1:1:1) and maintained in the green house for 4 weeks. Subsequently these in vitro regenerated plantlets were transferred to field and were found to be similar to parents in all morphological characters.

KEYWORDS: Arachis Hypogaea, Cotyledonary Node, Multiple Shoots, In Vitro Regeneration, Plantlet Establishment ABBREVIATIONS
PGRs-Plant growth regulators, BAP-N6-Benzylaminopurine,IBA-Indole-3-butyric acid, KIN-Kinetin, NAA--

Naphthalene acetic acid, SIM-Shoot induction medium, SEM-Shoot elongation medium, RIM-Root induction medium.

INTRODUCTION
Groundnut (Arachis hypogea L.) is an oil, food and fodder crop which plays an important role in the agricultural economies of the semi-arid tropics. It contributes significantly to food security and alleviates poverty (Naidu et al., 1999). As a legume, it improves soil fertility by fixing atmospheric nitrogen and increases productivity for smallholder farmers of the semi-arid cereal cropping systems (Smatt, 1994; Giller et al., 2002).Groundnut seeds contain 44-56% oil and 22-30% protein on a dry seed basis (Savage and Keenan, 1994). Major constraint is a lack of resistant to biotic and abiotic stresses in cultivated groundnut. Biotechnological approaches such as gene transfer for enhanced disease and pest resistance offer opportunities for the rapid improvement of peanut. Recently, microprojectile bombardnlent of embryogenic cultures of peanut has been employed to develop transgenic plants (Ozias-Akins et al., 1993).

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Groundnut has proven to be a difficult crop to manipulate in in vitro and only a limited success of whole plant regeneration (Heatley et al., 1996; Ponsamuel et al., 1998), organogenesis (Narasimhulu et al., 1983; Mckently et al., 1990; Cheng et al., 1992; Eapan et al., 1993; Li et al., 1994; Venkatachalam et al., 1996) and somatic embryogenesis (Constable et al., 1984; Ozias et al. 1992; Baker et al., 1992,1995; Chengalrayan et al., 1994, 1997; Kanyand et al., 1997) has been achieved. An efficient in vitro shoot regeneration protocol compatible with genetic transformation methods would be very useful in the rapid development of transgenic peanut plants. Hence the present investigation has been undertaken to develop the efficient and reproducible regeneration protocol for direct shoot organogenesis, multiple shoot development and plantlet establishment without intervening callus phase from cotyledonary nodal (CN) explants of two ground nut cvs ICG 7827 and 13942.

MATERIAL AND METHODS

Plant Material Mature seeds of groundnut cvs ICG 7827 and 13942 obtained from the germplasm bank of ICRISAT, Patancheru, Hyderabad, were used. The seeds were rinsed under running tap water for 10-15 min followed by liquid detergent Tween20 (5%-v/v) for 5 min and it was repeated twice. Later these were washed with sterile distilled water thoroughly. The seeds were sterilized with 0.1% (w/v) HgCl2 for 8 min and followed by rinsing in sterilized distilled water for 3-4 times. These sterilized seeds were dried on sterile tissue paper and the cotyledons were separated to isolate the zygotic embryo. These zygotic embryos were inoculated on MS medium with B5 vitamins containing 2% sucrose. The cotyledonary nodes were excised from these 8 day old seedlings and used as explants. Culture Media and Culture Conditions The cotyledonary nodal (CN) explants were cultured on shoot induction medium (SIM) containing MS salts (Murashige and Skoog, 1962) with B5 vitamins (Gamborg et al., 1968), 3% sucrose, and varying concentrations (150mg/L) of 6-Benzylaminopurin (BAP), Kinetin (KIN) alone, BAP+KIN (0.5-25mg/L) and also in combination with 0.5 mg/L NAA (Tables 1-2). The pH of the medium was adjusted to 5.80.02 either with 0.1N NaOH or 0.1N HCl before addition of agar. The medium was solidified with 0.8% Difco bacto-agar and autoclaved at 121C under 15 psi for 15-20 min. All the cultures were incubated at 252C under 16 h day exposure to white light for 4-5 weeks for multiple shoots induction. Later shoots were separated and sub cultured on shoot elongation medium (SEM) containing MS salts+B5 vitamins, 3% (w/v) sucrose supplemented with 0.5 BAP for two to three passages of four weeks each. In Vitro Rooting and Plantlet Establishment The micro-shoots after elongation (3-5cm long) were transferred to root induction medium (RIM) fortified with different concentrations of NAA/IBA/ NAA+IBA (0.5-2.0 mg/L). The in vitro regenerated plantlets were taken out from the culture vessels and washed with sterile distilled water to remove the remains of agar. Later shifted to plastic cups containing soil rite and kept in the plant growth chamber for acclimatization. These were covered with polythene bag to maintain the RH (85-90%). After 4 weeks, the plantlets were transferred to plastic cups containing soil mixture of soil: sand: vermicompost (1:1:1) and maintained in the greenhouse.

RESULTS AND DISCUSSIONS

Cotyledonary nodal explants were cultured on SIM supplemented with various concentrations of BAP (150mg/L)/ KIN (1-50mg/L)/BAP+KIN, NAA (0.5mg/L)+BAP(1-50mg/L) /KIN(1-50mg/L)/BAP+KIN. Multiple shoot

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formation was observed in all the concentrations of cytokinins alone and also cytokinnin+auxin combinations used (Tables 1-2). Maximum frequency number of multiple shoot (47.30.57 shoots/explant) formation in cv ICG 13942 at 20mg/L BAP followed by cv ICG 7827 (45.40.35 shoots/explant) at 15mg/L BAP was observed (Fig.1a-c). Longer shoots (2.50.13cm) formation was also observed in cv ICG 7827 at 20mg/L KIN+0.5 mg/L NAA and in cultivar ICG 13942 (2.60.06cm) at 15mg/L BAP+15mg/L KIN+0.5 mg/LNAA (Figs. 2-3). Two genotypes exhibited a tendency to develop enhanced number of multiple shoots with axillary branches in all the concentrations of BAP/KIN alone. The highest multiplication rates and more percentage of responding cultures were observed at 20 mg/L BAP followed by 15 mg/L BAP as a sole plant growth regulator. The maximum shooting potential was found in ICG 13942 (i.e.47.30.57) followed by ICG 7827 (i.e. 45.40.35). A further increase in BAP and KIN concentrations led to decrease in multiplication potential. Even at lower concentrations of BAP/KIN and cytokinin+auxin combinations showed the less percentage of response with lower number of multiple shoot formation. BAP and KIN levels at 50mg/L alone and in combination with 0.5 NAA in two genotypes did not show any significant increase for the induction of multiple shoots besides that growth was found to be stunted and leaves were not opened properly. The micro-shoots were separated from all the cultures and subcultured on SEM for 3 passages (Fig.1d). These micro-shoots were elongated without any shoot buds and callus formation. These elongated micro-shoots were used for in vitro rooting. In Vitro Rooting and Plantlet Establishment In vitro rooting was established on RIM supplemented with various concentrations of NAA/IBA and combination of NAA+IBA in two cultivars of peanut (Table 3, Fig. 4). Roots were induced within 15-25 days in cv ICG 7827 and 30-35 days in cv ICG 13942. Maximum number of roots and more percentage of response (22.20.10 roots/shoot) was observed in cv ICG 13942 at 1mg/L NAA followed by ICG 7827 (19.20.08 roots/shoot) (Fig.1e-g). Longer roots (3.20.28) formation was also observed in cv ICG 13942 at 1mg/L NAA and in cv ICG 7827(3.10.12) at 0.5mg/L IBA+1.5 NAA. The percentage of response and maximum number of roots/shoot increased in concentration of single auxin alone. The in vitro regenerated plantlets were successfully acclimatized in plant growth chamber. Later these were shifted to greenhouse. The survival percentage was found to be 92% before being transferred to field. The in vitro regenerated plants were found to be normal flowering and fruiting as that of parents (Fig.1h-k). Our results are in agreement with the findings of Banejee et al., (2007). According to these authors the auxin:cytokinin ratio was crucial for the regeneration of multiple shoot buds formation in groundnut. In the present study, BAP and KIN alone were able to induce proliferation of multiple shoot buds in two cvs studied. Thus, the effect of plant growth regulators not only depends on the concentration applied but also on its interaction with the endogenous growth regulators (Roy and Banerjee, 2003). Levels of endogenous growth regulaors in the explants are influenced by the duration of light, its quality and the intensity and also by the environmental factors during the culture (Kefeli et al., 1968). Verma Aman et al (2009) reported the maximum multiple shoots from Cotyledonary node explnts in cv RG-141 at 15 mg/L BAP (i.e. 43.60.72) as we have observed in the present investigation. In conclusion, among the cytokinins BAP alone was found to be more effective than KIN and also in combination with NAA. It was also recorded that the cv ICG 13942 was more effective for multiple shoot formation than cv ICG 7827. The present findings are of great importance, since it has been described as a tri-directional (direct shoot formation, multiple shoot bud formation and axillary branching and) micropropagation technique in a single medium,

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which has importance in genetic transformation experiments. The present protocol can be used for the gene transfer experiments in peanut cvs ICG 7827 & 13942 which is a prerequisite for regeneration of transgenic plants in peanut.

ACKNOWLEDGEMENTS
We thank the ICRISAT, Patancheru, Hyderabad (AP), India for providing the germplasm (seeds) of two cvs ICG 7827 and ICG 13942.

REFERENCES
1. Baker, C.M. and Wetzstein, H.Y. (1992). Somatic embryogenesis and plant regeneration from leaflets of peanut, Arachis hypogaea. Plant Cell Reports. 11: 71-75. 2. Baker, C.M. and Wetzstein, H.Y. (1995). Repetitive somatic embryogenesis in peanut cotyledon cultures by continual exposure to 2,4-D. Plant Cell Tiss Org Cult. 40:249-254. 3. Banerjee, P., Maity, S., Maity, S.S. and Banerjee, N. (2007). Influence of genotype on in vitro multiplication potential of Arachis hypogaea L. Acta Bot Croat. 66(1): 15-23. 4. Cheng, M.D., His, C.H. and Phillips, G.C. (1992). In vitro regeneration of Valencia-type peanut (Arachis hypogaea L.) from cultured petioles, epicotyls sections and other seedling explants. Peanut Sci. 19:82-87. 5. Chengalrayan, K., Sathaye, S.S. and Hazra, S. (1994). Somatic embryogenesis from mature-derived leaflets of peanut (Arachis hypogaea L.). Plant Cell Reports. 13:578-581. 6. Chengalrayan, K., Mhaske, V.B. and Hazra, S. (1997). High-frequency conversion of abnormal peanut somatic embryos. Plant Cell Reports. 16:783-786. 7. Constable, F. (1984). Callus culture: induction and maintenance. In: Cell Culture and Somatic Cell Genetics of Plants, Eds., Vasil, I.K., Academic Press, New York, USA, pp.27-35. 8. Eapan, S. and George, L. (1993). Plant regeneration from leaf discs of groundnut and pigeonpea: Influence of benzyl adenine, Indole acetic acid and Indole acetic acid amino acid conjugates. Plant Cell Tiss Org Cult. 35:223-227. 9. Gamborg, O.L., Miller, R.A. and Ojima, K. (1968). Nutrient requirements of suspension cultures of soyabean root cells. Exp. Cell Res. 50: 151-158. 10. Giller, K.E., Cadish, G. and Palm, C. (2002). The North-South divide Organic wastes, or resources for nutrient management? Agronomy, 22:703-709. 11. Heatley, M.E. and Smith, R.H. (1996). Whole plant regeneration from the shoot apex of Arachis hypogaea L. In vitro Cell Dev Biol. 32:115-118. 12. Kanyand, M., Peterson, C.M. and Prakash, C.S. (1997). The differentiation of emergences into adventitious shoots in peanut Arachis hypogaea (L.). Plant Sci. 126:87-95. 13. Kefeli, V., Turetskaya, R., Kutacek, M., Tshumakosski, N. and Krupnikova, T. (1968). Isolation and some physiological properties of natural plant growth inhibitors. Biol plantarum. 10:205. 14. Li, Z., Jarret, R.L., Pittman, R.N. and Demski, J.W. (1994). Shoot organogenesis from cultured seed explants of peanut (Arachis hypogaea L.) using Thidiazuron. In vitro cell Dev Biol. 30:187-191. 15. Mckently, A.H., Moore, G.A. and Gardner, F.P. (1990). In vitro plant regeneration of peanut from seed explants. Crop Sci. 30:192-196. 16. Murashige, T. and Skoog, I. (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiol. 15: 473-497.

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17. Naidu, R.A., Bottenberg, H., Subrahmanyan, P., Kimmins, F.M., Robinson, D.J. and Thresh, J. (1999). Epidemiology of ground nut rosette virus disease, current status and future research needs. Ann Appl Biol. 132:525-548. 18. Narasimhulu, S.B. and Reddy, G.M. (1983). Plantlet regeneration from different callus cultures of Arachis hypogaea L., Plant Sci Lett. 31:147-153. 19. Ozias-Akins, P., Anderson, W.F. and Holbrook, C.C. (1992). Somatic embryogenesis in Arachis hypogaea L.: Genotype comparison. Plant Sci. 83:103-111. 20. Ozias-Akins, P., Schnall, J.A., Anderson, W.F., Singsit, C., Clemente, T.E., Adang,M.J. and Weissinger, A.K. (1993). Regeneration of transgenic peanut plants from stably transformed embryogenic callus. Plant Science. 93:185-194. 21. Ponsamuel, J., Huhman, D.V., Cassidy, B.G. and Post-Beittenmiller, D. (1998). In vitro regeneration via caulogenesis and brassin-induced shoot conversion of dormant buds from plumular explants of peanut (Arachis hypogaea L. cv Okrun). Plant Cell Reports. 17:373-378. 22. Roy, J. and Banerjee, N. (2003). Induction of callus and plant regeneration from shoot tip explants of Dendrobium fimbriatum Lindl. Var. oculatum Hk.F Sci Hort. 97:333-340. 23. Savage, G.P. and Keenan, J.I. (1994). The composition and nutritive value of groundnut kernels. In: Smart J (Eds). The Groundnut crop: Scientific Basis for Improvement, Champman and Hall, London, pp. 173-213. 24. Smatt, J. (1994). The groundnut in farming systems and the rural economy; a global view. Chapman and Hall, London, pp. 664-669. 25. Venkatachalam, P., Subramaniampillai, A. and Jayabalan, N. (1996). In vitro callus culture and plant regeneration from different explants of groundnut (Arachis hypogaea L.). Breed Sci. 46: 315-320. 26. Verma Aman., Malik, C.P., Gupta, V.K. and Sinsinwar,Y.K. (2009). Response of groundnut varieties to plant growth regulator (BAP) to induce direct Organogenesis. World Journal of Agriculture Sciences. 5(3): 313317. Table1: Effect of BAP/KIN/BAP+KIN on Multiple Shoot Induction from Cotyledonary Node Explants in Two cvs of Peanut cv ICG 7827
Concentration of PGRs (mg/L) Percentage of Response Average no. of Shoots/Explant SEa Average Length of Shoots(cm) SEa

cv ICG 13942
Percentage of response Average no. of Shoots/ ExplantSEa Average Length of Shoots (cm) Sea

BAP 01 05 10 15 20 25 30 40 50 KIN 01 05 10 15 20

73 82 86 98 91 90 81 78 75 68 76 79 84 89

12.60.25 21.50.44 36.30.87 45.40.35 40.70.66 34.20.41 28.40.50 19.00.85 10.30.37 09.20.46 13.20.04 17.40.57 29.30.52 35.50.23

0.90.10 1.80.03 1.70.11 1.80.05 1.60.14 1.90.19 2.30.24 1.90.07 0.90.19 0.90.13 1.90.06 1.40.15 2.30.04 2.00.15

78 84 88 92 94 86 75 84 69 74 78 83 85 90

11.40.68 18.30.81 28.20.16 30.50.76 47.30.57 35.10.32 24.60.67 19.30.63 11.00.21 12.3 0.09 18.00.12 21.90.87 33.30.23 39.20.48

1.20.12 1.70.07 1.40.09 2.10.18 2.00.14 2.20.24 1.70.06 1.90.10 0.60.14 1.00.10 1.30.16 1.40.39 1.90.28 2.00.14

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25 95 30 88 40 76 50 68 BAP+KIN 0.5 0.5 58 2.5 2.5 63 5.0 5.0 71 10.0 10.0 79 15.0 15.0 70 20.0 20.0 66 25.0 25.0 54 a MeanStandard Error

42.30.91 28.40.72 14.50.84 12.40.53 12.30.32 17.50.44 26.60.04 37.20.25 23.00.43 19.80.87 13.40.49

Table 1-Contd., 2.10.19 1.80.04 1.40.07 0.80.19 0.70.08 1.50.19 2.20.01 32.60.04 39.70.82 27.60.57 1.10.21

85 87 75 62 45 56 63 68 71 60 56

32.50.87 29.50.44 24.00.76 19.00.31 14.00.43 18.80.09 25.80.62 32.60.04 39.70.82 27.60.57 16.00.02

2.40.09 1.80.03 1.20.19 0.70.20 0.60.19 1.20.04 1.00.21 2.20.06 2.00.52 1.80.24 1.10.04

Table 2: Effect of 0.5mg/L NAA+BAP/KIN/BAP+KIN on Multiple Shoot Induction from Cotyledonary Node Explants in Two cvs of Peanut cv ICG 7827
Concentration of PGRs (mg/L) NAA+BAP Percentage of Response Average no. of Shoots/Explant SEa Average Length of Shoots(cm) SEa

cv ICG 13942
Percentage of Response Average no. of Shoots/ ExplantSEa Average Length of Shoots (cm) Sea

0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5
a

01 05 10 15 20 25 30 40 50 01 05 10 15 20 25 30 40 50 0.5 2.5 05 10 15 20 25 0.5 2.5 05 10 15 20 25

45 53 61 66 75 78 71 65 51 47 55 61 73 81 87 80 80 65 58 61 74 74 65 54

07.80.38 14.20.54 22.30.67 28.00.42 29.70.66 27.60.81 23.40.12 20.00.85 13.30.57 04.60.26 11.20.24 17.80.57 21.30.22 23.40.35 26.60.81 26.60.81 14.20.54 09.40.63 05.30.52 11.20.54 28.60.68 17.40.65 15.00.03 10.80.67 06.40.89

0.70.08 1.00.10 1.80.15 1.80.13 1.60.14 1.90.13 2.30.24 1.80.07 1.10.19 0.90.12 1.90.16 1.60.05 2.30.04 2.50.33 2.00.12 2.20.14 1.80.07 1.10.09 1.20.08 1.40.09 1.10.21 2.10.34 2.10.34 1.50.46 1.30.21

54 57 63 69 76 70 64 58 43 66 69 71 79 88 77 71 65 62 69 71 71 80 75 60 55

08.30.48 18.80.71 26.20.96 30.50.76 28.70.32 25.30.56 21.50.67 18.00.63 12.00.41 03.20.29 09.60.02 15.80.47 20.20.86 24.70.68 22.50.07 19.50.34 14.00.56 11.00.31 09.70.53 12.80.89 18.01.02 23.60.04 22.00.20 18.60.97 11.70.42

0.90.12 1.70.17 1.60.05 2.20.18 2.10.04 2.40.14 1.50.06 1.10.09 0.60.11 1.00.10 1.60.16 1.40.39 1.80.28 2.00.14 2.20.09 1.80.03 2.00.19 0.90.20 1.50.09 1.60.14 1.90.21 2.50.16 2.40.32 1.90.54 1.20.04

NAA+KIN

NAA+ BAP+KIN

48

MeanStandard Error

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Table 3: Effect of NAA/IBA/NAA+IBA on In vitro Root Induction from Elongated Shoots of Peanut
Concentration of PGRs (mg/L) Average no. of Shoots/Explant SEa Average Length of Shoots(cm) SEa Percentage of response Average no. of Shoots/ ExplantSEa Average Length of Shoots (cm) Sea

Percentage of Response

NAA+BAP 0.5 1.0 1.5 2.0 IBA 0.5 1.0 1.5 2.0 NAA+ BAP 0.5 0.5 0.5 0.5 1.0 1.5

55 72 65 54 60 95 81 62 52 73 65 48 42 65

8.2 0.09 16.4 0.45 10.3 0.02 4.50.56 7.40.23 19.20.08 12.0 0.44 5.60.35 5.1 0.50 16.30.61 8.40.10 6.0 0.63 5.10.06 10.60.19 8.30.75 4.40.59

2.60.04 3.0 0.10 2.7 0.34 2.40.09 2.80.09 3.40.62 3.00.53 2.10.43 2.70.18 3.00.10 2.40.59 1.90.15 2.70.12 2.90.43 3.10.12 2.70.26

65 98 81 73 53 71 88 55 41 78 53 41 48 73 66 48

7.30.45 22.20.10 14.50.88 7.30.70 6.60.10 18.00.23 10.30.43 6.70.09 7.00.43 16.00.05 10.40.03 5.60.18 6.20.12 10.40.56 6.00.79 4.30.11

2.20.10 3.20.28 2.90.41 2.70.40 2.50.14 3.00.14 2.70.34 2.00.29 2.60.16 2.90.45 2.00.53 2.00.29 2.10.13 2.90.43 3.00.54 2.40.40

0.5 2.0 IBA+NAA 0.5 0.5 0.5 1.0 0.5 1.5

a

0.5 2.0 MeanStandard Error

60 38

Figure 1: a-h: In Vitro Regeneration and Plantlet Establishment from Cotyledonary Node Explants in Two cvs of Peanut

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a)

Germination of zygotic embryo on MS salts with B5 Vitamins medium.

b-c) Induction of multiple shoots from cotyledonary node explants on SIM with 25 mg/L KN in cv. ICG 7827 & 20 mg/L BAP in cv. ICG 13942 after 4 weeks of inoculation. d) Elongation of multiple shoots of cv ICG 7827 on SEM.

e-f) In vitro rooting of shoots on RIM in cvs ICG 7827& 13942 respectively (Note profuse rhizogenesis) g) Plantlet showing with lengthy roots prior to acclimatization. h-i) Acclimatization of plantlet in a plastic pot containing soilrite. j) Plants are shifted to plastic pot containing soilmix and maintained in green house.

h) Plants growing in the research field (one week after transplantation).

Figure 2: Effect of BAP/KIN/ BAP+KIN on Percentage of Shoot Induction and Average No. of Shoots/Explant from Cotyledonary Node Explants in Two cvs of Peanut

Figure 3: Effect of NAA+BAP/ NAA+KIN/NAA+BAP+KIN on Percentage of Shoot Induction and Average no. of Shoots/Explants from Cotyledonary Node Explants in Two cvs of Peanut

Figure 4: Effect of NAA/IBA/ NAA+IBA/IBA+NAA on Percentage of Root Induction and Average No. of Roots/Shoot from Cotyledonary Node Explants in Two cvs of Peanut